US 2005O281.883A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2005/0281883 A1 Daniloff et al. (43) Pub. Date: Dec. 22, 2005

(54) COMPOSITIONS AND SYSTEMS FOR Publication Classification FORMING CROSSLINKED BOMATERLALS AND ASSOCATED METHODS OF (51) Int. Cl...... A61K 9/14 PREPARATION AND USE (52) U.S. Cl...... 424/489 (76) Inventors: George Y. Daniloff, Mountain View, CA (US); Louis C. Sehl, Redwood (57) ABSTRACT City, CA (US); Olof Mikael Trollsas, San Jose, CA (US); Jacqueline Schroeder, Boulder Creek, CA (US); David M. Gravett, Vancouver (CA); Crosslinkable compositions are provided that readily Philip M. Toleikis, Vancouver (CA) crosslink in Situ to provide crosslinked biomaterials. The Correspondence Address: composition contains at least two biocompatible, non-im REED INTELLECTUAL PROPERTY LAW munogenic components having reactive groups thereon, GROUP with the functional groups Selected So as to enable inter 1400 PAGE MILL ROAD reaction between the components, i.e., crosslinking. In one PALO ALTO, CA 94304-1124 (US) embodiment, a first component has nucleophilic groups and a Second component has electrophilic groups. Additional (21) Appl. No.: 11/118,088 components may have nucleophilic or electrophilic groups. Methods for preparing and using the compositions are also (22) Filed: Apr. 28, 2005 provided as are kits for delivery of the compositions. Exem Related U.S. Application Data plary uses for the crosslinked compositions include tissue augmentation, biologically active agent delivery, bioadhe (60) Provisional application No. 60/566,569, filed on Apr. Sion, and prevention of adhesions following Surgery or 28, 2004. injury. Patent Application Publication Dec. 22, 2005 Sheet 1 of 2 US 2005/0281883 A1

FIG ... 1

Patent Application Publication Dec. 22, 2005 Sheet 2 of 2 US 2005/0281883 A1

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FIG. 3 US 2005/0281883 A1 Dec. 22, 2005

COMPOSITIONS AND SYSTEMS FOR FORMING by the instant invention, which is a mixture of two compo CROSSLINKED BOMATERIALS AND nents, each component having a core Substituted with reac ASSOCATED METHODS OF PREPARATION AND tive groups, where the reactive groups on one component are USE capable of reacting with the reactive groups on the other component. The components are essentially non-reactive in TECHNICAL FIELD a dry environment, and upon reaction form a three-dimen 0001. This invention relates generally to compositions Sional matrix. and Systems for forming crosslinked biomaterials, to the crosslinked biomaterials prepared thereby, and to methods SUMMARY OF THE INVENTION of using Such compositions as, for example, bioadhesives for 0008 One aspect of the invention relates to a homoge tissue augmentation, for prevention of Surgical adhesions, neous dry powder composition comprised of a first com for coating Surfaces of Synthetic implants, as drug delivery ponent having a core Substituted with m nucleophilic matrices, for ophthalmic applications, for Orthopedic appli groups, where me2; and a Second component having a core cations, as Sealants, as hemostats, and in other applications, Substituted with n electrophilic groups, where ne2 and as discussed herein and/or as appreciated by one of ordinary m+n>4, wherein the nucleophilic and electrophilic groups skill in the art. are non-reactive in a dry environment but are rendered reactive upon exposure to an aqueous environment Such that BACKGROUND OF THE INVENTION the components inter-react in the aqueous environment to 0002 Much work has been done in developing bioadhe form a three-dimensional matrix. A pharmaceutically sive materials. U.S. Pat. No. 5,162,430 to Rhee et al. acceptable carrier may also be included. describes the use of collagen-Synthetic polymer conjugates 0009. In one embodiment of the homogeneous dry pow prepared by covalently binding collagen to Synthetic hydro der composition, the nucleophilic and electrophilic groups philic polymerS Such as various derivatives of polyethylene undergo a nucleophilic Substitution reaction, a nucleophilic glycol. In a related patent, U.S. Pat. No. 5,328,955 to Rhee addition reaction, or both. The nucleophilic groups may be et al., various activated forms of polyethylene glycol and selected from -NH, -NHR', -N(R'), -SH, -OH, various linkages are described, which can be used to produce COOH, -CH-OH, -H, -PH, -PHR', -P(R'), collagen-Synthetic polymer conjugates having a range of -NH-NH, -CO-NH-NH2, and -CHN, where R' physical and chemical properties. U.S. Pat. No. 5,324,775 to is a hydrocarbyl group, and each R' may be the same or Rhee et al. also describes Synthetic hydrophilic polyethylene different. The electrophilic groups may be Selected from glycol conjugates, but the conjugates involve naturally CO-Cl, -(CO)-O-(CO)-R (where R is an alkyl occurring polymerS Such as polysaccharides. group), -CH=CH-CH=O and -CH=CH 0003) EP 0732 109 A1 to Rhee discloses a crosslinked C(CH.)=O, halo, -N=C=O, -N=C=S, biomaterial composition that is prepared using a hydropho -SOCH=CH, -O(CO)-C=CH, -O(CO)- bic crosslinking agent, or a mixture of hydrophilic and C(CH)=CH, -S-S-(CHN), -O(CO)- hydrophobic crosslinking agents, where the preferred hydro C(CHCH)=CH, -CH=CH-C=NH, -COOH, phobic crosslinking agents include hydrophobic polymers -(CO)O-N(COCH), -CHO, -(CO)O- that contain, or can be chemically derivatized to contain, two N(COCH)-S(O), OH, and -NCCOCH). or more Succinimidyl groups. 0010. In another embodiment of the homogeneous dry 0004 U.S. Pat. No. 5,580,923 to Yeung et al. discloses powder composition, the nucleophilic groups are amino Surgical adhesive material that comprises a Substrate mate groupS and the electrophilic groups are amine-reactive rial and an anti-adhesion binding agent. The Substrate mate groups. The amine-reactive groups may contain an electro rial is preferably collagen and the binding agent preferably philically reactive carbonyl group Susceptible to nucleo comprises at least one tissue-reactive functional group and at philic attack by a primary or Secondary amine. The amine least one Substrate-reactive functional group. reactive groups may be Selected from carboxylic acid esters, acid chloride groups, anhydrides, ketones, aldehydes, halo, 0005 U.S. Pat. No. 5,614.587 to Rhee et al. describes isocyanato, thioisocyanato, epoxides, activated hydroxyl bioadhesives that comprise collagen that is crosslinked using groups, olefins, carboxyl, Succinimidyl ester, SulfoSuccinim a multifunctionally activated Synthetic hydrophilic polymer. idyl ester, maleimido, epoxy, and etheneSulfonyl. 0006 U.S. Pat. No. 5,874,500 to Rhee et al. describes a crosslinked polymer composition that comprises one com 0011. In yet another embodiment of the homogeneous dry ponent having multiple nucleophilic groups and another powder composition, the nucleophilic groups are Sulfhydryl component having multiple electrophilic groups. Covalent groupS and the electrophilic groups are Sulfhydryl-reactive bonding of the nucleophilic and electrophilic groups forms groups. The Sulflhydryl-reactive groups may be Selected So as to form a thioester, imido-thioester, thioether, or disulfide a three dimensional matrix that has a variety of medical uses linkage upon reaction with the Sulfhydryl groups. Where the including tissue adhesion, Surface coatings for Synthetic Sulflhydryl-reactive groups form a disulfide linkage, they implants, and drug delivery. More recent developments may have the structure -S-S-Ar where Ar is a substi include the addition of a third component having either tuted or urisubstituted nitrogen-containing heteroaromatic nucleophilic or electrophilic groups, as is described in U.S. moiety or a non-heterocyclic aromatic group Substituted Pat. No. 6,458,889 to Trollsas et al. with an electron-withdrawing moiety. Where the sulfhydryl 0007. However, in spite of the advances in the art, there reactive groups form a thioether linkage, they may be remains a need for improved crosslinked biomaterials that Selected from maleimido, Substituted maleimido, haloalkyl, are easy to use and Store. This need, as well as others, is met epoxy, imino, aziridino, olefins, and C.B-unsaturated alde US 2005/0281883 A1 Dec. 22, 2005 hydes and ketones. The Sulfhydryl-reactive groups may be hydrophilic polymer may be selected from poly(methylox Selected from mixed anhydrides, ester derivatives of phos azoline) and poly(ethyloxazoline). phorus, ester derivatives of p-nitrophenol, p-nitrothiophenol and pentafluorophenol; esters of Substituted hydroxy 0016. Where the core is a hydrophobic polymer selected, lamines, including N-hydroxyphthalimide esters, N-hydrox the core may be selected from polylactic acid and polygly ySuccinimide esters, N-hydroxySulfoSuccinimide esters, and colic acid. N-hydroxyglutarimide esters, esters of 1-hydroxybenzotria 0017 Where the core is a C- hydrocarbyl, the core may zole, 3-hydroxy-3,4-dihydro-benzotriazin-4-one; 3-hy be selected from alkanes, diols, polyols, and polyacids. droxy-3,4-dihydro-quinazoline-4-One, carbonylimidazole 0018 Where the core is a heteroatom-containing C derivatives, acid chlorides, ketenes, and isocyanates. hydrocarbyl, the core may be selected from di- and poly 0012. In still another embodiment of the homogeneous electrophiles. dry powder composition, the number of nucleophilic groups 0019. In another embodiment of the homogeneous dry in the mixture is approximately equal to the number of powder composition, the first component has the Structure of electrophilic groups in the mixture. For example, the ratio of moles of nucleophilic groups to moles of electrophilic formula (I) groups may be about 2:1 to 1:2, with a ratio of 1:1 preferred. 0013 In a further embodiment of the homogeneous dry 0020 and the second component has the structure of powder composition, the core is Selected from hydrophilic formula (II) polymers, hydrophobic polymers, amphiphilic polymers, C. hydrocarbyls, and heteroatom-containing C-1 hydro 0021 wherein m and n are integers from 2-12 and carbyls. m+n>4; R and R' are independently selected from 0.014 Where the core is a hydrophilic polymer, the core hydrophilic polymers, hydrophobic polymers, may be a Synthetic or naturally occurring hydrophilic poly amphiphilic polymers, C. hydrocarbyls, and het mer. The hydrophilic polymer may be a linear, branched, eroatom-containing C. hydrocarbyls, X is a dendrimeric, hyperbranched, or Star polymer. The hydro nucleophilic group; Y is an electrophilic group; L' philic polymer may be selected from polyalkylene oxides, and L are linking groups; and p and q are integers polyols; poly(oxyalkylene)-Substituted diols and polyols; from 0-1. The components may inter-react to form polyoxyethylated Sorbitol; polyoxyethylated glucose, poly covalent bonds, noncovalent bonds, or both. Nonco (acrylic acids) and analogs and copolymers thereof; poly Valent bonds include ionic bonds, hydrogen bonds, maleic acids; polyacrylamides; poly(olefinic alcohols); or the association of hydrophobic molecular Seg poly(N-vinyl lactams); polyoxazolines; polyvinylamines; ments. In one preferred embodiment, all of the and copolymers thereof. The hydrophilic polymer may also molecular segments are the same. be selected from proteins, carboxylated polysaccharides, 0022. The homogeneous dry powder composition may aminated polysaccharides, and activated polysaccharides, further comprise a biologically active agent with or without Such as, for example, collagen and glycosaminoglycans. a pharmaceutically acceptable carrier. The pharmaceutically 0.015 Where the hydrophilic polymer is a polyalkylene acceptable carrier may be a micelle, a microSphere, or a oxide or polyols, the hydrophilic polymer may be Selected nanosphere. from polyethylene glycol and poly(ethylene oxide)- 0023. Where the pharmaceutically acceptable carrier is a poly(propylene oxide) copolymers. Where the hydrophilic microSphere or a nanosphere, the pharmaceutically accept polymer is a polyols, the hydrophilic polymer may be able carrier may be a degradable polymer, Such as a poly Selected from glycerol, polyglycerol and propylene glycol. ester, and the polyester may be a glycolide/lactide copoly Where the hydrophilic polymer is a poly(oxyalkylene)- mer. The degradable polymer may also be comprised of Substituted polyol, the hydrophilic polymer may be Selected residues of one or more monomerS Selected from the group from mono-, di- and tri-polyoxyethylated glycerol, mono consisting of lactide, lactic acid, glycolide, glycolic acid, and di-polyoxyethylated propylene glycol, and mono- and e-caprolactone, gamma-caprolactone, hydroxyvaleric acid, di-polyoxyethylated trimethylene glycol. Where the hydro hydroxybutyric acid, beta-butyrolactone, gamma-butyrolac philic polymer is a poly(acrylic acid), analog or copolymer tone, gamma-Valerolactone, y-decanolactone, Ö-decanolac thereof, the hydrophilic polymer may be selected from tone, trimethylene carbonate, 1,4-dioxane-2-one or 1,5-di poly(acrylic acid), poly(methacrylic acid), poly(hydroxyeth oxepan-2one.). ylmethacrylate), poly(hydroxyethylacrylate), poly(methyla lkylsulfoxide acrylates), and poly(methylalkylsulfoxide 0024. The homogeneous dry powder composition may methacrylates). Where the hydrophilic polymer is a poly further comprise a biologically active agent. acrylamide, the hydrophilic polymer may be Selected from 0025. In one embodiment of the invention, the homoge polyacrylamide, poly(methacrylamide), poly(dimethylacry neous dry powder composition further comprises a biologi lamide), poly(N-isopropylacrylamide), and copolymers cally active agent that is an anti-fibrotic agent. AS used in the thereof. Where the hydrophilic polymer is a poly(olefinic homogeneous dry powder composition, the anti-fibrotic alcohol), the hydrophilic polymer may be selected from agent may be used to inhibit any of the following: cell poly(vinyl alcohols) and copolymers thereof. Where the regeneration, angiogenesis, fibroblast migration, fibroblast hydrophilic polymer is a poly(N-vinyl lactam), the hydro proliferation, deposition of extracellular matrix, tissue philic polymer may be selected from poly(Vinyl pyrroli remodeling, adenosine deaminase, purine ring Synthesis, dones), poly(Vinyl caprolactams), and copolymers thereof. dihydrofolate reduction, ribonucleotide Synthesis or func Where the hydrophilic polymer is a polyoxazoline, the tion, thymidine monophosphate Synthesis or function, DNA US 2005/0281883 A1 Dec. 22, 2005

Synthesis, protein Synthesis, and microtubule function. The rollide antibiotic; a GPIb/IIIa receptor antagonist; an endot anti-fibrotic agent may also be used to block thymidine helin receptor antagonist; a peroxisome proliferator-acti monophosphate, to cause DNA damage, and to cause DNA Vated receptor agonist, an estrogen receptor agent, a adduct formation. Somastostatin analogue; a neurokinin 1 antagonist; a neuro kinin 3 antagonist; a VLA-4 antagonist; an osteoclast inhibi 0026. Any of the following anti-fibrotic agents may be tor; a DNA topoisomerase ATP hydrolyzing inhibitor; an used in the homogeneous dry powder composition: an angiotensin 1 converting enzyme inhibitor; an angiotensin II angiogenesis inhibitor, a 5-lipoxygenase inhibitor or antago antagonist, an enkephalinase inhibitor; a peroxisome prolif nist; a chemokine receptor antagonist; a cell cycle inhibitor; erator-activated receptor gamma agonist insulin Sensitizer, a a taxane, an anti-microtubule agent, paclitaxel, an analogue protein kinase C inhibitor; a ROCK (rho-associated kinase) or derivative of paclitaxel, a Vinca alkaloid; camptothecin or inhibitor; a CXCR3 inhibitor; Itk inhibitor; a cytosolic an analogue or derivative thereof; a podophyllotoxin, phospholipase A-alpha inhibitor, a PPAR agonist, an wherein the podophyllotoxin may be an etoposide or an immunosuppreSSant; an Erb inhibitor; an apoptosis agonist; analogue or derivative thereof; an anthracycline, wherein the a lipocortin agonist, a VCAM-1 antagonist; a collagen anthracycline may be doxorubicin or an analogue or deriva antagonist, an alpha 2 integrin antagonist, a TNF alpha tive thereof or the anthracycline may be mitoxantrone or an analogue or derivative thereof; a platinum compound; a inhibitor; a nitric oxide inhibitor; and a cathepsin inhibitor. nitroSourea; a nitroimidazole; a folic acid antagonist; a 0027. In another embodiment of the invention, the homo cytidine analogue; a pyrimidine analogue, a fluoropyrimi geneous dry powder composition further comprises a bio dine analogue; a purine analogue; a nitrogen mustard or an logically active agent that is a fibrosing agent. AS used in the analogue or derivative thereof; a hydroxyurea; a mytomicin homogeneous dry powder composition, the anti-fibrotic or an analogue or derivative thereof, an alkyl Sulfonate, a agent may be used to promote any of the following, regen benzamide or an analogue or derivative thereof, a nicotina eration; angiogenesis, fibroblast migration; fibroblast prolif mide or an analogue or derivative thereof; a halogenated eration; deposition of extracellular matrix (ECM); and tissue Sugar or an analogue or derivative thereof; a DNA alkylating remodeling. The fibrosing agent may also be used as an agent; an anti-microtubule agent; a topoisomerase inhibitor; arterial vessel wall irritant. a DNA cleaving agent; an antimetabolite; a nucleotide interconversion inhibitor; a hydroorotate dehydrogenase 0028. Fibrosing agents that may be used in the homoge inhibitor; a DNA intercalation agent; an RNA synthesis neous dry powder composition may be or may be comprised inhibitor; a pyrimidine synthesis inhibitor; a cyclin depen of Silk; silkworm Silk; Spider Silk; recombinant Silk, raw silk, dent protein kinase inhibitor; an epidermal growth factor hydrolyzed silk, acid-treated Silk, acylated Silk; mineral kinase inhibitor; an elastase inhibitor, a factor Xa inhibitor; particles, talc, chitosan, polylysine, fibronectin; bleomycin; a farnesyltransferase inhibitor; a fibrinogen antagonist, a or CTGF. The fibrosing agent may also be in the form of a guanylate cyclase Stimulant; a heat shock protein 90 antago particulate, which may be a biodegradable particulate or a nist, which may be a geldanamycin or an analogue or non-biodegradable particulate. Biodegradable particulates derivative thereof, a guanylate cyclase Stimulant; a HMG may be comprised of a material Selected from the group CoA reductase inhibitor, which may be Simvastatin or an consisting of polyester, polyanhydride, poly(anhydride analogue or derivative thereof; an IKK2 inhibitor; an IL-1 ester), poly(ester-amide), poly(ester-urea), polyorthoester, antagonist, an ICE antagonist; an IRAKantagonist; an IL-4 polyphosphoester, polyphosphazine, polycyanoacrylate, agonist; an immunomodulatory agent, Sirolimus or an ana collagen, chitosan, hyaluronic acid, chromic cat gut, algi logue or derivative thereof; everolimus or an analogue or nate, Starch, cellulose and cellulose ester. Non-biodegrad derivative thereof, tacrolimus or an analogue or derivative able particulates may be comprised of a material Selected thereof; biolmus or an analogue or derivative thereof, tres from the group consisting of polyester, polyurethane, Sili perimus or an analogue or derivative thereof; auranofin or an cone, polyethylene, polypropylene, polystyrene, polyacry analogue or derivative thereof., 27-0-demethylrapamycin or late, polymethacrylate, and Silk. Examples of preferred an analogue or derivative thereof, gusperimus or an ana particulates may be a particulate form of a member Selected logue or derivative thereof; pimecrolimus or an analogue or from the group consisting of Silk, talc, Starch, glass, Silicate, derivative thereof; ABT-578 or an analogue or derivative Silica, calcium phosphate, calcium Sulfate, calcium carbon thereof, an inosine monophosphate dehydrogenase ate, hydroxyapatite, Synthetic mineral, polymethylmethacry (IMPDH) inhibitor, which may be mycophenolic acid or an late, Silver nitrate, ceramic and other inorganic particles. analogue or derivative thereof or 1-alpha-25 dihydroxy 0029. In a further embodiment of the homogeneous dry Vitamin D or an analogue or derivative thereof, a leukot powder composition, the biologically active agent promotes riene inhibitor; an MCP-1 antagonist; an MMP inhibitor; an bone growth. Within this embodiment, the fibrosing agent NF kappa B inhibitor, which may be Bay 11-7082; an NO may promote the bone growth. Fibrosing agents that may antagonist; a p38 MAP kinase inhibitor, which may be SB promote bone growth may include a bone morphogenic 202190; a phosphodiesterase inhibitor; a TGFbeta inhibitor; protein and an osteogenic growth factor, the latter which a thromboxane A2 antagonist; a TNF alpha antagonist; a may be Selected from transforming growth factor, platelet TACE inhibitor; a tyrosine kinase inhibitor; vitronectin derived growth factor, and fibroblast growth factor. inhibitor; a fibroblast growth factor inhibitor; a protein kinase inhibitor; a PDGF receptor kinase inhibitor; an endot 0030. In another embodiment of the invention, the homo helial growth factor receptor kinase inhibitor; a retinoic acid geneous dry powder composition with a fibrosing agent receptor antagonist; a platelet derived growth factor receptor further comprises a pharmaceutical agent that induces Scle kinase inhibitor, a fibrinogen antagonist; an antimycotic rosis (a Sclerosant), wherein the Sclerosant may be a Surfac agent, Sulconizole, a bisphosphonate; a phospholipase A1 tant or it may be selected from the group consisting of inhibitor; a histamine H1/H2/H3 receptor antagonist; a mac ethanol, dimethyl Sulfoxide, Sucrose, Sodium chloride, dex US 2005/0281883 A1 Dec. 22, 2005

trose, glycerin, minocycline, tetracycline, doxycycline, poli acid residues Selected from the group consisting of amino docanol, Sodium tetradecyl Sulfate, Sodium morrhuate, and acids comprising primary amine groups and amino acids Sotradecol. comprising thiol groups, the Second component comprises a polyethylene glycol moiety, the electrophilic groups are 0031. In a further embodiment of the invention, the Succinimidyl moieties, and each of the first and Second homogeneous dry powder composition with a fibrosing crosslinkable components is biocompatible, Synthetic, and agent further comprises an inflammatory cytokine, which nonimmunogenic, and further whererin crosslinking of the may be selected from the group consisting of TGFB, PDGF, composition results in a biocompatible, nonimmunogenic, VEGF, bFGF, TNFo, NGF, GM-CSF, IGF-a, IL-1, IL-1-?3, IL-8, IL-6, and growth hormone. crosslinked matrix. 0032. In still another embodiment of the invention, the 0040 Any of the following are preferred embodiments of homogeneous dry powder composition with a fibrosing the croSSlinkable composition described immediately above: agent further comprises an agent that Stimulates cell prolif m>3, m=3, m=4, n=4, the electrophilic groups are Succin eration, which may be selected from the group consisting of imidyl moieties, all n are identical, and all m are identical. dexamethasone, isotretinoin (13-cis retinoic acid), 17-f- 0041. In one preferred embodiment, the selected amino estradiol, estradiol, 1-O-25 dihydroxyvitamin D, diethyl acid residues are lysine. Within this embodiment, any of the Stibesterol, cyclosporine A, L-NAME, all-trans retinoic acid following is preferred: m>3, m=3, m=4, n=4, the electro (ATRA), and analogues and derivatives thereof. philic groups are Succinimidyl moieties, all n are identical, 0033. In a further embodiment of the homogeneous dry and all m are identical. powder composition, the biologically active agent is mixed 0042. In another preferred embodiment, the selected with the first and Second components to form a mixture. amino acid residues are cysteine. Within this embodiment, 0034. In another embodiment of the homogeneous dry any of the following is preferred: m>3, m=3,m=4, n=4, the powder composition, the biologically active agent is chemi electrophilic groups are Succinimidyl moieties, all n are cally coupled to the first component or to the Second identical, and all m are identical. component. 0043 Still another aspect of the invention relates to a 0.035 Another aspect of the invention relates to a crosslinkable composition comprised of: (a) a first crosslink crosslinkable composition comprised of: (a) a first crosslink able component having m nucleophilic groups, wherein able component having m nucleophilic groups, wherein m22; and (b) a second crosslinkable component having in me2; and (b) a Second crosslinkable component having in electrophilic groups capable of reaction with the m nucleo electrophilic groups capable of reaction with the m nucleo philic groups to form covalent bonds, wherein ne2 and philic groups to form covalent bonds, wherein ne2 and m+ne5, the first component comprises two or more amino m+ne5, the first component comprises two or more amino acid residues Selected from the group consisting of amino acid residues Selected from the group consisting of amino acids comprising primary amine groups and amino acids acids comprising primary amine groupS and amino acids comprising thiol groups, the Second component comprises a comprising thiol groups, the Second component comprises a multifunctionally activated polyethylene glycol, and each of polyethylene glycol moiety, and each of the first and Second the first and Second crosslinkable components is biocom crosslinkable components is biocompatible, Synthetic, and patible, Synthetic, and nonimmunogenic, and further nonimmunogenic, and further wherein crosslinking of the wherein crosslinking of the composition results in a bio composition results in a biocompatible, nonimmunogenic, compatible, nonimmunogenic, crosslinked matrix. crosslinked matrix. 0044 Any of the following are preferred embodiments of 0036) Any of the following are preferred embodiments of the croSSlinkable composition described immediately above: the crosslinkable composition described immediately above: m>3, m=3, m=4, n=4, the electrophilic groups are Succin m>3, m=3, m=4, n=4, the electrophilic groups are Succin imidyl moieties, all n are identical, all m are identical, the imidyl moieties, all n are identical, and all m are identical. multifunctionally activated polyethylene glycol is tetrafunc 0037. In one preferred embodiment, the selected amino tionally activated polyethylene glycol, and the multifunc acid residues are lysine. Within this embodiment, any of the tionally activated polyethylene glycol is a Star-branched following is preferred: m>3, m=3, m=4, n=4, the electro polyethylene glycol. philic groups are Succinimidyl moieties, all n are identical, 0045. In one preferred embodiment, the selected amino and all m are identical. acid residues are lysine. Within this embodiment, any of the 0.038. In another preferred embodiment, the selected following is preferred: m>3, m=3, m=4, n=4, the electro amino acid residues are cysteine. Within this embodiment, philic groups are Succinimidyl moieties, all n are identical, any of the following is preferred: m>3, m=3,m=4, n=4, the all m are identical, and the multifunctionally activated electrophilic groups are Succinimidyl moieties, all n are polyethylene glycol is tetrafunctionally activated polyethyl identical, and all m are identical. ene glycol or the multifunctionally activated polyethylene glycol is a Star-branched polyethylene glycol. 0039. Yet another aspect of the invention relates to a crosslinkable composition comprised of: (a) a first crosslink 0046. In another preferred embodiment, the selected able component having m nucleophilic groups, wherein amino acid residues are cysteine. Within this embodiment, me2; and (b) a Second crosslinkable component having in any of the following is preferred: m>3, m=3,m=4, n=4, the electrophilic groups capable of reaction with the m nucleo electrophilic groups are Succinimidyl moieties, all n are philic groups to form covalent bonds, wherein ne2 and identical, all m are identical, and the multifunctionally m+ne5, the first component comprises two or more amino activated polyethylene glycol is tetrafunctionally activated US 2005/0281883 A1 Dec. 22, 2005 polyethylene glycol or the multifunctionally activated poly 11.0 to the homogeneous Solution to form a mixture; and (c) ethylene glycol is a Star-branched polyethylene glycol. placing the mixture into contact with tissue and allowing a 0047 Another aspect of the invention relates to a method three-dimensional matrix to form on the tissue. A preferred of forming a three-dimensional matrix comprising the Steps composition for use in this method is the homogeneous dry of: (a) providing a composition of the invention; and (b) powder composition. rendering the nucleophilic and electrophilic groups reactive by exposing the composition to an aqueous environment to 0052 A further aspect of the invention relates to a effect inter-reaction; wherein said exposure comprises: (i) method of forming a three-dimensional matrix on a Surface dissolving the composition in a first buffer Solution having of a device comprising the Steps of: (a) providing a com a pH within the range of about 1.0 to 5.5 to form a position of the invention; and (b) rendering the nucleophilic homogeneous Solution, and (ii) adding a second buffer and electrophilic groups reactive by exposing the composi solution having a pH within the range of about 6.0 to 11.0 tion to an aqueous environment to effect inter-reaction; to the homogeneous Solution; and (c) allowing a three wherein said exposure comprises: (i) dissolving the compo dimensional matrix to form. A preferred composition for use Sition in a first buffer Solution having a pH within the range in this method is the homogeneous dry powder composition. of about 1.0 to 5.5 to form a homogeneous solution, and (ii) The three-dimensional matrix of the invention described adding a Second buffer Solution having a pH within the range immediately above may be formed without input of any of about 6.0 to 11.0 to the homogeneous solution; and external energy or by polymerization. applying the homogeneous Solution to a Surface of a device; and allowing the three-dimensional matrix to form. A pre 0.048. In a preferred embodiment, the pH of the first ferred composition for use in this method is the homoge buffer solution is selected to retard the reactivity of the neous dry powder composition. nucleophilic groups on the first component by rendering the nucleophilic groups relatively non-nucleophilic. In this pre 0053 Another aspect of the invention relates to a method ferred embodiment, the second buffer Solution neutralizes of preventing Scarring in the vicinity of a medical implant the effect of the first buffer solution, so that the nucleophilic comprising the steps of: (a) providing a composition of the groups of the first component regain their nucleophilic invention; (b) rendering the nucleophilic and electrophilic character and inter-react with the electrophilic groups of the groups reactive by exposing the composition to an aqueous Second component. environment to effect inter-reaction; wherein Said exposure comprises: (i) dissolving the composition in a first buffer 0049. In another preferred embodiment, the composition, solution having a pH within the range of about 1.0 to 5.5 to first buffer Solution and second buffer Solution are housed form a homogeneous Solution, and (ii) adding a second Separately in a multiple-compartment Syringe System having buffer solution having a pH within the range of about 6.0 to a multiple barrels, a mixing head, and an exit orifice, Step 11.0 to the homogeneous Solution to form a mixture; and (b)(i) comprises adding the first buffer solution to the barrel applying the mixture to a Surface of a medical implant and housing the composition to dissolve the composition and allowing a three-dimensional matrix to form on the Surface form a homogeneous Solution, and extruding the homoge of the medical implant; and (d) placing the medical implant neous Solution into the mixing head; step (b)(ii) comprises into an animal host, wherein release of the anti-fibrotic agent Simultaneously extruding the Second buffer Solution into the from the composition inhibits Scarring in the animal host. In mixing head; and step (c) further comprises extruding the one embodiment, the anti-fibrotic agent is released into resulting composition through the orifice onto a Surface. tissue in the vicinity of the implant after deployment of the 0050 Yet another aspect of the invention relates to a implant. A preferred composition for use in this method is method of Sealing tissue of a patient comprising the Steps of: the homogeneous dry powder composition with an anti (a) providing a composition of the invention; (b) rendering fibrotic agent. the nucleophilic and electrophilic groups reactive by expos 0054 Yet another aspect of the invention relates to a ing the composition to an aqueous environment to effect method of promoting Scarring in the vicinity of a medical inter-reaction; wherein said exposure comprises: (i) dissolv implant comprising the steps of: (a) providing a composition ing the composition in a first buffer Solution having a pH of the invention; (b) rendering the nucleophilic and electro within the range of about 1.0 to 5.5 to form a homogeneous philic groups reactive by exposing the composition to an Solution, and (ii) adding a Second buffer Solution having a aqueous environment to effect inter-reaction, wherein Said pH within the range of about 6.0 to 11.0 to the homogeneous exposure comprises: (i) dissolving the composition in a first Solution to form a mixture; and (c) placing the mixture into buffer solution having a pH within the range of about 1.0 to contact with tissue and allowing a three-dimensional matrix 5.5 to form a homogeneous Solution, and (ii) adding a to form and Seal the tissue. A preferred composition for use Second buffer Solution having a pH within the range of about in this method is the homogeneous dry powder composition. 6.0 to 11.0 to the homogeneous Solution; and (c) applying 0051 Still another aspect of the invention relates to a the mixture to a Surface of a medical implant and allowing method of preventing adhesions between tissueS of a patient a three-dimensional matrix to form on the Surface of the comprising the steps of: (a) providing a composition of the medical implant; and (d) placing the medical implant into an invention; (b) rendering the nucleophilic and electrophilic animal host, wherein release of the fibrotic agent from the groups reactive by exposing the composition to an aqueous matrix inhibits Scarring in the animal host. In a preferred environment to effect inter-reaction; wherein Said exposure embodiment, the the fibrotic agent is released into tissue in comprises: (i) dissolving the composition in a first buffer the vicinity of the implant after deployment of the implant. solution having a pH within the range of about 1.0 to 5.5 to A preferred composition for use in this method is the form a homogeneous Solution, and (ii) adding a second homogeneous dry powder composition with a fibrosing buffer solution having a pH within the range of about 6.0 to agent. US 2005/0281883 A1 Dec. 22, 2005

0.055 A further aspect of the invention relates to a kit for buffer or as a solution with the second buffer. The kit may use in medical applications, comprising: (a) a homogeneous further comprise a pharmaceutically acceptable carrier as a dry powder composition comprised of: (i) a first component fourth component. The biologically active agent is packaged having a core Substituted with m nucleophilic groups, where with the pharmaceutically acceptable carrier. me2; and (ii) a second component having a core Substituted with n electrophilic groups, where ne2 and m+n>4, wherein 0060 Yet another embodiment of the kit for use in the nucleophilic and electrophilic groups are non-reactive in medical applications further comprises living cells or genes, a dry environment but are rendered reactive upon exposure and the medical application involves delivering the living to an aqueous environment Such that the components inter cells or genes. react in the aqueous environment to form a three-dimen 0061. Other medical applications that the kit may be used sional matrix; (b) a first buffer solution having a pH within for include adhering or Sealing biological tissue, bioadhe the range of about 1.0 to 5.5; and (c) a second buffer solution Sion, ophthalmic applications, tissue augmentation, adhe having a pH within the range of about 6.0 to 11.0; wherein Sion prevention, forming a Synthetic implant or coating a each component is packaged Separately and admixed imme Synthetic implant, treatment of aneurysms, and laparoscopic diately prior to use. It is preferred of course that prior to use, procedures. each component is in a separate Sterile package. 0062 Still another aspect of the invention relates to a kit 0056. Another aspect of the invention relates to a kit for for use in for use in medical applications, comprising: (a) a use in medical applications, comprising: (a) a composition first component having a core Substituted with m nucleo of the invention; (b) a first buffer solution having a pH philic groups, where me2; (b) a Second component having within the range of about 1.0 to 5.5; and (c) a second buffer a core Substituted with n electrophilic groups, where ne2 solution having a pH within the range of about 6.0 to 11.0, and m+n>4; (c) a first buffer solution having a pH within the wherein each component is packaged separately and range of about 1.0 to 5.5; and (d) a second buffer solution admixed immediately prior to use. A preferred composition having a pH within the range of about 6.0 to 11.0, wherein of the invention for use in this kit is the homogeneous dry the nucleophilic and electrophilic groups are non-reactive in powder composition. It is preferred that each component of a dry environment but are rendered reactive upon exposure the kit is in a separate Sterile package. to an aqueous environment Such that the components inter react in the aqueous environment to form a three-dimen 0057 The kit may further comprise a delivery device, Sional matrix and further wherein each component is pack which in one embodiment, may be a multi-compartment aged separately and admixed immediately prior to use. device. A preferred multi-compartment device of the inven tion is a multiple-compartment Syringe System having mul 0063. These and other aspects of the present invention are tiple barrels, a mixing head, and an exit orifice. Where the described in detail below. kit is a multiple-compartment Syringe System, the homoge neous dry powder composition, the first buffer Solution, and BRIEF DESCRIPTION OF THE DRAWINGS the Second buffer Solution are housed Separately in the 0064 FIG. 1 depicts a preferred multi-compartment multiple-compartment Syringe System. Syringe device of the present invention. 0.058. In another embodiment of the invention, the deliv 0065 FIGS. 2 and 3 schematically illustrate an embodi ery device is a pressurized delivery System. A preferred ment of the pressurized delivery device of the present preSSurized delivery System comprises: a plurality of fluid invention that includes a cap having an interior diffuser component inlets each adapted to communicate with a Surface and a lumen assembly for delivering fluid compo Source of different fluid components, at least one carrier fluid nents and a pressurized carrier fluid to the diffuser Surface. inlet adapted to communicate with a Source of a pressurized FIG. 1 depicts the device in exploded view and FIG. 2 carrier fluid; a diffuser Surface located downstream from the depicts the interior diffuser Surface of the cap. plurality of fluid component inlets and the at least one carrier fluid inlet; and an outlet extending through the diffuser DETAILED DESCRIPTION OF THE Surface, wherein the diffuser Surface is adapted to receive INVENTION fluid components thereon and has a shape effective to direct and maintain each received fluid component in a different 0.066 I. Definitions and Nomenclature flow path toward the outlet for mixing and dispensing 0067. Before describing the present invention in detail, it therethrough by the pressurized carrier fluid from the at least is to be understood that unless otherwise indicated this one carrier fluid inlet. Within this embodiment, a preferred invention is not limited to particular compositional forms, preSSurized carrier fluid is pressurized air and the preferred crosslinkable components, crosslinking techniques, or meth fluid components are the first buffer solution and the second ods of use, as Such may vary. It is also to be understood that buffer Solution of the invention. the terminology used herein is for the purpose of describing 0059 Another embodiment of the kit for use in medical particular embodiments only, and is not intended to be applications further comprises a biologically active agent limiting. and the medical application involves delivering the biologi 0068. It must be noted that, as used in this specification cally active agent. The biologically active agent may be and the appended claims, the Singular forms “a,”“an,” and packaged with the homogeneous dry powder composition “the' include plural referents unless the context clearly and may further comprise a pharmaceutically acceptable dictates otherwise. Thus, for example, "a multifunctional carrier packaged with the biologically active agent and the compound” refers not only to a Single multifunctional com homogeneous dry powder composition. The biologically pound but also to a combination of two or more of the same active agent may also be packaged as a Solution with the first or different multifunctional compounds, “a reactive group' US 2005/0281883 A1 Dec. 22, 2005

refers to a combination of reactive groups as well as to a ), as well as the thiol analogs thereof. All Suitable polymers Single reactive group, and the like. herein are biocompatible and non-immunogenic. The poly mers can be degradable or non-degradable. In a preferred 0069. Unless defined otherwise, all technical and scien mode, the polymers will be essentially non-degradable in tific terms used herein have the meaning commonly under Vivo over a period of at least Several months. stood by one of ordinary skill in the art to which the invention pertains. Although any methods and materials 0073. The term “synthetic' is used to refer to polymers, Similar or equivalent to those described herein may be useful compounds and other Such materials that are “chemically in the practice or testing of the present invention, preferred Synthesized.” For example, a Synthetic material in the methods and materials are described below. All patents, present compositions may have a molecular Structure that is patent applications and other publications mentioned herein identical to a naturally occurring material, but the material are incorporated herein by reference. Specific terminology perse, as incorporated in the compositions of the invention, of particular importance to the description of the present has been chemically Synthesized in the laboratory or indus invention is defined below. trially. “Synthetic' materials also include semi-synthetic materials, i.e., naturally occurring materials, obtained from 0070 The terms “inter-react” and “inter-reaction” as used a natural Source, that have been chemically modified in Some herein refer to the formation of covalent bonds, noncovalent way. Generally, however, the Synthetic materials herein are bonds, or both. The term thus includes crosslinking, which purely Synthetic, i.e., they are neither Semi-Synthetic nor involves both intermolecular crosslinks and optionally have a structure that is identical to that of a naturally intramolecular crosslinks as well, arising from the formation occurring material. of covalent bonds. Entanglement is another example of non-covalent bonding that may result after inter-reaction 0074 The term “effective amount” refers to the amount between two or more reactive groups. Covalent bonding of composition required in order to obtain the effect desired. between two reactive groups may be direct in which case an For example, a “tissue growth-promoting amount of a atom in reactive group is directly bound to an atom in the composition refers to the amount needed in order to Stimu other reactive group or it may be indirect through a linking late tissue growth to a detectable degree. TiSSue, in this group. Noncovalent bonds include ionic (electrostatic) context, includes connective tissue, bone, cartilage, epider bonds, hydrogen bonds, or the association of hydrophobic mis and dermis, blood, and other tissues. The actual amount molecular Segments, which may be the same or different. A that is determined to be an effective amount will vary crosslinked matrix may, in addition to covalent bonds, also depending on factorS Such as the size, condition, Sex and age include Such intermolecular and/or intramolecular noncova of the patient and can be more readily determined by the lent bonds. caregiver. 0071. When referring to polymers, the terms “hydro 0075. The term “in situ” as used herein means at the site philic' and “hydrophobic' are generally defined in terms of of administration. Thus, compositions of the invention can an HLB value, i.e., a hydrophilic lipophilic balance. A high be injected or otherwise applied to a specific Site within a HLB value indicates a hydrophilic compound, while a low patient's body, e.g., a Site in need of augmentation, and HLB value characterizes a hydrophobic compound. HLB allowed to crosslink at the site of injection. Suitable sites values are well known in the art, and generally range from will generally be intradermal or Subcutaneous regions for 1 to 18. Preferred multifunctional compound cores are augmenting dermal Support, at a bone fracture Site for bone hydrophilic, although as long as the multifunctional com repair, within Sphincter tissue for Sphincter augmentation pound as a whole contains at least one hydrophilic compo (e.g., for restoration of continence), within a wound or nent, crosslinkable hydrophobic components may also be Suture, to promote tissue regrowth; and within or adjacent to present. vessel anastomoses, to promote vessel regrowth. 0.072 The term “polymer' is used not only in the con 0076. The term “acqueous medium' includes solutions, ventional Sense to refer to molecules composed of repeating Suspensions, dispersions, colloids, and the like containing monomer units, including homopolymers, block copoly water. The term "aqueous environment’ means an environ mers, random copolymers, and graft copolymers, but also ment containing an aqueous medium. Similarly, the term refers to polyfunctional Small molecules that do not contain “dry environment’ means an environment that does not repeating monomer units but are "polymeric' in the Sense of contain an aqueous medium. being "polyfunctional, i.e., containing two or more func 0077. The terms “active agent,”“biologically active tional groups. Accordingly, it will be appreciated that when agent,”“therapeutic agent,”“pharmacologically active the term “polymer' is used, difunctional and polyfunctional agent,” and "drug are used interchangeably herein to refer Small molecules are included. Such moieties include, by way of example: the difunctional electrophiles disuccinimidyl to a chemical material or compound Suitable for adminis suberate (DSS), bis(sulfosuccinimidyl) suberate (BS), tration to a patient and that induces a desired effect. The dithiobis(Succinimidylpropionate) (DSP), bis(2-succinimi terms include agents that are therapeutically effective as well dooxy-carbonyloxy) ethylsulfone (BSOCOES), 3,3'-dithio as prophylactically effective. Also included are derivatives bis(sulfosuccinimidylpropionate (DTSSP); and the di- and and analogs of those compounds or classes of compounds polyfunctional nucleophiles ethylenediamine (HN Specifically mentioned that also induce the desired effect. CH-CH-NH), tetramethylene diamine (HN-CH 0078. As used herein the terms “active agent,”“biologi -NH), pentamethylene diamine (cadaverine) (HN cally active agent,”“therapeutic agent,”“pharmacologically CH-NH), hexamethylene diamine (HN-CH active agent,” and "drug” refer to an organic molecule that NH), bos(2-aminoethyl)amine (HN-CH-CH-NH) exerts biological effects in Vivo. For purposes of this dis ), and tris (2-aminoethyl)amine (N-CH-CH-NH) cussion, the term “biologically active agent' is used, with US 2005/0281883 A1 Dec. 22, 2005

the understanding that the use of this term does not exclude with disease, delivering therapeutic agents, and/or repairing the application to the remaining terms. Examples of bio or replacing or augmenting damaged or diseased organs and logically active agents include, by way of example and not tissues. While normally composed of biologically compat limitation, enzymes, receptor antagonists or agonists, hor ible Synthetic materials (e.g., medical-grade stainless Steel, mones, growth factors, autogenous bone marrow, antibiot titanium and other metals, polymerS Such as polyurethane, ics, antimicrobial agents and antibodies. The term biologi Silicon, PLA, PLGA and other materials) that are exogenous, cally active agent is also intended to encompass various cell Some medical devices and implants include materials types and genes that can be incorporated into the composi derived from animals (e.g., "Xenografts' Such as whole tions of the invention. Other examples of biologically active animal organs, animal tissueS Such as heart Valves, naturally agents include those that inhibit fibrosis and those that occurring or chemically-modified molecules Such as col promote fibrosis. In certain embodiments, a biologically lagen, hyaluronic acid, proteins, carbohydrates and others), active agent may promote adhesion between a tissue and a human donors (e.g., “allografts' Such as whole organs; Substrate (e.g., a Surface of a medical device). tissues Such as bone grafts, skin grafts and others), or from 0079. “Fibrosis,”“scarring,” or “fibrotic response” refers the patients themselves (e.g., “autografts. Such as Saphenous to the formation of fibrous tissue in response to injury or vein grafts, skin grafts, tendon/ligament/muscle transplants). medical intervention. Therapeutic agents which promote 0082. With regard to nomenclature pertinent to molecular (also referred to interchangeably herein as “induce,”“stimu Structures, the following definitions apply: late,”“cause,” and the like) fibrosis or Scarring are referred to interchangeably herein as “fibrosis-inducing agents,”“S- 0083) The term “alkyl” as used herein refers to a carring agents,"fibrosing agents," adhesion-inducing branched or unbranched Saturated hydrocarbon group typi agents,” and the like, where these agents do So through one cally although not necessarily containing 1 to about 24 or more mechanisms including: inducing or promoting carbon atoms, Such as methyl, ethyl, n-propyl, isopropyl, angiogenesis, Stimulating migration or proliferation of con n-butyl, isobutyl, t-butyl, octyl, decyl, and the like, as well nective tissue cells (Such as fibroblasts, Smooth muscle cells, as cycloalkyl groupS Such as cyclopentyl, cyclohexyl and the vascular Smooth muscle cells), inducing ECM production, like. Generally, although again not necessarily, alkyl groups and/or promoting tissue remodeling. Therapeutic agents herein contain 1 to about 12 carbon atoms. The term “lower which inhibit fibrosis or scarring are referred to herein as alkyl intends an alkyl group of one to six carbon atoms, "fibrosis-inhibiting agents,”“anti-Scarring agents,” and the preferably one to four carbon atoms. “Substituted alkyl refers to alkyl substituted with one or more substituent like, where these agents inhibit fibrosis through one or more groups. “Alkylene,”“lower alkylene,” and “substituted alky mechanisms including: inhibiting angiogenesis, inhibiting lene' refer to divalent alkyl, lower alkyl, and substituted migration or proliferation of connective tissue cells (such as alkyl groups, respectively. fibroblasts, Smooth muscle cells, vascular Smooth muscle cells), reducing ECM production, and/or inhibiting tissue 0084. The term “aryl” as used herein, and unless other remodeling. wise Specified, refers to an aromatic Substituent containing a single aromatic ring (monocyclic) or multiple aromatic 0080 “Sclerosing” refers to a tissue reaction in which an rings that are fused together, linked covalently, or linked to irritant is applied locally to a tissue which results in an a common group Such as a methylene or ethylene moiety. inflammatory reaction and is followed by Scar tissue forma The common linking group may also be a carbonyl as in tion at the Site of irritation. A pharmaceutical agent that benzophenone, an oxygen atom as in diphenylether, or a induces or promotes Sclerosis is referred to as a “Sclerosant,” nitrogen atom as in diphenylamine. Preferred aryl groups or a “Sclerosing agent.” Representative examples of Sclero contain one aromatic ring or two fused or linked aromatic Sants include ethanol, dimethyl Sulfoxide, Surfactants (e.g., rings, e.g., phenyl, naphthyl, biphenyl, diphenylether, diphe TRITON X, Sorbitan monolaurate, Sorbitan sesquioleate, nylamine, benzophenone, and the like. “Substituted aryl' glycerol monoStearate and polyoxyethylene, polyoxyethyl refers to an aryl moiety Substituted with one or more ene cetyl ether, and the like), Sucrose, Sodium chloride, Substituent groups, and the terms "heteroatom-containing dextrose, glycerin, minocycline, tetracycline, doxycycline, aryl' and “heteroaryl” refer to aryl in which at least one polidocanol, Sodium tetradecyl Sulfate, Sodium morrhuate, carbon atom is replaced with a heteroatom. The terms ethanolamine, phenol, Sarapin and Sotradecol. “arylene' and “substituted arylene' refer to divalent aryland 0081) “Anti-microtubule agents' should be understood to Substituted aryl groupS as just defined. include any protein, peptide, chemical, or other molecule which impairs the function of microtubules, for example, 0085. The term "heteroatom-containing” as in a "heteroa through the prevention or Stabilization of polymerization. tom-containing hydrocarbyl group' refers to a molecule or Compounds that Stabilize polymerization of microtubules molecular fragment in which one or more carbon atoms is are referred to herein as “microtubule Stabilizing agents. A replaced with an atom other than carbon, e.g., nitrogen, wide variety of methods may be utilized to determine the oxygen, Sulfur, phosphorus or Silicon. anti-microtubule activity of a particular compound, includ 0086) “Hydrocarbyl” refers to univalent hydrocarbyl ing for example, assays described by Smith et al. (Cancer radicals containing 1 to about 30 carbon atoms, preferably 1 Lett 79(2):213–219, 1994) and Mooberry et al., (Cancer Lett. to about 24 carbon atoms, most preferably 1 to about 12 96(2):261-266, 1995). The terms “medical device,”“im carbon atoms, including branched or unbranched, Saturated plant,”“medical implant, and the like are used Synony or unsaturated Species, Such as alkyl groups, alkenyl groups, mously to refer to any object that is designed to be placed aryl groups, and the like. The term “lower hydrocarbyl” partially or wholly within a patient’s body for one or more intends a hydrocarbyl group of one to six carbon atoms, therapeutic or prophylactic purposes Such as for restoring preferably one to four carbon atoms. The term “hydrocar physiological function, alleviating Symptoms associated bylene' intends a divalent hydrocarbyl moiety containing 1 US 2005/0281883 A1 Dec. 22, 2005

to about 30 carbon atoms, preferably 1 to about 24 carbon composition and the three-dimensional matrix formed there atoms, most preferably 1 to about 12 carbon atoms, includ from. The biological System can be a biological tissue, and ing branched or unbranched, Saturated or unsaturated Spe in a preferred embodiment, is living tissue. cies, or the like. The term “lower hydrocarbylene' intends a hydrocarbylene group of one to Six carbon atoms, preferably 0092. The resulting three-dimensional matrix is useful in one to four carbon atoms. “Substituted hydrocarbyl” refers a variety of contexts, and is particularly useful as a bioma to hydrocarbyl Substituted with one or more substituent terial for medial applications, Such as for bioadhesion, groups, and the terms "heteroatom-containing hydrocarbyl” delivery of biologically active agents, tissue augmentation, and “heterohydrocarbyl” refer to hydrocarbyl in which at tissue Sealing, vascular Sealing, needle hole Sealing, hemo least one carbon atom is replaced with a heteroatom. Simi Stasis, the prevention of adhesions following a Surgical larly, “substituted hydrocarbylene' refers to hydrocarbylene procedure or injury, and So forth. Substituted with one or more Substituent groups, and the 0093. The core and reactive groups can also be selected terms "heteroatom-containing hydrocarbylene' and "hetero So as to provide components that have one of more of the hydrocarbylene' refer to hydrocarbylene in which at least following features: are biocompatible, are non-immuno one carbon atom is replaced with a heteroatom. If not genic, and do not leave any toxic, inflammatory or immu otherwise indicated, “hydrocarbyl” indicates unsubstituted nogenic reaction products at the Site of administration. hydrocarbyl, Substituted hydrocarbyl, heteroatom-contain Similarly, the core and reactive groups can also be Selected ing hydrocarbyl, and Substituted heteroatom-containing So as to provide a resulting matrix that has one or more of hydrocarbyl. Unless otherwise indicated, the terms “hydro these features. carbyl” and “hydrocarbylene' include substituted hydrocar byl and Substituted hydrocarbylene, heteroatom-containing 0094. In one embodiment of the invention, Substantially hydrocarbyl and heteroatom-containing hydrocarbylene, immediately or immediately upon exposure to the aqueous and Substituted heteroatom-containing hydrocarbyl and Sub environment, the reactive groups on the components of the Stituted heteroatom-containing hydrocarbylene, respec composition begin to inter-react and form a three-dimen tively. sional matrix. The term “substantially immediately” is intended to mean within less than five minutes, preferably 0087. By “substituted” as in “substituted hydrocarbyl, within less than two minutes, and the term “immediately' is “substituted alkyl,” and the like, as alluded to in some of intended to mean within less than one minute, preferably the aforementioned definitions, is meant that in the hydro within less than 30 seconds. Typically, the three-dimensional carbyl, alkyl, or other moiety, at least one hydrogen atom composition will be completely formed within about 30 bound to a carbon atom is replaced with one or more minutes. Substituents that are functional groups Such as alkoxy, hydroxy, halo, nitro, and the like. Unless otherwise indi 0095. In one embodiment, the components and the result cated, it is to be understood that specified molecular Seg ing matrix are not Subject to enzymatic cleavage by matrix ments can be Substituted with one or more Substituents that metalloproteinases Such as collagenase, and are therefore do not compromise a compounds utility. For example, not readily degradable in Vivo. Further, the composition may “Succinimidyll” is intended to include unsubstituted Succin be readily tailored, in terms of the Selection and quantity of imidyl as well as SulfoSuccinimidyl and other Succinimidyl each component, to enhance certain properties, e.g., com groupS Substituted on a ring carbon atom, e.g., with alkoxy pression Strength, Swellability, tack, hydrophilicity, optical Substituents, polyether Substituents, or the like. clarity, and the like. 0088) II. The Components 0096. The homogeneous dry powder composition of the 0089. In accordance with the present invention, a com present invention is comprised of a first component having position is provided that contains at least two biocompatible, a core Substituted with nucleophilic groupS and a Second non-immunogenic components having reactive groups component having a core Substituted with electrophilic thereon, with the functional groups Selected So as to enable groups. The nucleophilic and electrophilic groups are non inter-reaction between the components, i.e., croSSlinking to reactive with one another when the first and Second com form a three-dimensional matrix. Each component has a core ponents are admixed in a dry environment but are rendered Substituted with reactive groups. Typically, the composition reactive upon exposure to an aqueous environment Such that will contain a first component having a core Substituted with the components inter-react in the aqueous environment to nucleophilic groupS and a Second component having a core form a three-dimensional matrix. In order for a three dimen Substituted with electrophilic groups. The invention also sional matrix to be formed, there is preferably plurality of encompasses compositions having more than two compo reactive groups present in each of the first and Second nents, where additional components may have nucleophilic components. In a preferred embodiment, one component has or electrophilic groups. a core Substituted with m nucleophilic groups, where me2, 0090 The reactive groups are selected so that the com and the other component has a core Substituted with n ponents are essentially non-reactive in an dry environment. electrophilic groups, where ne2 and m+n>4. Upon exposure to an aqueous environment, the components 0097 Thus, in one embodiment, the composition can be are rendered reactive and a plurality of components are then described as having components of formulas (I) and (II): able to inter-react in the aqueous environment to form a three-dimensional matrix. This matrix is preferably formed without input of any external energy, for example, at room temperature or at Slightly elevated temperature. 0098 wherein m and n are integers from 2-12 and 0.091 The composition is particularly suitable for appli m+n>4; R and R' are independently selected from hydro cation involving contact between a biological System and the philic polymers, hydrophobic polymers, amphiphilic poly US 2005/0281883 A1 Dec. 22, 2005

merS, C2-1 hydrocarbyls, and heteroatom-containing C2 reaction, a nucleophilic addition reaction, or both. The term hydrocarbyls, X is a nucleophilic group; Y is an electrophilic “electrophilic' refers to a reactive group that is Susceptible group; L' and L are linking groups; and p and q are integers to nucleophilic attack, i.e., Susceptible to reaction with an from 0-1. When p is 0, then a Specific nucleophilic group is incoming nucleophilic group. Electrophilic groups herein directly attached to the core R, however when p is 1, then a are positively charged or electron-deficient, typically elec Specific nucleophilic group is attached indirectly to the core tron-deficient. The term “nucleophilic' refers to a reactive via a linker group L. Each X group may be the Same or group that is electron rich, has an unshared pair of electrons different, and each Y group may be the same or different. acting as a reactive Site, and reacts with a positively charged 0099 Any additional components would have a formula or electron-deficient Site. such as Z-(L)l-R," where s is an integer from 2-6; R" is 0107 X may be virtually any nucleophilic group, so long Selected from hydrophilic polymers, hydrophobic polymers, as reaction can occur with the electrophilic group Y and also amphiphilic polymers, C-1 hydrocarbyls, and heteroatom with Z, when Z is present and is electrophilic. Analogously, containing C-1 hydrocarbyls, Z is a nucleophilic or elec Y may be virtually any electrophilic group, So long as trophilic group; L is a linking group; and r is an integer reaction can take place with X and also with Z, when Z is from 0-1. present and nucleophilic. The only limitation is a practical one, in that reaction between X and Y (and Z when present), 0100. In the components of formulas (I) and (II), each should be fairly rapid and take place automatically upon Side chain typically has one reactive group; howeverowever, admixture with an aqueous medium, without need for input the invention also encompasses components where the side of any external energy, e.g., heat, or potentially toxic or chains can contain more than one reactive group. Thus, for non-biodegradable reaction catalysts or other chemical example, the first component may have the formula (I): reagents. It is also preferred although not essential that reaction occur without need for ultraViolet or other radiation. 0101 where: a and b are integers from 0-1; c is an integer In one embodiment, the reactions between X and Y (and Z from 2-6; R" is selected from hydrophilic polymers, hydro when present), are complete in under 60 minutes, preferably phobic polymers, amphiphilic polymers, C. hydrocarbyls, under 30 minutes. Most preferably, the reaction occurs in and heteroatom-containing C-1 hydrocarbyls, X and X" about 5 to 15 minutes or less. are electrophilic groups; and Land L are linking groups. X 0.108 Examples of nucleopbilic groups suitable as X and X" may be the same or different. include, but are not limited to, -NH, -NHR', -N(R'), 0102) The components are either commercially available -SH, -OH, -COOH, -CH-OH, -H, -PH, or are readily Synthesized by techniques that are well known -PHR', -P(R), -NH-NH, -CO-NH-NH, in the art from commercially available materials. —CSH, N, etc. wherein R is a hydrocarbyl group and each R" may be the same or different. R is typically alkyl or 0103 1. Reactive Groups monocyclic aryl, preferably alkyl, and most preferably lower 0104 Prior to use, the composition is stored in a dry alkyl. Organometallic moieties are also useful nucleophilic environment that insures that the components remain essen groups for the purposes of the invention, particularly those tially non-reactive until use. Upon exposure to an aqueous that act as carbanion donors. Examples of organometallic environment, the reactive groups on the components are moieties include: Grignard functionalities -RMgHal rendered reactive and a plurality of components will then wherein R is a carbon atom (substituted or unsubstituted), inter-react to form the desired matrix. The dry composition and Hal is halo, typically bromo, iodo or chloro, preferably is preferably Stored under an inert atmosphere So that the bromo; and lithium-containing functionalities, typically components do not react with Oxygen. alkyllithium groups, Sodium-containing functionalities. 0105. In general, the concentration of the components 0109) It will be appreciated by those of ordinary skill in will be in the range of about 1 to 50 wt %, generally about the art that certain nucleophilic groups must be activated 2 to 40 wt %. The preferred concentration will depend on a with a base So as to be capable of reaction with an electro number of factors, including the type of component (i.e., philic group. For example, when there are nucleophilic type of molecular core and reactive groups), its molecular Sulfhydryl and hydroxyl groups in the multifunctional com weight, and the end use of the resulting three-dimensional pound, the compound must be admixed with an aqueous matrix. For example, use of higher concentrations of the base in order to remove a proton and provide an -ST or components, or using highly functionalized components, -O Species to enable reaction with the electrophilic group. will result in the formation of a more tightly crosslinked Unless it is desirable for the base to participate in the network, producing a Stiffer, more robust composition, Such reaction, a non-nucleophilic base is preferred. In Some as for example a gel. In general, the mechanical properties embodiments, the base may be present as a component of a of the three-dimensional matrix should be similar to the buffer Solution. Suitable bases and corresponding crosslink mechanical properties of the tissue to which the matrix (or ing reactions are described herein. matrix-forming components) will be applied. Thus, when 0110. The selection of electrophilic groups provided on the matrix will be used for an orthopedic application, the gel the multifunctional compound, must be made So that reac matrix should be relatively firm, e.g., a firm gel; however, tion is possible with the Specific nucleophilic groups. Thus, when the matrix will be used on Soft tissue, as for example when the X reactive groups are amino groups, the Y groups in tissue augmentation, the gel matrix should be relatively are Selected So as to react with amino groups. Analogously, Soft, e.g., a Soft gel. when the X reactive groups are Sulfhydryl moieties, the 0106 The reactive groups are electrophilic and nucleo corresponding electrophilic groups are Sulfhydryl-reactive philic groups, which undergo a nucleophilic Substitution groups, and the like. In general, examples of electrophilic US 2005/0281883 A1 Dec. 22, 2005

groups suitable as Y include, but are not limited to, -CO Sulfhydryl moiety. Such reactive groups include those that Cl, -(CO)-O-(CO)-R (where R is an alkyl group), form thioester linkages upon reaction with a Sulflhydryl CH=CH-CH=O and -CH=CH-C(CH.)=O, halo, group, such as those described in WO 00/62827 to Wallace N=C=O, -N=C=S, -SOCH=CH, -O(CO) et al. AS explained in detail therein, Sulflhydryl reactive C=CH, -O(CO)-C(CH)=CH, -S-S-(CHN), groups include, but are not limited to: mixed anhydrides, –O(CO)-C(CHCH)=CH, -CH=CH-C=NH, ester derivatives of phosphorus, ester derivatives of p-nitro -COOH, -(CO)O-N(COCH), —CHO, -(CO)O- phenol, p-nitrothiophenol and pentafluorophenol; esters of N(COCH-)-S(O).OH, and -NCCOCH). Substituted hydroxylamines, including N-hydroxyphthalim 0111 When X is amino (generally although not neces ide esters, N-hydroxysuccinimide esters, N-hydroxysulfos Sarily primary amino), the electrophilic groups present on Y luccinimide esters, and N-hydroxyglutarimide esters, esters are amine-reactive groups. Exemplary amine-reactive of 1-hydroxybenzotriazole; 3hydroxy-3,4-dihydro-benzotri groups include, by way of example and not limitation, the aZin-4-one, 3-hydroxy-3,4-dihydro-quinazoline-4-One, car following groups, or radicals thereof. (I) carboxylic acid bonylimidazole derivatives, acid chlorides, ketenes, and esters, including cyclic esters and “activated” esters; (2) acid isocyanates. With these Sulfhydryl reactive groups, auxiliary chloride groups (-CO-Cl); (3) anhydrides (-(CO)-O- reagents can also be used to facilitate bond formation, e.g., (CO)-R, where R is an alkyl group); (4) ketones and 1-ethyl-3-3-dimethylaminopropylcarbodiimide can be aldehydes, including C.f3-unsaturated aldehydes and ketones used to facilitate coupling of Sulfhydryl groups to carboxyl Such as -CH=CH-CH=O and -CH=CH containing groups. C(CH)=O; (5) halo groups; (6) isocyanate group 0116. In addition to the sulfhydryl reactive groups that (-N=C=O); (7) thioisocyanato group (-N=C=S); (8) form thioester linkages, various other Sulfhydryl reactive epoxides; (9) activated hydroxyl groups (e.g., activated with functionalities can be utilized that form other types of conventional activating agents Such as carbonyldiimidazole linkages. For example, compounds that contain methyl or Sulfonyl chloride); and (10) olefins, including conjugated imidate derivatives form imido-thioester linkages with Sulf olefins, such as ethenesulfonyl (-SOCH=CH-) and hydryl groups. Alternatively, Sulfhydryl reactive groups can analogous functional groups, including acrylate be employed that form disulfide bonds with sulfhydryl (-O(CO)-C=CH-), methacrylate (-O(CO)- groups, Such groups generally have the Structure -S-S- C(CH)=CH-), ethyl acrylate (-O(CO)- Ar where Ar is a Substituted or unsubstituted nitrogen C(CH2CH)=CH-), and ethyleneimino (-CH=CH containing heteroaromatic moiety or a non-heterocyclic aro C=NH). matic group Substituted with an electron-withdrawing 0112 In one embodiment the amine-reactive groups con moiety, Such that Ar may be, for example, 4-pyridinyl, tain an electrophilically reactive carbonyl group Susceptible o-nitrophenyl, m-nitrophenyl, p-nitrophenyl, 2,4-dinitrophe to nucleophilic attack by a primary or Secondary amine, for nyl, 2-nitro-4-benzoic acid, 2-nitro-4-pyridinyl, etc. In Such example the carboxylic acid esters and aldehydes noted instances, auxiliary reagents, i.e., mild oxidizing agents. Such above, as well as carboxyl groups (-COOH). as hydrogen peroxide can be used to facilitate disulfide bond 0113 Since a carboxylic acid group per Se is not Suscep formation. tible to reaction with a nucleophilic amine, components 0117 Yet another class of sulfhydryl reactive groups containing carboxylic acid groups must be activated So as to forms thioether bonds with Sulfhydryl groups. Such groups be amine-reactive. Activation may be accomplished in a include, inter alia, maleimido, Substituted maleimido, variety of ways, but often involves reaction with a suitable haloalkyl, epoxy, imino, and aziridino, as well as olefins hydroxyl-containing compound in the presence of a dehy (including conjugated olefins) Such as ethenesulfonyl, ethe drating agent Such as dicyclohexylcarbodiimide (DCC) or neimino, acrylate, methacrylate, and C.f3-unsaturated alde dicyclohexylurea (DHU). For example, a carboxylic acid hydes and ketones. can be reacted with an alkoxy-Substituted N-hydroxy-Suc 0118 When X is -OH, the electrophilic functional cinimide or N-hydroxySulfoSuccinimide in the presence of groups on the remaining component(s) must react with DCC to form reactive electrophilic groups, the N-hydrox hydroxyl groups. The hydroxyl group may be activated as ySuccinimide ester and the N-hydroxySulfoSuccinimide described above with respect to carboxylic acid groups, or it ester, respectively. Carboxylic acids may also be activated may react directly in the presence of base with a Sufficiently by reaction with an acyl halide Such as an acyl chloride (e.g., reactive electrophilic group Such as an epoxide group, an acetyl chloride), to provide a reactive anhydride group. In a aZiridine group, an acyl halide, an anhydride, and So forth. further example, a carboxylic acid may be converted to an 0119 When X is an organometallic nucleophilic group acid chloride group using, e.g., thionyl chloride or an acyl Such as a Grignard functionality or an alkyllithium group, chloride capable of an exchange reaction. Specific reagents Suitable electrophilic functional groups for reaction there and procedures used to carry out Such activation reactions with are those containing carbonyl groups, including, by will be known to those of ordinary skill in the art and are way of example, ketones and aldehydes. described in the pertinent texts and literature. 0.120. It will also be appreciated that certain functional 0114. Accordingly, in one embodiment, the amine-reac groups can react as nucleophilic or as electrophilic groups, tive groups are Selected from Succinimidyl ester depending on the Selected reaction partner and/or the reac (-O(CO)-N(COCH)), SulfoSuccinimidyl eSter tion conditions. For example, a carboxylic acid group can (-O(CO)-N(COCH)--S(O)-OH), maleimido act as a nucleophilic group in the presence of a fairly Strong (-N(COCH)2), epoxy, isocyanato, thioisocyanato, and base, but generally acts as an electrophilic group allowing ethenesulfonyl. nucleophilic attack at the carbonyl carbon and concomitant 0115) Analogously, when X is sulfhydryl, the electro replacement of the hydroxyl group with the incoming philic groups present on Y are groups that react with a nucleophilic group. US 2005/0281883 A1 Dec. 22, 2005

0121 These, as well as other embodiments are illustrated tion. For example, linking groups can be used to promote below, where the covalent linkages in the matrix that result hydrolysis, to discourage hydrolysis, or to provide a Site for upon covalent binding of Specific nucleophilic reactive enzymatic degradation. groups to Specific electrophilic reactive groups on the mul tifunctional compound include, Solely by way of example, 0.126 Examples of linking groups that provide hydrolyz the following: able sites, include, inter alia: ester linkages, anhydride

TABLE 1. Representative Nucleophilic Groups Representative Electrophilic Groups (X) (Y) Resulting Linkage -NH. -O-(CO)-O-N(COCH), -NH-(CO)-O- succinimidyl carbonate terminus -SH -O-(CO)-O-N(COCH), -S-(CO)-O- -OH -O-(CO)-O-N(COCH), -O-(CO)- -NH. -O(CO)-CH=CH, -NH-CHCH-(CO)-O- acrylate terminus -SH -O-(CO)-CH=CH, -S-CHCH-(CO)-O- -OH -O-(CO)-CH=CH, -O-CHCH-(CO)-O- -NH. -O(CO)-(CH-)-CO-N(COCH), -NH-(CO)-(CH)-(CO)-O- succinimidylglutarate terminus -SH -O(CO)-(CH-)-CO-N(COCH), -S-(CO)-(CH)-(CO)-O- -OH -O(CO)-(CH-)-CO-N(COCH), -O-(CO)-(CH)-(CO)-O- -NH. -O-CH-CO-N(COCH), -NH-(CO)-CH-O- succinimidyl acetate terminus -SH -O-CH-CO-N(COCH), -S-(CO)-CH-O- -OH -O-CH-CO-N(COCH), -O-(CO)-CH-O- -NH. -O-NH(CO)-(CH)-CO-N(COCH), NH-(CO)-(CH)-(CO)-NH-O succinimidyl succinimide terminus -SH -O-NH(CO)-(CH)-CO-N(COCH) -S-(CO)-(CH)-(CO)-NH-O- -OH -O-NH(CO)-(CH)-CO-N(COCH) -O-(CO)-(CH)-(CO)-NH-O- -NH. -O-(CH)-CHO -NH-(CO)-(CH)-O- propionaldehyde terminus -NH. O -NH-CH-CH(OH)-CH-O- and / V -NCH-CH(OH)-CH-O- -O-CH-CH-CH glycidyl ether terminus -NH. -O-(CH)-N=C=O -NH-(CO)-NH-CH-O- (isocyanate terminus) -NH -SO-CH=CH, -NH-CHCH-SO vinyl sulfone terminus -SH -SO2-CH=CH, -S-CHCH-SO

0122) The homogeneous dry powder may be compres linkages, Such as those obtained by incorporation of glut Sion molded into a thin sheet or membrane, which can then arate and Succinate; Ortho ester linkages; Ortho carbonate be sterilized using gamma or e-beam irradiation. The result linkages Such as trimethylene carbonate, amide linkages, ing dry membrane or sheet can be cut to the desired size or phosphoester linkages, C-hydroxy acid linkages, Such as chopped into Smaller Size particulates. those obtained by incorporation of lactic acid and glycolic 0123 2. Linking Groups acid; lactone-based linkages, Such as those obtained by incorporation of caprolactone, Valerolactone, Y-butyrolac 0.124. The reactive groups may be directly attached to the tone and p-dioxanone; and amide linkages Such as in a core, or they may be indirectly attached through a linking dimeric, oligomeric, or poly(amino acid) segment. group, with longer linking groups also termed “chain Examples of non-degradable linking groups include Succin extenders.” In the formulas (I) and (II) shown above, the imide, propionic acid and carboxymethylate linkages. See, optional linker groups are represented by L' and Lif, wherein for example, WO 99/07417 to Coury et al. Examples of the linking groups are present when p and q are equal to 1. enzymatically degradable linkages include Leu-Gly-Pro Ala, which is degraded by collagenase, and Gly-Pro-LyS, 0.125 Suitable linking groups are well known in the art. which is degraded by plasmin. See, for example, WO 97/22371 to Rhee et al. Linking groups are useful to avoid Steric hindrance problems that can 0127. Linking groups can also be included to enhance or Sometimes associated with the formation of direct linkages SuppreSS the reactivity of the various reactive groups. For between molecules. Linking groupS may additionally be example, electron-withdrawing groups within one or two used to link Several multifunctional compounds together to carbons of a Sulfhydryl group would be expected to diminish make larger molecules. In one embodiment, a linking group its effectiveness in coupling, due to a lowering of nucleo can be used to alter the degradative properties of the philicity. Carbon-carbon double bonds and carbonyl groups compositions after administration and resultant gel forma will also have such an effect. Conversely, electron-with US 2005/0281883 A1 Dec. 22, 2005 drawing groups adjacent to a carbonyl group (e.g., the containing C-1 hydrocarbyl. The heteroatom-containing reactive carbonyl of glutaryl-N-hydroxySuccinimidyl) C. hydrocarbyl can have 1 or 2 heteroatoms selected from would increase the reactivity of the carbonyl carbon with N, O and S. In a preferred embodiment, the molecular core respect to an incoming nucleophilic group. By contrast, is a Synthetic hydrophilic polymer. Sterically bulky groups in the vicinity of a reactive group can be used to diminish reactivity and thus reduce the coupling 0133 A. Hydrophilic Polymers rate as a result of Steric hindrance. 0134) The term “hydrophilic polymer” as used herein 0128 By way of example, particular linking groups and refers to a polymer having an average molecular weight and corresponding formulas are indicated in Table 2: composition that naturally renders, or is Selected to render the polymer as a whole “hydrophilic.” Preferred polymers TABLE 2 are highly pure or are purified to a highly pure State Such that the polymer is or is treated to become pharmaceutically Linking group Component structure pure. Most hydrophilic polymers can be rendered water Soluble by incorporating a Sufficient number of oxygen (or less frequently nitrogen) atoms available for forming hydro gen bonds in aqueous Solutions. 0.135 Synthetic hydrophilic polymers may be homopoly mers, block copolymers, random copolymers, or graft copolymers. In addition, the polymer may be linear or branched, and if branched, may be minimally to highly branched, dendrimeric, hyperbranched, or a Star polymer. O-(CO)-(CH) The polymer may include biodegradable Segments and blocks, either distributed throughout the polymer's molecu (CO)-O-(CH2) lar Structure or present as a Single block, as in a block copolymer. Biodegradable Segments are those that degrade So as to break covalent bonds. Typically, biodegradable Segments are Segments that are hydrolyzed in the presence of water and/or enzymatically cleaved in situ. Biodegradable Segments may be composed of Small molecular Segments Such as ester linkages, anhydride linkages, ortho ester link ages, ortho carbonate linkages, amide linkages, phosphonate 0129. In Table 2, X is generally in the range of 1 to about linkages, etc. Larger biodegradable “blocks' will generally 10; R is generally hydrocarbyl, typically alkyl or aryl, be composed of oligomeric or polymeric Segments incorpo preferably alkyl, and most preferably lower alkyl; and R is rated within the hydrophilic polymer. Illustrative oligomeric hydrocarbylene, heteroatom-containing hydrocarbylene, and polymeric Segments that are biodegradable include, by Substituted hydrocarbylene, or Substituted heteroatom-con way of example, poly(amino acid) segments, poly(orthoe taining hydrocarbylene) typically alkylene or arylene (again, Ster) segments, poly(orthocarbonate) segments, and the like. optionally Substituted and/or containing a heteroatom), pref erably lower alkylene (e.g., methylene, ethylene, n-propy 0.136 Synthetic hydrophilic polymers that are useful lene, n-butylene, etc.), phenylene, or amidoalkylene (e.g., herein include, but are not limited to: polyalkylene oxides, particularly polyethylene glycol (PEG) and poly(ethylene -(CO)-NH-CH). oxide)-poly(propylene oxide) copolymers, including block 0130. Other general principles that should be considered and random copolymers, polyols Such as glycerol, polyg with respect to linking groups are as follows. If a higher lycerol (PG) and particularly highly branched polyglycerol, molecular weight multifunctional compound is to be used, it propylene glycol, poly(oxyalkylene)-substituted diols, and will preferably have biodegradable linkages as described poly(oxyalkylene)-substituted polyols such as mono-, di above, so that fragments larger than 20,000 mol. wt. are not and tri-polyoxyethylated glycerol, mono- and di-polyoxy generated during resorption in the body. In addition, to ethylated propylene glycol, and mono- and di-polyoxyethy promote water miscibility and/or solubility, it may be lated trimethylene glycol, polyoxyethylated Sorbitol, poly desired to add Sufficient electric charge or hydrophilicity. oxyethylated glucose, poly(acrylic acids) and analogs and Hydrophilic groups can be easily introduced using known copolymers thereof, Such as polyacrylic acid per Se, poly chemical Synthesis, So long as they do not give rise to methacrylic acid, poly(hydroxyethylmethacrylate), poly(hy unwanted Swelling or an undesirable decrease in compres droxyethylacrylate), poly(methylalkylsulfoxide methacry Sive Strength. In particular, polyalkoxy Segments may lates), poly(methylalkylsulfoxide acrylates) and copolymers weaken gel Strength. of any of the foregoing, and/or with additional acrylate Species Such as aminoethyl acrylate and mono-2-(acryloxy)- 0131) 3. The Core ethyl Succinate; polymaleic acid; poly(acrylamides) Such as 0132) The “core” of each component is comprised of the polyacrylamide per se, poly(methacrylamide), poly(dim molecular structure to which the reactive groups are bound. ethylacrylamide), poly(N-isopropyl-acrylamide), and The molecular core can be a polymer, which includes copolymers thereof; poly(olefinic alcohols) Such as poly(Vi Synthetic polymers and naturally occurring polymers. The nyl alcohols) and copolymers thereof; poly(N-vinyl lactams) polymers can be hydrophilic, hydrophobic, or amphiphilic. Such as poly(Vinyl pyrrolidones), poly(N-vinyl caprolac The molecular core can also be a low molecular weight tams), and copolymers thereof; polyoxazolines, including component Such as a C- hydrocarbyl or a heteroatom poly(methyloxazoline) and poly(ethyloxazoline); and poly US 2005/0281883 A1 Dec. 22, 2005

Vinylamines, as well as copolymers of any of the foregoing. albumins, globulins, fibrinogen, fibrin and thrombin, with It must be emphasized that the aforementioned list of collagen particularly preferred; carboxylated polysaccha polymers is not exhaustive, and a variety of other Synthetic rides Such as polymannuronic acid and polygalacturonic hydrophilic polymerS may be used, as will be appreciated by acid; aminated polysaccharides, particularly the glycosami those skilled in the art. noglycans, e.g., hyaluronic acid, chitin, chondroitin Sulfate A, B, or C, keratin Sulfate, keratosulfate and heparin; and 0.137 Those of ordinary skill in the art will appreciate activated polysaccharides Such as dextran and Starch deriva that Synthetic polymerS Such as polyethylene glycol cannot tives. Collagen and glycosaminoglycans are preferred natu be prepared practically to have exact molecular weights, and rally occurring hydrophilic polymers for use herein. that the term “molecular weight” as used herein refers to the weight average molecular weight of a number of molecules 0.141. The term “collagen' as used herein refers to all in any given Sample, as commonly used in the art. Thus, a forms of collagen, including those, which have been pro sample of PEG 2,000 might contain a statistical mixture of cessed or otherwise modified. Thus, collagen from any polymer molecules ranging in weight from, for example, Source may be used in the compositions of the invention; for 1,500 to 2,500 daltons with one molecule differing slightly example, collagen may be extracted and purified from from the next over a range. Specification of a range of human or other mammalian Source, Such as bovine or molecular weights indicates that the average molecular porcine corium and human placenta, or may be recombi weight may be any value between the limits specified, and nantly or otherwise produced. The preparation of purified, may include molecules outside those limits. Thus, a molecu Substantially non-antigenic collagen in Solution from bovine lar weight range of about 800 to about 20,000 indicates an skin is well known in the art. For example, U.S. Pat. No. average molecular weight of at least about 800, ranging up 5,428,022 to Palefsky et al. discloses methods of extracting to about 20 kDa. and purifying collagen from the human placenta, and U.S. Pat. No. 5,667,839 to Berg discloses methods of producing 0.138. Other suitable synthetic hydrophilic polymers recombinant human collagen in the milk of transgenic include chemically Synthesized polypeptides, particularly animals, including transgenic cows. Non-transgenic, recom polynucleophilic polypeptides that have been Synthesized to binant collagen expression in yeast and other cell lines) is incorporate amino acids containing primary amino groups described in U.S. Pat. No. 6,413,742 to Olsen et al., U.S. Pat. (Such as lysine) and/or amino acids containing thiol groups No. 6,428,978 to Olsen et al., and U.S. Pat. No. 6,653,450 (Such as cysteine). Poly(lysine), a Synthetically produced to Berg et al. polymer of the amino acid lysine (145 MW), is particularly preferred. Poly(lysine)S have been prepared having any 0.142 Collagen of any type, including, but not limited to, where from 6 to about 4,000 primary amino groups, corre types I, II, III, IV, or any combination thereof, may be used sponding to molecular weights of about 870 to about 580, in the compositions of the invention, although type I is 000. Poly(lysine)s for use in the present invention preferably generally preferred. Either atelopeptide or telopeptide-con have a molecular weight within the range of about 1,000 to taining collagen may be used; however, when collagen from about 300,000, more preferably within the range of about a Source, Such as bovine collagen, is used, atelopeptide 5,000 to about 100,000, and most preferably, within the collagen is generally preferred, because of its reduced range of about 8,000 to about 15,000. Poly(lysine)s of immunogenicity compared to telopeptide-containing col varying molecular weights are commercially available from lagen. Peninsula Laboratories, Inc. (Belmont, Calif.). 0.143 Collagen that has not been previously crosslinked 0.139. Although a variety of different synthetic hydro by methods Such as heat, irradiation, or chemical crosslink philic polymers can be used in the present compositions, as ing agents is preferred for use in the invention, although indicated above, preferred Synthetic hydrophilic polymers previously crosslinked collagen may be used. are PEG and PG, particularly highly branched PG. Various 0144) Collagens for use in the present invention are forms of PEG are extensively used in the modification of generally, although not necessarily, in aqueous Suspension at biologically active molecules because PEG lackS toxicity, a concentration between about 20 mg/ml to about 120 antigenicity, and immunogenicity (i.e., is biocompatible), mg/ml, preferably between about 30 mg/ml to about 90 can be formulated So as to have a wide range of Solubilities, and does not typically interfere with the enzymatic activities mg/ml. Although intact collagen is preferred, denatured and/or conformations of peptides. A particularly preferred collagen, commonly known as gelatin, can also be used. Synthetic hydrophilic polymer for certain applications is a Gelatin may have the added benefit of being degradable PEG having a molecular weight within the range of about faster than collagen. 100 to about 100,000, although for highly branched PEG, far 0145 Nonfibrillar collagen is generally preferred for use higher molecular weight polymers can be employed, up to in compositions of the invention, although fibrillar collagens 1,000,000 or more, providing that biodegradable sites are may also be used. The term “nonfibrillar collagen” refers to incorporated ensuring that all degradation products will have any modified or unmodified collagen material that is in a molecular weight of less than about 30,000. For most Substantially nonfibrillar form, i.e., molecular collagen that PEGs, however, the preferred molecular weight is about is not tightly associated with other collagen molecules So as 1,000 to about 20,000, more preferably within the range of to form fibers. Typically, a solution of nonfibrillar collagen about 7,500 to about 20,000. Most preferably, the polyeth is more transparent than is a Solution of fibrillar collagen. ylene glycol has a molecular weight of approximately Collagen types that are nonfibrillar (or microfibrillar) in 10,000. native form include types IV, VI, and VII. 0140 Naturally occurring hydrophilic polymers include, 0146 Chemically modified collagens that are in non but are not limited to: proteins Such as collagen, fibronectin, fibrillar form at neutral pH include Succinylated collagen US 2005/0281883 A1 Dec. 22, 2005 and methylated collagen, both of which can be prepared (block copolymer) to form the amphiphilic polymer core of according to the methods described in U.S. Pat. No. 4,164, the components. The hydrophilic and hydrophobic portions 559 to Miyata et al. Methylated collagen, which contains may include any of the aforementioned hydrophilic and reactive amine groups, is a preferred nucleophile-containing hydrophobic polymers. component in the compositions of the present invention. In 0153. Alternately, the amphiphilic polymer core can be a another aspect, methylated collagen is a component that is hydrophilic polymer that has been modified with hydropho present in addition to first and Second components in the bic moieties (e.g., alkylated PEG or a hydrophilic polymer matrix-forming reaction of the present invention. Methy modified with one or more fatty chains), or a hydrophobic lated collagen is described in, for example, in U.S. Pat. No. polymer that has been modified with hydrophilic moieties 5,614,587 to Rhee et al. (e.g., "PEGylated phospholipids Such as polyethylene gly 0147 Collagens for use in the compositions of the colated phospholipids). present invention may start out in fibrillar form, then can be 0154 D. Low Molecular Weight Components rendered nonfibrillar by the addition of one or more fiber disassembly agent. The fiber disassembly agent must be O155 As indicated above, the molecular core of the present in an amount Sufficient to render the collagen Sub component can also be a low molecular weight compound, stantially nonfibrillar at pH 7, as described above. Fiber defined herein as being a C- hydrocarbyl or a heteroatom disassembly agents for use in the present invention include, containing C-1 hydrocarbyl, which contains 1 to 2 heteroa without limitation, various biocompatible alcohols, amino toms selected from N, O, S and combinations thereof. Such acids, inorganic Salts, and carbohydrates, with biocompat a molecular core can be Substituted with any of the reactive ible alcohols being particularly preferred. Preferred biocom groups described herein. patible alcohols include glycerol and propylene glycol. 0156 Alkanes are suitable C. hydrocarbyl molecular Non-biocompatible alcohols, Such as ethanol, methanol, and cores. Exemplary alkanes, for Substituted with a nucleo isopropanol, are not preferred for use in the present inven philic primary amino group and a Y electrophilic group, tion, due to their potentially deleterious effects on the body of the patient receiving them. Preferred amino acids include include, ethyleneamine (HN-CHCH-Y), tetramethyl arginine. Preferred inorganic Salts include Sodium chloride eneamine (HN-(CH)-Y), pentamethyleneamine and potassium chloride. Although carbohydrates, Such as (HN-(CHS)-Y), and hexamethyleneamine (HN various Sugars including Sucrose, may be used in the practice (CH)-Y). of the present invention, they are not as preferred as other O157 Low molecular weight diols and polyols are also types of fiber disassembly agents because they can have Suitable C-1 hydrocarbyls and include trimethylolpropane, cytotoxic effects in vivo. di(trimethylol propane), pentaerythritol, and diglycerol. Polyacids are also Suitable C. hydrocarbyls, and include 0148 Fibrillar collagen is less preferred for use in the trimethylolpropane-based tricarboxylic acid, di(trimethylol compositions of the present invention; however, as disclosed propane)-based tetracarboxylic acid, heptanedioic acid, in U.S. Pat. No. 5,614,587 to Rhee et al., fibrillar collagen, octanedioic acid (Suberic acid), and hexadecanedioic acid or mixtures of nonfibrillar and fibrillar collagen, may be (thapsic acid). preferred for use in adhesive compositions intended for long-term persistence in Vivo. 0158 Low molecular weight di- and poly-electrophiles are Suitable heteroatom-containing C-1 hydrocarbyl 0149 B. Hydrophobic Polymers molecular cores. These include, for example, disuccinimidyl 0150. The core of the components may also comprise a Suberate (DSS), bis(sulfosuccinimidyl) suberate (BS), hydrophobic polymer, including low molecular weight poly dithiobis(succinimidylpropionate) (DSP), bis(2-succinimi functional Species, although for most uses hydrophilic poly dooxycarbonyloxy) ethyl sulfone (BSOCOES), and 3,3'- mers are preferred. Generally, “hydrophobic polymers' dithiobis(sulfosuccinimidylpropionate (DTSPP), and their herein contain a relatively Small proportion of oxygen and/or analogs and derivatives. Low-molecular weight materials nitrogen atoms. Preferred hydrophobic polymers for use in comprising a plurality of acrylate moieties are present in one the invention generally have a carbon chain that is no longer aspect of the invention. Low molecular weight materials than about 14 carbons. Polymers having carbon chains comprising a plurality of thiol groups are present in another Substantially longer than 14 carbons generally have very aspect of the present invention. poor Solubility in aqueous Solutions and, as Such, have very 0159. 4. Preparation long reaction times when mixed with aqueous Solutions of Synthetic polymers containing multiple nucleophilic groups. 0160 The components are readily synthesized to contain Thus, use of short-chain oligomers can avoid Solubility a hydrophilic, hydrophobic or amphiphilic polymer core or related problems during reaction. Polylactic acid and polyg a low molecular weight core, functionalized with the desired lycolic acid are examples of two particularly Suitable hydro functional groups, i.e., nucleophilic or electrophilic groups, phobic polymers. which enable crosslinking. For example, preparation of first and Second components having a polyethylene glycol (PEG) 0151. C. Amphiphilic Polymers core is discussed below and in the examples; however, it is 0152 Generally, amphiphilic polymers have a hydro to be understood that the following discussion is for pur philic portion and a hydrophobic (or lipophilic) portion. The poses of illustration and analogous techniques may be hydrophilic portion can be at one end of the core and the employed with other polymers. hydrophobic portion at the opposite end, or the hydrophilic 0.161 With respect to PEG, first of all, various function and hydrophobic portions may be distributed randomly alized PEGs have been used effectively in fields such as (random copolymer) or in the form of Sequences or grafts protein modification (see Abuchowski et al., Enzymes as US 2005/0281883 A1 Dec. 22, 2005

Drugs, John Wiley & Sons: New York, N.Y. (1981) pp. 0168 Suitable tensile strength enhancers for use with the 367-383; and Dreborg et al. (1990) Crit. Rev. Therap. Drug multifunctional compound of the present invention include, Carrier Syst. 6:315), peptide chemistry (see Mutter et al., inter alia, collagen fibers, Synthetic polymer fibers, as well The Peptides, Academic: New York, N.Y. 2:285-332; and as other organic tensile Strength enhancers and inorganic Zalipsky et al. (1987) Int. J. Peptide Protein Res. 30:740), tensile Strength enhancers. The Synthetic polymer fibers can and the Synthesis of polymeric drugs (see Zalipsky et al. be either degradable or non-degradable. In a preferred (1983) Eur. Polym. J. 19:1177; and Ouchi et al. (1987) J. embodiment, the Synthetic polymeric fibers are degradable. Macromol. Sci. Chem. A24: 1011). These degradable polymer fibers may comprise polyesters, polyamides, poly(ortho esters), poly(anhydrides), poly 0162 Functionalized forms of PEG, including multi (phosphazines), poly(urethanes), poly(carbonates) and poly functionalized PEG, are commercially available, and are (dioxanones) as well as copolymers and blends thereof. A also easily prepared using known methods. For example, See particularly useful tensile Strength enhancer is Vicry1E Chapter 22 of Poly(ethylene Glycol) Chemistry: Biotechni (90:10 copolymer of glycolide and lactide). The use of cal and Biomedical Applications, J. Milton Harris, ed., tensile Strength enhancers, which are part of the broader Plenum Press, NY (1992). category of “fillers,” are well known. For example, Silicone 0163 Multi-functionalized forms of PEG are of particu gums, when croSS-linked with peroxides, are weak gels with lar interest and include, PEG succinimidylglutarate, PEG tensile strength on the order of only about 34 N/cm. When Succinimidyl propionate, Succinimidyl butylate, PEG Suc Suitably compounded with reinforcing fillers, the tensile cinimidyl acetate, PEG succinimidyl succinamide, PEG Strength of these gums may increase as much as fifty-fold. succinimidyl carbonate, PEG propionaldehyde, PEG gly Lichtenwalner et al., Eds., Encyclopedia of Polymer Science cidyl ether, PEG-isocyanate, and PEG-vinylsulfone. Many and Technology, Vol. 12, p. 535, John Wiley, New York, Such forms of PEG are described in U.S. Pat. Nos. 5,328,955 1970. Suitable tensile strength enhancers are those that have and 6,534,591, both to Rhee et al. Similarly, various forms inherent high tensile Strength and also can interact by of multi-amino PEG are commercially available from covalent or non-covalent bonds with the three-dimensional Sources such as PEG Shop, a division of Sun Bio of South matrix. The tensile strength enhancer should bond to the Korea (www.sunbio.com), Nippon Oil and Fats (Yebisu matrix, either mechanically or covalently, in order to provide Garden Place Tower, 20-3 Ebisu 4-chome, Shibuya-ku, tensile Support. Tensile Strengths of polyglycolide resorb Tokyo), Nektar Therapeutics (San Carlos, Calif., formerly able sutures are approximately 89,000 N/cm°; that of col Shearwater Polymers, Huntsville, Ala.) and from Hunts lagen fibers is 5000-10,000 N/cm (Tsuruta and Hayashi, man's Performance Chemicals Group (Houston,Tex.) under Eds., Biomedical Applications of Polymeric Materials, CRC the name Jeffamine(E) polyoxyalkyleneamines. Multi-amino Press, Boca Raton, Fla., 1993). PEGs useful in the present invention include the Jeffamine 0169. The components can also be prepared to contain diamines (“D” series) and triamines (“T” series), which contain two and three primary amino groups per molecule. various imaging agents Such as iodine, water Soluble iodo Analogous poly(sulfhydryl) PEGs are also available from derivative, or barium Sulfate, or fluorine, in order to aid Nektar Therapeutics, e.g., in the form of pentaerythritol Visualization of the compositions after administration via poly(ethylene glycol) ether tetra-sulfhydryl (molecular X-ray or '' F-MRI, respectively. weight 10,000). 0170 For use in tissue adhesion as discussed below, it may also be desirable to incorporate proteins Such as albu 0164 Reaction with succinimidyl groups to convert ter min, fibrin or fibrinogen into the multifunctional compound minal hydroxyl groups to reactive esters is one technique for to promote cellular adhesion. preparing a core with electrophilic groups Suitable for reac tion with nucleophilic groupS. Such as primary amines, thiols, 0171 In addition, the introduction of hydrocolloids such and hydroxyl groups. Other agents to convert hydroxyl as carboxymethylcellulose may promote tissue adhesion groups include carbonyldiimidazole and Sulfonyl chloride; and/or swellability. however, as discussed herein, a wide variety of electrophilic 0172 The crosslinkable composition may be comprised groups may be advantageously employed for reaction with of (a) a first crosslinkable component having m nucleophilic corresponding nucleophilic groups. Examples of Such elec groups, wherein me2; and (b) a second crosslinkable com trophilic groups include acid chloride groups, anhydrides, ponent having n electrophilic groups capable of reaction ketones, aldehydes, isocyanate, isothiocyanate, epoxides, with the m nucleophilic groups to form covalent bonds, and olefins, including conjugated olefins Such as ethene wherein ne2 and m+ne5, the first component comprises sulfonyl (-SOCH=CH-) and analogous functional two or more amino acid residues Selected from the group groupS. consisting of amino acids comprising primary amine groups and amino acids comprising thiol groups, the Second com 0165) III. The Compositions ponent comprises a polyethylene glycol moiety, and each of 0166 The components of the invention can be included the first and Second crosslinkable components is biocom in a pharmaceutical composition. A pharmaceutically patible, Synthetic, and nonimmunogenic, and further acceptable carrier may also be included. wherein crosslinking of the composition results in a bio 0167. In order to enhance matrix strength, it may be compatible, nonimmunogenic, crosslinked matrix. generally desirable to add a “tensile Strength enhancer to 0173 Any of the following are preferred embodiments of the composition. Such tensile Strength enhancers preferably the croSSlinkable composition described immediately above: comprise micron-size, preferably 5 to 40 microns in diam m>3, m=3, m=4, n=4, the electrophilic groups are Succin eter and 20 to 5000 microns in length, high tensile strength imidyl moieties, all n are identical, and all m are identical. fibers, usually with glass transition temperatures well above 0.174. In one preferred embodiment, the selected amino 37o C. acid residues are lysine. Within this embodiment, any of the US 2005/0281883 A1 Dec. 22, 2005 following is preferred: m>3, m=3, m=4, n=4, the electro 0182. In one preferred embodiment, the selected amino philic groups are Succinimidyl moieties, all n are identical, acid residues are lysine. Within this embodiment, any of the and all m are identical. following is preferred: m>3, m=3, m=4, n=4, the electro 0.175. In another preferred embodiment, the selected philic groups are Succinimidyl moieties, all n are identical, amino acid residues are cysteine. Within this embodiment, all m are identical, and the multifunctionally activated any of the following is preferred: m>3, m=3,m=4, n=4, the polyethylene glycol is tetrafunctionally activated polyethyl electrophilic groups are Succinimidyl moieties, all n are ene glycol or the multifunctionally activated polyethylene identical, and all m are identical. glycol is a Star-branched polyethylene glycol. 0176) The crosslinkable composition may also be com 0183 In another preferred embodiment, the selected prised of a crosslinkable composition comprised of: (a) a amino acid residues are cysteine. Within this embodiment, first crosslinkable component having m nucleophilic groups, any of the following is preferred: m>3, m=3,m=4, n=4, the wherein me2; and (b) a second crosslinkable component electrophilic groups are Succinimidyl moieties, all n are having n electrophilic groups capable of reaction with the m identical, all m are identical, and the multifunctionally nucleophilic groups to form covalent bonds, wherein ne2 activated polyethylene glycol is tetrafunctionally activated and m+ne5, the first component comprises two or more polyethylene glycol or the multifunctionally activated poly amino acid residues Selected from the group consisting of ethylene glycol is a Star-branched polyethylene glycol. amino acids comprising primary amine groups and amino 0.184 The crosslinkable composition may also be com acids comprising thiol groups, the Second component com prised of: (a) a first crosslinkable component having m prises a polyethylene glycol moiety, the electrophilic groups nucleophilic groups, wherein me2; and (b) a second are Succinimidyl moieties, and each of the first and Second crosslinkable component having n electrophilic groups crosslinkable components is biocompatible, Synthetic, and capable of reaction with the m nucleophilic groups to form nonimmunogenic, and further whererin crosslinking of the covalent bonds, wherein ne2 and m+ne5, the first compo composition results in a biocompatible, nonimmunogenic, nent comprises two or more amino acid residues Selected crosslinked matrix. from the group consisting of lysine and cysteine, the Second 0177. Any of the following are preferred embodiments of component comprises a polyethylene glycol moiety, and the crosslinkable composition described immediately above: each of the first and Second crosslinkable components is m>3, m=3, m=4, n=4, the electrophilic groups are Succin biocompatible, Synthetic, and nonimmunogenic, and imidyl moieties, all n are identical, and all m are identical. crosslinking of the composition results in a biocompatible, nonimmunogenic, croSSlinked matrix. 0178. In one preferred embodiment, the selected amino acid residues are lysine. Within this embodiment, any of the 0185. Any of the following are preferred embodiments of following is preferred: m>3, m=3, m=4, n=4, the electro the croSSlinkable composition described immediately above: philic groups are Succinimidyl moieties, all n are identical, m>3, m=3, m=4, n=4, the electrophilic groups are Succin and all m are identical. imidyl moieties, all n are identical, all m are identical, the first component consists of three lysine residues, and the first 0179. In another preferred embodiment, the selected component consists of three cysteine residues. amino acid residues are cysteine. Within this embodiment, any of the following is preferred: m>3, m=3,m=4, n=4, the 0186 The crosslinkable composition may also be com electrophilic groups are Succinimidyl moieties, all n are prised of: (a) a first crosslinkable component having m identical, and all m are identical. nucleophilic groups, wherein me2; and (b) a second crosslinkable component having n electrophilic groups 0180. The crosslinkable composition may also be com capable of reaction with the m nucleophilic groups to form prised of: (a) a first crosslinkable component having m covalent bonds, wherein ne2 and m+ne5, the first compo nucleophilic groups, wherein me2; and (b) a second nent comprises two or more lysine residues, the Second crosslinkable component having n electrophilic groups component comprises a polyethylene glycol moiety, and capable of reaction with the m nucleophilic groups to form each of the first and Second crosslinkable components is covalent bonds, wherein ne2 and m+ne5, the first compo biocompatible, Synthetic, and nonimmunogenic, and nent comprises two or more amino acid residues Selected crosslinking of the composition results in a biocompatible, from the group consisting of amino acids comprising pri nonimmunogenic, croSSlinked matrix. mary amine groupS and amino acids comprising thiol groups, the Second component comprises a multifunction 0187. Any of the following are preferred embodiments of ally activated polyethylene glycol, and each of the first and the croSSlinkable composition described immediately above: Second crosslinkable components is biocompatible, Syn m>3, m=3, m=4, n=4, the electrophilic groups are Succin thetic, and nonimmunogenic, and further wherein crosslink imidyl moieties, all n are identical, all m are identical, and ing of the composition results in a biocompatible, nonim the first component consists of three lysine residues. munogenic, crosslinked matrix. 0188 The crosslinkable composition may also be com 0181 Any of the following are preferred embodiments of prised of: (a) a first crosslinkable component having m the crosslinkable composition described immediately above: nucleophilic groups, wherein me2; and (b) a second m>3, m=3, m=4, n=4, the electrophilic groups are Succin crosslinkable component having n electrophilic groups imidyl moieties, all n are identical, all m are identical, the capable of reaction with the m nucleophilic groups to form multifunctionally activated polyethylene glycol is tetrafunc covalent bonds, wherein ne2 and m+ne5, the first compo tionally activated polyethylene glycol, and the multifunc nent consists of lysine residues, the Second component tionally activated polyethylene glycol is a Star-branched comprises a polyethylene glycol moiety, and each of the first polyethylene glycol. and Second crosslinkable components is biocompatible, Syn US 2005/0281883 A1 Dec. 22, 2005 thetic, and nonimmunogenic, and crosslinking of the com 0196. The crosslinkable composition may also be com position results in a biocompatible, nonimmunogenic, prised of: (a) a first crosslinkable component having m crosslinked matrix. nucleophilic groups, wherein me2; and (b) a second crosslinkable component having n electrophilic groups 0189 Any of the following are preferred embodiments of capable of reaction with the m nucleophilic groups to form the crosslinkable composition described immediately above: covalent bonds, wherein ne2 and m+ne5, the first compo m>3, m=3, m=4, n=4, the electrophilic groups are Succin nent comprises two or more lysine residues, the Second imidyl moieties, all n are identical, all m are identical, and component comprises a polyethylene glycol moiety, the the first component consists of three lysine residues. electrophilic groups are Succinimidyl moieties, and each of 0190. Another aspect of the invention relates to a the first and Second crosslinkable components is biocom crosslinkable composition comprised of: (a) a first crosslink patible, Synthetic, and nonimmunogenic, and crosslinking of able component having m nucleophilic groups, wherein the composition results in a biocompatible, nonimmuno me2; and (b) a Second crosslinkable component having in genic, crosslinked matrix. electrophilic groups capable of reaction with the m nucleo philic groups to form covalent bonds, wherein ne2 and 0.197 Any of the following are preferred embodiments of m+ne5, the first component comprises two or more cysteine the croSSlinkable composition described immediately above: residues, the Second component comprises a polyethylene m>3, m=3,m=4, n=4, all n are identical, all m are identical, glycol moiety, and each of the first and Second crosslinkable and the first component consists of three lysine residues. components is biocompatible, Synthetic, and nonimmuno 0198 The crosslinkable composition may also be com genic, and crosslinking of the composition results in a prised of: (a) a first crosslinkable component having m biocompatible, nonimmunogenic, crosslinked matrix. nucleophilic groups, wherein me2; and (b) a second 0191) Any of the following are preferred embodiments of crosslinkable component having n electrophilic groups the crosslinkable composition described immediately above: capable of reaction with the m nucleophilic groups to form m>3, m=3, m=4, n=4, the electrophilic groups are Succin covalent bonds, wherein ne2 and m+ne5, the first compo imidyl moieties, all n are identical, all m are identical, and nent consists of lysine residues, the Second component the first component consists of three cysteine residues. comprises a polyethylene glycol moiety, the electrophilic groups are Succinimidyl moieties, and each of the first and 0.192 The crosslinkable composition may also be com Second crosslinkable components is biocompatible, Syn prised of: (a) a first crosslinkable component having m thetic, and nonimmunogenic, and crosslinking of the com nucleophilic groups, wherein me2; and (b) a second position results in a biocompatible, nonimmunogenic, crosslinkable component having n electrophilic groups crosslinked matrix. capable of reaction with the m nucleophilic groups to form covalent bonds, wherein ne2 and m+n 25, wherein the first 0199 Any of the following are preferred embodiments of component consists of cysteine residues, the Second com the croSSlinkable composition described immediately above: ponent comprises a polyethylene glycol moiety, and each of m>3, m=3,m=4, n=4, all n are identical, all m are identical, the first and Second crosslinkable components is biocom and the first component consists of three lysine residues. patible, Synthetic, and nonimmunogenic, and crosslinking of 0200. The crosslinkable composition may also be com the composition results in a biocompatible, nonimmuno prised of: (a) a first crosslinkable component having m genic, crosslinked matrix. nucleophilic groups, wherein me2; and (b) a second 0193 Any of the following are preferred embodiments of crosslinkable component having n electrophilic groups the crosslinkable composition described immediately above: capable of reaction with the m nucleophilic groups to form me3, m=3, m=4, n=4, the electrophilic groups are Succin covalent bonds, wherein ne2 and m+ne5, the first compo imidyl moieties, all n are identical, all m are identical, and nent comprises two or more cysteine residues, the Second the first component consists of three cysteine residues. component comprises a polyethylene glycol moiety, the electrophilic groups are Succinimidyl moieties, and each of 0194 The crosslinkable composition may also be com the first and Second crosslinkable components is biocom prised of: (a) a first crosslinkable component having m patible, Synthetic, and nonimmunogenic, and crosslinking of nucleophilic groups, wherein me2; and (b) a second crosslinkable component having n electrophilic groups the composition results in a biocompatible, nonimmuno capable of reaction with the m nucleophilic groups to form genic, crosslinked matrix. covalent bonds, wherein ne2 and m+ne5, the first compo 0201 Any of the following are preferred embodiments of nent comprises two or more amino acid residues Selected the croSSlinkable composition described immediately above: from the group consisting of lysine and cysteine, the Second m>3, m=3,m=4, n=4, all n are identical, all m are identical, component comprises a polyethylene glycol moiety, the and the first component consists of three cysteine residues. electrophilic groups are Succinimidyl moieties, and each of 0202) The crosslinkable composition may also be com the first and Second crosslinkable components is biocom prised of: (a) a first crosslinkable component having m patible, Synthetic, and nonimmunogenic, and crosslinking of nucleophilic groups, wherein me2; (b) a second crosslink the composition results in a biocompatible, nonimmuno able component having n electrophilic groups capable of genic, crosslinked matrix. reaction with the m nucleophilic groups to form covalent 0.195 Any of the following are preferred embodiments of bonds, wherein ne2 and m+ne5, the first component con the crosslinkable composition described immediately above: Sists of cysteine residues, the Second component comprises m>3, m=3,m=4, n=4, all n are identical, all m are identical, a polyethylene glycol moiety, the electrophilic groups are the first component consists of three lysine residues, and the Succinimidyl moieties, and each of the first and Second the first component consists of three cysteine residues. crosslinkable components is biocompatible, Synthetic, and US 2005/0281883 A1 Dec. 22, 2005

nonimmunogenic, and croSSlinking of the composition buffer solution having a pH within the range of about 6.0 to results in a biocompatible, nonimmunogenic, crosslinked 11.0 to form a reactive solution. For example, 0.375 grams matrix. of the dry powder can be combined with 0.75 grams of the 0203) Any of the following are preferred embodiments of acid buffer to provide, after mixing, a homogeneous Solu the crosslinkable composition described immediately above: tion, where this Solution is combined with 1.1 grams of the m>3, m=3,m=4, n=4, all n are identical, all m are identical, basic buffer to provide a reactive mixture that substantially and the first component consists of three cysteine residues. immediately forms a three-dimensional matrix. 0204 IV. Administration and Formation of the Three 0211 1. Buffers Dimensional Matrix 0212. The buffer solutions are aqueous and can be any pharmaceutically acceptable basic or acid composition. The 0205 The invention is also directed at a method of term "buffer is used in a general Sense to refer to an acidic formulating an in-situ curing composition. This method or basic aqueous Solution, where the Solution may or may involves a delayed activation/triggering/initiation of the not be functioning to provide a buffering effect (i.e., resis reaction between the reactive groups, generating a cured tance to change in pH upon addition of acid or base) in the composition with consistent and uniform Strength. compositions of the present invention. 0206. The composition may be administered before, dur 0213 Acidic buffer solutions having a pH within the ing or after the components inter-react in the aqueous range of about 1.0 to 5.5, include by way of illustration and environment to form a three-dimensional matrix. Certain not limitation, Solutions of citric acid, hydrochloric acid, uses, which are discussed in greater detail below, Such as phosphoric acid, Sulfuric acid, AMPSO (3-(1,1-dimethyl tissue augmentation, may require the matrix to be formed 2-hydroxyethyl)amino2-hydroxy-propane-Sulfonic acid), before administration, whereas other applications, Such as acetic acid, lactic acid, and combinations thereof. In a tissue adhesion, require the compositions to be administered preferred embodiment, the acidic buffer Solution is a Solu before the inter-reaction has reached “equilibrium.” The tion of citric acid, hydrochloric acid, phosphoric acid, Sul point at which inter-reaction has reached equilibrium is furic acid, and combinations thereof. defined herein as the point at which the composition no longer feels tacky or Sticky to the touch. 0214) Regardless of the precise acidifying agent, the acidic buffer preferably has a pH such that it retards the 0207. The composition of the present invention is gen reactivity of the nucleophilic groups on the first component. erally delivered to the site of administration in such a way For example, a pH of 2.1 is generally Sufficient to retard the that the individual reactive groups of the compounds are nucleophilicity of thiol groups. A lower pH is typically exposed to the aqueous environment for the first time at the preferred when the first component contains amine groups as Site of administration, or immediately preceding adminis the nucleophilic groups. In general, the acidic buffer is an tration. Thus, the composition is preferably delivered to the acidic Solution that, when contacted with nucleophilic Site of administration using an apparatus that allows the groups that are present as part of the first component, renders composition to be delivered in an dry environment, where those nucleophilic groups relatively non-nucleophilic. the compounds are essentially non-reactive. 0215. An exemplary acidic buffer is a solution of hydro 0208. In one embodiment of the invention, a three-di chloric acid, having a concentration of about 6.3 mM and a mensional matrix is formed by the steps of: (a) providing a pH in the range of 2.1 to 2.3. This buffer may be prepared composition of the invention; (b) rendering the nucleophilic by combining concentrated hydrochloric acid with water, and electrophilic groups reactive by exposing the composi i.e., by diluting concentrated hydrochloric acid with water. tion to an aqueous environment to effect inter-reaction; Similarly, this buffer A may also be conveniently prepared wherein said exposure comprises: (i) dissolving the compo by diluting 1.23 grams of concentrated hydrochloric acid to Sition in a first buffer Solution having a pH within the range a volume of 2 liters, or diluting 1.84 grams of concentrated of about 1.0 to 5.5 to form a homogeneous solution, and (ii) hydrochloric acid to a volume to 3 liters, or diluting 2.45 adding a Second buffer Solution having a pH within the range grams of concentrated hydrochloric acid to a Volume of 4 of about 6.0 to 11.0 to the homogeneous solution; and (c) liters, or diluting 3.07 grams concentrated hydrochloric acid allowing a three-dimensional matrix to form. Typically, the to a Volume of 5 liters, or diluting 3.68 grams of concen matrix is formed, e.g., by polymerization, without input of trated hydrochloric acid to a volume to 6 liters. For safety any external energy. reasons, the concentrated acid is preferably added to water. 0209 The first and second components of the composi 0216 Basic buffer solutions having a pH within the range tion are typically combined in amounts Such that the number of about 6.0 to 11.0, include by way of illustration and not of nucleophilic groups in the mixture is approximately equal limitation, Solutions of glutamate, acetate, carbonate and to the number of electrophilic groups in the mixture. AS used carbonate Salts (e.g., Sodium carbonate, Sodium carbonate in this context, the term “approximately refers to a 2:1 to monohydrate and Sodium bicarbonate), borate, phosphate 1:2 ratio of moles of nucleophilic groups to moles of and phosphate Salts (e.g., monobasic Sodium phosphate electrophilic groups. A 1:1 molar ratio of nucleophilic to monohydrate and dibasic Sodium phosphate), and combina electrophilic groups is generally preferred. tions thereof. In a preferred embodiment, the basic buffer 0210. The first and second components are blended Solution is a Solution of carbonate Salts, phosphate Salts, and together to form a homogeneous dry powder. This powder is combinations thereof. then combined with a buffer solution having a pH within the 0217. In general, the basic buffer is an aqueous solution range of about 1.0 to 5.5 to form a homogeneous acidic that neutralizes the effect of the acidic buffer, when it is aqueous Solution, and this Solution is then combined with a added to the homogeneous Solution of the first and Second US 2005/0281883 A1 Dec. 22, 2005 components and the acid buffer, So that the nucleophilic 0224. The homogeneous dry powder composition and the groups of the first component regain their nucleophilic two buffer solutions may be conveniently formed under character (that has been masked by the action of the acidic aseptic conditions by placing each of the three ingredients buffer), thus allowing the nucleophilic groups to inter-react (dry powder, acidic buffer solution and basic buffer solution) with the electrophilic groups of the Second component. into Separate Syringe barrels. For example, the composition, first buffer Solution and second buffer Solution can be housed 0218. An exemplary basic buffer is an aqueous solution Separately in a multiple-compartment Syringe System having of carbonate and phosphate Salts. This buffer may be pre a multiple barrels, a mixing head, and an exit orifice. The pared by combining a base Solution with a Salt Solution. The first buffer solution can be added to the barrel housing the Salt Solution may be prepared by combining 34.7 g of composition to dissolve the composition and form a homo monobasic Sodium phosphate monohydrate, 49.3 g of geneous Solution, which is then extruded into the mixing Sodium carbonate monohydrate, and Sufficient water to head. The second buffer solution can be simultaneously provide a solution volume of 2 liter. Similarly, a 6 liter extruded into the mixing head. Finally, the resulting com Solution may be prepared by combining 104.0 g of monoba position can then be extruded through the orifice onto a sic Sodium phosphate monohydrate, 147.94 g of Sodium Surface. carbonate monohydrate, and Sufficient water to provide 6 liter of the salt solution. The basic buffer may be prepared by 0225. For example, the syringe barrels holding the dry combining 7.2 g of sodium hydroxide with 180.0 g of water. powder and the basic buffer may be part of a dual-Syringe The basic buffer is typically prepared by adding the base System, e.g., a double barrel Syringe as described in U.S. Pat. Solution as needed to the Salt Solution, ultimately to provide No. 4,359,049 to Redl et al. In this embodiment, the acid a mixture having the desired pH, e.g., a pH of 9.65 to 9.75. buffer can be added to the syringe barrel that also holds the dry powder, So as to produce the homogeneous Solution. In 0219. In general, the basic species present in the basic other words, the acid buffer may be added (e.g., injected) buffer should be sufficiently basic to neutralize the acidity into the Syringe barrel holding the dry powder to thereby provided by the acidic buffer, but should not be so nucleo produce a homogeneous Solution of the first and Second philic itself that it will react substantially with the electro components. This homogeneous Solution can then be philic groups of the Second component. For this reason, extruded into a mixing head, while the basic buffer is relatively “Soft' baseS Such as carbonate and phosphate are simultaneously extruded into the mixing head. Within the preferred in this embodiment of the invention. mixing head, the homogeneous Solution and the basic buffer 0220 To illustrate the preparation of a three-dimensional are mixed together to thereby form a reactive mixture. matrix of the present invention, one may combine an admix Thereafter, the reactive mixture is extruded through an ture of a first component (e.g., a polyethyleneglycol core orifice and onto a Surface (e.g., tissue), where a film is with four nucleophilic thiol groups, Such as pentaerythritol formed, which can function as a Sealant or a barrier, or the tetrakismercaptoethyl poly(oxyethylene) ether (“HS like. The reactive mixture begins forming a three-dimen PEG”) available from Aldrich Chemical Co. (Milwaukee, Sional matrix immediately upon being formed by the mixing Wis.), and a second component (e.g., a polyethyleneglycol of the homogeneous solution and the basic buffer in the core with four electrophilic N-hydroxySuccinimide groups, mixing head. Accordingly, the reactive mixture is preferably Such as pentaerythritol tetrakis 1-(1'-oxo-5-Succimidylpen extruded from the mixing head onto the tissue very quickly tanoate)-2-poly(oxyethylene) ether (“NHS-PEG,” 10,000 after it is formed so that the three-dimensional matrix forms MW, available from Aldrich Chemical Co.), with a first, on, and is able to adhere to, the tissue. acidic, buffer (e.g., an acid Solution, e.g., a dilute hydro 0226 AS preferred embodiment of the multi-compart chloric acid Solution) to form a homogeneous Solution. This ment Syringe System of the present invention is shown in homogeneous Solution is mixed with a Second, basic, buffer FIG. 1. The device is comprised of three syringes, two (e.g., a basic Solution, e.g., an aqueous Solution containing housing each of the two buffers of the present invention with phosphate and carbonate Salts) whereupon the first and the third Syringe housing the dry powder composition 1. The Second components Substantially immediately inter-react two Syringes housing the Solutions 1 are pre-assembled into with one another to form a three-dimensional matrix. a Syringe housing 2 with a transfer port closure 3 attached to 0221) 2. Delivery Systems the housing assembly 2 to allow mixing of the dry powder into the correct Syringe. A Syringe clip 4 is attached to the 0222 A. Multi-Compartment Devices plunger rod of the Syringe that does not require mixing with the dry powder composition. 0223 Suitable delivery systems for the homogeneous dry powder composition and the two buffer Solutions may 0227 Other systems for combining two reactive liquids involve a multi-compartment device, where one or more are well known in the art, and include the Systems described compartments contain the powder and one or more com in U.S. Pat. No. 6,454,786 to Holm et al.; U.S. Pat. No. partments contain the buffer Solutions needed to provide for 6,461,325 to Delmotte et al., U.S. Pat. No. 5,585,007 to the aqueous environment, So that the composition is exposed Antanavich et al.; U.S. Pat. No. 5,116,315 to Capozzi et al.; to the aqueous environment as it leaves the compartment. U.S. Pat. No. 4,631,055 to Redl et al.; and U.S. Patent Many devices that are adapted for delivery of multi-com Application Publication No. 2004/0068266 to Delmotte. ponent tissue Sealants/hemostatic agents are well known in the art and can also be used in the practice of the present 0228 B. Pressurized Delivery Devices invention. Alternatively, the composition can be delivered 0229. Other delivery systems for dispensing the multi using any type of controllable extrusion System, or it can be component compositions of the invention may include pres delivered manually in the form of a dry powder, and exposed Surized delivery devices, examples of which are described in to the aqueous environment at the Site of administration. commonly owned co-pending U.S. patent application Ser. US 2005/0281883 A1 Dec. 22, 2005

No. 10/957,493, filed on Oct. 1, 2004, and entitled “Mixing distal end 14 of the cap 10. The cap 10 is shown as having and Dispensing Fluid Components of a Multicomponent a cylindrical exterior surface 16 and an interior surface 18 Composition.” Such a pressurized delivery device may that terminates at opening 20, but additional cap shapes are include a diffuser Surface having an outlet extending there also suitable for use with the pressurized delivery device of through that is positioned downstream from a plurality of the present invention. As shown in FIG. 3, the interior inlets. While at least one inlet is adapted to communicate surface 18 of the cap 10 at end 14 serves to receive fluid with a Source of a pressurized carrier fluid, each of a components thereon. plurality of inlets is adapted to communicate with a Source 0232 Also provided is a generally elongate cylindrical of a different fluid component. Using this device, the dry connector 30 in the form of a unitary member having a first powder solution is premixed with the first buffer to form a terminus 32 and a second terminus 34. A plurality of inlet homogeneous Solution as previously described and this lumens 36A, 36B, and 36C traversing the length of the Solution is Subsequently communicated with a first fluid connector defined by termini 32 and 34. As depicted, the component. The Second fluid component will communicate connector 30 is detached from the cap 10. Inlet lumens 36A with the second buffer solution previously described. Once and 36B each communicate at the second terminus 34 with the diffuser surface receives fluid components from the a different Source of a fluid component, e.g., the first buffer inlets, each received fluid component is pushed toward the mixed with the dry powder in one Source and the Second outlet for mixing and dispensing therethrough by the pres buffer in the other source (not shown). Similarly, inlet Surized carrier fluid, typically a gas Such as air, from the lumens 36C are provided fluid communication at the second carrier fluid inlet. The diffuser Surface and the inlets may terminus 34 with a Source of pressurized carrier gas (not represent components of a mixing nozzle. shown). The carrier fluid inlet lumens 36C define a plane 0230. In general, there are two categories of gas enhanced that is perpendicular to a plane defined by the fluid compo nozzles for dispensing reactive components of a multicom nent inlet lumens 36A and 36B. As depicted in FIGS. 2 and ponent composition-those that involve internal mixing and 3, the first terminus 32 of the connector 30 has dimensions those that involve external mixing. When the diffuser Sur Suitable for forming a fluid-tight Seal against the interior face is a part of a nozzle, the nozzle may be considered an surface 18 of the cap 10 at its proximal end 20. internal-mixing nozzle. Unlike other internal-mixing tech nologies, the internal-mixing nozzle of the pressurized 0233. In operation, the cap 10 is placed over the first delivery device of the present invention provides Several terminus 32 of the connector 30 Such that the carrier fluid features that Serve individually and collectively to eliminate inlet lumens 36C are aligned with outlet orifice 12. In clogging. For example, a diffuser Surface typically has a addition, each of a plurality of different fluid component shape effective to direct and maintain each received fluid Sources is provided fluid communication with the fluid component in a different flow path on the diffuser surface component inlet lumens 36A and 36B and at least one source toward the outlet for mixing therein and dispensing there of pressurized carrier gas is provided fluid communication through. Due to the minimal residence time of the mixture with the carrier fluid inlet 36C. within the nozzle, reactive components do not have time to 0234. As discussed above, the interior surface 18 of the Set and clog the nozzle before the mixture is forced out of the cap 10 at distal end 14 serves as a diffuser surface 18 that is nozzle by the pressurized carrier fluid. In addition, the outlet adapted to receive fluid components thereon. AS depicted in may be aligned with any or all of the carrier fluid inlets that FIG. 2, the diffuser Surface 18 exhibits two-fold axial may be present in the nozzle to direct the pressurized carrier Symmetry. The dotted lines indicate the position of lumens fluid in a manner that enhances fluid component mixing and 36A, 36B, and 36C relative to the diffuser surface 18. to expel the mixture in a jet like manner. AS the orientation Similarly, in FIG. 2, the dashed lines shown within the of the diffuser Surface relative to the inlets affects the connector 30 indicate the separate flow paths of the fluid performance of the device, the diffuser Surface may be components emerging from the fluid component inlet permanently affixed or immobilized with respect to the lumens 36A and 36B, respectively, and directed by the inlets; however, when the diffuser Surface is detachable from diffuser surface 18 in a generally inward direction toward the the inlets, the nozzle may be disassembled to facilitate center outlet orifice 12. Once the fluid components reach cleaning and/or replacement of parts. For example, the outlet orifice 12, pressurized gas from carrier fluid lumens diffuser surface may be replaceable/and or disposable. Nev 36C mix the fluid components and force the mixture out of ertheless, when the pressurized delivery device of the the outlet orifice 12. present invention has diffuser Surface that is detachable from 0235. In the pressurized delivery device of the present the inlets, the device may be constructed to allow assembly invention, the geometries of and Spatial relationships of the components in only configurations that align the between the various components of diffuser surface 18 diffuser surface to the inlets such that the performance of the represent an important aspect of the preSSurized delivery device is optimized. device. For example, the pressurized delivery device may be 0231 FIGS. 2 and 3 illustrate an example of the pres used to carry out mixing of a plurality of reactive compo surized delivery device of the present invention in the form nents. Typically, nozzles for mixing reactive components are of a nozzle that includes all of the above-discussed features of the external mixing category because previously known which Serve eliminate the problems associated with nozzle internal mixing designs are prone to clogging. Clogging clogging. AS is the case with all figures referenced herein, in results when reactive components are mixed prior to intro which like parts are referenced by like numerals, FIGS. 2 duction to the gas Stream. In contrast, the pressurized and 3 are not necessarily to Scale, and certain dimensions delivery device provides a high-pressure area between the may be exaggerated for clarity of presentation. AS depicted inlets 36A-36C and the diffuser Surface 18 that serves to mix in FIG. 2, the nozzle 1 includes a cap 10 having a slot reactive fluids while Simultaneously forcing the mixture out shaped outlet orifice 12 that extends through the center of the of the orifice 12. US 2005/0281883 A1 Dec. 22, 2005 22

0236. In addition, the diffuser surface 18 is located down inlets are positioned Such that a high-pressure Zone is stream from the inlets 36A-36C and is effective to direct created between the component inlets and the diffuser Sur fluid components toward the outlet for mixing and dispens face while a comparatively low-pressure Zone is created ing therethrough by a preSSurized carrier fluid; thus, the downstream from the outlet. “Dead space” that serve to trap diffuser Surface 1-8 should exhibit an appropriate shape to residue is generally avoided. As a result, a fluid mixture is carry out its desired function while minimizing the odds of forced through the outlet in a jet-like fashion, thereby device clogging. For example, while the diffuser Surface 18 reducing any potential or actual buildup of residue that Serve depicted in FIGS. 2 and 3 is located within a cylindrical cap to clog the pressurized delivery device. 10 having a flat exterior circular end Surface and contains a 0241. In general, any of a number of carrier fluids may be centrally located slot-shaped orifice 12, Such geometry is not employed with the preSSurized delivery device of the present required. invention. For example, the carrier fluid may be gaseous 0237 As depicted in FIGS. 2 and 3, the exterior surface and/or liquid in nature. Typically, however, the carrier fluid of the cap is parallel to the diffuser surface. While such a is chemically inert with respect to the fluid components. parallel configuration of the exterior Surface of the cap is Suitable inert gases include, without limitation, air, carbon preferred, it is understood that it is merely exemplary and dioxide, nitrogen, argon, helium, gaseous perfluorinated not a requirement of the pressurized delivery System of the alkanes and ethers, gaseous chlorofluorocarbons and the present invention. Similarly, while both FIGS. 2 and 3 like. Suitable inert liquids include, without limitation, pol depict caps that exhibit axial Symmetry, Such axial Symmetry ySiloxanes, perfluroinated polyethers, and the like. PreSSur is merely preferred and not required. Where the caps of the ized air represents an economical and practical carrier fluid present invention are Symmetrical, the Symmetry may be for use with the pressurized delivery device. Equipment axial or mirror Symmetry. It is expected that variations on asSociated with pressurized air is well known in the art and diffuser Surface shapes and nozzle configurations may be may include pressurized tanks or cylinders as well as developed through routine experimentation. With respect to compressors. In Some instances, one or more check valves, the inlets, the pressurized delivery device of the present e.g., one-way valves, may be provided to prevent back flow invention generally requires a plurality of fluid component of fluid component resulting from pressure buildup associ inlets for communication with an equal or leSS number of ated with the use of the pressurized delivery device. Such Sources of fluid components. While a single carrier fluid check valves may be positioned upstream from the diffuser inlet may be provided, the pressurized delivery device Surface, e.g., within lumens associated with the inlets. Such typically provides a plurality of carrier fluid inlets. Often the check valves are particularly useful when inlet lumens are carrier fluid inlets are provided communication with a single Short, e.g., about 2 to about 5 centimeters in length, because Source of carrier fluid via a splitter or manifold, though a the potential for back flow tends to be inversely proportional plurality of carrier fluid Sources may be advantageously used to the length of the lumens; however, check valves may be as well in certain instances. employed with longer lumens as well. 0242. The portions of the device that contact multicom 0238. In addition, inlets are typically each located at the ponent composition and the fluid components thereofshould terminus of a corresponding lumen. In Some instances, the be inert and preferably repellant to the materials contacted. lumens may coextend through an elongate cylindrical con Thus, portions of the device that contact the fluids in nector 30, as depicted in FIG. 2 (with 36A-36C depicting operation should be Selected according to the fluids them the lumens). Alternatively, the lumens may extend through Selves. For example, the device or components thereof may Separate tubes. Furthermore, tubes and/or tubing members be made from a plastic Such as polycarbonates, polyure may be constructed to form a lumen assembly. For example, thane, polyesters, acrylics, ABS polymers, polysulfone, and various lengths of multilumen delivery tubing for use in the like. Adhesion inhibiting coatings Such as polysiloxanes, Specific Surgical or non-Surgical applications. Particularly in perfluorinated polymers, and the like may be used as well. laparoscopic applications, it may be useful to employ flex Thus, the diffuser Surface is typically inert and optionally ible tubing. The tubing Serves to maintain Separation of the repelling to the fluid components. Similar, lumen Surfaces two fluid components and provide a pathway for the delivery that may contact the fluid components or the carrier fluid are of pressurized gas to the diffuser Surface. typically inert and optionally repelling to the corresponding 0239). Additional features also serve to enhance the mix fluid as well. ing and delivery performance of the pressurized delivery 0243 The pressurized delivery device of the present device of the present invention. AS discussed above, two or invention is particularly useful for dispensing multicompo more fluid components may be individually delivered nent compositions. While Some gaseous components may be through inlets to impinge upon the diffuser Surface. Typi used, the pressurized delivery device is particularly useful cally, the components first impinge upon the diffuser plate for liquids. Thus, at least one fluid component is usually a near the outlet to reduce the residence time of the compo liquid. Often, each fluid component includes a liquid. For nents in the device. Any of a number of means may be used example, the pressurized delivery device is useful to dis to provide motive force to introduce fluid components pense composition Such as fluid mixtures, wherein the through the inlets and toward the outlet. Exemplary motive mixing of a plurality of fluids results in an increase in force means include pumps, compressors, pistons, and the Viscosity Sufficient to impair mixture flow. Such composi like. tions may be formed from fluid components that are chemi 0240 Then, as the diffuser plate directs the components cally reactive with respect to each other. In Some instances, toward the outlet 12, and the pressurized carrier fluid Simul a crosslinking agent may be provided. taneously provides a force to mix and expel the components 0244. In practice, then, a diffuser Surface having an outlet through the outlet. Accordingly, one or more the carrier fluid extending therethrough Such that the diffuser Surface is US 2005/0281883 A1 Dec. 22, 2005 23 downstream from a plurality of fluid component inlets and is attached to the housing assembly to allow mixing of the at least one carrier fluid inlet. A different fluid component is dry powders into the correct Syringe. A clip is attached to the directed from each of the fluid component inlets toward the plunger rod of the Syringe that does not require mixing with diffuser Surface. In Some instances, fluid components are the dry powders. The Second pouch is a powder component directed at Substantially the same flow rate toward the pouch, which consists of a Syringe containing the dry diffuser Surface. Alternatively, the fluid components are powder(s) and a dessicant package. The third pouch is an directed at different flow rates toward the diffuser Surface. applicator pouch, which contains two applicators. Typically, the flow rate of the carrier fluid is higher than that for the fluid components. The diffuser Surface maintains and 0251 To use the preferred kit of FIG. 1, each pouch is directs each received fluid component in a different flow opened using aseptic techniques and the contents of each path toward the outlet. Pressurized carrier fluid from the at pouch are transferred into a sterile field. In the sterile field, least one carrier fluid inlet is also directed through the outlet, the liquid and powder components are prepared as follows. thereby mixing the fluid components present at the outlet Without removing the Syringe clip, the luer cap on the and dispensing the composition through the outlet. transfer port closure is removed. The cap is removed from the powder Syringe and the powder Syringe is connected to 0245) V. Kits the opening of the transfer port closure. The liquid is transferred into the powder by forcefully depressing the 0246 The compositions of the invention can also be plunger. The contents between the two Syringes are mixed packaged in kits and used in a variety of medical applica back and forth between the two syringes until the Solid is tions. The kit would include buffer Solutions, as well as completely dissolved (e.g., 18-20 times). The entire content written or otherwise illustrated instructions for use. Atypical is then pushed into the Syrinting contained in the Syringe kit for use in medical applications, comprises: (a) a homo housing. The powder Syringe is disengaged by detaching the geneous dry powder composition comprised of: (i) a first transfer port closure by grasping the powder Syringe barrel; component having a core Substituted with m nucleophilic pressing the levers on the Syringe housing, and pulling both groups, where me2; and (ii) a Second component having a the empty powder Syringe and transfer port closure from the core Substituted with n electrophilic groups, where ne2 and housing. To expel all air from the Syringe, the Syringe tips m+ne 4, wherein the nucleophilic and electrophilic groups are held up, the Syringe plungers are leveled, the Syringe clip are non-reactive in a dry environment but are rendered is rotated to connect to the other plunger; and holding the reactive upon exposure to an aqueous environment Such that Syringe upright, all air is expelled from the Syringe. AS a the components inter-react in the aqueous environment to final step, the applicator is Snapped onto the end of the form a three-dimensional matrix; (b) a first buffer solution having a pH within the range of about 1.0 to 5.5; and (c) a Syringe housing making the composition ready to use. A Second buffer Solution having a pH within the range of about clear gel should be seen approximately three minutes fol 6.0 to 11.0, wherein each component is packaged Separately lowing the mixing of the components. and admixed immediately prior to use. AS is evident to those 0252) In another embodiment of the kit, a pressurized of ordinary skill in the art, prior to use, each component delivery device is included in the kit. AS previously should remain in a separate Sterile package. described, the pressurized delivery device of the present 0247. In another embodiment, the kit can further com invention includes a plurality of fluid component inlets each prise a delivery System that will allow the composition to be adapted to communicate with a Source of different fluid delivered as a Spray. The Spray can be generated by manually components, at least one carrier fluid inlet adapted to mixing the components and passing them through a Spray communicate with a Source of a pressurized carrier fluid; a diffuser surface located downstream from the plurality of nozzle. The Spray generation can also be accomplished by fluid component inlets and the at least one carrier fluid inlet; using a flow of gas (for example, air, nitrogen, carbon and an outlet extending through the diffuser Surface, wherein dioxide). the diffuser Surface is adapted to receive fluid components 0248 Kits contemplated under the present invention will thereon and has a shape effective to direct and maintain each preferably include a delivery System for the compositions of received fluid component in a different flow path toward the the present invention. Delivery devices that may be included outlet for mixing and dispensing therethrough by the pres in the kits will preferably be one of the multi-component Surized carrier fluid from the at least one carrier fluid inlet. Syringe device and/or the pressurized delivery devices described herein. 0253 Kits contemplated under the present invention are not limited to the devices described herein and may also 0249. In one embodiment of the kit, a multi-component include any other suitable delivery device known in the art Syringe device is included in the kit. AS previously of drug delivery. described, the multi-component Spray device may be a multiple-compartment Syringe System having multiple bar 0254 Exemplary medical applications involve, by way of rels, a mixing head, and an exit orifice, wherein the dry illustration and not limitation, adhering or Sealing biological powder composition, the first buffer, and the second buffer tissue, delivering a biologically active agent (in which case, are housed Separately in the multiple-compartment Syringe the kit would further comprise a biologically active agent, for example, mixed with the components to form a homo System. geneous mixture or packaged separately), delivering cells 0250 FIG. 1 describes a preferred embodiment of the and genes (in which case, the kit would further comprise the multi-compartment device. When provided in a kit, the living cells or genes, for example, mixed with the compo device is provided with three pouches. The first pouch is a nents to form a homogeneous mixture or packaged Sepa liquid components pouch, which consists of two Syringes rately), bioadhesion, in ophthalmic applications, for tissue that are preassembled into a housing. A transfer port closure augmentation, for adhesion prevention, forming Synthetic US 2005/0281883 A1 Dec. 22, 2005 24 implants and coating Synthetic implants, and for the treat tissue may be rapidly pressed against the first tissue or 2) by ment of aneurysms. In a preferred embodiment, the mixture bringing the tissues in close juxtaposition and then applying of the biologically active agents with the components is a the compositions. In addition, the compositions can be used homogeneous mixture; however, this feature is not required. to fill Spaces in Soft and hard tissues that are created by Whether packaged together or Separately, each of the com disease or Surgery. ponents and the biological agent should be in Sterile pack 0261) Therefore, one embodiment of the invention is a ages prior to use. method of Sealing tissue of a patient comprising the Steps of: 0255 For purposes of description only, the surgical use of (a) providing a composition of the invention; (b) rendering the multi-compartment Syringe kit of FIG. 1 is described. As the nucleophilic and electrophilic groups reactive by expos a preliminary Step, blood circulation to the Surgical Site is ing the composition to an aqueous environment to effect restored to expand the graft by unclamping the Site and after inter-reaction; wherein said exposure comprises: (i) dissolv circulation is restored, the Site is reclamped to Stop circula ing the composition in a first buffer Solution having a pH tion. ExceSS blood is aspirated and all Surfaces are air dried within the range of about 1.0 to 5.5 to form a homogeneous prior to application of the composition. Holding the appli Solution, and (ii) adding a Second buffer Solution having a cator approximately 3 cm from the site (touching the Site or pH within the range of about 6.0 to 11.0 to the homogeneous holding more than 6 cm from the site is not recommended), Solution to form a mixture; and (c) placing the mixture into Sealant is forcibly applied to the Site. To enhance mixing, the contact with tissue and allowing a three-dimensional matrix applicator is moved quickly along the anastomotic Site. If the to form and Seal the tissue. In another embodiment, the composition is to be applied to more than one site, the compositions can be applied in conjunction with an applicator tip should be wiped with gauze and the device implanted medical device Such that it prevents the leakage of should be set upright to prevent clogging. If the composition gases, liquids or Solids from the device or from the device does not gel within 30 Seconds, i.e., the composition remains tissue interface. For example, following the implantation of watery on the site, the site should be flushed with Saline and a vascular graft (either Synthetic or biological), there is often the material aspirated. If the treated Site fails to Seal, the leakage of blood through the Suture holes in the graft or at Surface should be blotted dry, reclamping the vessel may be the interface between the graft and the tissue. The compo required to dry the field for reapplication of the composition. Sition of the invention can be applied to this area to prevent If the applicator becomes clogged, it should be replaced with further blood leakage. a new applicator. 0262. In certain aspects of the invention, the composition may be further combined with a fibrosing agent to further 0256 VI. Uses enhance the properties of the Sealant or adhesive. In one 0257 The compositions of the present invention can be aspect of the present invention, a fibrosing (i.e., Scarring) used in a variety of different applications. In general, the agent can be included in a polymeric Sealant Spray which compositions can be adapted for use in any tissue engineer Solidifies into a film or coating to promote fibrosis and Seal ing application where Synthetic gel matrices are currently air leaks. being utilized. For example, the compositions are useful as 0263. In one illustrative application, a fibrosing agent tissue Sealants, vascular Sealants, in tissue augmentation, in may be included with the polymer composition for use as a tissue repair, as hemostatic agents, in preventing tissue pulmonary Sealant during open or endoscopic lung reduction adhesions, in providing Surface modifications, and in drug/ Surgeries, for example, to Seal off pulmonary bullae in open cell/gene delivery applications and may be used in a variety and endoscopic lung destruction procedures. The addition of of open, endocopic, and laparoscopic Surgical procedures. a fibrosis-inducing agent to a pulmonary Sealant can induce One of skill in the art can easily determine the appropriate the formation of a Stable, fibrous Scar that permanently Seals administration protocol to use with any particular composi the parietal Surface of the lung at the Surgical location (or the tion having a known gel Strength and gelation time. A more alveolar Surface of the lung if delivered endoscopically detailed description of Several Specific applications is given during lung reduction Surgery), reduces hospitalization time below. and prevents recurrence of the air leak. Clinically a fibrosis inducing pulmonary Sealant can be useful to improve the 0258 1. Tissue Sealants and Adhesives outcomes in open lung Surgery, endoscopic lung reduction 0259. In one application, the compositions described Surgery for emphysema (severe COPD), esophageal leaks herein can be used for medical conditions that require a after endoscopy or resection, complications of treatment of coating or Sealing layer to prevent the leakage of gases, other intra-thoracic malignancies, pleural effusion, hae liquid or Solids. mothorax, pneumothorax, chylothorax, complications of 0260 The method entails applying the composition to the aspiration, and tuberculosis. damaged tissue or organ to Seal 1) vascular and or other 0264. It should be apparent to one of skill in the art that tissues or organs to stop or minimize the flow of blood; 2) potentially any adhesion or fibrosis-inducing agents thoracic tissue to stop or minimize the leakage of air; 3) described above may be utilized alone, or in combination gastrointestinal tract or pancreatic tissue to Stop or minimize with the present composition, in the practice of this embodi the leakage of fecal or tissue contents; 4) bladder or urethra ment. Exemplary fibrosing agents for use in Sealants and to Stop or minimize the leakage of urine; 5) dura to stop or adhesive include talc, Silk, wool, chitosan, polylysine, minimize the leakage of CSF; and 6) skin or Serosal tissue fibronectin, bleomycin, and connective tissue growth factor to Stop the leakage of Serosal fluid. These compositions may (CTGF), as well as analogues and derivatives of the afore also be used to adhere tissues together Such as Small vessels, mentioned. nerves or dermal tissue. The compositions can be used 1) by 0265. The exact dose administered can vary with the applying them to the Surface of one tissue and then a Second composition of the Sealant or adhesive; however, certain US 2005/0281883 A1 Dec. 22, 2005 25 principles can be applied in the application of this art. Drug of the Surface area of the lung to which drug is applied) dose can be calculated as a function of dose per unit area (of should fall within the range of 0.05 ug-10 ug per mm of the amount of the Sealant being applied), total drug dose Surface area coated. In another embodiment, polylysine administered can be measured and appropriate Surface con should be applied to a lung surface at a dose of 0.05 centrations of active drug can be determined. Regardless of tag/mm-10 ug/mm of Surface area coated. AS specific the method of incorporation of the drug into the Sealant or (polymeric and non-polymeric) drug delivery vehicles and adhesive, the exemplary fibrosing agents, used alone or in Specific pulmonary Sealants can release polylysine at differ combination, should be administered under the following ing rates, the above dosing parameters should be utilized in dosing guidelines: combination with the release rate of the drug from the 0266 Utilizing talc as an exemplary fibrosis-inducing pulmonary Sealant Such that a minimum concentration of agent, the total dose of talc delivered from a Sealant, or 0.01 nM to 1000 uM of polylysine is delivered to the tissue. coated onto the Surface of a lung, should not exceed 100 mg 0270 Utilizing fibronectin as an exemplary fibrosis-in (range of 1 lug to 100 mg). In one embodiment, the total ducing agent, the total dose of fibronectin delivered from a amount of talc released from the Sealant should be in the pulmonary Sealant, or coated onto the Surface of a lung, range of 10 ug to 50 mg. The dose per unit area (i.e., the should not exceed 100 mg (range of 1 ug to 100 mg). In one dosage of talc as a function of the Surface area of the lung embodiment, the total amount of fibronectin released from to which drug is applied) should fall within the range of 0.05 the sealant should be in the range of 10 ug to 50 mg. The tug-10 ug per mm of Surface area coated. In another embodi dose per unit area (i.e., the dosage of fibronectin as a ment, talc should be applied to a lung Surface at a dose of function of the Surface area of the lung to which drug is 0.05 ug/mm-10 ug/mm of surface area coated. applied) should fall within the range of 0.05 lug-10 ug per 0267 Utilizing silk as an exemplary fibrosis-inducing mm of Surface area coated. In another embodiment, agent, the total dose of Silk delivered from a pulmonary fibronectin should be applied to a lung Surface at a dose of Sealant, or coated onto the Surface of a lung, should not 0.05ug/mm-10 ug/mm of Surface area coated. AS specific exceed 100 mg (range of 1 ug to 100 ug). In one embodi (polymeric and non-polymeric) drug delivery vehicles and ment, the total amount of Silk released from the Sealant Specific pulmonary Sealants can release fibronectin at dif should be in the range of 10 ug to 50 mg. The dose per unit fering rates, the above dosing parameters should be utilized area (i.e., the dosage of Silk as a function of the Surface area in combination with the release rate of the drug from the of the lung to which drug is applied) should fall within the Sealant Such that a minimum concentration of 0.01 nM to range of 0.05 ug-10 ug per mm of Surface area coated. In 1000 uM of fibronectin is delivered to the tissue. another embodiment, Silk should be applied to a lung Surface at a dose of 0.05 ug/mM-10 ug/mm of surface area coated. 0271 Utilizing bleomycin as an exemplary fibrosis-in AS specific (polymeric and non-polymeric) drug delivery ducing agent, the total dose of bleomycin delivered from a vehicles and Specific pulmonary Sealants can release Silk at pulmonary Sealant, or coated onto the Surface of a lung, differing rates, the above dosing parameters should be should not exceed 100 mg (range of 0.01ug to 100 mg). In one embodiment, the total amount of bleomycin released utilized in combination with the release rate of the drug from from the sealant should be in the range of 0.010 ug to 50 mg. the Sealant Such that a minimum concentration of 0.01 nM The dose per unit area (i.e., the dosage of bleomycin as a to 1000 uM of silk is delivered to the tissue. function of the Surface area of the lung to which drug is 0268. Utilizing chitosan as an exemplary fibrosis-induc applied) should fall within the range of 0.005ug-10 ug per ing agent, the total dose of chitosan delivered from a mm of Surface area coated. In another embodiment, bleo pulmonary Sealant, or coated onto the Surface of a lung, mycin should be applied to a lung surface at a dose of 0.005 should not exceed 100 mg (range of 1 ug to 100 mg). In one g/mm-10 ug/mm of Surface area coated. AS specific (poly embodiment, the total amount of chitosan released from the meric and non-polymeric) drug delivery vehicles and spe Sealant should be in the range of 10 ug to 50 mg. The dose cific pulmonary Sealants can release bleomycin at differing per unit area (i.e., the dosage of chitosan as a function of the rates, the above dosing parameters should be utilized in Surface area of the lung to which drug is applied) should fall combination with the release rate of the drug from the within the range of 0.05 ug-10 ug per mm of Surface area Sealant Such that a minimum concentration of 0.001 nM to coated. In another embodiment, chitosan should be applied 1000 uM of bleomycin is delivered to the tissue. to a lung Surface at a dose of 0.05 ug/mm-10 ug/mm of Surface area coated. AS Specific (polymeric and non-poly 0272) Utilizing CTGF as an exemplary fibrosis-inducing meric) drug delivery vehicles and specific pulmonary Seal agent, the total dose of CTGF delivered from a pulmonary ants can release chitosan at differing rates, the above dosing Sealant, or coated onto the Surface of a lung, should not parameters should be utilized in combination with the exceed 100 mg (range of 0.01 ug to 100 mg). In one release rate of the drug from the Sealant Such that a minimum embodiment, the total amount of CTGF released from the concentration of 0.01 nM to 1000 uM of chitosan is deliv sealant should be in the range of 0.10 ug to 50 mg. The dose ered to the tissue. per unit area (i.e., the dosage of CTGF as a function of the Surface area of the lung to which drug is applied) should fall 0269. Utilizing polylysine as an exemplary fibrosis-in within the range of 0.005 ug-10 ug per mm of Surface area ducing agent, the total dose of polylysine delivered from a coated. In another embodiment, CTGF should be applied to pulmonary Sealant, or coated onto the Surface of a lung, a lung surface at a dose of 0.005 ug/mm°-10 ug/mm of should not exceed 100 mg (range of 1 ug to 100 mg). In one Surface area coated. AS Specific (polymeric and non-poly embodiment, the total amount of polylysine released from meric) drug delivery vehicles and specific pulmonary Seal the sealant should be in the range of 10 ug to 50 mg. The ants can release CTGF at differing rates, the above dosing dose per unit area (i.e., the dosage of polylysine as a function parameters should be utilized in combination with the US 2005/0281883 A1 Dec. 22, 2005 26 release rate of the drug from the Sealant Such that a minimum dose of 0.0001 ug/mm-200 ug/mm. Minimum concentra concentration of 0.001 nM to 1000 uM of CTGF is delivered tion of 10'-10M of proliferative agent is to be main to the tissue. tained on the device Surface. 0273 The fibrosing agent (e.g., talc, Silk, chitosan, polyl 0276 2. Biologically Active Agent Delivery ysine, fibronectin, bleomycin, CTGF) may be released from 0277. The compositions may also be used for localized the pulmonary Sealant Such that fibrosis in the tissue is delivery of various drugs and other biologically active promoted for a period ranging from Several hours to Several agents. The biologically active agent can either be admixed months. For example, the fibrosing agent may be released in with the compositions of the invention or be chemically effective concentrations for a period ranging from 1 hour-30 coupled to one of the individual components in the compo days. It should be readily evident given the discussions Sition, e.g., by attachment to one of the reactive groups. For provided herein that analogues and derivatives of the fibroS example, processes for covalently binding biologically ing agent (e.g., analogues and derivatives of talc, silk, active agents Such as growth factors using functionally chitosan, polylysine, fibronectin, bleomycin, CTGF, as pre activated polyethylene glycols are described in U.S. Pat. No. viously described) with similar functional activity can be 5,162,430 to Rhee et al. Such compositions preferably utilized for the purposes of this invention; the above dosing include linkages that can be easily biodegraded, for example parameters are then adjusted according to the relative as a result of enzymatic degradation, resulting in the release potency of the analogue or derivative as compared to the of the active agent into the target tissue, where it will exert parent compound (e.g., a compound twice as potent as the its desired therapeutic effect. agent is administered at half the above parameters, a com 0278 In certain aspects, the biologically active agent pound half as potent as the agent is administered at twice the may be incorporated with a polymeric or non-polymeric above parameters, etc.). carrier to facilitate the incorporation of the agent into the composition. In certain aspects, the carrier may facilitate 0274. Optionally, the sealant may alone or additionally Sustained release of the agent from the composition over a comprise an inflammatory cytokine (e.g., TGFB, PDGF, prolonged period of time (e.g., over the course of Several VEGF, bFGF, TNFo, NGF, GM-CSF, IGF-a, IL-1, IL-1-?3, days, weeks, or months). For many embodiments, localized IL-8, IL-6, and growth hormone) or an analogue or deriva delivery as well as localized Sustained delivery of the agent tive thereof. Inflammatory cytokines are to be used in may be desired. For example, a therapeutic agent may be formulations at concentrations that range from 0.0001 tug/ml admixed with, blended with, conjugated to, or, otherwise to approximately 20 mg/ml depending on the Specific clini modified to contain a polymeric composition (which may be cal application, formulation type (e.g., gel, liquid, Solid, either biodegradable or non-biodegradable) or non-poly Semi-solid), formulation chemistry, duration of required meric composition in order to release the therapeutic agent application, type of medical device interface and formula over a period of time. tion volume and or Surface area coverage required. Prefer ably, the inflammatory cytokine is released in effective 0279 Representative examples of biodegradable poly concentrations for a period ranging from 1-180 days. The merS Suitable for the delivery of therapeutic agents include total dose for a single application is typically not to exceed albumin, collagen, gelatin, hyaluronic acid, Starch, cellulose 500 mg (range of 0.0001 ug to 100 mg); preferred 0.001 ug and cellulose derivatives (e.g., regenerated cellulose, meth to 50 mg. When used as a device coating, the dose is per unit ylcellulose, hydroxypropylcellulose, hydroxypropylmethyl area of 0.0001 ug-500 ug per mm’; with a preferred dose of cellulose, carboxymethylcellulose, cellulose acetate phtha 0.001 tug/mm-200 ug/mm. Minimum concentration of late, cellulose acetate Succinate, 10-10 g/ml of inflammatory cytokine is to be maintained hydroxypropylmethylcellulose phthalate), casein, dextrans, polysaccharides, fibrinogen, poly(ether ester) multiblock on the device Surface. copolymers, based on poly(ethylene glycol) and poly(buty 0275. Furthermore, the sealant may alone or additionally lene terephthalate), tyrosine-derived polycarbonates (e.g., comprise an agent that Stimulates cellular proliferation. U.S. Pat. No. 6,120,491), poly(hydroxyl acids), poly(D.L- Examples include: dexamethasone, isotretinoin (13-cis ret lactide), poly(D.L-lactide-co-glycolide), poly(glycolide), inoic acid), 17-f-estradiol, estradiol, 1-O-25 dihydroxyvita poly(hydroxybutyrate), polydioxanone, poly(alkylcarbon min D, diethylstibesterol, cyclosporine A, L-NAME, all ate) and poly(orthoesters), polyesters, poly(hydroxyvaleric trans retinoic acid (ATRA), and analogues and derivatives acid), polydioxanone, polyesters, poly(malic acid), poly(tar thereof. Doses used are those concentrations which are tronic acid), poly(acrylamides), polyanhydrides, polyphos demonstrated to Stimulate cell proliferation. The prolifera phaZenes, poly(amino acids), poly(alkylene oxide)-poly(es tive agents are to be used in formulations at concentrations ter) block copolymers (e.g., X-Y, X-Y-X, Y-X-Y, that range from 0.0000001 to 25 mg/ml depending on the R-(Y-X), or R-(X-Y), where X is a polyalkylene Specific clinical application, formulation type (e.g., gel, oxide (e.g., poly(ethylene glycol, poly(propylene glycol) liquid, Solid, Semi-solid), formulation chemistry, duration of and block copolymers of poly(ethylene oxide) and poly(pro required application, type of medical device interface and pylene oxide) (e.g., PLURONIC and PLURONIC R series formulation volume and or Surface area coverage required. of polymers from BASF Corporation, Mount Olive, N.J.) Preferably, the proliferative agent is released in effective and Y is a polyester, where the polyester may comprise the concentrations for a period ranging from 1-180 days. The residues of one or more of the monomerS Selected from total dose for a single application is typically not to exceed lactide, lactic acid, glycolide, glycolic acid, e-caprolactone, 500 mg (range of 0.0001 ug to 200 mg); preferred 0.001 ug gamma-caprolactone, hydroxyvaleric acid, hydroxybutyric to 100 mg. When used as a device coating, the dose is per acid, beta-butyrolactone, gamma-butyrolactone, gamma unit area of 0.00001 ug-500 ug per mm; with a preferred Valerolactone, y-decanolactone, 8-decanolactone, trimethyl US 2005/0281883 A1 Dec. 22, 2005 27 ene carbonate, 1,4-dioxane-2-one or 1,5-dioxepan-2One Valeric acid, hydroxybutyric acid, beta-butyrolactone, (e.g., PLGA, PLA, PCL, polydioxanone and copolymers gamma-butyrolactone, gamma-Valerolactone, y-decanolac thereof) and R is a multifunctional initiator), and the copoly tone, Ö-decanolactone, trimethylene carbonate, 1,4-dioxane mers as well as blends thereof (see generally, Illum, L., 2-one or 1,5-dioxepan-2One and R is a multifunctional Davids, S. S. (eds.) “Polymers in Controlled Drug Delivery” initiator), Silicone rubbers, poly(styrene)block-poly(isobu Wright, Bristol, 1987; Arshady, J. Controlled Release 17:1- tylene)-block-poly(styrene), poly(acrylate) polymers and 22, 1991; Pitt, Int. J. Phar. 59:173-196, 1990; Holland et al., blends, admixtures, or co-polymers of any of the above. J. Controlled Release 4:155-0180, 1986). Other preferred polymers include collagen, poly(alkylene 0280 Representative examples of non-degradable poly oxide)-based polymers, polysaccharides Such as hyaluronic merS Suitable for the delivery of therapeutic agents include acid, chitosan and fucans, and copolymers of polysaccha poly(ethylene-co-Vinyl acetate) (“EVA) copolymers, aro rides with degradable polymers. matic polyesters, such as poly(ethylene terephthalate), sili cone rubber, acrylic polymers (polyacrylate, polyacrylic 0282. Other representative polymers capable of Sustained acid, polymethylacrylic acid, polymethylmethacrylate, poly localized delivery of therapeutic agents described herein (butyl methacrylate)), poly(alkylcynoacrylate) (e.g., poly include carboxylic polymers, polyacetates, polycarbonates, (ethylcyanoacrylate), poly(butylcyanoacrylate) poly(hexyl polyethers, polyethylenes, polyvinylbutyrals, polysilanes, cyanoacrylate) poly(octylcyanoacrylate)), acrylic resin, polyureas, polyoxides, polystyrenes, polysulfides, poly Sul polyethylene, polypropylene, polyamides (nylon 6.6), poly fones, polysulfonides, polyvinylhalides, pyrrollidones, rub urethanes (e.g., CHRONOFLEX AL and CHRONOFLEX bers, thermal-setting polymers, croSS-linkable acrylic and AR (both from CardioTech International, Inc., Woburn, methacrylic polymers, ethylene acrylic acid copolymers, Mass.), TECOFLEX, and BIONATE (Polymer Technology Styrene acrylic copolymers, Vinyl acetate polymers and Group, Inc., Emeryville, Calif.)), poly(ester urethanes), copolymers, Vinyl acetal polymers and copolymers, epoxies, poly(ether urethanes), poly(ester-urea), polyethers (poly melamines, other amino resins, phenolic polymers, and (ethylene oxide), poly(propylene oxide), polyoxyalkylene copolymers thereof, water-insoluble cellulose ester poly ether block copolymers based on ethylene oxide and pro mers (including cellulose acetate propionate, cellulose pylene oxide such as the PLURONIC polymers (e.g., F-127 acetate, cellulose acetate butyrate, cellulose nitrate, cellulose or F87) from BASF Corporation (Mount Olive, N.J.), and acetate phthalate, and mixtures thereof), polyvinylpyrroli poly(tetramethylene glycol), Styrene-based polymers (poly done, polyethylene glycols, polyethylene oxide, polyvinyl styrene, poly(styrene Sulfonic acid), poly(styrene)-block alcohol, polyethers, polysaccharides, hydrophilic polyure poly(isobutylene)-block-poly(styrene), poly(styrene)-poly thane, polyhydroxyacrylate, dextran, Xanthan, hydroxypro (isoprene) block copolymers), and vinyl polymers pyl cellulose, and homopolymers and copolymers of N-Vi (polyvinylpyrrolidone, poly(Vinyl alcohol), poly(Vinyl nylpyrrolidone, N-vinylactam, N-vinyl butyrolactam, acetate phthalate) as well as copolymers and blends thereof. N-Vinyl caprolactam, other vinyl compounds having polar Polymers may also be developed which are either anionic pendant groups, acrylate and methacrylate having hydro (e.g., alginate, carrageenan, carboxymethyl cellulose, poly philic esterifying groups, hydroxyacrylate, and acrylic acid, (acrylamido-2-methyl propane Sulfonic acid) and copoly and combinations thereof; cellulose esters and ethers, ethyl mers thereof, poly(methacrylic acid and copolymers thereof cellulose, hydroxyethyl cellulose, cellulose nitrate, cellulose and poly(acrylic acid) and copolymers thereof, as well as acetate, cellulose acetate butyrate, cellulose acetate propi blends thereof, or cationic (e.g., chitosan, poly-L-lysine, onate, natural and Synthetic elastomers, rubber, acetal, Sty polyethylenimine, and poly(allyl amine)) and blends thereof rene polybutadiene, acrylic resin, polyvinylidene chloride, (see generally, Dunn et al., J. Applied Polymer Sci. 50:353 polycarbonate, homopolymers and copolymers of vinyl 365, 1993; Cascone et al., J. Materials Sci. Materials in compounds, polyvinylchloride, and polyvinylchloride Medicine 5:770-774, 1994; Shiraishi et al., Biol. Pharm. acetate. Bull. 16(11): 1164-1168, 1993; Thacharodi and Rao, Int'l J. 0283 Representative examples of patents relating to Pharm. 120:115-118, 1995; Miyazaki et al., Int'l J. Pharm. drug-delivery polymers and their preparation include PCT 118:257-263, 1995). Publication Nos. WO 98/19713, WO 01/17575, WO 0281. Some examples of preferred polymeric carriers for 01/41821, WO 01/41822, and WO 01/15526 (as well as the the practice of this invention include poly(ethylene-co-Vinyl corresponding U.S. applications), U.S. Pat. Nos. 4,500,676; acetate), polyurethanes, poly (D.L-lactic acid) oligomers 4,582,865; 4,629,623; 4,636,524; 4,713,448; 4,795,741; and polymers, poly (L-lactic acid) oligomers and polymers, 4,913,743; 5,069,899; 5,099,013; 5,128,326; 5,143,724; poly (glycolic acid), copolymers of lactic acid and glycolic 5,153,174, 5,246,698; 5,266,563; 5,399,351; 5,525,348; acid, copolymers of lactide and glycolide, poly (caprolac 5,800,412; 5,837,226; 5,942.555; 5,997.517; 6,007,833; tone), poly (Valerolactone), polyanhydrides, copolymers of 6,071.447; 6,090,995; 6,106,473; 6,110,483; 6,121,027; poly (caprolactone) or poly (lactic acid) with a polyethylene 6,156,345; 6,214,901; 6,368,611; 6,630,155; 6,528,080; glycol (e.g., MePEG), block copolymers of the form X-Y, RE37,950; 6,46,1631; 6,143,314; 5,990,194; 5,792.469; X-Y-X, Y-X-Y, R-(Y-X), or R-(X-Y), where 5,780,044; 5,759,563; 5,744,153; 5,739,176; 5,733,950; X is a polyalkylene oxide (e.g., poly(ethylene glycol, 5,681873; 5,599,552; 5,340,849; 5,278,202; 5,278,201; poly(propylene glycol) and block copolymers of poly(eth 6,589,549; 6,287,588; 6,201,072; 6,117,949; 6,004,573; ylene oxide) and poly(propylene oxide) (e.g., PLURONIC 5,702.717; 6,413,539; 5,714,159; 5,612,052; and U.S. and PLURONIC R series of polymers from BASF Corpo Patent Application Publication Nos. 2003/0068377, 2002/ ration, Mount Olive, N.J.) and Y is a polyester, where the 0192286, 2002/0076441, and 2002/0090398. polyester may comprise the residues of one or more of the monomerS Selected from lactide, lactic acid, glycolide, gly 0284. It should be obvious to one of skill in the art that colic acid, e-caprolactone, gamma-caprolactone, hydroxy the polymers as described herein can also be blended or US 2005/0281883 A1 Dec. 22, 2005 28 copolymerized in various compositions as required to fies into a film or coating for use as a device/implant Surface deliver therapeutic doses of biologically active agents. coating or to line the tissueS of the implantation Site. Such 0285) Drug delivery vehicles may take a variety of forms. Sprays may be prepared from microSpheres of a wide array For example, the carrier may be in the form of microSpheres of sizes, including for example, from 0.1 um to 3 um, from (e.g., PLGA, PLLA, PDLLA, PCL, gelatin, polydioxanone, 10 um to 30 tim, and from 30 um to 100 um. poly(alkylcyanoacrylate)), nanospheres (PLGA, PLLA, 0288. In one aspect, biologically active agents Such as PDLLA, PCL, gelatin, polydioxanone, poly(alkylcy growth factors or fibrosis-inducing agents may be delivered anoacrylate)) (See, e.g., Hagan et al., Proc. Intern. Symp. from the composition to a local tissue site in order to Control Rel. Bioact. Mater 22, 1995; Kwon et al., Pharm facilitate Scar formation, tissue healing, and/or regeneration. Res. 12(2):192-195; Kwon et al., Pharm Res. 10(7):970-974; Thus, in one aspect, a method is provided for delivering a Yokoyama et al., J. Contr. Rel. 32:269-277, 1994; Grefet al., biologically active agent, where the composition also Science 263:1600-1603, 1994; Bazile et al., J. Pharm. Sci. includes the biologically active agent (e.g., a fibrosing 84:493-498, 1994), emulsions (see, e.g., Tarr et al., Pharm agent) to be delivered, and steps (a) and (b) are as described Res. 4: 62-165, 1987), microemulsions, micelles (SDS, for the method of sealing tissue. Step (c) would involve block copolymers of the form X-Y, Y-X-Y, R-(Y- allowing a three-dimensional matrix to form and delivering X), R-(X-Y), and X-Y-X (where X in a polyalkylene the biologically active agent. oxide (e.g., poly(ethylene glycol, poly(propylene glycol) and block copolymers of poly(ethylene oxide) and poly(pro 0289. As described above, the composition may include pylene oxide) (e.g., PLURONIC and PLURONIC R series an agent that promotes fibrosis. Compositions that include a of polymers from BASF Corporation, Mount Olive, N.J.) fibrosis-inducing agent may be used in a variety of appli and Y is a biodegradable polyester, where the polyester may cations, including, without limitation, tissue augmentation, comprise the residues of one or more of the monomers bone growth, treatment of aneurysms, filling and blocking of Selected from lactide, lactic acid, glycolide, glycolic acid, Voids in the body, medical devices coatings, and for use in e-caprolactone, gamma-caprolactone, hydroxyvaleric acid, Sealant compositions. hydroxybutyric acid, beta-butyrolactone, gamma-butyrolac 0290. In certain embodiments, the fibrosis or adhesion tone, gamma-Valerolactone, y-decanolactone, Ö-decanolac inducing agent is Silk. Silk refers to a fibrous protein, and tone, trimethylene carbonate, 1,4-dioxane-2-one or 1,5-di may be obtained from a number of Sources, typically Spiders oxepan-2One (e.g., PLG-PEG-PLG) and R is a and silkworms. Typical silks contain about 75% of actual multifunctional initiator), and Zeolites. fiber, referred to as fibroin, and about 35% sericin, which is a gummy protein that holds the filaments together. Silk 0286. Other types of carriers that may utilized to contain filaments are generally very fine and long-as much as and deliver therapeutic agents described herein include: 300-900 meters long. There are several species of domesti cyclodextrins, Such as hydroxypropyl cyclodextrin (Cserhati cated Silkworm that are used in commercial Silk production, and Hollo, Int. J. Pharm. 108:69-75, 1994), liposomes (see, however, Bombyx mori is the most common, and most Silk e.g., Sharma et al., Cancer Res. 53:5877-5881, 1993; comes from this source. Other Suitable silkworms include Sharma and Straubinger, Pharm. Res. 11(60):889-896, Philosamia cynthia ricini, Antheraea yamamai, Antheraea 1994; WO 93/18751; U.S. Pat. No. 5,242,073), liposome/gel pernyi, and Antheraea my litta. Spider Silk is relatively more (WO 94/26254), nanocapsules (Bartoli et al., J. Microen difficult to obtain, however, recombinant techniques hold capsulation 7(2):191-197, 1990), implants (Jampel et al., promise as a means to obtain Spider Silk at economical prices Invest. Ophthalm. Vis. Science 34(11):3076-3083, 1993; (see, e.g., U.S. Pat. Nos. 6,268,169; 5,994,099; 5,989,894; Walter et al., Cancer Res. 54:22017-2212, 1994), nanopar and 5,728,810, which are exemplary only). Biotechnology ticles (Violante and Lanzafame PAACR), nanoparticles has allowed researchers to develop other Sources for Silk modified (U.S. Pat. No. 5,145,684), nanoparticles (surface production, including animals (e.g., goats) and vegetables modified) (U.S. Pat. No. 5,399,363), micelles such as are described in Alkan-Onyiuksel et al., Pharm. Res. 11(2):206 (e.g., potatoes). Silk from any of these Sources may be used 212, 1994), micelle (surfactant) (U.S. Pat. No. 5,403,858), in the present invention. synthetic phospholipid compounds (U.S. Pat. No. 4,534, 0291. A commercially available silk protein is available 899), gas borne dispersion (U.S. Pat. No. 5,301,664), liquid from Croda, Inc., of Parsippany, N.J., and is Sold under the emulsions, foam, Spray, gel, lotion, cream, ointment, dis trade names CROSILK LIQUID (silk amino acids), CRO persed vesicles, particles or droplets Solid- or liquid-aero SILK 10,000 (hydrolyzed silk), CROSILK POWDER (pow sols, microemulsions (U.S. Pat. No. 5,330,756), polymeric dered silk), and CROSILKQUAT (cocodiammonium shell (nano- and micro-capsule) (U.S. Pat. No. 5,439,686), hydroxypropyl Silk amino acid). Another example of a and implants (U.S. Pat. No. 4,882,168). commercially available silk protein is SERICIN, available 0287. Within certain aspects of the present invention, from Peritapharm, LTD, a division of Kordia, BV, of the therapeutic agents may be fashioned in the form of micro Netherlands. Further details of such silk protein mixtures Spheres, microparticles and/or nanoparticles having any size can be found in U.S. Pat. No. 4,906,460, to Kim, et al., ranging from 50 nm to 500 um, depending upon the par assigned to Sorenco. Silk useful in the present invention ticular use. These compositions can be formed by Spray includes natural (raw) silk, hydrolyzed silk, and modified drying methods, milling methods, coacervation methods, Silk, i.e., Silk that has undergone a chemical, mechanical, or W/O emulsion methods, W/O/W emulsion methods, and Vapor treatment, e.g., acid treatment or acylation (See, e.g., Solvent evaporation methods. In other aspects, these com U.S. Pat. No. 5,747,015). positions can include microemulsions, emulsions, liposomes 0292 Raw silk is typically twisted into a strand suffi and micelles. Compositions comprising a drug loaded car ciently Strong for weaving or knitting. Four different types rier may also be readily applied as a "spray”, which Solidi of Silk thread may be produced by this procedure: organzine, US 2005/0281883 A1 Dec. 22, 2005 29 crepe, tram and thrown Singles. Organzine is a thread made tensin II, growth hormone (GH), and Synthetic peptides, by giving the raw Silk a preliminary twist in one direction analogues or derivatives of these factors are also Suitable for and then twisting two of these threads together in the release from Specific implants and devices to be described opposite direction. Crepe is Similar to organzine but is later. Other examples include CTGF (connective tissue twisted to a much greater extent. Twisting in only one growth factor); inflammatory microcrystals (e.g., crystalline direction two or more raw silk threads makes tram. Thrown minerals such as crystalline Silicates); bromocriptine, meth Singles are individual raw Silk threads that are twisted in ylsergide, methotrexate, chitosan, N-carboxybutyl chitosan, only one direction. Any of these types of Silk threads may be carbon tetrachloride, thioacetamide, fibrosin, ethanol, bleo used in the present invention. mycin, naturally occurring or Synthetic peptides containing 0293. The silk used in the present invention may be in the Arg-Gly-Asp (RGD) sequence, generally at one or both any suitable form that allows the silk to be joined with the termini (see e.g., U.S. Pat. No. 5,997.895), and tissue medical implant, e.g., the Silk may be in thread or powder adhesives, Such as cyanoacrylate and crosslinked poly(eth based forms. The silk can be prepared in the powdered form ylene glycol)-methylated collagen compositions, Such as by several different methods. For example the silk can be described below. milled (e.g., cryomill) into a powdered form. Alternatively 0296 Other examples of fibrosis-inducing agents include the silk can be dissolved in a suitable solvent (e.g., HFIP or agents that promote bone growth, Such as for example bone 9M LiBr) and then sprayed (electrospray, spray dry) or morphogenic proteins. Examples of bone morphogenic pro added to a non-Solvent to produce a powder. Furthermore, teins include the following: BMP-2, BMP-3, BMP-4, BMP the Silk may have any molecular weight, where various 5, BMP-6 (Vgr-1), BMP-7 (OP-1), BMP-8, BMP-9, BMP molecular weights are typically obtained by the hydrolysis 10, BMP-11, BMP-12, BMP-13, BMP-14, BMP-15, and of natural Silk, where the extent and harshness of the BMP-16. Of these, BMP-2, BMP-3, BMP-4, BMP-5, BMP hydrolysis conditions determines the product molecular 6, and BMP-7. Bone morphogenic proteins are described, weight. For example, the Silk may have an average (number for example, in U.S. Pat. Nos. 4,877,864; 5,013,649; 5,661, or weight) molecular weight of about 200 to 5,000. See, e.g., 007; 5,688,678; 6,177,406; 6,432,919; and 6,534,268, and JP-B-59-29199 (examined Japanese patent publication) for a Wozney, J. M., et al. (1988) Science, 242(4885): 1528-1534. description of conditions that may be used to hydrolyze Silk. 0297 Other representative examples of fibrosis-inducing 0294. A discussion of silk may be found in the following agents include components of extracellular matrix (e.g., documents, which are exemplary only: Hinman, M. B., et al. fibronectin, fibrin, fibrinogen, collagen (e.g., bovine col “Synthetic spider silk: a modular fibre”Trends in Biotech lagen), fibrillar and non-fibrillar collagen, adhesive glyco nology, 2000, 18(9) 374-379; Vollrath, F. and Knight, D. P. proteins, proteoglycans (e.g., heparin Sulfate, chondroitin “Liquid crystalline spinning of spider silk’Nature, 2001, Sulfate, dermatan Sulfate), hyaluronan, Secreted protein 410(6828) 541-548; and Hayashi, C. Y., et al. “Hypotheses acidic and rich in cysteine (SPARC), thrombospondins, that correlate the Sequence, Structure, and mechanical prop tenacin, and cell adhesion molecules (including integrins, erties of spider silk proteins' Int. J. Biol. Macromolecules, Vitronectin, fibronectin, laminin, hyaluronic acid, elastin, 1999, 24(2-3), 265-270; and U.S. Pat. No. 6,427.933. bitronectin), proteins found in basement membranes, and 0295) Other representative examples of fibrosis and adhe fibrosin) and inhibitors of matrix metalloproteinases, Such as Sion-inducing agents include irritants (e.g., talc, talcum TIMPs (tissue inhibitors of matrix metalloproteinases) and powder, copper, metallic beryllium (or its oxides), wool Synthetic TIMPs, e.g., marimistat, batimistat, doxycycline, (e.g., animal wool, wood wool, and Synthetic wool), quartz tetracycline, minocycline, TROCADE, Ro-1130830, CGS dust, Silica, crystalline Silicates), polymers (e.g., polylysine, 27023A, and BMS-275291. polyurethanes, poly(ethylene terephthalate), polytetrafluo 0298 Within various embodiments of the invention, a roethylene (PTFE), poly(alkylcyanoacrylates), and poly composition incorporates a compound which acts to Stimu (ethylene-co-Vinylacetate)); vinyl chloride and polymers of late cellular proliferation. In certain embodiments, a com Vinyl chloride; peptides with high lysine content; growth position may incorporate a compound which acts to Stimu factors and inflammatory cytokines involved in angiogen late cellular proliferation in addition to a fibrosing agent. esis, fibroblast migration, fibroblast proliferation, ECM syn Representative examples of agents that Stimulate cellular thesis and tissue remodeling, Such as epidermal growth proliferation include, pyruvic acid, naltrexone, leptin, factor (EGF) family, transforming growth factor-O. (TGF-C), D-glucose, insulin, amlodipine, alginate oligosaccharides, transforming growth factor-B (TGF-B-1, TGF-B-2, TGF-B- minoxidil, dexamethasone, isotretinoin (13-cis retinoic 3), platelet-derived growth factor (PDGF), fibroblast growth acid), 17-B-estradiol, estradiol, 1-C-25 dihydroxyvitamin factor (acidic-aFGF; and basic-bFGF), fibroblast stimulating D, diethylstibesterol, cyclosporine A, L-NAME (L-NG factor-1, activins, vascular endothelial growth factor nitroarginine methyl ester (hydrochloride)), all-trans retinoic (including VEGF-2, VEGF-3, VEGF-A, VEGF-B, VEGF acid (ATRA), and analogues and derivatives thereof. Other C, placental growth factor-PIGF), angiopoietins, insulin examples of agents that Stimulate cellular proliferation like growth factors (IGF), hepatocyte growth factor (HGF), include: Sphingosine 1-phosphate receptor agonist (e.g., connective tissue growth factor (CTGF), myeloid colony FTY-720 (1,3-propanediol, 2-amino-2-(2-(4-octylphenyl Stimulating factors (CSFs), monocyte chemotactic protein, )ethyl)-hydrochloride; immunostimulants, Such as Imupe granulocyte-macrophage colony-stimulating factors (GM done (methanone, 5-amino-2-(4-methyl-1-piperidinyl)phe CSF), granulocyte colony-stimulating factor (G-CSF), mac nyl(4-chlorophenyl)-, DIAPEP227 synthetic peptide rophage colony-stimulating factor (M-CSF), erythropoietin, (Peptor Ltd., Israel)); and nerve growth factor agonist, e.g., interleukins (particularly IL-1, IL-8, and IL-6), tumor necro NG-012 (5H,9H,13H,21H,25H,-dibenzoku 1.5,9,15,19 sis factor-O. (TNF-C), nerve growth factor (NGF), pentaoxacyclotetracosin-5,9,13.21,25-pentone, 7,8,11,12, interferon-Clinterferon-3-histamine, endothelin-1, angio 15,16,23,24,27,28-decahydro-2,4,18,20-tetrahydroxy-11 US 2005/0281883 A1 Dec. 22, 2005 30

(hydroxymethyl)-7,15,23.27-tetramethyl-, NG-121, SS-701 tively charged compounds. Similarly, if a molar excess of (2,2':6'2"-terpyridine, 4'-(4-methylphenyl)-, trihydrochlo electrophilic groups are used, the resulting matrix has a net ride, AMPAlex (piperidine, 1-(6-quinoxalinylcarbonyl)-, negative charge and can be used to ionically bind and deliver RGH-2716 (8-4,4-bis(4-fluorophenyl)butyl-3-(1,1-dim positively charged compounds. Examples of negatively and ethylethyl)-4-methylene-1-Oxa-3,8-diaza-Spiro4.5decan positively charged compounds that can be delivered from 2-one, and TDN-345 (1-oxa-3,8-diazaspiro4.5decan-2- these matrices include various drugs, cells, proteins, and One, 8-4.4-bis(4-fluorophenyl)butyl-3-(1,1- polysaccharides. Negatively charged collagens, Such as Suc dimethylethyl)-4-methylene-). cinylated collagen, and glycosaminoglycan derivatives Such 0299 Particularly useful biologically active agents for as Sodium hyaluronate, keratan Sulfate, keratosulfate, use in the compositions of the present invention are cytok Sodium chondroitin Sulfate A, Sodium dermatan Sulfate B, ines, which are biologically active molecules including Sodium chondroitin Sulfate C, heparin, esterified chondroitin growth factors and active peptides, which aid in healing or Sulfate C, and esterified heparin, can also be effectively regrowth of normal tissue. The function of cytokines is incorporated into the matrix as described above. Positively two-fold: 1) they can incite local cells to produce new charged collagens, Such as methylated collagen, and gly collagen or tissue, or 2) they can attract cells to the Site in cosaminoglycan derivatives Such as esterified deacetylated need of correction. AS Such, cytokines, as well as appropriate hyaluronic acid, esterified deacetylated desulfated chon combinations of cytokines, Serve to encourage “biological droitin Sulfate A, esterified deacetylated desulfated chon anchoring of an implant within the host tissue, by facili droitin Sulfate C, deacetylated desulfated keratan Sulfate, tating the regrowth and remodeling of the implant into deacetylated desulfated keratosulfate, esterified desulfated normal bone tissue. Cytokines may also be used in the heparin, and chitosan, can also be Similarly incorporated. treatment of wounds. Examples of cytokines include, by 0302) In another aspect, biologically active agents such way of illustration and not limitation, transforming growth as fibrosis-inhibiting agents may be delivered from the factors (TGFs); fibroblast growth factors (FGFs), including composition to a local tissue site in order to inhibit Scar both acidic FGF and basic FGF; platelet derived growth formation, tissue healing, and/or regeneration. Thus, in one factors (PDGFs) such as PDGF-AA, PDGF-AB, and PDGF aspect, a method is provided for delivering a biologically BB; epidermal growth factors (EGFs); connective tissue active agent, where the composition also includes the bio activated peptides (CTAPs); colony Stimulating factors logically active agent (e.g., a fibrosis-inhibiting agent) to be (CSFs); erythropoietin (EPO); nerve growth factor (NGF); delivered, and steps (a) and (b) are as described for the osteogenic factors, B-thromboglobulin; tumor necrosis fac method of sealing tissue. Step (c) would involve allowing a tors (TNFs); interleukins; interferons (IFNs); bone morpho three-dimensional matrix to form to deliver the biologically genic protein (BMP); and biologically active analogs, frag active agent. Compositions that include a fibrosis-inhibiting ments, and derivatives of such growth factors. Members of agent may be used in a variety of applications, including, the transforming growth factor (TGF) Supergene family, without limitation, Surgical adhesion prevention and medical which are multifunctional regulatory proteins, are particu devices coatings. Numerous therapeutic compounds have larly preferred. Members of the TGF supergene family been identified that are of utility in the invention including: include TGF-C. and the beta transforming growth factors (for example, TGF-B, 1, TGF-B2, TGF-B3); bone morphogenetic 0303 3. Angiogenesis Inhibitors proteins (for example, BMP-1, BMP-2, BMP-3, BMP-4, 0304. In one embodiment, the pharmacologically active BMP-5, BMP-6, BMP-7, BMP-8, BMP-9); heparin-binding compound is an angiogenesis inhibitor, Such as, for example, growth factors, e.g., FGFs, EGFs, PDGFs; insulin-like 2-ME (NSC-659853), PI-88 (D-mannose), O-6-O- growth factors (IGFs); inhibins such as Inhibin A and phosphono-alpha-D-mannopyranosyl-(1-3)-O-alpha-D- Inhibin B; growth differentiating factors, e.g., GDF-1); and mannopyranosyl-(1-3)-O-alpha-D-mannopyranosyl-(1-3)- activins Such as Activin A, Activin B, Activin AB. Growth O-alpha-D-mannopyranosyl-(1-2)-hydrogen Sulphate), factors can be isolated from native or natural Sources, Such thalidomide (1H-isoindole-1,3 (2H)-dione, 2-(2,6-dioxo-3- as from mammalian cells, or can be prepared Synthetically, piperidinyl)-), CDC-394, CC-5079, ENMD-0995 (S-3- such as by recombinant DNA techniques or by various amino-phthalidoglutarimide), AVE-8062A, Vatalanib, chemical processes. In addition, analogs, fragments, or SH-268, halofuginone hydrobromide, atiprimod dimaleate derivatives of these factors can be used, provided that they (2-azaspivo4.5decane-2-propanamine, N,N-diethyl-8,8- exhibit at least some of the biological activity of the native dipropyl, dimaleate), ATN-224, CHIR-258, combretastatin molecule. For example, analogs can be prepared by expres A-4 (phenol, 2-methoxy-5-2-(3,4,5-trimethoxyphe Sion of genes altered by Site-specific mutagenesis or other nyl)ethenyl-, (Z)-), GCS-100LE, or an analogue or deriva genetic engineering techniques. tive thereof. 0300. By varying the relative molar amounts of the 0305. Other examples of angiogenesis inhibitors for use different the reactive groups on the components, it is poS in the compositions of the invention include: 2-methoX Sible to alter the net charge of the resulting three-dimen yestradiol, A6, ABT-510, ABX-IL8, actimid, Ad5FGF-4, Sional matrix, in order to prepare a matrix for the delivery of AG3340, alpha5beta1 integrin antibody, AMG001, anecor a charged compound Such as a protein or ionizable drug. AS tave acetate, angiocol, angiogenix, angiostatin, angiozyme, Such, the delivery of charged proteins or drugs, which would antiangiogenic antithrombin3, anti-VEGF, anti-VEGF Mab, normally diffuse rapidly out of a neutral carrier matrix, can aplidine, aptosyn, ATN-161, avastin, AVE8062A, Bay be controlled. 12-9566, benefin, BioBypass CAD, MS275291, CAI, car 0301 For example, if a molar excess of nucleophilic boxymidotriazole, CC 4047, CC 5013, CC7085, CDC801, groups are used, the resulting matrix has a net positive Celebrex, CEP-7055, CGP-41251/PKC412, cilengitide, charge and can be used to ionically bind and deliver nega CM101, col-3, combretastatin, combretastatin A4P, CP-547, US 2005/0281883 A1 Dec. 22, 2005

632, CP-564, 959, Del-1, dexrazoxane, didemnin B, tech Inc. Annapolis, Md.), Medi-522, metaret (Suramin), DMXAA, EMD 121974, endostatin, FGF (AGENT 3), neoretna, neovastat (AEtema Laboratories), NM-3, NPe6, flavopiridol, GBC-100, genistein concentrated polysaccha NV1 FGF (Gencel/Aventis), octreotide, oltipraz, paclitaxel ride, green tea extract, HIF-1alpha, human chorio-gonadot (e.g., taxol, docetaxel, or paxene), pegaptainib Sodium (Eye rophin, IM862, INGN 201, interferon alpha-2a, interleukin tech), penicillamine, pentosan polysulphate, PI-88, prinom 12, iressa, ISV-120, LY317615, LY-333531, Mab hull 591 astat (Agouron Pharmaceuticals), PSK, psorvastat, PTK787/ DOTA-90 Yttrium, marimastat, Medi-522, metaret, ZK222584, ranibizumab (Lucentis, Genentech), razoxane, neoretna, neovastat, NM-3, NPe6, NVIFGF, octreotide, olt replistatatin (Platelet factor-4), revimid, RhuMab, ipraz, paclitaxel, pegaptainib Sodium, penicillamine, pen Ro317453, squalamine (Magainin Pharmaceuticals, Inc. Plymouth Meeting, Pa.), SU101 (Sugen Inc. Redwood City, tosan polysulphate, prinomastat, PSK, psorvastat, PTK787/ Calif.), SU11248, SU5416 (Sugen), SU6668 (Sugen), ZK222584, ranibizumab, raZOxane, replistatatin, revimid, tamoxifen, tecogalan Sodium, temptostatin, tetrathiomol, RhuMab, Ro317453, squalamine, SU101, SU11248, tetrathiomolybdate, SU5416, SU6668, tamoxifen, tecogalan sodium, temptosta tin, tetrathiomol, tetrathiomolybdate, thalomid, TNP-470, 0306 Anti-angiogensis compounds found in vivo may be UCN-01, VEGF, VEGF trap, Vioxx, vitaxin, vitaxin-2, used in the compositions and methods described including ZD6126, ZD6474, angiostatin (plasminogen fragment), a angiostatin (plasminogen fragment), metalloproteinase TIMPs, antiangiogenic antithrombin III, pigment epithelial inhibitors (TIMPs), antiangiogenic antithrombin III derived factor (PEDF), canstatin, placental ribonuclease (aaATIII), pigment epithelial-derived factor (PEDF), cansta inhibitor, cartilage-derived inhibitor (CDI), plasminogen tin, placental ribonuclease inhibitor, cartilage-derived activator inhibitor, CD59 complement fragment, platelet inhibitor (CDI), plasminogen activator inhibitor, CD59 factor-4, endostatin (collagen XVIII fragment), prolactin 16 complement fragment, platelet factor-4 (PF4), endostatin kD fragment, fibronectin fragment, proliferin-related pro (collagen XVIII fragment), prolactin 16 kD fragment, tein, gro-beta, a retinoid, a heparinase, tetrahydrocortisol-S, fibronectin fragment, proliferin-related protein, gro-beta, heparin hexasaccharide fragment, thrombospondin-1, retinoids, heparinases, tetrahydrocortisol-S, heparin hexas human chorionic gonadotropin, transforming growth factor accharide fragment, thrombospondin-1, human chorionic beta, interferon alpha, interferon beta, or interferon gamma, gonadotropin (hCG), transforming growth factor-beta, inter tumistatin, interferon inducible protein, vasculoStatin, inter feron alpha/beta/gamma, tumistatin, interferon inducible leukin-12, vasostatin (calreticulin fragment), kringle 5 (plas protein (IP-10), vasculostatin, interleukin-12 (IL-12), vasos minogen fragment), angioarrestin, or 2-methoxyestradiol. tatin (calreticulin fragment), kringle 5 (plasminogen frag Angiogenesis inhibitors also include antagonists of angio ment), angioarrestin, and 2-methoxyestradiol. genin, placental growth factor, angiopoietin-1, platelet-de 0307 Compounds that inhibit, block, or antagonize the rived endothelial cell growth factor, Del-1, platelet-derived angiogenic activity of the following Species in Vivo may be growth factor-BB, aFGF, bFGF, pleiotrophin, follistatin, used in the methods and compositions described herein proliferin, granulocyte colony-stimulating factor, transform including angiogenin, placental growth factor, angiopoietin ing growth factor-alpha, hepatocyte growth factor, trans 1, platelet-derived endothelial cell growth factor (PD forming growth factor-beta, interleukin-8, tumor necrosis ECGF), Del-1, platelet-derived growth factor-BB (PDGF factor-alpha, leptin, Vascular endothelial growth factor, mid BB), fibroblast growth factors: acidic (aFGF) and basic kine, progranulin, 2-methoxyestradiol (PANZEM) (bFGF), pleiotrophin (PTN), follistatin, proliferin, granulo (EntreMed), A6, ABT-510, ABX-IL8 (Abgenix), actimid, cyte colony-stimulating factor (G-CSF), transforming Ad5FGF-4 (Collateral Therapeutics), AG3340 (Agouron growth factor-alpha (TGF-alpha), hepatocyte growth factor Pharmaceuticals Inc. LaJolla, Calif.), alpha5beta1 integrin (HGF)/scatter factor (SF), transforming growth factor-beta antibody, AMG001 (AnCes/Daichi Pharmaceuticals), (TGF-beta), interleukin-8 (IL-8), tumor necrosis factor-al anecortave acetate (Retaane, Alcon), angiocol, angiogenix pha (TNF-alpha), leptin, vascular endothelial growth factor (Endovasc Ltd), angiostatin (EntreMed), angiozyme, anti (VEGF)/vascular permeability factor (VPF), midkine, and angiogenic antithrombin 3 (Genzyme Molecular Oncology), progranulin. anti-VEGF (Genentech), anti-VEGF Mab, aplidine, aptosyn, ATN-161, avastin (bevacizumab), AVE8062A, Bay 12-9566 0308 Other examples of angiogenesis inhibitors for use (Bayer Corp. West Haven, Conn.), benefin, BioBypass CAD in the present compositions include 2-methoxyestradiol, (VEGF-121) (GenVec), MS275291, CAI (carboxy-amido prinomastat, batimastat, BAY 12-9566, carboxyamidotriaz imidazole), carboxymidotriazole, CC 4047 (Celgene), CC ole, CC-1088, dextromethorphan acetic, dimethylxan 5013 (Celgene), CC7085, CDC 801 (Celgene), Celebrex thenone acetic acid, EMD 121974, endostatin, IM-862, (Celecoxib), CEP-7055, CGP-41251/PKC412, cilengitide, marimastat, matrix metalloproteinase, penicillamine, CM 101 (Carbomed Brentwood, Term.), col-3 (CollaGeneX PTK787/ZK 222584, RPI.4610, squalamine, squalamine Pharmaceuticals Inc. Newton, Pa.), combretastatin, combre lactate, SU5416, (..+-)-thalidomide, S-thalidomide, R-tha tastatin A4P (Oxigene/Bristol-Myers Squibb), CP-547, 632, lidomide, TNP-470, combretastatin, paclitaxel, tamoxifen, CP-564,959, Del-1 (VLTS-589) (Valentis), dexrazoxane, COL-3, neovastat, BMS-275291, SU6668, interferon-alpha, didemnin B, DMXAA, EMD 121974, endostatin anti-VEGF antibody, Medi-522 (Vitaxin II), CAI, celecoxib, (EntreMed), FGF (AGENT 3) (Berlex (Krannert Institute of Interleukin-12, IM862, Amilloride, Angiostatin. RTM. Pro Cardiology)), flavopiridol, GBC-100, genistein concentrated tein, Angiostatin K1-3, Angiostatin K1-5, Captopril, DL polysaccharide, green tea extract, HIF-1alpha (Genzyme), alpha-Difluoromethylomithine, DL-alpha-Difluoromethylo human chorio-gonadotrophin, IM862 (Cytran), INGN 201, mithine HCl, His-Tag.RTM. Endostatin.T.M. Protein, interferon alpha-2a, interleukin-12, iressa, ISV-120 (Batim Fumagillin, Herbimycin A, 4-Hydroxyphenylretinamide, astat), LY317615, LY-333531 (Eli Lilly and Company), Mab gamma-interferon, Juglone, Laminin, Laminin Hexapeptide, huJ591-DOTA-90 Yttrium (90Y), marimastat (British Bio Laminin Pentapeptide, Lavendustin A, Medroxyprogester US 2005/0281883 A1 Dec. 22, 2005 32 one, Medroxyprogesterone Acetate, Minocycline, Minocy examples of Such agents include taxanes (e.g., paclitaxel cline HCl, Placental Ribonuclease Inhibitor, Suramin, (discussed in more detail below) and docetaxel) (Schiff et Sodium Salt Suramin, Human Platelet Thrombospondin, al., Nature 277:665-667, 1979; Long and Fairchild, Cancer Tissue Inhibitor of Metalloproteinase 1, Neutrophil Granu Research 54:4355-4361, 1994; Ringel and Horwitz, J. Natl locyte Tissue Inhibitor of Metalloproteinase 1, and Rheu Cancer Inst. 83(4):288-291, 1991; Pazdur et al., Cancer matoid Synovial Fibroblast Tissue Inhibitor of Metallopro Treat. Rev. 19(40):351-386, 1993), etanidazole, nimorazole teinase 2. (B. A. Chabner and D. L. Longo. Cancer Chemotherapy and 0309 4.5-Lipoxygenase Inhibitors and Antagonists Biotherapy-Principles and Practice. Lippincott-Raven 0310. In another embodiment, the pharmacologically Publishers, New York, 1996, p. 554), perfluorochemicals active compound is a 5-lipoxygenase inhibitor or antagonist with hyperbaric oxygen, transfusion, erythropoietin, (e.g., Wy-50295 (2-naphthaleneacetic acid, alpha-methyl-6- BW12C, nicotinamide, hydralazine, BSO, WR-2721, IudR, (2-quinolinylmethoxy)-, (S)-), ONO-LP-269 (2,11,14-eico DUdR, etanidazole, WR-2721, BSO, mono-Substituted satrienamide, N-(4-hydroxy-2-(1H-tetrazol-5-yl)-8-quinoli keto-aldehyde compounds (L. G. Egyud. Keto-aldehyde nyl)-, (E.Z.Z)-), licofelone (1H-pyrrolizine-5-acetic acid, amine addition products and method of making Same. U.S. 6-(4-chlorophenyl)-2,3-dihydro-2,2-dimethyl-7-phenyl-), Pat. No. 4,066,650, Jan. 3, 1978), nitroimidazole (K. C. CMI-568 (urea, N-butyl-N-hydroxy-N-(4-(3-(methylsulfo Agrawal and M. Sakaguchi. Nitroimidazole radioSensitizers nyl)-2-propoxy-5-(tetrahydro-5-(3,4,5-trimethoxyphenyl)- for Hypoxic tumor cells and compositions thereof. U.S. Pat. 2-furanyl)phenoxy)butyl)-trans-), IP-751 ((3R,4R)-(delta No. 4,462.992, Jul. 31, 1984), 5-substituted-4-nitroimida 6)-THC-DMH-11-oic acid), PF-5901 (benzenemethanol, Zoles (Adams et al., Int. J. Radiat. Biol. Relat. Stud. Phys., alpha-pentyl-3-(2-quinolinylmethoxy)-), LY-293111 (ben Chem. Med. 40(2):153-61, 1981), SR-2508 (Brown et al., zoic acid, 2-(3-(3-((5-ethyl-4-fluoro-2-hydroxy(1,1'-biphe Int. J. Radiat. Oncol, Biol. Phys. 7(6):695-703, 1981), nyl)-4-yl)oxy)propoxy)-2-propylphenoxy)-), RG-5901-A 2H-isoindolediones (J. A. Myers, 2H-Isoindolediones, the (benzenemethanol, alpha-pentyl-3-(2-quinolinylmethoxy), synthesis and use as radiosensitizers. U.S. Pat. No. 4,494, hydrochloride), rilopirox (2(1H)-pyridinone, 6-((4-(4-chlo 547, Jan. 22, 1985), chiral (((2-bromoethyl)-amino)methyl)- rophenoxy)phenoxy)methyl)-1-hydroxy-4-methyl-), nitro-1H-imidazole-1-ethanol (V. G. Beylin, et al., Process L-674636 (acetic acid, ((4-(4-chlorophenyl)-1-(4-(2-quino for preparing chiral (((2-bromoethyl)-amino)methyl)-nitro linylmethoxy)phenyl)butyl)thio)-AS)), 7-((3-(4-methoxy 1H-imidazole-1-ethanol and related compounds. U.S. Pat. tetrahydro-2H-pyran-4-yl)phenyl)methoxy)-4-phenylnaph No. 5,543,527, Aug. 6, 1996; U.S. Pat. No. 4,797.397; Jan. tho(2,3-c)furan-1 (3H)-one, MK-886 (1H-indole-2- 10, 1989; U.S. Pat. No. 5,342.959, Aug. 30, 1994), nitroa propanoic acid, 1-((4-chlorophenyl)methyl)-3-((1,1- niline derivatives (W. A. Denny, et al. Nitroaniline deriva dimethylethyl)thio)-alpha, alpha-dimethyl-5-(1- tives and the use as anti-tumor agents. U.S. Pat. No. 5,571, methylethyl)-), quiflapon (1H-indole-2-propanoic acid, 845, Nov. 5, 1996), DNA-affinic hypoxia selective 1-((4-chlorophenyl)methyl)-3-((1,1-dimethylethyl)thio)-al cytotoxins (M. V. Papadopoulou-Rosenzweig. DNA-affinic pha, alpha-dimethyl-5-(2-quinolinylmethoxy)-), quiflapon hypoxia selective cytotoxins. U.S. Pat. No. 5,602,142, Feb. (1H-Indole-2-propanoic acid, 1-((4-chlorophenyl)methyl)- 11, 1997), halogenated DNA ligand (R. F. Martin. Haloge 3-((1,1-dimethylethyl)thio)-alpha, alpha-dimethyl-5-(2- nated DNA ligand radioSensitizers for cancer therapy. U.S. quinolinylmethoxy)-), docebenone (2,5-cyclohexadiene-1, Pat. No. 5,641,764, Jun. 24, 1997), 1.2.4 benzotriazine 4-dione, 2-(12-hydroxy-5,10-dodecadiynyl)-3,5,6- oxides (W. W. Lee et al. 1,2,4-benzotriazine oxides as trimethyl-), zileuton (urea, N-(1-benzo(b)thien-2-ylethyl)- radioSensitizers and Selective cytotoxic agents. U.S. Pat. No. N-hydroxy-), or an analogue or derivative thereof). 5,616,584, Apr. 1, 1997; U.S. Pat. No. 5,624,925, Apr. 29, 1997; Process for Preparing 1,2,4-Benzotriazine oxides. U.S. 0311 5. Chemokine Receptor Antagonists CCR (1,3, and Pat. No. 5,175,287, Dec. 29, 1992), nitric oxide (J. B. 5) Mitchell et al., Use of Nitric oxide releasing compounds as 0312. In another embodiment, the pharmacologically hypoxic cell radiation sensitizers. U.S. Pat. No. 5,650,442, active compound is a chemokine receptor antagonist which Jul. 22, 1997), 2-nitroimidazole derivatives (M. J. Suto et al. inhibits one or more subtypes of CCR (1, 3, and 5) (e.g., 2-Nitroimidazole derivatives useful as radiosensitizers for ONO-4128 (14.9-triazaspiro(5.5)undecane-2,5-dione, hypoxic tumor cells. U.S. Pat. No. 4,797.397, Jan. 10, 1989; 1-butyl-3-(cyclohexylmethyl)-9-((2,3-dihydro-1,4-benzo T. Suzuki. 2-Nitroimidazole derivative, production thereof, dioxin-6-yl)methyl-), L-381, CT-112 (L-arginine, L-threo and radioSensitizer containing the same as active ingredient. nyl-L-threonyl-L-seryl-L-glutaminyl-L-Valyl-L-arginyl-L- U.S. Pat. No. 5,270,330, Dec. 14, 1993; T. Suzuki et al. prolyl-), AS-900004, SCH-C, ZK-811752, PD-172084, 2-Nitroimidazole derivative, production thereof, and radi UK-427857, SB-380732, vMIP II, SB-265610, DPC-168, oSensitizer containing the Same as active ingredient. U.S. TAK-779 (N,N-dimethyl-N-(4-(2-(4-methylphenyl)-6,7-di Pat. No. 5,270,330, Dec. 14, 1993; T. Suzuki. 2-Nitroimi hydro-5H-benzocyclohepten-8-ylcarboxamido)benyl)tet dazole derivative, production thereof and radioSensitizer rahydro-2H-pyran-4-aminium chloride), TAK-220, KRH containing the same as active ingredient; Patent EP 0513 1120), GSK766994, SSR-150106, or an analogue or 351 B 1, Jan. 24, 1991), fluorine-containing nitroazole derivative thereof). Other examples of chemokine receptor derivatives (T. Kagiya. Fluorine-containing nitroazole antagonists include a-Immunokine-NNS03, BX-471, CCX derivatives and radioSensitizer comprising the Same. U.S. 282, Sch-350634; Sch-351125; Sch-417690; SCH-C, and Pat. No. 4,927,941, May 22, 1990), copper (M. J. Abrams. analogues and derivatives thereof. Copper Radiosensitizers. U.S. Pat. No. 5,100,885, Mar. 31, 1992), combination modality cancer therapy (D. H. Picker et 0313 6. Cell Cycle Inhibitors al. Combination modality cancer therapy. U.S. Pat. No. 0314. In another embodiment, the pharmacologically 4,681,091, Jul. 21, 1987). 5-CldC or (d)HU or 5-halo-2'- active compound is a cell cycle inhibitor. Representative halo-2'-deoxy-cytidine or -uridine derivatives (S. B. Greer. US 2005/0281883 A1 Dec. 22, 2005 33

Method and Materials for Sensitizing neoplastic tissue to concurrent external beam radiotherapy prolongs Survival in radiation. U.S. Pat. No. 4,894,364 Jan. 16, 1990), platinum metastatic brain tumor models. Cancer Research complexes (K. A. Skov. Platinum Complexes with one 56(22):5217-5223, 1996) and paclitaxel (Tishler R. B. et al. radiosensitizing ligand. U.S. Pat. No. 4,921,963. May 1, Taxol: a novel radiation Sensitizer. International Journal of 1990; K. A. Skov. Platinum Complexes with one radiosen Radiation Oncology and Biological Physics 22(3):613-617, sitizing ligand. Patent EP 0287317 A3), fluorine-containing 1992). nitroazole (T. Kagiya, et al. Fluorine-containing nitroazole 0315) A number of the above-mentioned cell cycle inhibi derivatives and radioSensitizer comprising the Same. U.S. tors also have a wide variety of analogues and derivatives, Pat. No. 4,927,941. May 22, 1990), benzamide (W. W. Lee. including, but not limited to, cisplatin, cyclophosphamide, Substituted Benzamide Radiosensitizers. U.S. Pat. No. misonidazole, tiripaZamine, nitroSourea, mercaptopurine, 5,032,617, Jul. 16, 1991), autobiotics (L. G. Egyud. Auto methotrexate, flurouracil, epirubicin, doxorubicin, Vindesline biotics and the use in eliminating nonself cells in Vivo. U.S. and etoposide. Analogues and derivatives include Pat. No. 5,147,652. Sep. 15, 1992), benzamide and nicoti (CPA)Pt(DOLYM) and (DACH)Pt(DOLYM) cisplatin namide (W. W. Lee et al. Benzamide and Nictoinamide (Choi et al., Arch. Pharmacal Res. 22(2):151-156, 1999), Radiosensitizers. U.S. Pat. No. 5,215,738, Jun. 1, 1993), Cis-(PtCl(4.7-H-5-methyl-7-oxo)1,2,4(triazolo(1.5-a)pyri acridine-intercalator (M. Papadopoulou-Rosenzweig. Acri midine)) (Navarro et al., J. Med. Chem. 41(3):332-338, dine Intercalator based hypoxia Selective cytotoxins. U.S. 1998), (Pt(cis-1,4-DACH)(trans-Cla)(CBDCA)). 4MeOH Pat. No. 5,294,715, Mar. 15, 1994), fluorine-containing cisplatin (Shamsuddin et al., Inorg. Chem. 36(25):5969 nitroimidazole (T. Kagiya et al. Fluorine containing 5971, 1997), 4-pyridoxate diammine hydroxy platinum nitroimidazole compounds. U.S. Pat. No. 5,304,654, Apr. (Tokunaga et al., Pharm. Sci. 3(7):353–356, 1997), Pt(II) . . 19, 1994), hydroxylated texaphyrins (J. L. Sessler et al. . Pt(II) (Pt(NHCHN(C(CH2)(CH)))) (Navarro et al., Hydroxylated texaphrins. U.S. Pat. No. 5,457,183, Oct. 10, Inorg. Chem. 35(26):7829-7835, 1996), 254-S cisplatin ana 1995), hydroxylated compound derivative (T. Suzuki et al. logue (Koga et al., Neurol. Res. 18(3):244-247, 1996), Heterocyclic compound derivative, production thereof and o-phenylenediamine ligand bearing cisplatin analogues radioSensitizer and antiviral agent containing Said derivative (Koeckerbauer & Bednarski, J. Inorg. Biochem. 62(4):281 as active ingredient. Publication Number 011106775 A 298, 1996), trans,cis-(Pt(OAc)I (en)) (Kratochwil et al., J. (Japan), Oct. 22, 1987; T. Suzuki et al. Heterocyclic com Med. Chem. 39(13):2499-2507, 1996), estrogenic 1,2-dia pound derivative, production thereof and radioSensitizer, rylethylenediamine ligand (with Sulfur-containing amino antiviral agent and anti cancer agent containing said deriva acids and glutathione) bearing cisplatin analogues (Bednar tive as active ingredient. Publication Number 01139596 A ski, J. Inorg. Biochem. 62(1):75, 1996), cis-1,4-diaminocy (Japan), Nov. 25, 1987; S. Sakaguchi et al. Heterocyclic clohexane cisplatin analogues (Shamsuddin et al., J. Inorg. compound derivative, its production and radioSensitizer Biochem. 61(4):291-301, 1996), 5' orientational isomer of containing Said derivative as active ingredient; Publication cis-(Pt(NH)(4-aminoTEMP-O){d(GpG)}) (Dunham & Number 63170375 A (Japan), Jan. 7, 1987), fluorine con Lippard, J. Am. Chem. Soc. 117(43): 10702-12, 1995), taining 3-nitro-1,2,4-triazole (T. Kagitani et al. Novel fluo chelating diamine-bearing cisplatin analogues (Koecker rine-containing 3-nitro-1,2,4-triazole and radioSensitizer bauer & Bednarski, J. Pharm. Sci. 84(7):819-23, 1995), containing same compound. Publication Number 02076861 1,2-diarylethyleneamine ligand-bearing cisplatin analogues A (Japan), Mar. 31, 1988), 5-thiotretrazole derivative or its (Otto et al., J. Cancer Res. Clin. Oncol. 121(1):31-8, 1995), salt (E. Kano et al. Radiosensitizer for Hypoxic cell. Pub (ethylenediamine)platinum(II) complexes (Pasini et al., J. lication Number 61010511 A (Japan), Jun. 26, 1984), Chem. Soc., Dalton Trans. 4:579-85, 1995), CI-973 cisplatin Nitrothiazole (T. Kagitani et al. Radiation-sensitizing agent. analogue (Yang et al., Int. J. Oncol. 5(3):597-602, 1994), Publication Number 61167616 A (Japan) Jan. 22, 1985), cis-diamminedichloroplatinum(II) and its analogues cis-1,1- imidazole derivatives (S. Inayma et al. Imidazole derivative. cyclobutanedicarbosylato(2R)-2-methyl-1,4-butanediam Publication Number 6203767 A (Japan) Aug. 1, 1985; mineplatinum(II) and cis-diammine(glycolato)platinum Publication Number 62030768 A (Japan) Aug. 1, 1985; (Claycamp & Zimbrick, J. Inorg. Biochem., 26(4):257-67, Publication Number 62030777 A (Japan) Aug. 1, 1985), 1986; Fan et al., Cancer Res. 48(11):3135-9, 1988; Heiger 4-nitro-1,2,3-triazole (T. Kagitani et al. Radiosensitizer. Bernays et al., Biochemistry 29(36):8461-6, 1990; Kikkawa Publication Number 620395.25 A (Japan), Aug. 15, 1985), et al., J. Exp. Clin. Cancer Res. 12(4):233-40, 1993; Murray 3-nitro-1,2,4-triazole (T. Kagitani et al. Radiosensitizer. et al., Biochemistry 31(47): 11812-17, 1992; Takahashi et al., Publication Number 62138427 A (Japan), Dec. 12, 1985), Cancer Chemother. Pharmacol. 33(1):31-5, 1993), cis Carcinostatic action regulator (H. Amagase. Carcinostatic amine-cyclohexylamine-dichloroplatinum(II) (Yoshida et action regulator. Publication Number 63099017 A (Japan), al., Biochem. Pharmacol. 48(4):793-9, 1994), gem-diphos Nov. 21, 1986),4,5-dinitroimidazole derivative (S. Inayama. phonate cisplatin analogues (FR 2683529), (meso-1,2-bis(2, 4,5-Dinitroimidazole derivative. Publication Number 6-dichloro-4-hydroxyplenyl)ethylenediamine) dichloro 63310873 A (Japan) Jun. 9, 1987), nitrotriazole Compound platinum(II) (Bednarski et al., J. Med. Chem. 35(23):4479 (T. Kagitanil Nitrotriazole Compound. Publication Number 85, 1992), cisplatin analogues containing a tethered dansyl 07149737 A (Japan) Jun. 22, 1993), cisplatin, doxorubin, group (Hartwig et al., J. Am. Chem. Soc. 114(21):8292-3, misonidazole, mitomycin, tiripaZamine, nitroSourea, mer 1992), platinum(II) polyamines (Siegmann et al., Inorg. captopurine, methotrexate, flurouracil, bleomycin, Vincris Met.-Containing Polym. Mater., (Proc. Am. Chem. Soc. Int. tine, carboplatin, epirubicin, doxorubicin, cyclophospha Symp), 335-61, 1990), cis-(3H)dichloro(ethylenediamine mide, Vindesine, etoposide (I. F. Tannock. Review Article: )platinum(II) (Eastman, Anal. Biochem. 197(2):311-15, Treatment of Cancer with Radiation and Drugs. Journal of 1991), trans-diamminedichloroplatinum(II) and cis Clinical Oncology 14(12):3156-3174, 1996), camptothecin (Pt(NH)-(N-cytosine)Cl) (Bellon & Lippard, BiophyS. (Ewend M. G. et al. Local delivery of chemotherapy and Chem. 35(2-3): 179-88, 1990), 3H-cis-1,2-diaminocyclohex US 2005/0281883 A1 Dec. 22, 2005 34 anedichloroplatinum(II) and 3H-cis-1,2-diaminocyclohex 58(2):153-162, 1999), anthracycline disaccharide doxorubi anemalonatoplatinum (II) (Oswald et al., ReS. Commun. cin analogue (Pratesi et al., Clin. Cancer Res. 4(11):2833 Chem. Pathol. Pharmacol. 64(1):41-58, 1989), diaminocar 2839, 1998), N-(trifluoroacetyl)doxorubicin and 4'-O- boxylatoplatinum (EPA 296321), trans-(D,1)-1,2-diami acetyl-N-(trifluoroacetyl)doxorubicin (Berube & Lepage, nocyclohexane carrier ligand-bearing platinum analogues Synth. Commun. 28(6):1109-1116, 1998), 2-pyrrolinodoxo (Wyrick & Chaney, J. Labelled Compa. Radiopharm. rubicin (Nagy et al., Proc. Nat'l Acad. Sci. U.S.A. 25(4):349-57, 1988), aminoalkylaminoanthraquinone-de 95(4): 1794-1799, 1998), disaccharide doxorubicin ana rived cisplatin analogues (Kitov et al., Eur: J. Med. Chem. logues (Arcamone et al., J. Nat'l Cancer Inst. 89(16): 1217 23(4):381-3, 1988), spiroplatin, carboplatin, iproplatin and 1223, 1997), 4-demethoxy-7-O-(2,6-dideoxy-4-O-(2,3,6- JM40 platinum analogues (Schroyen et al., Eur: J. Cancer trideoxy-3-amino-O-L-lyxo-hexopyranosyl)-O-L-lyxo Clin. Oncol. 24(8): 1309-12, 1988), bidentate tertiary hexopyranosyl)adriamicinone doxorubicin disaccharide diamine-containing cisplatinum derivatives (Orbell et al., analogue (Monteagudo et al., Carbohydr. Res. 300(1):11-16, Inorg. Chim. Acta 152(2):125-34, 1988), platinum(II), plati 1997), 2-pyrrolinodoxorubicin (Nagy et al., Proc. Natl num(IV) (Liu & Wang, Shandong Yike Daxue Xuebao Acad. Sci. U.S.A. 94(2):652-656, 1997), morpholinyl doxo 24(1):35-41, 1986), cis-diammine(1,1-cyclobutanedicar rubicin analogues (Duran et al., Cancer Chemother. Phar boxylato-)platinum(II) (carboplatin, JM8) and ethylenedi macol. 38(3):210-216, 1996), enaminomalonyl-O-alanine ammine-malonatoplatinum(II) (JM40) (Begg et al., Radio doxorubicin derivatives (Seitz et al., Tetrahedron Lett. ther. Oncol. 9(2): 157-65, 1987), JM8 and JM9 cisplatin 36(9): 1413-16, 1995), cephalosporin doxorubicin deriva analogues (Harstrick et al., Int. J. Androl. 10(1); 139-45, tives (Vrudhula et al., J. Med. 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Acta 73(4):912-15, 1990), Cancer 65(5):703-7, 1992), N-hydroxysuccinimide ester 1,3,2-dioxa- and -oxazaphosphorinane cyclophosphamide doxorubicin derivatives (Demant et al., Biochim. BiophyS. analogues (Yang et al., Tetrahedron 44(20):6305-14, 1988), Acta 1118(1):83-90, 1991), polydeoxynucleotide doxorubi C5-Substituted cyclophosphamide analogues (Spada, Uni cin derivatives (Ruggiero et al., Biochim. BiophyS. Acta versity of Rhode Island Dissertation, 1987), tetrahydroox 1129(3):294-302, 1991), morpholinyl doxorubicin deriva azine cyclophosphamide analogues (Valente, University of tives (EPA 434960), mitoxantrone doxorubicin analogue Rochester Dissertation, 1988), phenylketone cyclophospha (Krapcho et al., J. Med. Chem. 34(8):2373-80. 1991), mide analogues (Hales et al., Teratology 39(1):31-7, 1989), AD198 doxorubicin analogue (Traganos et al., Cancer Res. phenylketophosphamide cyclophosphamide analogues 51(14):3682-9, 1991), 4-demethoxy-3'-N-trifluoroacetyl (Ludeman et al., J. Med. Chem. 29(5):716-27, 1986), ASTA doxorubicin (Horton et al., Drug Des. Delivery 6(2):123-9, Z-7557 cyclophosphamide analogues (Evans et al., Int. J. 1990), 4'-epidoxorubicin (Drzewoski et al., Pol. J. Pharma Cancer 34(6):883-90, 1984), 3-(1-oxy-2,2,6,6-tetramethyl col. Pharm. 40(2): 159-65, 1988; Weenen et al., Eur: J. 4-piperidinyl)cyclophosphamide (Tsui et al., J. Med. Chem. Cancer Clin. Oncol. 20(7):919-26, 1984), alkylating cyano 25(9): 1106-10, 1982), 2-oxobis(2-?-chloroethylamino)-4-, morpholino doxorubicin derivative (Scudder et al., J. Natl 6-dimethyl-1,3,2-oxazaphosphorinane cyclophosphamide Cancer Inst. 80(16): 1294–8, 1988), deoxydihydroiodooxo (Carpenter et al., Phosphorus Sulfur 12(3):287-93, 1982), rubicin (EPA 275966), adriblastin (Kalishevskaya et al., 5-fluoro- and 5-chlorocyclophosphamide (Foster et al., J. Vestn. Mosk. Univ., 16(Biol. 1):21-7, 1988), 4'-deoxydoxo Med. Chem. 24(12): 1399-403, 1981), cis- and trans-4-phe rubicin (Schoelzel et al., Leuk. Res. 10(12): 1455-9, 1986), nylcyclophosphamide (Boyd et al., J. Med. Chem. 4-demethyoxy-4'-O-methyldoxorubicin (Giuliani et al., 23(4):372-5, 1980), 5-bromocyclophosphamide, 3,5-dehy Proc. Int. Congr. Chemother. 16:285-70-285-77, 1983), drocyclophosphamide (Ludeman et al., J. Med. Chem. 3'-deamino-3'-hydroxydoxorubicin (Horton et al., J. Anti 22(2): 151-8, 1979), 4-ethoxycarbonyl cyclophosphamide biot. 37(8):853-8, 1984), 4-demethyoxy doxorubicin ana analogues (Foster, J. Pharm. Sci. 67(5):709-10, 1978), ary logues (Barbieri et al., Drugs Exp. Clin. Res. 10(2):85-90, laminotetrahydro-2H-1,3,2-oxazaphosphorine 2-oxide 1984), N-L-leucyl doxorubicin derivatives (Trouet et al., cyclophosphamide analogues (Hamacher, Arch. Pharm. Anthracyclines (Proc. Int. Symp. Tumor Pharmacother.), (Weinheim, Ger) 310(5):J,428-34, 1977), NSC-26271 179-81, 1983), 3'-deamino-3'-(4-methoxy-1-piperidinyl) cyclophosphamide analogues (Montgomery & Struck, Can doxorubicin derivatives (U.S. Pat. No. 4,314,054), cer Treat. Rep. 60(4):J381-93, 1976), benzo annulated 3'-deamino-3'-(4-mortholinyl) doxorubicin derivatives (U.S. cyclophosphamide analogues (Ludeman & Zon, J. Med. Pat. No. 4,301.277), 4'-deoxydoxorubicin and 4'-O-methyl Chem. 18(12):J1251-3, 1975), 6-trifluoromethylcyclophos doxorubicin (Giuliani et al., Int. J. Cancer 27(1):5-13, phamide (Farmer & Cox, J. Med. Chem. 18(11):J1106-10, 1981), aglycone doxorubicin derivatives (Chan & Watson, J. 1975), 4-methylcyclophosphamide and 6-methycyclophos Pharm. Sci 67(12):1748-52, 1978), SM 5887 (Pharma phamide analogues (Cox et al., Biochem. Pharmacol. Japan 1468:20, 1995), MX-2 (Pharma Japan 1420:19, 24(5):J599-606, 1975); FCE 23762 doxorubicin derivative 1994), 4'-deoxy-13(S)-dihydro-4'-iododoxorubicin (EP (Quaglia et al., J. Liq. Chromatogr. 17(18):3911-3923, 275966), morpholinyl doxorubicin derivatives (EPA 1994), annamycin (Zou et al., J. Pharm. 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No. 4,585,859), 27:681-9, 1982), TA-077 (Fujimoto & Ogawa, Cancer 3'-deamino-3'-(4-methoxy-1-piperidinyl) doxorubicin Chemother: Pharmacol. 9(3):134-9, 1982), gentianose derivatives (U.S. Pat. No. 4,314,054) and 3-deamino-3-(4- nitrosourea derivatives (JP 82 80396), CNCC, RFCNU, morpholinyl) doxorubicin derivatives (U.S. Pat. No. 4,301, RPCNU AND chlorozotocin (CZT) (Marzin et al., INSERM 277); 4,5-dimethylmisonidazole (Born et al., Biochem. Symp., 19(Nitrosoureas Cancer Treat.):165-74, 1981), thio Pharmacol. 43(6):1337-44, 1992), azo and azoxy misonida colchicine nitroSourea analogues (George, Shih Ta Hsuch zole derivatives (Gattavecchia & Tonelli Int. J. Radiat. Biol. Pao (Taipei) 25:355-62, 1980), 2-chloroethyl-nitrosourea Relat. Stud. Phys., Chem. Med. 45(5):469-77, 1984); (Zeller & Eisenbrand, Oncology 38(1):39–42, 1981), ACNU, RB90740 (Wardman et al., Br. J. Cancer, 74 Suppl. 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Med Chem. 23(3):324-6, 1980), ergoline nitrosourea derivatives (Miyahara et al., Heterocycles nitrosourea derivatives (Crider et al., J. Med. Chem. 39(1):361-9, 1994), ACNU (Matsunaga et al., Immunophar 22(1):32-5, 1979), glucopyranose nitrosourea derivatives macology 23(3):199-204, 1992), tertiary phosphine oxide (JP 78 95917), 1-(2-chloroethyl)-3-cyclohexyl-1-ni nitroSourea derivatives (Guguva et al., Pharmazie trosourea (Farmer et al., J. Med. Chem. 21(6):514-20, 1978), 46(8):603, 1991), sulfamerizine and sulfamethizole 4-(3-(2-chloroethyl)-3-nitroSoureid-O)-cis-cyclohexanecar nitrosourea derivatives (Chiang et al., Zhonghua Yaozue boxylic acid (Drewinko et al., Cancer Treat. Rep. Zazhi 43(5):401-6, 1991), thymidine nitrosourea analogues 61(8):J1513-18, 1977), RPCNU (ICIG 1163) (Larnicol et (Zhang et al., Cancer Commun. 3(4): 119-26, 1991), 1,3- al., Biomedicine 26(3):J176-81, 1977), IOB-252 (Sorodocet bis(2-chloroethyl)-1-nitroSourea (August et al., Cancer ReS. al., Rev. Roum. Med., Virol. 28(1):J 55-61, 1977), 1,3-bis(2- 51(6):1586-90, 1991), 2.2,6,6-tetramethyl-1-oxopiperidi chloroethyl)-1-nitrosourea (BCNU) (Siebert & Eisenbrand, unium nitrosourea derivatives (U.S.S.R. 1261253), 2- and Mutat. Res. 42(1):J45-50, 1977), 1-tetrahydroxycyclopen 4-deoxy Sugar nitrosourea derivatives (U.S. Pat. No. 4,902, tyl-3-nitroso-3-(2-chloroethyl)-urea (U.S. Pat. No. 4,039, 791), nitroxyl nitrosourea derivatives (U.S.S.R. 1336489), 578), d-1-1-(B-chloroethyl)-3-(2-oxo-3-hexahydroazepi fotemustine (Boutin et al., Eur: J. Cancer Clin. Oncol. nyl)-1-nitrosourea (U.S. Pat. No. 3,859,277) and gentianose 25(9): 1311-16, 1989), pyrimidine (II) nitrosourea deriva nitrosourea derivatives (JP 57080396); 6-S-aminoacy tives (Wei et al., Chung-hua Yao Hsuch Tsa Chih 41(1):19 loxymethyl mercaptopurine derivatives (Harada et al., 26, 1989), CGP 6809 (Schieweck et al., Cancer Chemother. Chem. Pharm. 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Zh. 15(8):65-7, 1981); indoline ring and a modified ySucrose (NS-1C) and 6'-((((2-chloroethyl)nitroSoami ornithine or glutamic acid-bearing methotrexate derivatives no)carbonyl)amino)-6'-deoxysucrose (NS-1D) nitrosourea (Matsuoka et al., Chem. Pharm. Bull. 45(7): 1146-1150, derivatives (Tanoh et al., Chemotherapy (Tokyo) 33(11):969 1997), alkyl-substituted benzene ring C bearing methotrex 77, 1985), CNCC, RFCNU and chlorozotocin (Mena et al., ate derivatives (Matsuoka et al., Chem. Pharm. Bull. Chemotherapy (Basel) 32(2): 131-7, 1986), CNUA (Edan 44(12):2287-2293, 1996), benzoxazine or benzothiazine ami et al., Chemotherapy (Tokyo) 33(5):455-61, 1985), moiety-bearing methotrexate derivatives (Matsuoka et al., J. 1-(2-chloroethyl)-3-isobutyl-3-(B-maltosyl)-1-nitrosourea Med. Chem. 40(1):105-111, 1997), 10-deazaminopterin ana (Fujimoto & Ogawa, Jpn. J. Cancer Res. (Gann) 76(7):651 logues (DeGraw et al., J. Med. 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Biol, 163(Folyl Antifolyl 3, 1991), f.y-uethano methotrexate analogues (Rosowsky et al., Pteridines 2(3):133-9, 1991), 10-deazaminopterin (10 Polyglutamates): 95-100, 1983), 3',5'-dichloromethotrexate EDAM) analogue (Braakhuis et al., Chem. Biol. Pteridines, (Rosowsky & Yu, J. Med. Chem. 26(10): 1448-52, 1983), Proc. Int. Symp. Pteridines Folic Acid Deriv., 1027-30, diaZOketone and chloromethylketone methotrexate ana 1989), Y-tetrazole methotrexate analogue (Kalman et al., logues (Gangjee et al., J. Pharm. Sci. 71(6):717-19, 1982), Chem. Biol. Pteridines, Proc. Int. Symp. Pteridines Folic 10-propargylaminopterin and alkyl methotrexate homologs Acid Deriv., 1154-7, 1989), N-(L-C-aminoacyl) methotrex (Piper et al., J. Med. Chem. 25(7):877-80, 1982), lectin ate derivatives (Cheung et al., Heterocycles 28(2):751-8, derivatives of methotrexate (Lin et al., JNCI 66(3):523-8, 1989), meta and ortho isomers of aminopterin (Rosowsky et 1981), polyglutamate methotrexate derivatives (Galivan, al., J. Med. Chem. 32(12):2582, 1989), hydroxymethyl Mol. Pharmacol. 17(1):105-10, 1980), halogentated meth methotrexate (DE 267495), Y-fluoromethotrexate (McGuire otrexate derivatives (Fox, JNCI 58(4):J955-8, 1977), et al., Cancer Res. 49(16):4517-25, 1989), polyglutamyl 8-alkyl-7,8-dihydro analogues (Chaykovsky et al., J. Med. methotrexate derivatives (Kumar et al., Cancer ReS. Chem. 20(10):J1323-7, 1977), 7-methyl methotrexate 46(10):5020-3, 1986), gem-diphosphonate methotrexate derivatives and dichloromethotrexate (Rosowsky & Chen, J. analogues (WO 88/06158), C- and Y-substituted methotrex Med. Chem. 17(12):J1308-11, 1974), lipophilic methotrex ate analogues (Tsushima et al., Tetrahedron 44(17):5375-87, ate derivatives and 3',5'-dichloromethotrexate (Rosowsky, J. 1988), 5-methyl-5-deaza methotrexate analogues (U.S. Pat. Med. Chem. 16(10):J1190-3, 1973), deaza amethopterin No. 4,725,687), N8-acyl-NO-(4-amino-4-deoxypteroyl)-L- analogues (Montgomery et al., Ann. N.Y. Acad. Sci. ornithine derivatives (Rosowsky et al., J. Med. Chem. 186:J227-34, 1971), MX068 (Pharma Japan, 1658:18, 1999) 31(7): 1332-7, 1988), 8-deaza methotrexate analogues and cysteic acid and homocysteic acid methotrexate ana (Kuehl et al., Cancer Res. 48(6): 1481-8, 1988), acivicin logues (EPA 0142220); N3-alkylated analogues of 5-fluo methotrexate analogue (Rosowsky et. al., J. Med. Chem. 30(8): 1463-9, 1987), polymeric platinol methotrexate rouracil (Kozai et al., J. Chem. Soc., Perkin Trans. derivative (Carraher et al., Polym. Sci. Technol. (Plenum), 1(19):3145-3146, 1998), 5-fluorouracil derivatives with 1,4- 35(Ady: Biomed Polym.):311-24, 1987), methotrexate-Y- oxaheteroepane moieties (Gomez et al., Tetrahedron dimyristoylphophatidylethanolamine (Kinsky et al., Bio 54(43): 13295-13312, 1998), 5-fluorouracil and nucleoside chim. Biophys. Acta 917(2):211-18, 1987), methotrexate analogues (Li, Anticancer Res. 17(1A):21-27, 1997), cis polyglutamate analogues (Rosowsky et al., Chem. Biol. and trans-5-fluoro-5,6-dihydro-6-alkoxyuracil (Van der Wilt Pteridines, Pteridines Folid Acid Deriv., Proc. Int. Symp. et al., Br. J. Cancer 68(4):702-7, 1993), cyclopentane 5-fluo Pteridines Folid Acid Deriv.: Chem. Biol. Clin. Aspects: rouracil analogues (Hronowski & Szarek, Can. J. Chem. 985-8, 1986), poly-y-glutamyl methotrexate derivatives 70(4): 1162-9, 1992), A-OT-fluorouracil (Zhang et al., Zong (Kisliuk et al., Chem. Biol. Pteridines, Pteridines Folid Acid guo Yiyao Gongye Zazhi 20(11):513-15, 1989), N4-tri Deriv., Proc. Int. Symp. Pteridines Folid Acid Deriv.: methoxybenzoyl-5'-deoxy-5-fluorocytidine and 5'-deoxy-5- Chem., Biol. Clin. Aspects: 989-92, 1986), deoxyuridylate fluorouridine (Miwa et al., Chem. Pharm. Bull. 38(4):998 methotrexate derivatives (Webber et al., Chem. Biol. Pte 1003, 1990), 1-hexylcarbamoyl-5-fluorouracil (Hoshi et al., ridines, Pteridines Folid Acid Deriv., Proc. Int. Symp. Pte J. Pharmacobio-Dun. 3(9):478-81, 1980; Maehara et al., ridines Folid Acid Deriv.: Chem. Biol. Clin. Aspects: 659 Chemotherapy (Basel) 34(6):484-9, 1988), B-3839 (Prajda 62, 1986), iodoacetyl lysine methotrexate analogue et al., In Vivo 2(2):151-4, 1988), uracil-1-(2-tetrahydrofu (Delcamp et al., Chem. Biol. Pteridines, Pteridines Folid ryl)-5-fluorouracil (Anai et al., Oncology 45(3): 144-7, Acid Deriv., Proc. Int. Symp. Pteridines Folid Acid Deriv.: 1988), 1-(2'-deoxy-2'-fluoro-B-D-arabinofuranosyl)-5-fluo Chem. Biol. Clin. Aspects: 807-9, 1986), 2, omega.-diami rouracil (Suzuko et al., Mol. Pharmacol. 31(3):301-6, 1987), noalkanoid acid-containing methotrexate analogues doxifluridine (Matuura et al., Oyo Yakuri 29(5):803-31, (McGuire et al., Biochem. Pharmacol. 35(15):2607-13, 1985), 5'-deoxy-5-fluorouridine (Bollag & Hartmann, Eur: J. 1986), polyglutamate methotrexate derivatives (Kamen & Winick, Methods Enzymol. 122 (Vitam. Coenzymes, Pt. Cancer 16(4):427-32, 1980), 1-acetyl-3-O-toluyl-5-fluorou G):339-46, 1986), 5-methyl-5-deaza analogues (Piper et al., racil (Okada, Hiroshima J. Med. Sci. 28(1):49-66, 1979), J. Med. Chem. 29(6):1080-7, 1986), quinazoline methotrex 5-fluorouracil-m-formylbenzene-Sulfonate (JP 55059173), ate analogue (Mastropaolo et al., J. Med. Chem. 29(1): 155 N'-(2-furanidyl)-5-fluorouracil (JP 53149985) and 1-(2-tet 8, 1986), pyrazine methotrexate analogue (Lever & Vestal, rahydrofuryl)-5-fluorouracil (JP52089680); 4'-epidoxorubi J. Heterocycl. Chem. 22(1):5-6, 1985), cysteic acid and cin (Lanius, Adv. Chemother. Gastrointest. Cancer, (Int. homocysteic acid methotrexate analogues (U.S. Pat. No. Symp.), 159-67, 1984); N-substituted deacetylvinblastine 4,490,529), Y-tert-butyl methotrexate esters (Rosowsky et amide (vindesine) sulfates (Conrad et al., J. Med. Chem. al., J. Med. Chem. 28(5):660-7, 1985), fluorinated methotr 22(4):391-400, 1979); and Cu(II)-VP-16 (etoposide) com exate analogues (Tsushima et al., Heterocycles 23(1):45-9, plex (Tawa et al., Bioorg. Med. Chem. 6(7): 1003-1008, 1985), folate methotrexate analogue (Trombe, J. Bacteriol. 1998), pyrrollecarboxamidino-bearing etoposide analogues 160(3):849-53, 1984), phosphonoglutamic acid analogues (Ji et al., Bioorg. Med. Chem. Lett. 7(5):607-612, 1997), US 2005/0281883 A1 Dec. 22, 2005 37

4B-amino etoposide analogues (Hu, University of North 9-deoxotaxol, 7-deoxy-9-deoxotaxol, 10-desacetoxy-7- Carolina Dissertation, 1992), Y-lactone ring-modified ary deoxy-9-deoxotaXol, Derivatives containing hydrogen or lamino etoposide analogues (Zhou et al., J. Med. Chem. acetyl group and a hydroxy and tert-butoxycarbonylamino, 37(2):287-92, 1994), N-glucosyl etoposide analogue (Allevi et al., Tetrahedron Lett. 34(45):7313-16, 1993), etoposide sulfonated 2'-acryloyltaxol and sulfonated 2'-O-acyl acid A-ring analogues (Kadow et al., BioOrg. Med. Chem. Lett. taxol derivatives, SuccinyltaXol, 2-y-aminobutyryltaXol for 2(1):17-22, 1992), 4'-deshydroxy-4'-methyl etoposide mate, 2-acetyl taxol, 7-acetyl taxol, 7-glycine carbamate (Saulnier et al., Bioorg. Med. Chem. Lett. 2(10): 1213-18, taxol. 2'-OH-7-PEG(5000) carbamate taxol, 2-benzoyl and 1992), pendulum ring etoposide analogues (Sinha et al., Eur: J. Cancer 26(5):590-3, 1990) and E-ring desoxy etoposide 2,7-dibenzoyl taxol derivatives, other prodrugs (2'-acetyl analogues (Saulnier et al., J. Med. Chem. 32(7): 1418-20, taxol; 2,7-diacetyltaxol; 2'Succinyltaxol; 2'-(beta-alanyl)- 1989). taxol); 2'gamma-aminobutyryl taxol formate; ethylene gly 0316. Within one preferred embodiment of the invention, colderivatives of 2'-succinyl taxol; 2'-glutaryltaxol; 2'-(N.N- the cell cycle inhibitor is paclitaxel, a compound which dimethylglycyl) taxol; 2'-(2-(N.N- disrupts mitosis (M-phase) by binding to tubulin to form dimethylamino)propionyl)taxol; 2'orthocarboxybenzoyl abnormal mitotic Spindles or an analogue or derivative taxol; 2'aliphatic carboxylic acid derivatives of taxol, Pro thereof. Briefly, paclitaxel is a highly derivatized diterpenoid drugs {2'(N,N-diethylaminopropionyl)taxol. 2'(N,N-dim (Wani et al., J. Am. Chem. Soc. 93:2325, 1971) which has ethylglycyl)taxol, 7(N,N-dimethylglycyl)taxol, 2,7-di-(N, been obtained from the harvested and dried bark of Taxus N-dimethylglycyl)taxol, 7(N.N- brevifolia (Pacific Yew) and Taxomyces andreanae and diethylaminopropionyl)taxol, 2,7-di(N.N- Endophytic fungus of the Pacific Yew (Stierle et al., Science diethylaminopropionyl)taxol. 2'-(L-glycyl)taxol, 7-(L- 60:214-216, 1993). “Paclitaxel” (which should be under glycyl)taxol, 2,7-di(L-glycyl)taxol, 2-(L-alanyl)taxol, stood herein to include formulations, prodrugs, analogues 7-(L-alanyl)taxol, 2,7-di(L-alanyl)taxol, 2-(L-leucyl)taxol, and derivatives such as, for example, TAXOL (Bristol Myers Squibb, New York, N.Y., TAXOTERE (Aventis Phar 7-(L-leucyl)taxol; 2,7-di(L-leucyl)taxol. 2'-(L-isoleucyl maceuticals, France), docetaxel, 10-desacetyl analogues of )taxol, 7-(L-isoleucyl)taxol, 2,7-diCL-isoleucyl)taxol, 2-(L- paclitaxel and 3N-desbenzoyl-3'N-t-butoxy carbonyl ana valyl)taxol, 7-(L-valyl)taxol, 27-di(L-valyl)taxol. 2'-(L- logues of paclitaxel) may be readily prepared utilizing phenylalanyl)taxol, 7-(L-phenylalanyl)taxol, 2,7-di(L- techniques known to those skilled in the art (see, e.g., Schiff phenylalanyl)taxol, 2-(L-prolyl)taxol, 7-(L-prolyl)taxol, et al., Nature 277:665-667, 1979; Long and Fairchild, Can 2,7-di(L-prolyl)taxol. 2'-(L-lysyl)taxol, 7-(L-lysyl)taxol, cer Research 54:4355-4361, 1994; Ringel and Horwitz, J. 2,7-di(L-lysyl)taxol, 2’-(L-glutamyl)taxol, 7-(L-glutamyl Natl Cancer Inst. 83(4):288-291, 1991; Pazdur et al., Can )taxol, 2,7-di(L-glutamyl)taxol, 2-(L-arginyl)taxol, 7-(L- cer Treat. Rev. 19(4):351-386, 1993; WO 94/07882; WO arginyl)taxol, 2,7-di(L-arginyl)taxol, taxol analogues with 94/07881; WO 94/07880, WO 94/07876; WO 93/23555; modified phenylisoserine side chains, TAXOTERE, (N-de WO 93/10076; WO94/00156; WO 93/24476; EP 590267; benzoyl-N-tert-(butoxycaronyl)-10-deacetyltaxol, and tax WO 94/20089; U.S. Pat. Nos. 5,294,637; 5,283,253; 5,279, anes (e.g., baccatin III, cephalomannine, 10-deacetylbacca 949; 5,274,137; 5,202,448; 5,200,534, 5,229,529; 5,254, 580; 5,412,092; 5,395,850; 5,380,751; 5,350,866; 4,857, tin III, brevifoliol, yunantaXusin and taxusin); and other 653; 5,272,171; 5,411,984; 5,248,796; 5,248,796; 5,422, taxane analogues and derivatives, including 14-beta-hy 364; 5,300,638; 5,294,637; 5,362.831; 5,440,056; 4,814, droxy-10 deacetybaccatin III, debenzoyl-2-acyl paclitaxel 470; 5,278,324; 5,352,805; 5,411,984; 5,059,699; 4,942, derivatives, benzoate paclitaxel derivatives, phosphonooxy 184; Tetrahedron Letters 35(52):9709-9712, 1994; J. Med. and carbonate paclitaxel derivatives, Sulfonated 2'-acryloyl Chem. 35:4230-4237, 1992, J. Med. Chem. 34:992-998, taxol; Sulfonated 2'-O-acyl acid paclitaxel derivatives, 1991, J. Natural Prod. 57(10): 1404-1410, 1994, J. Natural 18-site-Substituted paclitaxel derivatives, chlorinated pacli Prod57(11):1580-1583, 1994; J. Am. Chem. Soc. 110:6558 taxel analogues, C4 methoxy ether paclitaxel derivatives, 6560, 1988), or obtained from a variety of commercial Sulfonamide taxane derivatives, brominated paclitaxel ana Sources, including for example, Sigma Chemical Co., St. logues, Girard taxane derivatives, nitrophenyl paclitaxel, Louis, Mo. (T7402-from Taxus brevifolia). 10-deacetylated Substituted paclitaxel derivatives, 14-beta 0317 Representative examples of paclitaxel derivatives hydroxy-10 deacetylbaccatin III taxane derivatives, C7 tax or analogues include 7-deoxy-docetaxol, 7,8-cyclopropatax ane derivatives, C10 taxane derivatives, 2-debenzoyl-2-acyl anes, N-Substituted 2-aZetidones, 6,7-epoxy paclitaxels, 6,7- taxane derivatives, 2-debenzoyl and -2-acyl paclitaxel modified paclitaxels, 10-desacetoxytaxol, 10-deacetyltaXol derivatives, taxane and baccatin III analogues bearing new (from 10-deacetylbaccatin III), phosphonooxy and carbon C2 and C4 functional groups, n-acyl paclitaxel analogues, ate derivatives of taxol, taxol 2,7-di(Sodium 1,2-benzene 10-deacetylbaccatin III and 7-protected-10-deacetylbaccatin dicarboxylate, 10-desacetoxy-11,12-dihydrotaxol-10, III derivatives from 10-deacetyl taxol A, 10-deacetyl taxol 12(18)-diene derivatives, 10-desacetoxytaxol, Protaxol (2'- B, and 10-deacetyl taxol, benzoate derivatives of taxol, and/or 7-O-ester derivatives), (2'-and/or 7-O-carbonate 2-aroyl-4-acyl paclitaxel analogues, orthro-ester paclitaxel derivatives), asymmetric Synthesis of taxol Side chain, fluoro analogues, 2-aroyl-4-acyl paclitaxel analogues and 1-deoxy taxols, 9-deoxotaxane, (13-acetyl-9-deoxobaccatine III, paclitaxel and 1-deoxy paclitaxel analogues. US 2005/0281883 A1 Dec. 22, 2005 38

0318. In one aspect, the cell cycle inhibitor is a taxane having the structure of formula (III):

(III)

0319 where the gray-highlighted portions may be sub 0322) wherein R, is Selected from hydrogen, alkyl, Stituted and the non-highlighted portion is the taxane core. phenyl, alkoxy, amino, phenoxy (Substituted or A side-chain (labeled “A” in the diagram) is desirably present in order for the compound to have good activity as unsubstituted); Rs is Selected from hydrogen, alkyl, a cell cycle inhibitor. Examples of compounds having this hydroxyalkyl, alkoxyalkyl, aminoalkyl, phenyl (Sub structure include paclitaxel (Merck Index entry 7117), doc stituted or unsubstituted), alpha or beta-naphthyl; etaxol (TAXOTERE, Merck Index entry 3458), and 3'-desphenyl-3'-(4-ntirophenyl)-N-debenzoyl-N-(t-butoxy and Ro is Selected from hydrogen, alkanoyl, Substi carbonyl)-10-deacetyltaxol. tuted alkanoyl, and aminoalkanoyl, where Substitu tions refer to hydroxyl, Sulthydryl, allalkoxyl, car 0320 In one aspect, Suitable taxanes Such as paclitaxel and its analogues and derivatives are disclosed in U.S. Pat. boxyl, halogen, thioalkoxyl, N,N-dimethylamino, No. 5,440,056 as having the structure of formula (IV): alkylamino, dialkylamino, nitro, and -OSOH, and/ or may refer to groups containing Such Substitutions, (IV) R is Selected from hydrogen or oxygen-containing groups, Such as hydrogen, hydroxyl, alkoyl, alkanoy loxy, aminoalkanoyloxy, and peptidyalkanoyloxy, R, is selected from hydrogen or oxygen-containing groups, Such as hydrogen, hydroxyl, alkoyl, alkanoy loxy, aminoalkanoyloxy, and peptidyalkanoyloxy,

O and may further be a silyl containing group or a Sulphur containing group; R is selected from acyl, alkyl, alkanoyl, aminoalkanoyl, peptidylalkanoyl 0321 wherein X may be oxygen (paclitaxel), hydrogen (9-deoxy derivatives), thioacyl, or dihydroxyl precursors; R' and aroyl; Rs is Selected from acyl, alkyl, alkanoyl, is selected from paclitaxel or TAXOTERE side chains or aminoalkanoyl, peptidylalkanoyl and aroyl; R is alkanoyl having the structure of formula (V) Selected from hydrogen or oxygen-containing groups, Such as hydrogen, hydroxyl alkoyl, alkanoy loxy, aminoalkanoyloxy, and peptidyalkanoyloxy. (V) 0323) In one aspect, the paclitaxel analogues and deriva tives useful as cell cycle inhibitors are disclosed in PCT International Patent Application No. WO 93/10076. As disclosed in this publication, the analogue or derivative should have a side chain attached to the taxane nucleus at ORo C, as shown in the Structure of formula VI, in order to confer antitumor activity to the taxane. US 2005/0281883 A1 Dec. 22, 2005 39

cell (NSC) lung, Small cell lung; breast, prostate; cervical; endometrial; head and neck cancers. 10 9 (VI) 0327 In another aspect, the anti-microtuble agent (micro tubule inhibitor) is albendazole (carbamic acid, 5-(propy lthio)-1H-benzimidazol-2-yl-, methyl ester), LY-355703 (1,4-dioxa-8,11-diazacyclohexadec-13-ene-2.5,9,12 13 tetrone, 10-(3-chloro-4-methoxyphenyl)methyl-6,6-dim ethyl-3-(2-methylpropyl)-16-(1S)-1-(2S,3R)-3-phenylox iranylethyl-, (3S,10R,13E,16S)-), Vindesline (Vincaleukoblastine, 3-(aminocarbonyl)-O4-deacetyl-3- 0324 WO 93/10076 discloses that the taxane nucleus de(methoxycarbonyl)-), or WAY-174286 may be Substituted at any position with the exception of the 0328. In another aspect, the cell cycle inhibitor is a Vinca existing methyl groups. The Substitutions may include, for alkaloid. Vinca alkaloids have the following general Struc example, hydrogen, alkanoyloxy, alkenoyloxy, aryloyloxy. ture of formulas (VI) and (VII). They are indole-dihydroin In addition, OXO groups may be attached to carbons labeled dole dimers.

(VI)

(VII) re dihydroindole

2, 4, 9, and/or 10. AS well, an OXetane ring may be attached 0329. As disclosed in U.S. Pat. Nos. 4,841,045 and at carbons 4 and 5. AS well, an oxirane ring may be attached 5,030,620, R, can be a formyl or methyl group or alternately to the carbon labeled 4. H. R can also be an alkyl group or an aldehyde-Substituted 0325 In one aspect, the taxane-based cell cycle inhibitor alkyl (e.g., CHCHO). R is typically a CH or NH group; useful in the present invention is disclosed in U.S. Pat. No. however it can be alternately substituted with a lower alkyl 5,440,056, which discloses 9-deoxo taxanes. These are ester or the ester linking to the dihydroindole core may be compounds lacking an OXO group at the carbon labeled 9 in substituted with C(O)-R where R is NH, an amino acid the taxane structure shown above (formula VI). The taxane ester or a peptide ester. R is typically C(O)CH, CH or H. ring may be substituted at the carbons labeled 1, 7 and 10 Alternately, a protein fragment may be linked by a bifunc (independently) with H, OH, O-R, or O-CO-R where R tional group, Such as maleoyl amino acid. R can also be is an alkyl or an aminoalkyl. AS well, it may be Substituted at carbons labeled 2 and 4 (independently) with aryol, substituted to form an alkyl ester which may be further alkanoyl, aminoalkanoyl or alkyl groups. The Side chain of Substituted. R may be -CH2- or a single bond. Rs and R. formula (V) may be substituted at R, and Rs (independently) may be H, OH or a lower alkyl, typically -CHCH with phenyl rings, Substituted phenyl rings, linear alkanes/ Alternatively R and R7 may together form an OXetane ring. alkenes, and groups containing H, O or N. Ro may be R, may alternately be H. Further substitutions include mol substituted with H, or a substituted or unsubstituted alkanoyl ecules wherein methyl groups are Substituted with other grOup. alkyl groups, and whereby unsaturated rings may be deriva tized by the addition of a Side group Such as an alkane, 0326 Taxanes in general, and paclitaxel is particular, is alkene, alkyne, halogen, ester, amide or amino group. considered to function as a cell cycle inhibitor by acting as an anti-microtubule agent, and more Specifically as a Stabi 0330 Exemplary Vinca alkaloids are vinblastine, Vincris lizer. These compounds have been shown useful in the tine, Vincristine Sulfate; Vindesline, and Vinorelbine, having treatment of proliferative disorders, including: non-Small the structures of formula (VIII): US 2005/0281883 A1 Dec. 22, 2005 40

(VIII)

R R Rs R Vinblastine: CH CH C(O)CH, OH CH, Vincristine: CHO CH C(O)CH, OH CH, Vindesline: CH NH, H OH CH, Winorelbine: CH CH CH H single bond

0331 Analogues typically require the Side group (shaded 0334 Exemplary camptothecin compounds include topo area) in order to have activity. These compounds are thought tecan, irinotecan (CPT-11), 9-aminocamptothecin, 21-lac to act as cell cycle inhibitors by functioning as anti-micro tam-20(S)-camptothecin, 10,11-methylenedioxycamptoth tubule agents, and more specifically to inhibit polymeriza ecin, SN-38, 9-nitrocamptothecin, tion. These compounds have been shown useful in treating 10-hydroxycamptothecin. Exemplary compounds have the proliferative disorders, including NSC lung, Small cell lung; structures of formula (X): breast, prostate; brain; head and neck; retinoblastoma; blad der; and penile cancers, and Soft tissue Sarcoma.

0332. In another aspect, the cell cycle inhibitor is a (X) camptothecin, or an analog or derivative thereof. Camptoth ecins have the following general structure of formula (IX):

(IX)

R R Rs Camptothecin: H H H Topotecan: OH (CH.)NHCH, H SN-38: OH H CHs OH X: O for most analogs, NH for 21-lactam analogs 0333. In this structure, X is typically O, but can be other 0335 Camptothecins have the five rings shown here. The groups, e.g., NH in the case of 21-lactam derivatives. R is ring labeled E must be intact (the lactone rather than typically H or OH, but may be other groups, e.g., a termi carboxylate form) for maximum activity and minimum nally hydroxylated C. alkane. R is typically H or an amino toxicity. These compounds are useful to as cell cycle inhibi containing group Such as (CH)NHCH, but may be other tors, where they can function as topoisomerase I inhibitors groups e.g., NO, NH, halogen (as disclosed in, e.g., U.S. and/or DNA cleavage agents. They have been shown useful Pat. No. 5,552,156) or a short alkane containing these in the treatment of proliferative disorders, including, for groups. R is typically H or a short alkyl Such as C.H. R. example, NSC lung, Small cell lung, and cervical cancers. is typically H but may be other groups, e.g., a methylene 0336. In another aspect, the cell cycle inhibitor is a dioxy group with R. podophyllotoxin, or a derivative or an analogue thereof. US 2005/0281883 A1 Dec. 22, 2005 41

Exemplary compounds of this type are etoposide or tenipo and 4,296,105, Rs may be OH or an ether linked alkyl group. side, which have the following structures of formula (XI): R may also be linked to the anthracycline ring by a group other than C(O), Such as an alkyl or branched alkyl group having the C(O) linking moiety at its end, Such as -CHCH(CH-X)C(O)-R, wherein X is H or an alkyl (XI) group (see, e.g., U.S. Pat. No. 4,215,062). R may alternately be a group linked by the functional group =N-NHC(O)- Y, where Y is a group Such as a phenyl or Substituted phenyl ring. Alternately Rs may have the following Structure of formula (XIII):

(XIII)

HC O

NH H3CO OCH R9 OH R10

R Etoposide CH 0342 in which Ro is OH either in or out of the plane of the ring, or is a Second Sugar moiety Such as R. Rio may be Teniposide H or form a Secondary amine with a group Such as an aromatic group, Saturated or partially Saturated 5 or 6 membered heterocyclic having at least one ring nitrogen (see U.S. Pat. No. 5,843.903). Alternately, R may be derived from an amino acid, having the Structure 0337 These compounds are thought to function as cell -C(O)CH(NHR)(R), in which R is H, or forms a C cycle inhibitors by being topoisomerase II inhibitors and/or membered alkylene with R. R. may be H, alkyl, ami by DNA cleaving agents. They have been shown useful as noalkyl, amino, hydroxy, mercapto, phenyl, benzyl or meth antiproliferative agents in, e.g., Small cell lung, prostate, and ylthio (see U.S. Pat. No. 4,296,105). brain cancers, and in retinoblastoma. 0338 Another example of a DNA topoisomerase inhibi 0343 Exemplary anthracyclines are doxorubicin, dauno tor is lurtotecan dihydrochloride (11H-1,4-dioxino 2,3-g rubicin, idarubicin, epirubicin, pirarubicin, Zorubicin, and pyrano.3',4':6.7indolizino1,2-biquinoline-9,12 (8H,14H)- carubicin. Suitable compounds have the Structures of for dione, 8-ethyl-2,3-dihydro-8-hydroxy-15-(4-methyl-1- mula (XIV): piperazinyl)methyl-, dihydrochloride, (S)-). 0339. In another aspect, the cell cycle inhibitor is an anthracycline. Anthracyclines have the following general (XIV) structure of formula (XII), where the R groups may be a variety of organic groups:

(XII)

HC O NH2 R3 R R Rs Doxorubicin: OCH CHOH OH out of ring plane 0340 According to U.S. Pat. No. 5,594,158, suitable R Epirubicin: OCH, CHOH OH in ring plane groups are: R is CH or CH-OH, R is daunosamine or H; (4 epimer of doxorubicin) R and R are independently one of OH, NO, NH, F, Cl, Daunorubicin: OCH CH OH out of ring plane Br, I, CN, H or groups derived from these; Rs 7 are all H or Idarubicin: H CH, OH out of ring plane Rs and R are H and R7 and Rs are alkyl or halogen, or Vice Pirarubicin OCH OH A. Zorubicin OCH =N-NHC(O)CHs B versa: R, and R are H and Rs and Rs are alkyl or halogen. Carubicin OH CH B 0341. According to U.S. Pat. No. 5,843.903, R may be a conjugated peptide. According to U.S. Pat. Nos. 4,215,062 US 2005/0281883 A1 Dec. 22, 2005

-continued -continued

(XIV) (XIV)

HC O

O NH2 HC R NH R3 2 R R Rs B:

R R Rs O / CH O A: OH NH2

O C- 0344) Other suitable anthracyclines are anthramycin, mitoxantrone, menogaril, nogalamycin, aclacinomycin A, olivomycin A, chromomycin A, and plicamycin having the StructureS:

Anthramycin

OH OH H HC N

Naa 21 NH2 O

OH O HN-1a OH NH

OH O HNS-1a OH NH Mitoxanitrone US 2005/0281883 A1 Dec. 22, 2005 43

-continued

OR CH O HO

O

HO

R R R R

Olivomycin A COCH(CH), CH COCH, H Chromomycin As COCH, CH COCH, CH Plicamycin H H H CH,

R R R

Menoganl H OCH H Nogalamycin O-sugar H COOCH,

Sugar:

CH HC O

HCO OCH CH US 2005/0281883 A1 Dec. 22, 2005 44

-continued

HC O N(CH3)2 O

HC O

OH O

HC O

O Aclacinomycin A

0345 These compounds are thought to function as cell cycle inhibitors by being topoisomerase inhibitors and/or by DNA cleaving agents. They have been shown useful in the treatment of proliferative disorders, including Small cell lung, breast, endometrial; head and neck, retinoblastoma; liver; bile duct; islet cell; and bladder cancers, and Soft tissue SCO. 0346. In another aspect, the cell cycle inhibitor is a platinum compound. In general, Suitable platinum com plexes may be of Pt(II) or Pt(IV) and have this basic structure of formula (XV):

(XV) Z1 R1-X Rs1NyPt Z2

0347 wherein X and Y are anionic leaving groups such 0349 Exemplary platinum compounds are cisplatin, car as Sulfate, phosphate, carboxylate, and halogen; R and R boplatin, oxaliplatin, and miboplatin having the Structures: are alkyl, amine, amino alkyl any may be further Substituted, and are basically inert or bridging groups. For Pt(II) com plexes Z and Z are non-existent. For Pt(IV) Z and Z. may NH be anionic groupS. Such as halogen, hydroxy, carboxylate, NH 's-n ester, sulfate or phosphate. See, e.g., U.S. Pat. Nos. 4,588, 3 NH, 831 and 4,250,198. CI-Pt-NH. O 0348 Suitable platinum complexes may contain multiple Pt atoms. See, e.g., U.S. Pat. Nos. 5,409,915 and 5,380,897. O For, example bisplatinum and triplatinum complexes of the Cisplatin Carboplatin type: US 2005/0281883 A1 Dec. 22, 2005 45

-continued -continued

OH O NH2 O. NH2 M / O Pt N OH

O. NH" 2 OH OH O N CH Oxaliplatin Nimustine Chlorozotocin

OH

O O OH O.\s NH2 . H OH O OH O/ HN

O HC n l NH Milboplatin Ninno Streptozocin 0350. These compounds are thought to function as cell cycle inhibitors by binding to DNA, i.e., acting as alkylating agents of DNA. These compounds have been shown useful 0352. Other suitable R groups include cyclic alkanes, in the treatment of cell proliferative disorders, including, alkanes, halogen Substituted groups, Sugars, aryl and het e.g., NSC lung, Small cell lung, breast, cervical; brain; head eroaryl groups, phosphonyl and Sulfonyl groups. AS dis and neck; esophageal, retinoblastom; liver; bile duct, blad closed in U.S. Pat. No. 4,367,239, R may suitably be CH-C(X)(Y)(Z), wherein X and Y may be the same or der; penile, and Vulvar cancers, and Soft tissue Sarcoma. different members of the following groups: phenyl, cycly 0351) In another aspect, the cell cycle inhibitor is a hexyl, or a phenyl or cyclohexyl group Substituted with nitroSourea. NitroSourease have the following general for groups Such as halogen, lower alkyl (C), trifluore methyl, mula (XVI), where typical R groups are shown below. cyano, phenyl, cyclohexyl, lower alkyloxy (C). Z has the following structure: -alkylene-N-RR, where R and R may be the same or different members of the following group: lower alkyl (C) and benzyl, or together R and R (XVI) O may form a Saturated 5 or 6 membered heterocyclic Such as pyrrolidine, piperidine, morfoline, thiomorfoline, N-lower Rsn 1 NH - alkyl piperazine, where the heterocyclic may be optionally Substituted with lower alkyl groups. N NoS. 0353 As disclosed in U.S. Pat. No. 6,096,923, R and R' of formula (XVI) may be the same or different, where each R Group: may be a Substituted or unsubstituted hydrocarbon having HC 1-10 carbons. Substitutions may include hydrocarbyl, halo, O ester, amide, carboxylic acid, ether, thioether and alcohol OH groups. As disclosed in U.S. Pat. No. 4,472.379, R of formula (XVI) may be an amide bond and a pyranose 1N1 OH O-CH structure (e.g., methyl 2-(N-(N-(2-chloroethyl)-N-nitroso OH Carmustine Ranimustine carbamoyl)-glycyl)amino-2'-deoxy-C-D-glucopyranoside). As disclosed in U.S. Pat. No. 4,150,146, R of formula (XVI) may be an alkyl group of 2 to 6 carbons and may be CH Substituted with an ester, Sulfonyl, or hydroxyl group. It may also be substituted with a carboxylic acid or CONH2 group. uralP OM No1Not, 0354) Exemplary nitrosoureas are BCNU (carmustine), Lomustine Fotemustine methyl-CCNU (semustine), CCNU (lomustine), ranimus tine, nimustine, chlorozotocin, fotemustine, and Streptozo cin, having the following Structures: US 2005/0281883 A1 Dec. 22, 2005 46

1N1 Carmustine

OH O-CH OH Ranimustine Lomustine

OH OH

Chlorozotocin

Fotemustine

0355 These nitrosourea compounds are thought to func 0357 Suitable nitroimidazole compounds are disclosed tion as cell cycle inhibitors by binding to DNA, that is, by in, e.g., U.S. Pat. Nos. 4,371,540 and 4,462,992. functioning as DNA alkylating agents. These cell cycle 0358 In another aspect, the cell cycle inhibitor is a folic inhibitors have been shown useful in treating cell prolifera acid antagonist, Such as methotrexate or derivatives or tive disorderS Such as, for example, islet cell; Small cell lung; analogues thereof, including edatrexate, trimetrexate, ralti melanoma, and brain cancers. trexed, piritrexim, denopterin, tomudeX, and pteropterin. Methotrexate analogues have the following general Structure 0356. In another aspect, the cell cycle inhibitor is a of formula (XVIII): nitroimidazole, where exemplary nitroimidazoles are met ronidazole, benznidazole, etanidazole, and misonidazole, having the structure of formula (XVII): (XVIII)

(XVII) r R1 "r\ { R2 R R Rs 0359 The identity of the R group may be selected from Metronidazole OH CH NO, Benzinidazole C(O)NHCH-benzyl NO, H organic groups, particularly those groups Set forth in U.S. Etanidazole CONHCHCH-OH NO, H Pat. Nos. 5,166,149 and 5,382,582. For example, R may be N, R may be N or C(CH), R and R may H or alkyl, e.g., CH, R may be a single bond or NR, where R is H or alkyl US 2005/0281883 A1 Dec. 22, 2005 47 group. Rses may be H, OCH, or alternately they can be 0360 wherein n=1 for methotrexate, n=3 for pteropterin. halogens or hydro groups. R, is a side chain of the general The carboxyl groups in the Side chain may be esterified or structure of formula (XIX): form a salt Such as a Zn "salt. Ro and Rio can be NH2 or may be alkyl substituted. (XIX) 0361 Exemplary folic acid antagonist compounds have O the structures of formulas (XX) and (XXI): NH HO

O O OH

(XX) R1 N NH2 n n Rs R4 2N2N R5 R6 R R0

R, R8

Ro R. R R R Rs Re R, Rs.

Methotrexete NH, N N H N(CH.) H H A (n = 1) H Edatrexate NH, N N H N(CHCH.) H H A (n = 1) H Tometrexate NH, N C(CH.) H NH H OCH, OCH OCH Pteropterin NH, N N H N(CH.) H H A (n = 3) H Denopterin OH N N CH N(CH.) H H A (n = 1) H Piritrexim NH, N C(CH.) H single OCH, H H OCH, H bond

A:

O

NH HO

O O OH

(XXI) Hooc1N O th Nn CH Hooc1N S N NH

Tomludex US 2005/0281883 A1 Dec. 22, 2005 48

0362. These compounds are thought to function as cell 0364. These compounds are thought to function as cell cycle inhibitors by Serving as antimetabolites of folic acid. cycle inhibitors as acting as antimetabolites of pyrimidine. They have been shown useful in the treatment of cell These compounds have been shown useful in the treatment proliferative disorders including, for example, Soft tissue of cell proliferative disorders including, for example, pan Sarcoma, Small cell lung, breast, brain, head and neck, creatic, breast, cervical, NSC lung, and bile duct cancers. bladder, and penile cancers. 0365. In another aspect, the cell cycle inhibitor is a 0363. In another aspect, the cell cycle inhibitor is a pyrimidine analogue. In one aspect, the pyrimidine ana cytidine analogue, Such as cytarabine or derivatives or logues have the general structure of formula (XXIII): analogues thereof, including enocitabine, FMdC ((E(-2'- deoxy-2-(fluoromethylene)cytidine), gemcitabine, 5-azac itidine, ancitabine, and 6-aZauridine. Exemplary compounds (XXIII) have the structure of formula (XXII): R6 R7 R8 YN R5 (XXII) RO 1. J Y R1 O N O R2 N21l R4 es ! R3 R' HO O 0366 wherein positions 2',3' and 5' on the Sugar ring (R, R R and Ra, respectively) can be H, hydroxyl, phosphoryl (see, e.g., U.S. Pat. No. 4,086,417) or ester (see, e.g., U.S. Pat. No. 3,894,000). Esters can be of alkyl, cycloalkyl, aryl OH R3 or heterocyclo/aryl types. The 2 carbon can be hydroxylated R R Rs R at either R or R', the other group is H. Alternately, the 2 Cytarabine H OH H CH carbon can be Substituted with halogens e.g., fluoro or Enocitabine C(O)(CH)2CH OH H CH difluoro cytidines Such as Gemcytabine. Alternately, the Gemcitabine H F F CH Sugar can be Substituted for another heterocyclic group Such Azacitidine H H OH N as a furyl group or for an alkane, an alkyl ether or an amide FMidC H CHF H CH linked alkane such as C(O)NH(CH-)-CH. The 2° amine can NH be substituted with an aliphatic acyl (R) linked with an amide (see, e.g., U.S. Pat. No. 3,991,045) or urethane (see, e.g., U.S. Pat. No. 3,894,000) bond. It can also be further Substituted to form a quaternary ammonium salt. Rs in the pyrimidine ring may be N or CR, where R is H, halogen N containing groups, or alkyl (See, e.g., U.S. Pat. No. 4,086, HO O 417). R and R, can together can form an oxo group or OS R=-NH-R and R7–H. Rs is Hor R, and Rs together can form a double bond or Rs can be X, where X is a structure of formula (XXIV): OH. H. Ancitabine (XXIV) n O O1. N1 N HO O 0367 Specific pyrimidine analogues are disclosed in U.S. Pat. No. 3,894,000 (see, e.g., 2'-O-palmityl-ara-cytidine, OH OH 3'-O-benzoyl-ara-cytidine, and more than 10 other 6-AZauridine examples); U.S. Pat. No. 3,991,045 (see, e.g., N4-acyl-1-B- D-arabinofuranosylcytosine, and numerous acyl groups derivatives as listed therein, Such as palmitoyl. US 2005/0281883 A1 Dec. 22, 2005 49

0368. In another aspect, the cell cycle inhibitor is a tive thereof, including 5-iododeoxyuridine (5-IudR), 5-bro fluoropyrimidine analogue, Such as 5-fluorouracil, or an modeoxyuridine (5-BudR), fluorouridine triphosphate analogue or derivative thereof, including carmofur, doxiflu (5-FUTP), and fluorodeoxyuridine monophosphate ridine, emitefur, tegafur, and floxuridine. Exemplary com (5-dFUMP). Exemplary compounds have the structures of pounds have the structures of formulas (XXV): formula (XXVI):

(XXV) O (XXVI) R2 F n N es is 1. R1 R R OH 5-Fluorouracil H H Carmofur C(O)NH(CH)-CH, H 5-Fluoro-2'-deoxyuridine: R = F Doxifluridine A1 H 5-Bromo-2'-deoxyuridine: R = Br Floxuridine A2 H 5-Iodoo-2'-deoxyuridine: R = I Emitefur CHOCHCH B Tegafur C H A1 0370. These compounds are thought to function as cell cycle inhibitors by Serving as antimetabolites of pyrimidine. These compounds have been shown useful in the treatment HO of cell proliferative disorderS Such as breast, cervical, non O melanoma skin, head and neck, esophageal, bile duct, pan creatic, islet cell, penile, and Vulvar cancers. 0371. In another aspect, the cell cycle inhibitor is a purine OH, OH analogue. Purine analogues have the following general A2 structure of formula (XXVII):

HO (XXVII) O H3 R2 X

OH R1ls CC)N B R

CN O 21 O O 0372 wherein X is typically carbon; R is H, halogen, S. amine or a Substituted phenyl; R is H, a primary, Secondary O N O or tertiary amine, a Sulfur containing group, typically -SH, an alkane, a cyclic alkane, a heterocyclic or a Sugar; R is H, a Sugar (typically a furanose or pyranose structure), a C Substituted Sugar or a cyclic or heterocyclic alkane or aryl group. See, e.g., U.S. Pat. No. 5,602,140 for compounds of this type. 0373) In the case of pentostatin, X-R2 is -CH-CH(OH)-. In this case a second carbon atom is O inserted in the ring between X and the adjacent nitrogen atom. The X-N double bond becomes a single bond. 0374 U.S. Pat. No. 5,446,139 describes suitable purine 0369 Other suitable fluoropyrimidine analogues include analogues of the type shown in the Structure of formula 5-FudR (5-fluoro-deoxyuridine), or an analogue or deriva (XXVIII): US 2005/0281883 A1 Dec. 22, 2005 50

nitrogens are present in ring A, one must be in the W position. If only one is present, it must not be in the Q (XXVIII) position. V and Q must not be simultaneously nitrogen. Z. R and Q must not be simultaneously nitrogen. IfZ is nitrogen, Y2V R is not present. Furthermore, R are independently one of | B \ H, halogen, C-7 alkyl, C-7 alkenyl, hydroxyl, mercapto, R1 s Rs C, alkylthio, C-, alkoxy, C-, alkenyloxy, aryl oxy, nitro, A. O primary, Secondary or tertiary amine containing group. Rs-s are H or up to two of the positions may contain indepen N") (r. dently one of OH, halogen, cyano, azido, Substituted amino, R2 R8 R7 Rs and R7 can together form a double bond. Y is H, a C-7 alkylcarbonyl, or a mono- di or triphosphate. O Ny 0376 Exemplary suitable purine analogues include 6-mercaptopurine, thiguanosine, thiamiprine, cladribine, fludaribine, tubercidin, puromycin, pentoxyfilline, where 0375 wherein N signifies nitrogen and V, W, X, Z can be these compounds may optionally be phosphorylated. Exem either carbon or nitrogen with the following provisos. Ring plary compounds have the structures of formulas (XXIX) A may have 0 to 3 nitrogen atoms in its structure. If two and (XXX):

(XXIX) R2 Ny R ls N R

R R Rs

6-Mercaptopurine H SH H Thioguanosine NH, SH B A: ON N B: HO y O H S N CH OH OH

Thiamiprine NH, A. H Cladribine Cl NH, B. Fludarabine F NH, B B2: HO B: HO O O H OH

OH OH

Puromycin H N(CH), B. B4: HO Tubercidin H NH, B O H NH2

O

(XXX) US 2005/0281883 A1 Dec. 22, 2005 51

-continued

CH O Ny "sn-n- N N O O CH Pentoxyfilline

0377 These compounds are thought to function as cell 0381 Examples of suitable nitrogen mustards are dis cycle inhibitors by Serving as antimetabolites of purine. closed in U.S. Pat. No. 3,808.297, wherein A is: 0378. In another aspect, the cell cycle inhibitor is a nitrogen mustard. Many Suitable nitrogen mustards are O (A) known and are Suitably used as a cell cycle inhibitor in the n p3 present invention. Suitable nitrogen mustards are also so known as cyclophosphamides. N 0379 A preferred nitrogen mustard has the general struc YR, ture of formula (XXXI): (XXXI) R1 0382 R are H or CHCHCl; R is H or oxygen 1 N N1)- containing groups Such as hydroperoxy; and R can where A is: be alkyl, aryl, heterocyclic. The cyclic moiety need (A) not be intact. See, e.g., U.S. Pat. Nos. 5,472,956, 4,908,356, 4,841,085 that describe the following 'NCso type of structure of formula (XXXII): N R2 (XXXII) R

0380 or -CH or other alkane, or chloronated alkane, typically CH-CH(CH) Cl, or a polycyclic group Such as B, or a Substituted phenyl Such as C or a heterocyclic group Such as D. 0383 wherein R is H or CHCHCl, and R are various Substituent groups.

(B) 0384 Exemplary nitrogen mustards include methylchlo roethamine, and analogues or derivatives thereof, including methylchloroethamine oxide hydrohchloride, novembichin, and mannomustine (a halogenated Sugar). Exemplary com pounds have the following Structures:

(C) Cl C O V / \ / V N CHCI N C CH (D) R Mechlorethanime Oxide HCI

R Mechlorethanime CH Novembichin CHCH(CH)Cl US 2005/0281883 A1 Dec. 22, 2005 52

0385) The nitrogen mustard may be cyclophosphamide, ifosfamide, perfosfamide, or torofosfamide, where these compounds have the structures of formula (XXXIII): (XXXVI)

N Cl (XXXIII) R1 \- R1 R2 R3. Cl On P 1. N-1- R R R S. Yo Chlorambucil CHCOOH H H N Melphalan COOH NH, H n R2 Chlornaphazine H together forms a benzene ring R

R R R 0388. The nitrogen mustard may be uracil mustard, Cyclophosphamide H CHCHCl H which has the structure of formula (XXXVII): Ifosfamide CHCHCl H H Perfosfamide CHCHCl H OOH (XXXVII) Torofosfamide CHCHCl CHCHCl H

0386 The nitrogen mustard may be estramustine, or an -- / \ analogue or derivative thereof, including phenesterine, pred O nimustine, and estramustine PO. Thus, Suitable nitrogen mustard type cell cycle inhibitors of the present invention have the structures of formulas (XXXIV) and (XXXV): 0389. The nitrogen mustards are thought to function as cell cycle inhibitors by Serving as alkylating agents for DNA. Nitrogen mustards have been shown useful in the (XXXIV) treatment of cell proliferative disorders including, for example, Small cell lung, breast, cervical, head and neck, ra prostate, retinoblastoma, and Soft tissue Sarcoma. O N 0390 The cell cycle inhibitor of the present invention N-> may be a hydroxyurea. Hydroxyureas have the following O general structure of formula (XXXVIII): HC (XXXVIII) R (O1, O R l O-X R N Estramustine OH R2 R1 Phenesterine C(CH)(CH)-CH(CH), (XXXV) 1c. 0391 Suitable hydroxyureas are disclosed in, for N example, U.S. Pat. No. 6,080,874, wherein R is a group n1no represented by the structure of formula (XXXIX): O O CHYo (XXXIX) O CH Prednimustine rN\, A. 2 S R2 R 0387. The nitrogen mustard may be chlorambucil, or an analogue or derivative thereof, including melphalan and chlormaphazine. Thus, Suitable nitrogen mustard type cell 0392 and R is an alkyl group having 1-4 carbons and R. cycle inhibitors of the present invention have the Structures is one of H, acyl, methyl, ethyl, and mixtures thereof, Such of formula (XXXVI): as a methylether. US 2005/0281883 A1 Dec. 22, 2005 53

0393. Other suitable hydroxyureas are disclosed in, e.g., proliferative disorderS Such as, for example, esophageal, U.S. Pat. No. 5,665,768, wherein R is a cycloalkenyl group, liver, bladder, and breast cancers. for example N-(3-(5-(4-fluorophenylthio)-furyl)-2-cyclo penten-1-yl)N-hydroxyurea; R is H or an alkyl group hav 04.01. In another aspect, the cell cycle inhibitor is an alkyl ing 1 to 4 carbons and R is H., X is H or a cation. Sulfonate, Such as buSulfan, or an analogue or derivative 0394 Other suitable hydroxyureas are disclosed in, e.g., thereof, Such as treoSulfan, improSulfan, pipOSulfan, and U.S. Pat. No. 4,299,778, wherein R is a phenyl group pipobroman. Exemplary compounds have the following Substituted with on or more fluorine atoms; R is a cyclo StructureS: propyl group; and R and X is H. 0395 Other suitable hydroxyureas are disclosed in, e.g., U.S. Pat. No. 5,066,658, wherein RandR together with the adjacent nitrogen form which is represented by the Structure of formula (XL): HC-S-O O-S-CH O O (XL) N (Cyn R Y - N Busulfan single bond 21 (CH2)m Improsulfan -CH-NH-CH

Piposulfan 0396 wherein m is 1 or 2, n is 0-2 and Y is an alkyl group. 0397. In one aspect, the hydroxy urea has the structure of formula (XLI):

(XLI)

HN ls NH on Pipobroman Hydroxyurea 0402. These compounds are thought to function as cell 0398 Hydroxyureas are thought to function as cell cycle cycle inhibitors by Serving as DNA alkylating agents. inhibitors by serving to inhibit DNA synthesis. 0403. In another aspect, the cell cycle inhibitor is a 0399. In another aspect, the cell cycle inhibitor is a benzamide. In yet another aspect, the cell cycle inhibitor is mytomicin, Such as mitomycin C, or an analogue or deriva tive thereof, Such as porphyromycin. Exemplary compounds a nicotinamide. These compounds have the basic structure of have the structures of formula (XLII): formula (XLIII):

(LXIII) (XLII)

04.04 wherein X is either O or S; A is commonly NH or it can be OH or an alkoxy group; B is N or C-R, where R is H or an ether-linked hydroxylated alkane Such as R OCHCH-OH, the alkane may be linear or branched and Mitomycin C H may contain one or more hydroxyl groups. Alternately, B Porphyromycin CH (N-methyl Mitomycin C) may be N-Rs in which case the double bond in the ring involving B is a single bond. Rs may be H, and alkyl or an aryl group (see, e.g., U.S. Pat. No. 4,258,052); R is H, OR, 0400. These compounds are thought to function as cell SR or NHR, where R is an alkyl group; and R is H, a cycle inhibitors by Serving as DNA alkylating agents. Mito lower alkyl, an ether linked lower alkyl such as O-Me or mycins have been shown useful in the treatment of cell -O-ethyl (see, e.g., U.S. Pat. No. 5,215,738). US 2005/0281883 A1 Dec. 22, 2005 54

04.05 Suitable benzamide compounds have the structures 04.09. In another aspect, the cell cycle inhibitor is a of formula (XLIV): halogenated Sugar, Such as mitolactol, or an analogue or derivative thereof, including mitobronitol and mannomus tine. Exemplary compounds have the Structures: (XLIV) X

Z. CHBr CHBr CH-NH2CH2CH2Cl 1N NH2 Y H OH HO H HO H N N 4 HO H HO H HO H Benzamides HO H H OH H OH X = O or S Y = H, OR, CH3, or acetoxy H OH H OH H OH Z = H, OR, SR, or NHR R = alkyl group CHBr CHBr CHNH2CH2CH2Cl Mitolactol Mitobronitol Mannomustine 0406 where additional compounds are disclosed in U.S. Pat. No. 5,215,738, (listing some 32 compounds). 0410. In another aspect, the cell cycle inhibitor is a diazo 04.07 Suitable nicotinamide compounds have the struc compound, Such as azaserine, or an analogue or derivative tures of formula (XLV): thereof, including 6-diazo-5-oxo-L-norleucine and 5-diaz ouracil (also a pyrimidine analog). Exemplary compounds have the structures of formula (XLVI): (XLV)

N NH2 (XLVI) 2 N Nicotinamides X = O or S Z = H, OR, SR, NHR R = alkyl group R R AZaserine O single bond 0408 where additional compounds are disclosed in U.S. 6-diazo-5-oxo- single bond CH, Pat. No. 5,215,738, L-norleucine

0411. Other compounds that may serve as cell cycle inhibitors according to the present invention are paZelliptine; R2 O O wortmannin; metoclopramide; RSU; buthionine sulfoxime; R2 N-P-NH ls o1Y tumeric; curcumin; AG337, a thymidylate synthase inhibi R2 tor; levamisole; lentinan, a polysaccharide; raZOxane, an N EDTA analogue; indomethacin, chlorpromazine, C. and B R2 interferon; MnBOPP; gadolinium texaphyrin, 4-amino-1.8- R R2 naphthalimide; staurosporine derivative of CGP; and R2 R2 SR-2508. 0412. Thus, in one aspect, the cell cycle inhibitor is a DNA alylating agent. In another aspect, the cell cycle HC A inhibitor is an anti-microtubule agent. In another aspect, the cell cycle inhibitor is a topoisomerase inhibitor. In another aspect, the cell cycle inhibitor is a DNA cleaving agent. In another aspect, the cell cycle inhibitor is an antimetabolite. In another aspect, the cell cycle inhibitor functions by O O YCH, inhibiting adenosine deaminase (e.g., as a purine analogue). Carboquone In another aspect, the cell cycle inhibitor functions by inhibiting purine ring Synthesis and/or as a nucleotide inter R R conversion inhibitor (e.g., as a purine analogue Such as BenZOdepa phenyl H mercaptopurine). In another aspect, the cell cycle inhibitor Meturedepa CH CH functions by inhibiting dihydrofolate reduction and/or as a Uredepa CH H thymidine monophosphate block (e.g., methotrexate). In another aspect, the cell cycle inhibitor functions by causing DNA damage (e.g., bleomycin). In another aspect, the cell US 2005/0281883 A1 Dec. 22, 2005 55 cycle inhibitor functions as a DNA intercalation agent and/or trideoxy-alpha-L-lyxo-hexopyranosyl)oxy)-7,8,9,10-tet RNA synthesis inhibition (e.g., doxorubicin, aclarubicin, or rahydro-6,11-trihydroxy-1-methoxy-, (8S-cis)-), gemcitab detorubicin (acetic acid, diethoxy-, 2-4-(3-amino-2,3,6- ine hydrochloride (cytidine, 2'-deoxy-2,2'-difluoro-, trideoxy-alpha-L-lyxo-hexopyranosyl)oxyl-1,2,3,4,6,11 monohydrochloride), nitacrine (1,3-propanediamine, N,N- hexahydro-2,5,12-trihydroxy-7-methoxy-6,11-dioxo-2- dimethyl-N'-(1-nitro-9-acridinyl)-), carboplatin (platinum, naphthacenyl)-2-oxoethyl ester, (2S-cis)-)). In another diammine(1,1-cyclobutanedicarboxylato(2-))-, (SP-4-2)-), aspect, the cell cycle inhibitor functions by inhibiting pyri altretamine (1,3,5-triazine-2,4,6-triamine, N.N.N',N',N',N- midine Synthesis (e.g., N-phosphonoacetyl-L-aspartate). In hexamethyl-), teniposide (furo(3',4':6,7)naphtho(2,3-d)-1,3- another aspect, the cell cycle inhibitor functions by inhib iting ribonucleotides (e.g., hydroxyurea). In another aspect, dioxol-6(5aH)-one, 5,8.8a,9-tetrahydro-5-(4-hydroxy-3,5- the cell cycle inhibitor functions by inhibiting thymidine dimethoxyphenyl)-9-((4,6-O-(2-thienylmethylene)-f-D- monophosphate (e.g., 5-fluorouracil). In another aspect, the glucopyranosyl)oxy)-, (5R-(5alpha,5af3.8aAlpha,9B(R)))- cell cycle inhibitor functions by inhibiting DNA synthesis ), eptaplatin (platinum, ((4R,5R)-2-(1-methylethyl)-1,3- (e.g., cytarabine). In another aspect, the cell cycle inhibitor dioxolane-4,5-dimethanamine-kappa N4.kappa functions by causing DNA adduct formation (e.g., platinum N5)(propanedioato(2-)-kappa O1, kappa O3)-, (SP-4-2)-), compounds). In another aspect, the cell cycle inhibitor amrubicin hydrochloride (5,12-naphthacenedione, 9-acetyl functions by inhibiting protein Synthesis (e.g., L-aspargin 9-amino-7-((2-deoxy-f-D-erythro-pentopyranosyl)oxy)-7, ase). In another aspect, the cell cycle inhibitor functions by 8,9,10-tetrahydro-6,11-dihydroxy-, hydrochloride, (7S-cis)- inhibiting microtubule function (e.g., taxanes). In another ), ifosfamide (2H-1,3,2-oxazaphosphorin-2-amine, N,3- aspect, the cell cycle inhibitor acts at one or more of the bis(2-chloroethyl)tetrahydro-2-oxide), cladribine steps in the biological pathway shown in FIG. 1. (adenosine, 2-chloro-2'-deoxy-), mitobronitol (D-mannitol, 1,6-dibromo-1,6-dideoxy-), fludaribine phosphate (9H-pu 0413 Additional cell cycle inhibitor S useful in the rin-6-amine, 2-fluoro-9-(5-O-phosphono-f-D-arabinofura present invention, as well as a discussion of the mechanisms of action, may be found in Hardman J. G., Limbird L. E. nosyl)-), enocitabine (docosanamide, N-(1-3-D-arabino Molinoff R. B., Ruddon R W., Gilman A. G. editors, furanosyl-1,2-dihydro-2-oxo-4-pyrimidinyl)-), Vindesine Chemotherapy of Neoplastic Diseases in Goodman and (Vincaleukoblastine, 3-(aminocarbonyl)-O4-deacetyl-3- Gilman's The Pharmacological Basis of Therapeutics Ninth de(methoxycarbonyl)-), idarubicin (5,12-naphthacenedione, Edition, McGraw-Hill Health Professions Division, New 9-acetyl-7-((3-amino-2,3,6-trideoxy-alpha-L-lyxo-hexopy York, 1996, pages 1225-1287. See also U.S. Pat. Nos. ranosyl)oxy)-7,8,9,10-tetrahydro-6,9,11-trihydroxy-, (7S 3,387,001; 3,808.297; 3,894,000; 3,991,045; 4,012,390; cis)-), Zinostatin (neocarzinostatin), Vincristine (Vincaleuko 4,057,548; 4,086,417; 4,144,237; 4,150,146; 4,210,584; blastine, 22-oxo-), tegafur (2,4(1H,3H)-pyrimidinedione, 4,215,062; 4,250,189; 4,258,052; 4,259,242; 4,296,105; 5-fluoro-1-(tetrahydro-2-furanyl)-), razoxane (2,6-pipera 4,299,778; 4,367,239; 4,374,414; 4,375.432; 4,472,379; Zinedione, 4,4'-(1-methyl-1,2-ethanediyl)bis-), methotrexate 4,588,831; 4,639,456; 4,767,855; 4,828,831; 4.841,045; (L-glutamic acid, N-(4-(((2,4-diamino-6-pteridinyl)methyl 4,841,085; 4,908,356; 4,923,876; 5,030,620; 5,034,320; )methylamino)benzoyl)-), raltitrexed (L-glutamic acid, 5,047,528; 5,066,658; 5,166,149; 5,190,929; 5,215,738; N-((5-(((1,4-dihydro-2-methyl-4-oxo-6-quinazolinyl)meth 5,292,731; 5,380,897; 5,382,582; 5,409,915; 5,440,056; yl)methylamino)-2-thienyl)carbonyl)-), oxaliplatin (plati 5,446,139; 5,472,956; 5,527,905; 5,552,156; 5,594,158; num, (1,2-cyclohexanediamine-N,N')(ethanedioato(2-)-O, 5,602,140; 5,665,768; 5,843,903; 6,080,874; 6,096,923; and O)-, (SP-4-2-(1R-trans))-), doxifluridine (uridine, 5'-deoxy REO30561. 5-fluoro-), mitolactol (galactitol, 1,6-dibromo-1,6-dideoxy ), piraubicin (5,12-naphthacenedione, 10-((3-amino-2,3,6- 0414. In another embodiment, the cell-cycle inhibitor is trideoxy-4-O-(tetrahydro-2H-pyran-2-yl)-alpha-L-lyxo camptothecin, mitoxantrone, etoposide, 5-fluorouracil, hexopyranosyl)oxy)-7,8,9,10-tetrahydro-6,8,11-trihydroxy doxorubicin, methotrexate, peloruside A, mitomycin C, or a 8-(hydroxyacetyl)-1-methoxy-, (8S-(8 alpha, 10 CDK-2 inhibitor or an analogue or derivative of any member alpha(S*)))-), docetaxel (2R,3S)-N-carboxy-3-phenylisos of the class of listed compounds. erine, N-tert-butyl ester, 13-ester with 53,20-epoxy-1,2 0415. In another embodiment, the cell-cycle inhibitor is alpha,4,73,103,13 alpha-hexahydroxytax-1-en-9-one 4-ac HTI-286, plicamycin; or mithramycin, or an analogue or etate 2-benzoate-), capecitabine (cytidine, 5-deoxy-5- derivative thereof. fluoro-N-((pentyloxy)carbonyl)-), cytarabine (2 (1H)-py rimidone, 4-amino-1-?3-D-arabino furanosyl-), valrubicin 0416) Other examples of cell cycle inhibitors also (pentanoic acid, 2-(1,2,3,4,6,11-hexahydro-2,5,12-trihy include, e.g., 7-hexanoyltaxol (QP-2), cytochalasin A, lan droxy-7-methoxy-6,11-dioxo-4-((2,3,6-trideoxy-3-((trifluo trunculin D, actinomycin-D, Ro-31-7453 (3-(6-nitro-1-me roacetyl)amino)-alpha-L-lyxo-hexopyranosyl)oxy)-2-naph thyl-3-indolyl)-4-(1-methyl-3-indolyl)pyrrole-2,5-dione), thacenyl)-2-oxoethyl ester (2S-cis)-), trofosfamide (3-2- PNU-151807, brostallicin, C2-ceramide, cytarabine ocfos (chloroethyl)-2-(bis(2-chloroethyl)amino)tetrahydro-2H-1, fate (2 (1H)-pyrimidinone, 4-amino-1-(5-O-(hydroxy(octa 3,2-oxazaphosphorin 2-oxide), prednimustine (pregna-1,4- decyloxy)phosphinyl)-3-D-arabinofuranosyl)-, monoso diene-3,20-dione, 21-(4-(4-(bis(2- dium salt), paclitaxel (5f,20-epoxy-1,2 alpha,4,7,103,13 chloroethyl)amino)phenyl)-1-oxobutoxy)-11,17 alpha-hexahydroxytaX-11-en-9-one-4,10-diacetate-2-ben dihydroxy-, (11B)-), lomustine (Urea, N-(2-chloroethyl)-N'- Zoate-13-(alpha-phenylhippurate)), doxorubicin (5,12-naph cyclohexyl-N-nitroSO-), epirubicin (5,12-naphthacenedione, thacenedione, 10-((3-amino-2,3,6-trideoxy-alpha-L-lyxo 10-((3-amino-2,3,6-trideoxy-alpha-L-arabino-hexopyrano hexopyranosyl)oxy)-7,8,9,10-tetrahydro-6,8,11-trihydroxy syl)oxy)-7,8,9,10-tetrahydro-6,8,11-trihydroxy-8-(hydroxy 8-(hydroxyacetyl)-1-methoxy-, (8S)-cis-), daunorubicin acetyl)-1-methoxy-, (8S-cis)-), or an analogue or derivative (5,12-naphthacenedione, 8-acetyl-10-((3-amino-2,3,6- thereof).

US 2005/0281883 A1 Dec. 22, 2005 59

0446. The structures of sirolimus, everolimus, and tac EP0184162B1 and U.S. Pat. No. 6,258,823) everolimus and rolimus are provided below in Table 3: derivatives thereof (e.g., U.S. Pat. No. 5,665,772). Further representative examples of Sirolimus analogues and deriva TABLE 3 tives include ABT-578 and others may be found in PCT Publication Nos. WO 97/10502, WO 96/41807, WO Name Code Name Company Structure 96/35423, WO 96/03430, WO 9600282, WO95/16691, WO Everolimus SAR-943 Novartis See below 9515328, WO95/07468, WO95/04738, WO95/04060, WO Sirolimus AY-22989 Wyeth See below RAPAMUNE NSC-22608O 94/25022, WO 94/21644, WO 94/18207, WO 94/10843, Rapamycin WO 94/09010, WO 94/04540, WO 94/02485, WO Tacrolimus FK506 Fujusawa See below 94/02137, WO 94/02136, WO 93/25533, WO 93/18043, WO 93/13663, WO 93/11130, WO 93/101.22, WO 1-1. 93/04680, WO 92/14737, and WO92/05179. Representative U.S. patents include U.S. Pat. Nos. 6,342,507; 5,985,890; 5,604,234, 5,597,715; 5,583,139; 5,563,172; 5,561,228; No 5,561,137; 5,541,193; 5,541,189; 5,534,632; 5,527,907; 5,484,799; 5,457,194, 5,457,182; 5,362,735; 5,324,644; 5,318,895; 5,310,903; 5,310,901; 5,258,389; 5.252,732; 5,247,076; 5,225,403; 5,221,625; 5,210,030; 5,208,241, 5,200,411; 5,198.421; 5,147,877; 5,140,018; 5,116,756; 5,109,112, 5,093,338; and 5,091,389. 0448. In one aspect, the fibrosis-inhibiting agent may be, e.g., rapamycin (Sirolimus), everolimus, biolimus, tresperi mus, auranofin, 27-0-demethylrapamycin, tacrolimus, gus perimus, pimecrolimus, or ABT-578.

0449 21. Inosine Monophosphate Dehydrogenase Inhibitors Everolimus 0450. In another embodiment, the pharmacologically active compound is an inoSine monophosphate dehydroge nase (IMPDH) inhibitor (e.g., mycophenolic acid, mycophe nolate mofetil (4-hexenoic acid, 6-(1,3-dihydro-4-hydroxy 6-methoxy-7-methyl-3-oxo-5-isobenzofuranyl)-4-methyl-, 2-(4-morpholinyl)ethyl ester, (E)-), ribavirin (1H-1,2,4-tria Zole-3-carboxamide, 1-3-D-ribofuranosyl-), tiazofurin (4-thiazolecarboxamide, 2-3-D-ribofuranosyl-), Viramidine, aminothiadiazole, thiophenfurin, tiazofurin) or an analogue ... I IO or derivative thereof. Additional representative examples are included in U.S. Pat. Nos. 5,536,747, 5,807,876, 5,932,600, 6,054,472, 6,128,582, 6,344,465, 6,395,763, 6,399,773, 6.420,403, 6,479,628, 6,498,178, 6.514,979, 6,518,291, 6,541,496, 6,596,747, 6,617,323, 6,624,184, Patent Appli cation Publication Nos. 2002/004.0022A1, 2002/ 0052513A1, 2002/0055483A1, 2002/0068346A1, 2002/

O111378A1, 2002/0111495A1, 2002/0123520A1, 2002/ 0143176A1, 2002/0147160A1, 2002/0161038A1, 2002/ O173491A1, 2002/0183315A1, 2002/0193612A1, 2003/ 0027845A1, 2003/0068302A1, 2003/0105073A1, 2003/ O130254A1, 2003/0143197A1, 2003/0144300A1, 2003/ O166201A1, 2003/0181497A1, 2003/0186974A1, 2003/ 0186989A1, 2003/0195202A1, and PCT Publication Nos. WO 0024725A1, WO 00/25780A1, WO 00/26197A1, WO 00/51615A1, WO 00/56331A1, WO 00/73288A1, WO 01/00622A1, WO 01/66706A1, WO 01/79246A2, WO 01/81340A2, WO 01/85952A2, WO 02/16382A1, WO 02/18369A2, WO 2051814A1, WO 2057287A2, WO2057425A2, WO 2060875A1, WO 2060896A1, WO 2060898A1, WO 2068058A2, WO 3020298A1, WO Sirolimus 3037349A1, WO 303.9548A1, WO 3045901A2, WO 3047512A2, WO 3053958A1, WO 3055447A2, WO 3059269A2, WO 3063573A2, WO 3087071A1, WO 0447 Further sirolimus analogues and derivatives 90/01545A1, WO 97/40028A1, WO 97/41211 A1, WO include tacrolimus and derivatives thereof (e.g., 98/40381A1, and WO99/55663A1).

US 2005/0281883 A1 Dec. 22, 2005 l)amino)(4-(6,7-dimethoxyquinazolin-4-yl)piperazinyl kinase inhibitor (e.g., leflunomide (4-isoxazolecarboxamide, )methane-1-thione), or an analogue or derivative thereof). 5-methyl-N-(4-(trifluoromethyl)phenyl)-, or an analogue or 0481 35. Protein Kinase Inhibitors derivative thereof). 0482 In another embodiment, the pharmacologically 0491) 40. Fibrinogen Antagonists active compound is a protein kinase inhibitor (e.g., 0492. In another embodiment, the pharmacologically KP-0201448, NPC15437 (hexanamide, 2,6-diamino-N-((1- active compound is a fibrinogen antagonist (e.g., picotamide (1-oxotridecyl)-2-piperidinyl)methyl)-), fasudil (1H-1,4-di (1,3-benzenedicarboxamide, 4-methoxy-N,N'-bis(3-pyridi azepine, hexahydro-1-(5-isoquinolinylsulfonyl)-), midos nylmethyl)-, or an analogue or derivative thereof). taurin (benzamide, N-(2,3,10,11,12,13-hexahydro-10 methoxy-9-methyl-1-oxo-9,13-epoxy-1H,9H-diindolo(1.2, 0493 41. Antimycotic Agents 3-gh:3',2,1'-lm)pyrrolo(3,4-j)(1.7)benzodiazonin-11-yl)-N- 0494. In another embodiment, the pharmacologically methyl-, (9Alpha,10B,11.f3,13Alpha)-).fasudil (1H-1,4- active compound is an antimycotic agent (e.g., miconazole, diazepine, hexahydro-1-(5-isoquinolinylsulfonyl)-, Sulconizole, parthenolide, rosconitine, nyStatin, isoconazole, dexniguldipine (3.5-pyridinedicarboxylic acid, 1,4-dihydro fluconazole, ketoconasole, imidazole, itraconazole, terpin 2,6-dimethyl-4-(3-nitrophenyl)-, 3-(4,4-diphenyl-1-pip afine, elonazole, bifonazole, clotrimazole, conazole, ter eridinyl)propyl methyl ester, monohydrochloride, (R)-), conazole (piperazine, 1-(4-((2-(2,4-dichlorophenyl)-2-(1H LY-317615 (1H-pyrole-2,5-dione, 3-(1-methyl-1H-indol-3- 1,2,4-triazol-1-ylmethyl)-1,3-dioxolan-4- yl)-4-1-1-(2-pyridinylmethyl)-4-piperidinyl)-1H-indol-3- yl)methoxy)phenyl)-4-(1-methylethyl)-, cis-), isoconazole yl-, monohydrochloride), perifosine (piperidinium, 4-hy (1-(2-(2-6-dichlorobenzyloxy)-2-(2-,4-dichlorophenyl droxy(octadecyloxy)phosphinyloxy)-1,1-dimethyl-, inner )ethyl)), griseofulvin (spiro(benzofuran-2 (3H), 1'-(2)cyclo salt), LY-333531 (9H,18H-5,21:12,17-dimethenodibenzo(e, hexane)-3,4'-dione, 7-chloro-2',4,6-trimeth-oxy-6'methyl-, k)pyrrolo(3,4-h)(1,4,13)oxadiazacyclohexadecine-18.20 (1'S-trans)-), bifonazole (1H-imidazole, 1-((1,1'-biphenyl)- (19H)-dione.9-(dimethylamino)methyl)-6,7,10,11-tetrahy 4-ylphenylmethyl)-), econazole nitrate (1-(2-((4-chlorophe dro-, (S)-), Kynac, SPC-100270 (1,3-octadecanediol, nyl)methoxy)-2-(2,4-dichlorophenyl)ethyl)-1H-imidazole 2-amino-, S-(R,R)-), Kynacyte, or an analogue or nitrate), croconazole (1H-imidazole, 1-(1-(2-((3-chlorophe derivative thereof). nyl)methoxy)phenyl)ethenyl)-), Sertaconazole (1H-Imida 0483 36. PDGF Receptor Kinase Inhibitors zole, 1-(2-((7-chlorobenzo(b)thien-3-yl)methoxy)-2-(2,4- dichlorophenyl)ethyl)-), omoconazole (1H-imidazole, 1-(2- 0484. In another embodiment, the pharmacologically (2-(4-chlorophenoxy)ethoxy)-2-(2,4-dichlorophenyl)-1- active compound is a PDGF receptor kinase inhibitor (e.g., methylethenyl)-, (Z)-), flutrimazole (1H-imidazole, 1-((2- RPR-127963E, or an analogue or derivative thereof). fluorophenyl)(4-fluorophenyl)phenylmethyl)-), fluconazole 0485 37. Endothelial Growth Factor Receptor Kinase (1H-1,2,4-triazole-1-ethanol, alpha-(2,4-difluorophenyl)-al Inhibitors pha-(1H-1,2,4-triazol-1-ylmethyl)-), neticonazole (1H-Imi dazole, 1-(2-(methylthio)-1-(2-(pentyloxy)phenyl)ethenyl)-, 0486 In another embodiment, the pharmacologically monohydrochloride, (E)-), butoconazole (1H-imidazole, active compound is an endothelial growth factor receptor 1-(4-(4-chlorophenyl)-2-((2,6-dichlorophenyl)thio)butyl)-, kinase inhibitor (e.g., CEP-7055, SU-0879 (E)-3-(3,5-di tert-butyl-4-hydroxyphenyl)-2-(aminothiocarbonyl)acry (+/-)-), clotrimazole (1-((2-chlorophenyl)diphenylmethyl)- lonitrile), BIBF-1000, AG-013736 (CP-868596), AMG-706, 1H-imidazole, or an analogue or derivative thereof). AVE-0005, NM-3 (3-(2-methylcarboxymethyl)-6-methoxy 0495 42. Bisphosphonates 8-hydroxy-isocoumarin), Bay-43-9006, SU-011248, or an 0496. In another embodiment, the pharmacologically analogue or derivative thereof). active compound is a bisphosphonate (e.g., clodronate, 0487 38. Retinoic Acid Receptor Antagonists alendronate, pamidronate, Zoledronate, or an analogue or 0488. In another embodiment, the pharmacologically derivative thereof). active compound is a retinoic acid receptor antagonist (e.g., 0497 43. Phospholipase A1 Inhibitors etarotene (Ro-15-1570) (naphthalene, 6-(2-(4-(ethylsulfo nyl)phenyl)-1-methylethenyl)-1,2,3,4-tetrahydro-1,1,4,4- 0498. In another embodiment, the pharmacologically tetramethyl-, (E)-), (2E,4E)-3-methyl-5-(2-((E)-2-(2,6,6-tri active compound is a phospholipase A1 inhibitor (e.g., methyl-1-cyclohexen-1-yl)ethenyl)-1-cyclohexen-1-yl)-2,4- ioteprednol etabonate (androsta-1,4-diene-17-carboxylic pentadienoic acid, tocoretinate (retinoic acid, 3,4-dihydro acid, 17-((ethoxycarbonyl)oxy)-11-hydroxy-3-oxo-, chlo 2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H-1- romethyl ester, (11.f3,17alpha)-, or an analogue or derivative benzopyran-6-yl ester, (2R*(4R*.8R)-(+)-), aliretinoin thereof). (retinoic acid, cis-9, trans-13-), beXarotene (benzoic acid, 4-(1-(5,6,7,8-tetrahydro-3,5,5,8,8-pentamethyl-2-naphtha 0499 44. Histamine H1/H2/H3 Receptor Antagonists lenyl)ethenyl)-), tocoretinate (retinoic acid, 3,4-dihydro-2, 0500. In another embodiment, the pharmacologically 5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H-1-ben active compound is a histamine H1, H2, or H3 receptor Zopyran-6-yl ester, I2R*(4R*,8R)-(+)-, or an analogue or antagonist (e.g., ranitidine (1,1-ethenediamine, N-(2-((5- derivative thereof). ((dimethylamino)methyl)-2-furanyl)methyl)thio)ethyl)-N'- methyl-2-nitro-), niperotidine (N-(2-((5-(dimethylami 0489) 39. Platelet Derived Growth Factor Receptor no)methyl)furfuryl)thio)ethyl)-2-nitro-N'-piperonyl-1,1- Kinase Inhibitors ethenediamine), famotidine (propanimidamide, 3-(((2- 0490. In another embodiment, the pharmacologically ((aminoiminomethyl)amino)-4-thiazolyl)methylthio)-N- active compound is a platelet derived growth factor receptor (aminosulfonyl)-), roxitadine acetate HCl (acetamide, US 2005/0281883 A1 Dec. 22, 2005 64

2-(acetyloxy)-N-(3-(3-(1-piperidinylmethyl)phenoxy)pro (2,4-thiazolidinedione, 5-((4-(2-(methyl-2-pyridinylamino pyl), monohydrochloride), lafutidine (acetamide, 2-((2-fura )ethoxy)phenyl)methyl)-, (Z)-2-butenedioate (1:1)), piogli nylmethyl)sulfinyl)-N-(4-(4-(1-piperidinylmethyl)-2-py taZone hydrochloride (2,4-thiazolidinedione, 5-((4-(2-(5- ridinyl)oxy)-2-butenyl)-, (Z)-), nizatadine (1,1- ethyl-2-pyridinyl)ethoxy)phenyl)methyl)-, ethenediamine, N-(2-(((2-(dimethylamino)methyl)-4- monohydrochloride (+/-)-), etofylline clofibrate (propanoic thiazolyl)methyl)thio)ethyl)N'-methyl-2-nitro-), elbrotidine acid, 2-(4-chlorophenoxy)-2-methyl-, 2-(1,2,3,6-tetrahydro (benzenesulfonamide, N-(((2-(((2-((aminoiminomethy 1,3-dimethyl-2,6-dioxo-7H-purin-7-yl)ethyl ester), etofi l)amino)-4-thiazoly)methyl)thio)ethyl)amino)methylene)-4- brate (3-pyridinecarboxylic acid, 2-(2-(4-chlorophenoxy)-2- bromo-), rupatadine (5H-benzo(5,6)cyclohepta(1,2-b)pyri methyl-1-oxopropoxy)ethyl ester), clinofibrate (butanoic dine, 8-chloro-6,11-dihydro-11-(1-((5-methyl-3- acid, 2,2'-(cyclohexylidenebis(4,1-phenyleneoxy)bis(2- pyridinyl)methyl)-4-piperidinylidene)-, trihydrochloride-), methyl-)), beZafibrate (propanoic acid, 2-(4-(2-((4-chlo fexofenadine HCl (benzeneacetic acid, 4-(1-hydroxy-4- robenzoyl)amino)ethyl)phenoxy)-2-methyl-), binifibrate (4(hydroxydiphenylmethyl)-1-piperidinyl)butyl)-alpha, (3-pyridinecarboxylic acid, 2-(2-(4-chlorophenoxy)-2-me alpha-dimethyl-, hydrochloride, or an analogue or derivative thyl-1-oxopropoxy)-1,3-propanediyl ester), or an analogue thereof). or derivative thereof). 0509. In one aspect, the pharmacologically active com 0501) 45. Macrollide Antibiotics pound is a peroxisome proliferator-activated receptor alpha 0502. In another embodiment, the pharmacologically agonist, such as GW-590735, GSK-677954, GSK501516, active compound is a macrollide antibiotic (e.g., dirithromy pioglitazone hydrochloride (2,4-thiazolidinedione, 5-4-2- cin (erythromycin, 9-deoxo-1-deoxy-9,11-(imino(2-(2- (5-ethyl-2-pyridinyl)ethoxyphenyl)methyl-, monohydro methoxyethoxy)ethylidene)oxy)-, (9S(R))-), flurithromycin ethylsuccinate (erythromycin, 8-fluoro-mono(ethylbutane chloride (+/-)-, or an analogue or derivative thereof). dioate) (ester)-), erythromycin Stinoprate (erythromycin, 0510 49. Estrogen Receptor Agents 2'-propanoate, compound with N-acetyl-L-cysteine (1:1)), 0511. In another embodiment, the pharmacologically clarithromycin (erythromycin, 6-O-methyl-), azithromycin active compound is an estrogen receptor agent (e.g., estra (9-deoxo-9a-aza-9a-methyl-9a-homoerythromycin-A), diol, 17-B-estradiol, or an analogue or derivative thereof). tellithromycin (3-de((2,6-dideoxy-3-C-methyl-3-O-methyl 0512 50. Somatostatin Analogues alpha-L-ribo-hexopyranosyl)oxy)-1,1,12-dideoxy-6-O-me thyl-3-oxo-12,11-(oxycarbonyl((4-(4-(3-pyridinyl)-1H-imi 0513. In another embodiment, the pharmacologically dazol-1-yl)butyl)imino))-), roXithromycin (erythromycin, active compound is a Somatostatin analogue (e.g., angio 9-(O-((2-methoxyethoxy)methyl)oxime)), rokitamycin (leu peptin, or an analogue or derivative thereof). comycin V, 4B-butanoate 3B-propanoate), RV-11 (erythro 0514) 51. Neurokinin 1 Antagonists mycin monopropionate mercaptosuccinate), midecamycin 0515. In another embodiment, the pharmacologically acetate (leucomycin V, 3B,9-diacetate 3,4B-dipropanoate), active compound is a neurokinin 1 antagonist (e.g., midecamycin (leucomycin V, 3,4B-dipropanoate), josamy GW-597599, lanepitant (1,4'-bipiperidine)-1'-acetamide, cin (leucomycin V, 3-acetate 4B-(3-methylbutanoate), or an N-(2-(acetyl((2-methoxyphenyl)methyl)amino)-1-(1H-in analogue or derivative thereof). dol-3-ylmethyl)ethyl)-(R)-), nolpitantium chloride (1-azoni abicyclo[2.2.2]octane, 1-2-3-(3,4-dichlorophenyl)-1-3- 0503 46. GPIIb IIIa Receptor Antagonists (1-methylethoxy)phenyl)acetyl-3-piperidinylethyl-4- 0504. In another embodiment, the pharmacologically phenyl-, chloride, (S)-), or Saredutant (benzamide, N-4-4- active compound is a GPIb IIIa receptor antagonist (e.g., (acetylamino)-4-phenyl-1-piperidinyl-2-(3,4- tirofiban hydrochloride (L-tyrosine, N-(butylsulfonyl)-O-(4- dichlorophenyl)butyl-N-methyl-, (S)-), or vofopiitant (4-piperidinyl)butyl)-, monohydrochloride-), eptifibatide (3-piperidinamine, N-2methoxy-5-5-(trifluoromethyl)- (L-cysteinamide, N6-(aminoiminomethyl)-N-2-(3-mer 1H-tetrazol-1-yl)phenyl)methyl)-2-phenyl-, (2S,3S)-, or an capto-1-oxopropyl)-L-lysylglycyl-L-alpha-aspartyl-L-tryp analogue or derivative thereof). tophyl-L-prolyl-, cyclic(1->6)-disulfide), xemilofiban 0516) 52. Neurokinin 3 Antagonist hydrochloride, or an analogue or derivative thereof). 0517. In another embodiment, the pharmacologically 0505 47. Endothelin Receptor Antagonists active compound is a neurokinin 3 antagonist (e.g., talnetant (4-quinolinecarboxamide, 3-hydroxy-2-phenyl-N-(1S)-1- 0506. In another embodiment, the pharmacologically phenylpropyl-, or an analogue or derivative thereof). active compound is an endothelin receptor antagonist (e.g., bosentan (benzenesulfonamide, 4-(1,1-dimethylethyl)-N-(6- 0518). 53. Neurokinin Antagonist (2-hydroxyethoxy)-5-(2-methoxyphenoxy)(2,2'-bipyrimi 0519 In another embodiment, the pharmacologically din)-4-yl)-, or an analogue or derivative thereof). active compound is a neurokinin antagonist (e.g., GSK 679769, GSK-823296, SR-489686 (benzamide, N-4-4- 0507 48. Peroxisome Proliferator-Activated Receptor (acetylamino)-4-phenyl-1-piperidinyl-2-(3,4-dichlorophe Agonists nyl)butyl-N-methyl-, (S)-), SB-223412; SB-235375 0508. In another embodiment, the pharmacologically (4-quinolinecarboxamide, 3-hydroxy-2-phenyl-N-(1S)-1- active compound is a peroxisome proliferator-activated phenylpropyl-), UK-226471, or an analogue or derivative receptor agonist (e.g., gemfibrozil (pentanoic acid, 5-(2,5- thereof). dimethylphenoxy)-2,2-dimethyl-), fenofibrate (propanoic 0520, 54. VLA-4 Antagonist acid, 2-(4-(4-chlorobenzoyl)phenoxy)-2-methyl-, 1-methyl 0521. In another embodiment, the pharmacologically ethyl ester), ciprofibrate (propanoic acid, 2-(4-(2,2-dichlo active compound is a VLA-4 antagonist (e.g., GSK683699, rocyclopropyl)phenoxy)-2-methyl-), rosiglitaZone maleate or an analogue or derivative thereof). US 2005/0281883 A1 Dec. 22, 2005

0522. 55. Osteoclast Inhibitor 7,8,12b-octahydro-6-oxo-, 4S-4 alpha, 7 alpha(R), 0523. In another embodiment, the pharmacologically 12b?3-), AVE-7688, or an analogue or derivative thereof). active compound is a osteoclast inhibitor (e.g., ibandronic 0532) 60. Peroxisome Proliferator-Activated Receptor acid (phosphonic acid, 1-hydroxy-3-(methylpentylamino Gamma Agonist Insulin Sensitizer )propylidenebis-), alendronate Sodium, or an analogue or 0533. In another embodiment, the pharmacologically derivative thereof). active compound is peroxisome proliferator-activated recep 0524) 56. DNA Topoisomerase ATP Hydrolysing Inhibi tor gamma agonist insulin Sensitizer (e.g., rosiglitaZone tOr maleate (2,4-thiazolidinedione, 5-(4-(2-(methyl-2-pyridi nylamino)ethoxy)phenyl)methyl)-, (Z)-2-butenedioate 0525) In another embodiment, the pharmacologically (1:1), fargilitazar (GI-262570, GW-2570, GW-3995, active compound is a DNA topoisomerase ATP hydrolysing GW-5393, GW-9765), LY-929, LY-519818, LY-674, or inhibitor (e.g., enoxacin (1,8-naphthyridine-3-carboxylic LSN-862), or an analogue or derivative thereof). acid, 1-ethyl-6-fluoro-1,4-dihydro-4-OXO-7-(1-piperazinyl)- ), levofloxacin (7H-Pyrido 1,2,3-de-1,4-benzoxazine-6- 0534 61. Protein Kinase C Inhibitor carboxylic acid, 9-fluoro-2,3-dihydro-3-methyl-10-(4-me 0535 In another embodiment, the pharmacologically thyl-1-piperazinyl)-7-oxo-, (S)-), ofloxacin (7H-pyrido1,2, active compound is a protein kinase C inhibitor, Such as 3-de-1,4-benzoxazine-6-carboxylic acid, 9-fluoro-2,3- ruboxistaurin mesylate (9H,18H-5,21:12,17-dimethenod dihydro-3-methyl-10-(4-methyl-1-piperazinyl)-7-oxo-, (+/- ibenzo(ek)pyrrolo(3,4-h)(1,4,13)oxadiazacyclohexadecine )-), pefloxacin (3-quinolinecarboxylic acid, 1-ethyl-6- 18.20(19H)-dione.9-(dimethylamino)methyl)-6,7,10,11 fluoro-1,4-dihydro-7-(4-methyl-1-piperazinyl)-4-oxo-), tetrahydro-, (S)-), Safingol (1,3-octadecanediol, 2-amino-, pipemidic acid (pyrido2,3-dipyrimidine-6-carboxylic acid, S-(R*,R)-), or enzastaurin hydrochloride (1H-pyrole-2, 8-ethyl-5,8-dihydro-5-oxo-2-(1-piperazinyl)-), pirarubicin 5dione, 3-(1-methyl-1H-indol-3-yl)-4-1-1-(2-pyridinylm (5,12-naphthacenedione, 10-3-amino-2,3,6-trideoxy-4-O- ethyl)-4-piperidinyl)-1H-indol-3-yl-, monohydrochloride), (tetrahydro-2H-pyran-2-yl)-alpha-L-lyxo-hexopyranosyl oxy-7,8,9,10-tetrahydro-6,8,11-trihydroxy-8-(hydroxy or an analogue or derivative thereof. acetyl)-1-methoxy-, 8S-8 alpha,10 alpha(S)-), 0536 62. ROCK (Rho-Associated Kinase) Inhibitors Sparfloxacin (3-quinolinecarboxylic acid, 5-amino-1-cyclo 0537. In another embodiment, the pharmacologically propyl-7-(3,5-dimethyl-1-piperazinyl)-6,8-difluoro-1,4-di active compound is a ROCK (rho-associated kinase) inhibi hydro-4-oxo-, cis-), AVE-6971, cinoxacin (1.3dioxolo4, tor, such as Y-27632, HA-1077, H-1152 and 4-1-(ami 5-gcinnoline-3-carboxylic acid, 1-ethyl-1,4-dihydro-4-oxo noalkyl)-N-(4-pyridyl)cyclohexanecarboxamide or an ana ), or an analogue or derivative thereof). logue or derivative thereof. 0526 57. Angiotensin I Converting Enzyme Inhibitor 0538 63. CXCR3 Inhibitors 0527. In another embodiment, the pharmacologically active compound is an angiotensin I converting enzyme 0539. In another embodiment, the pharmacologically inhibitor (e.g., ramipril (cyclopentabpyrrole-2-carboxylic active compound is a CXCR3 inhibitor such as T487, acid, 1-2-1-(ethoxycarbonyl)-3-phenylpropylamino)-1- TO906487 or analogue or derivative thereof. oxopropyloctahydro-, 2S-1R*(R)).2 alpha, 3af3, 6af3 0540) 64. ITK Inhibitors ), trandolapril (1H-indole-2-carboxylic acid, 1-2-(1-car boxy-3-phenylpropyl)amino-1-oxopropyloctahydro-, 2S 0541. In another embodiment, the pharmacologically 1R*(R*).2 alpha,3a alpha,7af3-), fasidotril (L-alanine, active compound is an Itkinhibitor such as BMS-509744 or N-(2S)-3-(acetylthio)-2-(1,3-benzodioxol-5-ylmethyl)-1- an analogue or derivative thereof. Oxopropyl-, phenylmethyl ester), cilaZapril (6H-pyridazino 0542) 65. Cytosolic Phospholipase Al-Alpha Inhibitors 1,2-a1,2diazepine-1-carboxylic acid, 9-1-(ethoxycar bonyl)-3-phenylpropylaminoloctahydro-10-oxo-, 1S-1 0543. In another embodiment, the pharmacologically alpha, 9 alpha(R)-), ramipril (cyclopentabpyrrole-2-car active compound is a cytosolic phospholipase A-alpha boxylic acid, 1-2-1-(ethoxycarbonyl)-3-phenylpropyl inhibitor such as efipladib (PLA-902) or analogue or deriva amino-1-oxopropyloctahydro-, 2S-1R*(R*)), 2 alpha, tive thereof. 3af33,6af3-, or an analogue or derivative thereof). 0544) 66. PPAR Agonist 0528 58. Angiotensin H Antagonist 0545. In another embodiment, the pharmacologically active compound is a PPAR Agonist (e.g., Metabolex ((-)- 0529. In another embodiment, the pharmacologically benzeneacetic acid, 4-chloro-alpha-3-(trifluoromethyl)- active compound is an angiotensin II antagonist (e.g., phenoxy-, 2-(acetylamino)ethyl ester), balaglitaZone (5-(4- HR-720 (1H-imidazole-5-carboxylic acid, 2-butyl-4-(meth (3-methyl-4-oxo-3,4-dihydro-quinazolin-2-yl-methoxy)- ylthio)-1-2'-(propylamino)carbonyl)aminosulfonyl 1, benzyl)-thiazolidine-2,4-dione), ciglitaZone (2,4- 1'-biphenyl-4-yl)methyl-, dipotassium Salt, or an analogue thiazolidinedione, 5-4-(1-methylcyclohexyl)methoxy or derivative thereof). phenyl)methyl]-), DRF-10945, farglitazar, GSK-677954, 0530) 59. Enkephalinase Inhibitor GW-409544, GW-501516, GW-590735, GW-590735, K-111, KRP-101, LSN-862, LY-519818, LY-674, LY-929, 0531. In another embodiment, the pharmacologically muraglitazar; BMS-298,585 (Glycine, N-(4-methoxyphe active compound is an enkephalinase inhibitor (e.g., Aventis noxy)carbonyl-N-(4-2-(5-methyl-2-phenyl-4-ox 100240 (pyrido 2,1-a2benzazepine-4-carboxylic acid, azolyl)ethoxyphenylmethyl-), netoglitazone; isaglitaZone 7-2-(acetylthio)-1-oxo-3-phenylpropylamino-1,2,3,4,6, (2,4-thiazolidinedione, 5-6-(2-fluorophenyl)methoxy-2- US 2005/0281883 A1 Dec. 22, 2005 66 naphthalenyl)methyl-), Actos AD-4833; U-72107A (2,4- droxyethyl)-3-methyl-), lufironil (2,4-Pyridinedicarboxam thiazolidinedione, 5-4-2-(5-ethyl-2-pyridinyl)ethoxy ide, N,N'-bis(2-methoxyethyl)-), or an analogue or deriva phenyl)methyl-, monohydrochloride (+/-)-), JTT-501; tive thereof). PNU-182716 (3.5-Isoxazolidinedione, 4-4-2-(5-methyl 2-phenyl-4-oxazolyl)ethoxyphenyl)methyl-), AVANDIA 0558 73. Alpha 2 Integrin Antagonist (from SB Pharmco Puerto Rico, Inc. (Puerto Rico); BRL 0559). In another embodiment, the pharmacologically 48482; BRL-49653; BRL-49653c; NYRACTA and Venvia active compound is an alpha 2 integrin antagonist (e.g., (both from (SmithKline Beecham (United Kingdom)); tesa E-7820, or an analogue or derivative thereof). glitazar ((2S)-2-ethoxy-3-4-2-4-(methylsulfonyl)oxy phenylethoxyphenylpropanoic acid), troglitazone (2,4- 0560) 74. TNF Alpha Inhibitor Thiazolidinedione, 5-4-(3,4-dihydro-6-hydroxy-2,5,7,8- 0561. In another embodiment, the pharmacologically tetramethyl-2H-1-benzopyran-2-yl)methoxyphenyl active compound is a TNF alpha inhibitor (e.g., ethyl methyl-), and analogues and derivatives thereof). pyruvate, GenZ-29155, lentinan (Ajinomoto Co., Inc. 0546 67. Immunosuppressants (Japan)), linomide (3-quinolinecarboxamide, 1,2-dihydro-4- hydroxy-N,1-dimethyl-2-oxo-N-phenyl-), UR-1505, or an 0547. In another embodiment, the pharmacologically analogue or derivative thereof). active compound is an immunosuppressant (e.g., batebulast (cyclohexanecarboxylic acid, 4-(aminoiminomethy 0562) 75. Nitric Oxide Inhibitor l)aminomethyl-, 4-(1,1-dimethylethyl)phenyl ester, trans 0563. In another embodiment, the pharmacologically ), cyclomunine, exalamide (benzamide, 2-(hexyloxy)-), active compound is a nitric oxide inhibitor (e.g., guanidi LYN-001, CCI-779 (rapamycin 42-(3-hydroxy-2-(hy oethyldisulfide, or an analogue or derivative thereof). droxymethyl)-2-methylpropanoate)), 1726; 1726-D; AVE 1726, or an analogue or derivative thereof). 0564) 76. Cathepsin Inhibitor 0548, 68. Erb Inhibitor 0565. In another embodiment, the pharmacologically active compound is a cathepsin inhibitor (e.g., SB-462795 or 0549. In another embodiment, the pharmacologically an analogue or derivative thereof). active compound is an Erb inhibitor (e.g., canertinib dihy drochloride (N-4-(3-(chloro-4-fluoro-phenylamino)-7-(3- 0566) 77. Delivery of Cells and Genes morpholin-4-yl-propoxy)-quinazolin-6-yl)-acrylamide 0567 The compositions of the invention can also be used dihydrochloride), CP-724714, or an analogue or derivative to deliver various types of living cells or genes to a desired thereof). site of administration in order to form new tissue. The term 0550 69. Apoptosis Agonist "genes' as used herein is intended to encompass genetic material from natural Sources, Synthetic nucleic acids, DNA, 0551. In another embodiment, the pharmacologically antisense-DNA and RNA. Thus, this aspect of the invention active compound is an apoptosis agonist (e.g., CEFLATO is a method for delivering living cells or genes, where the NIN (CGX-635) (from Chemgenex Therapeutics, Inc., composition also includes the cells of genes to be delivered, Menlo Park, Calif.), CHML, LBH-589, metoclopramide and steps (a) and (b) are as described for the method of (benzamide, 4-amino-5-chloro-N-2-(diethylamino)ethyl Sealing tissue. Step (c) would involve allowing a three 2-methoxy-), patupilone (4,17 dimensional matrix to form and delivering the cells or genes. dioxabicyclo(14.1.0)heptadecane-5,9-dione, 7,11-dihy droxy-8,8,10,12,16-pentamethyl-3-(1-methyl-2-(2-methyl 0568. When used to deliver cells, for example, mesen 4-thiazolyl)ethenyl, (1R,3S,7S,10R,11S,12S,16R)), AN-9; chymal Stem cells can be delivered to produce cells of the pivanex (butanoic acid, (2,2-dimethyl-1-oxopropoxy)m- Same type as the tissue into which they are delivered. ethyl ester), SL-100; SL-102; SL-11093; SL-11098; Mesenchymal stem cells are not differentiated and therefore SL-11099; SL-93; SL-98; SL-99, or an analogue or deriva can differentiate to form various types of new cells due to the tive thereof). presence of an active agent or the effects (chemical, physi cal, etc.) of the local tissue environment. Examples of 0552) 70. Lipocortin Agonist mesenchymal Stem cells include Osteoblasts, chondrocytes, 0553. In another embodiment, the pharmacologically and fibroblasts. Osteoblasts can be delivered to the site of a active compound is an lipocortin agonist (e.g., CGP-13774 bone defect to produce new bone; chondrocytes can be (9Alpha-chloro-6Alpha-fluoro-11B,17alpha-dihydroxy delivered to the Site of a cartilage defect to produce new 16Alpha-methyl-3-oxo-1,4-androstadiene-17f8-carboxylic cartilage, fibroblasts can be delivered to produce collagen acid-methylester-17-propionate), or analogue or derivative wherever new connective tissue is needed; neurectodermal cells can be delivered to form new nerve tissue, epithelial thereof). cells can be delivered to form new epithelial tissues, Such as 0554) 71. VCAM-1 Antagonist liver, pancreas, etc. 0555. In another embodiment, the pharmacologically 0569. The cells or genes may be either allogeneic or active compound is a VCAM-1 antagonist (e.g., DW-908e, Xenogeneic in origin. For example, the compositions can be or an analogue or derivative thereof). used to deliver cells or genes from other species that have been genetically modified. Because the composition of the 0556 72. Collagen Antagonist invention is not easily degraded in Vivo, cells and genes 0557. In another embodiment, the pharmacologically entrapped within the three-dimensional matrix will be iso active compound is a collagen antagonist (e.g., E-5050 lated from the patient's own cells and, as Such, will not (Benzenepropanamide, 4-(2,6-dimethylheptyl)-N-(2-hy provoke an immune response in the patient. In order to US 2005/0281883 A1 Dec. 22, 2005 67 entrap the cells or genes within this matrix, the cells or genes transplants). For example, the compositions of the invention are pre-mixed with the components in an initial environ can be used to adhere a donor cornea to the eye of a recipient ment. Upon eXposure to the aqueous environment, a three patient. AS used herein, the term "synthetic implant” refers dimensional matrix is formed, thereby entrapping the cells to any biocompatible material intended for implantation into or genes within the matrix. AS discussed above for biologi the body of a patient not encompassed by the above defi cally active agents, when used to deliver cells or genes, the nitions for native tissue and non-native tissue. Synthetic components may also contain biodegradable groups, or the implants include, for example, artificial blood vessels, heart composition may also contain biodegradable compounds, to Valves, artificial organs, bone prostheses, implantable lenti aid in controlled release of the cells or genes at the intended cules, vascular grafts, Stents, and Stent/graft combinations, site of delivery. etc. 0570) 78. Bioadhesives 0576 79. Ophthalmic Applications 0571 AS used herein, the terms “bioadhesive,”“biologi 0577 Because of their optical clarity, the compositions of cal adhesive,” and “Surgical adhesive” are used interchange the invention are particularly well Suited for use in oph ably to refer to biocompatible compositions capable of thalmic applications. See for example, Margalit et al. (2000) effecting temporary or permanent attachment between the “Bioadhesives for Intraocular Use'Retina 20:469-477. For Surfaces of two native tissues, or between a native tissue example, a Synthetic lenticule for correction of vision can be Surface and either a non-native tissue Surface or a Surface of attached to the Bowman's layer of the cornea of a patient's a Synthetic implant. eye using the methods of the present invention. In a manner similar to that described in U.S. Pat. No. 5,565,519 to Rhee 0572 In a general method for effecting the attachment of et al., the compositions can be molded into a desired a first Surface to a Second Surface, the composition of the lenticular shape, either during or after formation of the invention is applied to a first Surface, which is then contacted three-dimensional matrix. The resulting collagen lenticule with a Second Surface to effect adhesion therebetween. can then be attached to the Bowman's layer of a de Preferably, the composition is exposed to the aqueous envi epithelialized cornea of a patient's eye using the methods of ronment to initiate reaction among the reactive groups, then the present invention. By applying the composition to the delivered to the first Surface before Substantial reaction has anterior Surface of the cornea, then contacting the anterior occurred. The first Surface is then contacted with the Second Surface of the cornea with the posterior Surface of the Surface, preferably immediately, to effect adhesion. Thus, lenticule before Substantial reaction has occurred, reactive another embodiment of the invention is a method of bioad groups (e.g., electrophilic groups) on the components will hesion between two Surfaces, where steps (a) and (b) are as also covalently bind to collagen molecules in both the described for the method of Sealing tissue, and step (c) corneal tissue and the lenticule to firmly anchor the lenticule involves allowing a three-dimensional matrix to form and in place. Alternatively, the composition can be applied first adhere the Surfaces. to the posterior Surface of the lenticule, which is then 0573 The two surfaces may be held together manually, or contacted with the anterior Surface of the cornea. using other appropriate means, while the reaction is pro 0578 Thus, another embodiment of the invention is a ceeding to completion. Reaction is typically Sufficiently method of ophthalmic repair of the cornea, where steps (a) complete for adhesion to occur within about 5 to 60 minutes and (b) are as described for the method of Sealing tissue, and after exposure of the composition to the aqueous environ Step (c) involves allowing a three-dimensional matrix to ment; however, the time required for complete reaction to form and adhering a lenticule to the cornea. occur is dependent on a number of factors, including the type and molecular weight of each reactive component, the 0579. The compositions of the present invention are also degree of functionalization, and the concentration of the Suitable for use in Vitreous replacement. In addition, the components (i.e., higher concentrations result in faster reac compositions of the present invention may be used for the tion times). delivery of active agents. 0574. At least one of the first and second surfaces is 0580 80. Tissue Augmentation preferably a native tissue Surface. AS used herein, the term 0581. The compositions of the invention can also be used “native tissue' refers to biological tissues that are native to for augmentation of soft or hard tissue within the body of a the body of the patient being treated, and is intended to mammalian Subject. AS Such, they may be better than include biological tissues that have been elevated or currently marketed collagen-based materials for Soft tissue removed from one part of the body of a patient for implan augmentation, because they are leSS immunogenic and more tation to another part of the body of the same patient (Such persistent. Examples of Soft tissue augmentation applica as bone autografts, skin flap autografts, etc.). For example, tions include Sphincter (e.g., urinary, anal, esophageal) aug the compositions of the invention can be used to adhere a mentation and the treatment of rhytids and Scars. Examples piece of skin from one part of a patient's body to another part of hard tissue augmentation applications include the repair of the body, as in the case of a burn Victim. and/or replacement of bone and/or cartilaginous tissue. 0575. The other surface may be a native tissue surface, a Thus, another embodiment of the invention is a method of non-native tissue Surface, or a Surface of a Synthetic implant. tissue augmentation for a pre-selected Site, where steps (a) AS used herein, the term “non-native tissue' refers to and (b) are as described for the method of Sealing tissue, and biological tissues that have been removed from the body of Step (c) involves allowing a three-dimensional matrix to a donor patient (who may be of the same species or of a form at the pre-Selected Site. different species than the recipient patient) for implantation 0582 The compositions are particularly Suited for use as into the body of a recipient patient (e.g., tissue and organ a replacement material for Synovial fluid in Osteoarthritic US 2005/0281883 A1 Dec. 22, 2005 68 joints, Serving to reduce joint pain and improve joint func in GI Surgery (causing bowel obstruction), gynecological tion by restoring a Soft hydrogel network in the joint. The Surgery (causing pain and/or infertility), tendon repairs compositions can also be used as a replacement material for (causing shortening and flexion deformities), joint capsule the nucleus pulpOSuS of a damaged intervertebral disk. The procedures (causing capsular contractures), and nerve and nucleus pulpoSuS of the damaged disk is first removed, and muscle repair procedures (causing diminished or lost func the composition is then injected or otherwise introduced into tion). the center of the disk. The composition may either be 0587. The placement of medical devices and implants exposed to the aqueous environment prior to introduction also increases the risk that Surgical adhesions will occur. In into the disk, or allowed to inter-react in situ. addition to the above mechanisms, an implanted device can 0583. In a general method for effecting augmentation of trigger a "foreign body' response where the immune System tissue within the body of a mammalian Subject, the compo recognizes the implant as foreign and triggers an inflamma Sition is injected Simultaneously with exposure to the aque tory reaction that ultimately leads to Scar tissue formation. A ous environment, to a tissue Site in need of augmentation Specific form of foreign body reaction in response to medical through a Small-gauge (e.g., 25-32 gauge) needle. Once device placement is complete enclosure (“walling off) of inside the patient's body, the reactive groups on the com the implant in a capsule of Scar tissue (encapsulation). ponents inter-react with each other to form a three-dimen Fibrous encapsulation of implanted devices and implants Sional matrix in situ. In addition, in Some embodiments of can complicate any procedure, but breast augmentation and the invention, the reactive groups on the components can reconstruction Surgery, joint replacement Surgery, hernia react with body tissue to further enhance tissue augmenta repair Surgery, artificial vascular graft Surgery, Stent place tion. For example, Some of the reactive electrophilic groups ment, and neuroSurgery are particularly prone to this com may also react with primary amino groups only Sine residues plication. In each case, the implant becomes encapsulated by of collagen molecules within the patient's own tissue, pro a fibrous connective tissue capsule which compromises or Viding for “biological anchoring of the composition with impairs the function of the Surgical implant (e.g., breast the host tissue. implant, artificial joint, Surgical mesh, vascular graft, Stent or dural patch). 0584) 81. Adhesion Prevention 0588 Adhesions generally begin to form within the first 0585 Another use of the compositions of the invention is Several days after Surgery. Generally, adhesion formation is to coat tissues in order to prevent the formation of adhesions an inflammatory reaction in which factors are released, following Surgery or injury to internal tissueS or organs. increasing vascular permeability and resulting in fibrinogen Surgical adhesions are abnormal, fibrous bands of Scar tissue influx and fibrin deposition. This deposition forms a protein that can form inside the body as a result of the healing matrix that bridges the abutting tissues. Fibroblasts accu process that follows any open or minimally invasive Surgical mulate, attach to the protein matrix, deposit collagen and procedure including abdominal, gynecologic, cardiotho induce angiogenesis. If this cascade of events can be pre racic, Spinal, plastic, vascular, ENT, ophthalmologic, uro vented within 4 to 5 days following Surgery, then adhesion logic, neuro, or orthopedic Surgery. Surgical adhesions are formation may be inhibited. typically connective tissue Structures that form between adjacent injured areas within the body. Briefly, localized 0589 The compositions of the invention may be used to areas of injury trigger an inflammatory and healing response prevent adhesion formation in a wide variety of Surgical that culminates in healing and Scar tissue formation. If procedures including Spinal and neuroSurgical procedures Scarring results in the formation of fibrous tissue bands or (e.g., open Surgical resection of a ruptured lumbar disc or adherence of adjacent anatomical structures (that should be entrapped spinal nerve root (laminectomy); disectomies; and Separate), Surgical adhesion formation is said to have microlumbar disc excision (microdiscectomy)); gynecologi occurred. Adhesions can range from flimsy, easily Separable cal Surgical procedures (e.g., hysterectomy, myomectomy, Structures to dense, tenacious fibrous structures that can only endometriosis, infertility, birth control (e.g., tubal ligation), be separated by Surgical dissection. While many adhesions reversal of Sterilization, pain, dysmennorrhea, dysfunctional are benign, Some can cause Significant clinical problems and uterine bleeding, ectopic pregnancy, OVarian cysts, and are a leading cause of repeat Surgical intervention. gynecologic malignancies); abdominal Surgical procedures (e.g., hernia repair (abdominal, Ventral, inguinal, incisional), 0586 Since interventions involve a certain degree of bowel obstruction, inflammatory bowel disease (ulcerative trauma to the operative tissues, Virtually any procedure (no colitis, Crohn's disease), appendectomy, trauma (penetrat matter how well executed) has the potential to result in the ing wounds, blunt trauma), tumor resection, infections formation of clinically Significant adhesion formation. (abscesses, peritonitis), cholecystectomy, gastroplasty (bari Adhesions can be triggered by Surgical trauma Such as atric Surgery), esophageal and pyloric strictures, colostomy, cutting, manipulation, retraction or Suturing, as well as from diversion iliostomy, anal-rectal fistulas, hemorrhoidecto inflammation, infection (e.g., fungal or mycobacterium), mies, Splenectomy, hepatic tumor resection, pancreatitis, bleeding or the presence of a foreign body. Surgical trauma bowel perforation, upper and lower GI bleeding, and may also result from tissue drying, ischemia, or thermal ischemic bowel); cardiac Surgical procedure (e.g., transplant injury. Due to the diverse etiology of Surgical adhesions, the Surgery, vascular repair, coronary artery bypass grafting potential for formation exists regardless of whether the (CABG), congenital heart defects, and valve replacements, Surgery is done in a So-called minimally invasive fashion Staged procedures and reoperations (particularly repeat (e.g., catheter-based therapies, laparoscopy) or in a standard CABG Surgery)); orthopedic Surgical procedures (e.g., Sur open technique involving one or more relatively large inci gical interventions performed as a result of injury or trauma Sions. Although a potential complication of any Surgical (e.g., fractures (open and closed), sprains, joint dislocations, intervention, Surgical adhesions are particularly problematic crush injuries, ligament and muscle tears, tendon injuries, US 2005/0281883 A1 Dec. 22, 2005 69 nerve injuries, congenital deformities and malformations, from about 90 days to about 180 days. In one aspect, the total joint or partial joint replacement, and cartilage inju drug is released in effective concentrations for a period ries); and cosmetic or reconstructive Surgical procedure ranging from 1-90 dayS. (e.g., breast augmentation, breast reconstruction after cancer 0593. The exemplary anti-fibrosing agents, used alone or Surgery, craniofacial procedures, reconstruction after in combination, should be administered under the following trauma, congenital craniofacial reconstruction and oculo dosing guidelines. The total amount (dose) of anti-Scarring plastic Surgical procedures). agent in the composition can be in the range of about 0.01 tug-10 ug, or 10 ug-10 mg, or 10 mg-250 mg, or 250 0590 For certain applications compositions may be mg-1000 mg, or 1000 mg-2500 mg. The dose (amount) of include and/or release a therapeutic agent able to reduce anti-Scarring agent per unit area of Surface to which the Scarring (i.e., a fibrosis-inhibiting agent) at a Surgical Site, agent is applied may be in the range of about 0.01 ug/mm-1 Such as to prevent or inhibit the formation of post-operative tum, or 1 lug/mm-10 ug/mm, or 10 ug/mm-250 ug/mm. adhesions. Within one embodiment of the invention, com 250 ug/mm-1000 ug/mm, or 1000 ug/mm-2500 ug/mm. positions for the prevention of Surgical adhesions may 0594 Provided below are exemplary dosage ranges for include or be adapted to release an agent that inhibits one or various anti-Scarring agents that can be used in conjunction more of the five general components of the process of with compositions for treating or preventing Surgical adhe fibrosis (or Scarring), including: inflammatory response and Sions in accordance with the invention. (A) Cell cycle inflammation, migration and proliferation of connective inhibitors including doxorubicin and mitoxantrone. DOXO tissue cells (Such as fibroblasts or Smooth muscle cells), rubicin analogues and derivatives thereof: total dose not to formation of new blood vessels (angiogenesis), deposition exceed 25 mg (range of 0.1 ug to 25 mg); preferred 1 ug to of extracellular matrix (ECM), and remodeling (maturation 5 mg. Dose per unit area of 0.01 ug-100 ug per mm; and organization of the fibrous tissue). By inhibiting one or preferred dose of 0.1 ug/mm-10 ug/mm. Mitoxantrone and more of the components of fibrosis (or Scarring), the over analogues and derivatives thereof: total dose not to exceed growth of Scar tissue at a Surgical Site may be inhibited or 5 mg (range of 0.01 ug to 5 mg); preferred 0.1 ug to 1 mg. Dose per unit area of 0.01 ug-20 ug per mm; preferred dose reduced. of 0.05 ug/mm-3 ug/mm. (B) Cell cycle inhibitors includ 0591 Examples of fibrosis-inhibiting agents that may be ing paclitaxel and analogues and derivatives (e.g., doc combined with the present compositions to prevent the etaxel) thereof: total dose not to exceed 10 mg (range of 0.1 formation of adhesions include the following: cell cycle pig to 10 mg); preferred 1 lug to 3 mg. Dose per unit area of inhibitors including (A) anthracyclines (e.g., doxorubicin 0.1 ug-10 ug per mm ; preferred dose of 0.25 ug/mm°-5 and mitoxantrone), (B) taxanes (e.g., paclitaxel, TAXO Aug/mm. (C) Cell cycle inhibitors such as podophyllotoxins TERE and docetaxel), and (C) podophyllotoxins (e.g., eto (e.g., etoposide): total dose not to exceed 10 mg (range of poside); (D) immunomodulators (e.g., Sirolimus, everoli 0.1 ug to 10 mg); preferred 1 lug to 3 mg. Dose per unit area mus, tacrolimus); (E) heat shock protein 90 antagonists (e.g., of 0.1 ug-10 ug per mm; preferred dose of 0.25 ug/mm-5 geldanamycin, 17-AAG, 17-DMAG), (F) HMGCoA reduc Aug/mm. (D) Immunomodulators including sirolimus and tase inhibitors (e.g., Simvastatin); (G) inosine monophos everolimus. Sirolimus (i.e., rapamycin, RAPAMUNE): total phate dehydrogenase inhibitors (e.g., mycophenolic acid, dose not to exceed 10 mg (range of 0.1 ug to 10 mg); 1-alpha-25 dihydroxy vitamin D.); (H)NF kappa B inhibi preferred 10 ug to 1 mg. Dose per unit area of 0.1 ug-100 lig tors (e.g., Bay 11-7082); (I) antimycotic agents (e.g., Sul per mm'; preferred dose of 0.5 lug/mm°-10 ug/mm . Everoli conizole) and (J) p38 MAP kinase inhibitors (e.g., mus and derivatives and analogues thereof: total dose should not exceed 10 mg (range of 0.1 ug to 10 mg); preferred 10 SB202190), as well as analogues and derivatives of the tug to 1 mg. Dose per unit area of 0.1 ug-100 ug per mm of aforementioned. Surface area; preferred dose of 0.3 ug/mm-10 ug/mm. (E) 0592. The drug dose administered from the present com Heat shock protein 90 antagonists (e.g., geldanamycin) and positions for Surgical adhesion prevention will depend on a analogues and derivatives thereof: total dose not to exceed variety of factors, including the type of formulation, the 20 mg (range of 0.1 ug to 20 mg); preferred 1 lug to 5 mg. location of the treatment Site, and the type of condition being Dose per unit area of 0.1 ug-10 ug per mm; preferred dose treated; however, certain principles can be applied in the of 0.25 ug/mm-5 ug/mm. (F) HMGCoA reductase inhibi application of this art. Drug dose can be calculated as a tors (e.g., Simvastatin) and analogues and derivatives function of dose per unit area (of the treatment site), total thereof: total dose not to exceed 2000 mg (range of 10.0 ug drug dose administered can be measured and appropriate to 2000 mg); preferred 10 ug to 300 mg. Dose per unit area Surface concentrations of active drug can be determined. of 1.0 ug-1000 ug per mm; preferred dose of 2.5 lug/mm Drugs are to be used at concentrations that range from 500 ug/mm. (G) Inosine monophosphate dehydrogenase several times more than to 50%, 20%, 10%, 5%, or even less inhibitors (e.g., mycophenolic acid, 1-alpha-25 dihydroxy than 1% of the concentration typically used in a Single Vitamin D) and analogues and derivatives thereof: total Systemic dose application. In certain aspects, the anti-Scar dose not to exceed 2000 mg (range of 10.0 ug to 2000 mg); ring agent is released from the polymer composition in preferred 10 ug to 300 mg. Dose per unit area of 1.0 lug-1000 effective concentrations in a time period that may be mea tug per mm; preferred dose of 2.5 lug/mm-500 ug/mm. Sured from the time of infiltration into tissue adjacent to the (H)NF kappa B inhibitors (e.g., Bay 11-7082) and analogues device, which ranges from about less than 1 day to about 180 and derivatives thereof: total dose not to exceed 200 mg dayS. Generally, the release time may also be from about leSS (range of 1.0 ug to 200 mg); preferred 1 ug to 50 mg. Dose than 1 day to about 180 days; from about 7 days to about 14 per unit area of 1.0 ug-100 ug per mm; preferred dose of 2.5 days; from about 14 days to about 28 days; from about 28 tag/mm-50 ug/mm. (I) Antimycotic agents (e.g., Sulconi days to about 56 days; from about 56 days to about 90 days; Zole) and analogues and derivatives thereof; total dose not to US 2005/0281883 A1 Dec. 22, 2005 70 exceed 2000 mg (range of 10.0 ug to 2000 mg); preferred 10 may be applied during open or endoscopic gynecological tug to 300 mg. Dose per unit area of 1.0 ug-1000 ug per mm; Surgery to the tissue Surface of the pelvic Side wall, adnexa, preferred dose of 2.5 lug/mm-500 ug/mm and (J) p38 MAP uterus and any adjacent affected tissues during the Surgical kinase inhibitors (e.g., SB202190) and analogues and procedure or to the Surface of an implanted device or implant derivatives thereof: total dose not to exceed 2000 mg (range (e.g., genital-urinary Stents, bulking agents, Sterilization of 10.0 ug to 2000 mg); preferred 10 ug to 300 mg. Dose per devices (e.g., valves, clips and clamps), and tubal occlusion unit area of 1.0 ug-1000 ug per mm; preferred dose of 2.5 implants and plugs) and/or the tissue Surrounding the tug/mm-500 ug/mm. implant before, during, or after the Surgical procedure. 0595. In a general method for coating tissues to prevent 0601 For the prevention of adhesions associated with the formation of adhesions following Surgery, the composi abdominal Surgical procedures, the compositions alone or tion is exposed to the aqueous environment and a thin layer loaded with a therapeutic agent (e.g., a fibrosis-inhibiting of the composition is then applied to the tissues comprising, agent) may be applied during open, endoscopic, or laparo Surrounding, and/or adjacent to the Surgical Site before Scopic abdominal Surgery to the tissue Surface of the peri Substantial reaction has occurred. Application of the com toneal cavity, Visceral peritoneum, abdominal organs, position to the tissue Site may be by extrusion, brushing, abdominal wall and any adjacent affected tissues during the spraying (as described above), or by any other convenient Surgical procedure or to the Surface of an implanted device means. Thus, the invention also relates to a method of or implant and/or the tissue Surrounding the implant before, preventing adhesions between tissueS of a patient, where during, or after the Surgical procedure. Representative Steps (a) and (b) are as described for the method of Sealing examples of implants for use in abdominal procedures tissue, and Step (c) involves allowing a three-dimensional includes, without limitation, hernia meshes, restriction matrix to form on the tissue, and thus prevent tissue adhe devices for obesity, implantable Sensors, implantable pumps, SO. peritoneal dialysis catheters, peritoneal drug-delivery cath eters, GI tubes for drainage or feeding, portosystemic 0596 Following application of the composition to the shunts, shunts for ascites, gastroStomy or percutaneous Surgical Site, inter-reaction is allowed to continue in Situ feeding tubes, jejunostomy endoscopic tubes, colostomy prior to closure of the Surgical incision. Once the reaction devices, drainage tubes, biliary T-tubes, hemostatic has reached equilibrium, tissues that are brought into contact implants, enteral feeding devices, colonic and biliary Stents, with the coated tissues will not adhere thereto. The Surgical low profile devices, gastric banding implants, capsule endo Site can then be closed using conventional means Such as by Sutures, and So forth. Scopes, anti-reflux devices, and esophageal Stents. 0602 For the prevention of adhesions associated with 0597. In general, compositions that achieve complete cardiac Surgical procedures, the compositions alone or inter-reaction within a relatively short period of time (i.e., loaded with a therapeutic agent (e.g., a fibrosis-inhibiting 5-15 minutes following exposure of the multifunctional agent) may be applied during open or endoscopic heart compounds to the modified environment) are preferred for Surgery to the tissue Surface of the pericardium (or infiltrated use in the prevention of Surgical adhesions, So that the into the pericardial sac), heart, great vessels, pleura, lungs, Surgical Site may be closed relatively Soon after completion chest wall and any adjacent affected tissues during the of the Surgical procedure. Surgical procedure or to the Surface of an implanted device 0598 Certain surgical procedures may involve placement or implant and/or the tissue Surrounding the implant before, of a medical device or implant at the Surgical Site, in which during, or after the Surgical procedure. Representative case it may be desirable to apply the composition (with or examples of implants for use in cardiac procedures includes, without a therapeutic agent) to the Surface of the implant, to without limitation, heart Valves (porcine, artificial), ven the implant-tissue interface, and/or to tissue in the vicinity of tricular assist devices, cardiac pumps, artificial hearts, Stents, the implanted device to minimize the formation of post bypass grafts (artificial and endogenous), patches, cardiac operative Surgical adhesions, unwanted Scarring in the vicin electrical leads, defibrillators and pacemakers. ity of the implant, and encapsulation of the implant by a 0603 For the prevention of adhesions associated with fibrous connective tissue capsule. orthopedic Surgical procedures, the compositions alone or 0599 For the prevention of adhesions in spinal and loaded with a therapeutic agent (e.g., a fibrosis-inhibiting neuroSurgical procedures, the compositions alone or loaded agent) may be applied during open or arthroscopic ortho with a therapeutic agent (e.g., a fibrosis-inhibiting agent) pedic Surgery to the tissue Surface of the bone, joint, muscle, may be applied to the tissue Surface at a spinal or neuro tendon, ligament, cartilage and any adjacent affected tissues Surgical site or to the Surface of an implanted device (e.g., during the Surgical procedure or to the Surface of an dural patches, Spinal prostheses, artificial disc, rods, bone implanted orthopedic device or implant and/or the tissue fixation devices (e.g., anchoring plates and bone Screws), Surrounding the implant before, during, or after the Surgical injectable filling or bulking agents for discs, Spinal grafts, procedure. Representative examples of implants for use in Spinal nucleus implants intervertebral disc Spacers, fusion orthopedic procedures include plates, rods, Screws, pins, cages, or to implants placed in the brain, Such as drains, wires, total and partial joint prostheses (artificial hips, knees, shunts, drug-delivery pumps, or neurostimulation devices) shoulders, phalangeal joints), reinforcement patches, tissue and/or the tissue Surrounding the implant before, during, or fillers, Synthetic bone fillers, bone cement, Synthetic graft after the Surgical procedure. material, allograft material, autograft material, artificial discS, Spinal cages, and intermedulary rods. 0600 For the prevention of adhesions associated with gynecological procedures, the compositions alone or loaded 0604 For the prevention of adhesions associated with with a therapeutic agent (e.g., a fibrosis-inhibiting agent) cosmetic or reconstructive Surgical procedures, the compo US 2005/0281883 A1 Dec. 22, 2005

Sitions alone or loaded with a therapeutic agent (e.g., a around the device generally can help to Secure the device in fibrosis-inhibiting agent) may be applied during open or the implantation site. Device migration can result in device endoscopic cosmetic Surgery to the Soft tissue implant failure and, depending on the type and location of the device, Surface before, during, or after the implantation procedure or can lead to leakage, vessel occlusion, and/or damage to the to the Surface of the tissue of the implantation pocket Surrounding tissue. Incorporation of a fibrosis-inducing immediately prior to, or during implantation of the Soft agent with the compositions of the invention can provide an tissue implant. Representative examples of Soft tissue effective, long-lasting and biocompatible approach for implants for use in cosmetic, plastic, and reconstructive anchoring implantable medical devices into or onto biologi Surgical procedures include face, nose, breast, chin, but cal tissue. tocks, chest, lip and cheek implants) or to the Surface of the Soft tissue implant and/or the tissue Surrounding the implant 0610. In certain embodiments, the medical implant, when before, during, or after implantation of the Soft tissue placed in to a tissue, releases an agent that induces or implant. promotes adhesion between the implant and the tissue or a fibrotic reaction. In other embodiments, the medical implant 0605 82. Implants and Coating Material for Implants contains or is made of a fibrosing agent, but does not release 0606. The compositions of the invention can also be the fibrosing agent. In Such embodiments, the fibrosing formed as Solid implants, a term that is used herein to refer agent contained in the medical implant induces or promotes to any Solid object which is designed for insertion and use by direct contact of the agent to the tissue where the implant within the body, and includes bone and cartilage implants is placed. (e.g., artificial joints, retaining pins, cranial plates, and the 0611 Alternatively, or in addition, the tissue cavity into like, of metal, plastic and/or other materials), breast implants which the device or implant is placed can be treated with a (e.g., Silicone gel envelopes, foam forms, and the like), fibrosis-inducing agent prior to, during, or after the implan catheters and cannulas intended for long-term use (beyond tation procedure. This can be accomplished, for example, by about three days) in place, artificial organs and vessels (e.g., topical application of the composition comprising a fibroS artificial hearts, pancreases, kidneys, blood vessels, and the ing agent or by Spraying the composition into the anatomical like), drug delivery devices (including monolithic implants, Space where the device can be placed or at the interface pumpS and controlled release devices Such as Alzet(R) between the implant and the tissue Surface. minipumps (DURECT Corporation, Cupertino, Calif.), ste roid pellets for anabolic growth or contraception, and the 0612 Representative examples of medical implants of like), Sutures for dermal or internal use, periodontal mem particular utility for use in combination with a fibrosis branes, ophthalmic shields, corneal lenticules, and the like. inducing agent include, but are not restricted to, Orthopaedic implants (artificial joints, ligaments and tendons, Screws, 0607 Another use of the compositions is as a coating plates, and other implantable hardware), dental implants, material for an implant (e.g., Synthetic implants). intravascular implants (particularly arterial and venous 0608) 83. Medical Implants Combined with Fibrosing occlusion devices and implants, vascular destructive Agents implants), male and female contraceptive or sterilization devices and implants, Soft palate implants, embolization 0609. In one aspect, medical implants may contain and/or devices, Surgical meshes (e.g., hernia repair meshes, tissue are adapted to release an agent which induces or promotes Scaffolds), fistula treatments, and spinal implants (e.g., arti adhesion between the implant and tissue or a fibrotic reac ficial intervertebral discs, Stent grafts, Spinal fusion devices, tion. The clinical performance of numerous medical devices etc.). may be improved by anchoring the device effectively into the Surrounding tissue to provide either structural Support or 0613 AS medical implants are made in a variety of to facilitate Scarring and healing. Effective attachment of the configurations and sizes, the exact dose administered can device into the Surrounding tissue, however, is not always vary with the amount injected or with the device size, readily achieved. One reason for ineffective attachment is Surface area and design; however, certain principles can be that implantable medical devices generally are composed of applied in the application of this art. Drug dose can be materials that are highly biocompatible and designed to calculated as a function of dose per unit area (of the portion reduce the host tissue response. These materials (e.g., stain of the device being coated), total drug dose administered can leSS Steel, titanium based alloys, fluoropolymers, and ceram be measured, and appropriate Surface concentrations of ics) typically do not provide a good Substrate for host tissue active drug can be determined. It should be readily evident attachment and ingrowth during the Scarring process. AS a to one of skill in the art that any of the previously described result of poor attachment between the device and the host fibrosis inducing agents or derivatives or analogues thereof tissue, devices can have a tendency to migrate within the can be utilized with the present compositions without devi vessel or tissue in which they are implanted. The extent to ating from the Spirit and Scope of the invention. which a particular type of medical device can move or 0.614 Regardless of the method of application of the drug migrate after implantation depends on a variety of factors to the implant, the exemplary fibrosing agents, used alone or including the type and design of the device, the material(s) in combination, should be administered under the following from which the device is formed, the mechanical attributes dosing guidelines: (e.g., flexibility and ability to conform to the Surrounding geometry at the implantation site), the Surface properties, 0.615 Utilizing talc as an exemplary fibrosis-inducing and the porosity of the device or device surface. The agent, the total amount of talc delivered from an implant or tendency of a device to loosen after implantation also coated onto the surface of an implant should not exceed 100 depends on the type of tissue and the geometry at the mg (range of 1 ug to 100 mg). In one embodiment, the total treatment site, where the ability of the tissue to conform amount of talc released from the implant should be in the US 2005/0281883 A1 Dec. 22, 2005 72 range of 10 ug to 50 mg. The dose per unit area of the device embodiment, the total amount of bleomycin released from (i.e., the dosage of talc as a function of the Surface area of the implant should be in the range of 0.10 ug to 50 mg. The the portion of the device to which drug is applied and/or dose per unit area of the device (i.e., the dosage of bleomy incorporated) should fall within the range of 0.05 ug-10 ug cin as a function of the Surface area of the portion of the per mm of Surface area coated. In another embodiment, talc device to which drug is applied and/or incorporated) should should be applied to an implant surface at a dose of 0.05 fall within the range of 0.005ug-10 ug per mm of surface Aug/mm-10 ug/mm of Surface area coated. area coated. In another embodiment, bleomycin should be 0616) Utilizing silk as an exemplary fibrosis-inducing applied to an implant surface at a dose of 0.005 ug/mm-10 agent, the total amount of Silk delivered from an implant or tag/mm of Surface area coated. In one embodiment, bleo coated onto the surface of an implant should not exceed 100 mycin is released from the Surface of an implant Such that mg (range of 1 ug to 100 mg). In one embodiment, the total fibrosis in the tissue is promoted for a period ranging from amount of silk released from the prosthesis should be in the Several hours to Several months. range of 10 ug to 50 mg. The dose per unit area of the device 0621 Utilizing CTGF as an exemplary fibrosis-inducing (i.e., the dosage of silk as a function of the Surface area of agent, the total amount of CTGF delivered from an implant the portion of the device to which drug is applied and/or or coated onto the Surface of an implant should not exceed incorporated) should fall within the range of 0.05 ug-10 ug 100 mg (range of 0.01 ug to 100 mg). In one embodiment, per mm of Surface area coated. In another embodiment, silk the total amount of CTGF released from the implant should should be applied to an implant at a dose of 0.05ug/mm°-10 be in the range of 0.10 ug to 50 mg. The dose per unit area Aug/mm of Surface area coated. of the device (i.e., the dosage of CTGF as a function of the 0617 Utilizing chitosan as an exemplary fibrosis-induc Surface area of the portion of the device to which drug is ing agent, the total amount of chitosan delivered from an applied and/or incorporated) should fall within the range of implant or coated onto the Surface of an implant, should not 0.005 ug-10 ug per mm of Surface area coated. In another exceed 100 mg (range of 1 lug to 100 mg). In one embodi embodiment, CTGF should be applied to an implant surface ment, the total amount of chitosan released from the implant at a dose of 0.005 ug/mm-10 ug/mm of surface area should be in the range of 10 ug to 50 mg. The dose per unit coated. area of the device (i.e., the dosage of chitosan as a function 0622. The fibrosing agent (e.g., talc, Silk, chitosan, polyl of the Surface area of the portion of the device to which drug ysine, fibronectin, bleomycin, CTGF) may be released from is applied and/or incorporated) should fall within the range the Surface of the implant Such that fibrosis in the tissue is of 0.05 ug-10 ug per mm of Surface area coated. In another promoted for a period ranging from Several hours to Several embodiment, chitosan should be applied to an implant months. For example, the fibrosing agent may be released in Surface at a dose of 0.05 ug/mm-10 ug/mm of Surface area effective concentrations for a period ranging from 1 hour-30 coated. days. It should be readily evident given the discussions 0618. Utilizing polylysine as an exemplary fibrosis-in provided herein that analogues and derivatives of the fibroS ducing agent, the total amount polylysine delivered from an ing agent (e.g., analogues and derivatives of talc, silk, implant or coated onto the Surface of an implant should not chitosan, polylysine, fibronectin, bleomycin, CTGF, as pre exceed 100 mg (range of 1 lug to 100 mg). In one embodi viously described) with similar functional activity can be ment, the total amount of polylysine released from the utilized for the purposes of this invention; the above dosing implant should be in the range of 10 ug to 50 mg. The dose parameters are then adjusted according to the relative per unit area of the device (i.e., the dosage of polylysine as potency of the analogue or derivative as compared to the a function of the Surface area of the portion of the device to parent compound (e.g., a compound twice as potent as the which drug is applied and/or incorporated) should fall within agent is administered at half the above parameters, a com the range of 0.05 ug-10 ug per mm of Surface area coated. pound half as potent as the agent is administered at twice the In another embodiment, polylysine should be applied to an above parameters, etc.). implant surface at a dose of 0.05 ug/mm-10 ug/mm of 0623. As described above, the device may additionally Surface area coated. comprise an inflammatory cytokine (e.g., TGFB, PDGF, 0619 Utilizing fibronectin as an exemplary fibrosis-in VEGF, bFGF, TNFo, NGF, GM-CSF, IGF-a, IL-1, IL-1-?3, ducing agent, the total amount of fibronectin delivered from IL-8, IL-6, and growth hormone) and/or a bone morpho an implant or coated onto the Surface of an implant, should genic protein (BMP) (e.g., BMP-2, BMP-3, BMP-4, BMP-5, not exceed 100 mg (range of 1 lug to 100 mg). In one BMP-6, or BMP-7 or an analogue or derivative thereof). embodiment, the total amount of fibronectin released from 0624 Bone morphogenic protein(s) (e.g., BMP-2, BMP the prosthesis should be in the range of 10 ug to 50 mg. The 3, BMP-4, BMP-5, BMP-6, or BMP-7 or an analogue or dose per unit area of the device (i.e., the dosage of fibronec derivative thereof) are to be used in formulations at con tin as a function of the Surface area of the portion of the centrations that range from 0.001 ug/ml to approximately 20 device to which drug is applied and/or incorporated) should mg/ml depending on the Specific clinical application, for fall within the range of 0.05 ug-10 ug per mm of surface mulation type (e.g., gel, liquid, Solid, Semi-Solid), formula area coated. In another embodiment, talc should be applied tion chemistry, duration of required application, type of to an implant Surface at a dose of 0.05 ug/mm-10 ug/mm medical device interface and formulation Volume and or of Surface area coated. Surface area coverage required. Preferably, the bone mor 0620. Utilizing bleomycin as an exemplary fibrosis-in phogenic protein is released in effective concentrations for a ducing agent, the total amount of bleomycin delivered from period ranging from 1-180 days. The total dose for a single an implant, or coated onto the Surface of an implant, should application is typically not to exceed 500 mg (range of 0.001 not exceed 100 mg (range of 0.01 ug to 100 mg). In one Aug to 500 mg); preferred 1 lug to 250 mg. When used as a US 2005/0281883 A1 Dec. 22, 2005 73 device coating, the dose is per unit area of 0.001 ug-1000 ug the formation of post-operative Surgical adhesions, per mm’; with a preferred dose of 0.01 tug/mm-200 unwanted Scarring in the vicinity of the implant, and encap tug/mm. Minimum concentration of 10-10" M of bone Sulation of the implant by a fibrous connective tissue cap morphogenic protein is to be maintained on the device Sule. Surface. 0630. Examples of medical devices of particular utility 0625) Inflammatory cytokines are to be used in formula for use in combination with a fibrosis-inhibiting agent tions at concentrations that range from 0.0001 ug/ml to include, but are not restricted to vascular Stents, gastrointes approximately 20 mg/ml depending on the Specific clinical tinal Stents, tracheal/bronchial Stents, genital-urinary Stents, application, formulation type (e.g., gel, liquid, Solid, Semi ENT Stents, intraocular lenses, implants for hypertrophic Solid), formulation chemistry, duration of required applica Scars and keloids, vascular grafts, anastomotic connector tion, type of medical device interface and formulation Vol devices, Surgical adhesion barriers, glaucoma drainage ume and or Surface area coverage required. Preferably, the devices, film or mesh, prosthetic heart Valves, tympanos inflammatory cytokine is released in effective concentra tomy tubes, penile implants, endotracheal and tracheostomy tions for a period ranging from 1-180 days. The total dose tubes, peritoneal dialysis catheters, intracranial preSSure for a Single application is typically not to exceed 500 mg monitors, Vena cava filters, central venous catheters, Ven (range of 0.0001 ug to 100 mg); preferred 0.001 ug to 50 mg. tricular assist devices (e.g., LVAD’s), Spinal prostheses, and When used as a device coating, the dose is per unit area of gastrointestinal drainage tubes. 0.0001 ug-500 ug per mm’; with a preferred dose of 0.001 tug/mm-200 ug/mm. Minimum concentration of 10"-10-4 0631. In one aspect, the medical device may be an g/ml of inflammatory cytokine is to be maintained on the electrical device (e.g., a device having electrical components device Surface. that can be placed in contact with tissue in an animal host and can provide electrical excitation to nervous or muscular 0626 Furthermore, the device may alone or additionally tissue). Electrical devices can generate electrical impulses comprise an agent that Stimulates cellular proliferation. and may be used to treat many bodily dysfunctions and Examples include: dexamethasone, isotretinoin (13-cis ret disorders by blocking, masking, or Stimulating electrical inoic acid), 17-f-estradiol, estradiol, 1-O-25 dihydroxyvita Signals within the body. Electrical medical devices of par min D, diethylstibesterol, cyclosporine A, L-NAME, all ticular utility in the present invention include, but are not trans retinoic acid (ATRA), and analogues and derivatives restricted to, devices used in the treatment of cardiac rhythm thereof. Doses used are those concentrations which are abnormalities, pain relief, epilepsy, Parkinson's Disease, demonstrated to Stimulate cell proliferation. The prolifera movement disorders, obesity, depression, anxiety and hear tive agents are to be used in formulations at concentrations ing loSS. Other examples of electrical devices include neu that range from 0.1 ng/ml to 25 mg/ml depending on the rostimulator and neurostimulation devices (e.g., electrical Specific clinical application, formulation type, formulation devices for electrical excitation of the central, autonomic, or chemistry, duration of required application, type of medical peripheral nervous System), cardiac Stimulation device Such device interface and formulation volume and or Surface area as cardiac rhythm management devices, cardiac pacemak coverage required. Preferably, the proliferative agent is ers, implantable cardiac defibrillators (ICD) and other elec released in effective concentrations for a period ranging trical devices for electrical excitation of cardiac muscle from 1-180 days. The total dose for a single application is tissue (including the specialized cardiac muscle cells that typically not to exceed 500 mg (range of 0.0001 ug to 200 make up the conductive pathways of the heart). Electrical mg); preferred 0.001 ug to 100 mg. When used as a device devices also include electrical leads which are used as a coating, the dose is per unit area of 0.00001 ug-500 ug per conductor to carry electrical Signals from the generator to mm’; with a preferred dose of 0.0001 ug/mm-200 tug/mm . the tissues. The electrical lead may be a wire or other Minimum concentration of 10'-10 M of proliferative material that transmits electrical impulses from a generator agent is to be maintained on the device Surface. (e.g., pacemaker, defibrillator, or other neurostimulator). 0627 84. Medical Implants Combined with Fibrosis Electrical leads may be unipolar, in which they are adapted Inhibiting Agents to provide effective therapy with only one electrode. Multi polar leads are also available, including bipolar, tripolar and 0628. In another aspect, medical implants may be coated quadripolar leads. with, or otherwise adapted to release or incorporate an agent which inhibits the formation of reactive Scar tissue on, or 0632. In another aspect, the medical device may be an around, the Surface of the device or implant. Compositions implantable Sensor (i.e., a medical device that is implanted that include a fibrosis-inhibiting agent may be used in in the body to detect blood or tissue levels of a particular combination with a variety of medical implants to make chemical (e.g., glucose, electrolytes, drugs, hormones) and/ them resistant to overgrowth by inflammatory and fibrous or changes in body chemistry, metabolites, function, pres Scar tissue upon implantation. Compositions and methods Sure, flow, physical Structure, electrical activity or other are described for coating medical devices and implants with variable parameter). Representative examples of implant drug-delivery compositions Such that the pharmaceutical able Sensors include, blood/tissue glucose monitors, electro agent is delivered in therapeutic levels over a period Suffi lyte Sensors, blood constituent Sensors, temperature Sensors, cient to allow normal healing to occur. pH sensors, optical Sensors, amperometric Sensors, preSSure Sensors, biosensors, Sensing transponders, Strain Sensors, 0629. Upon implantation, excessive scar tissue growth activity Sensors and magnetoresistive Sensors. can occur around the all or parts of the implant, which can lead to a reduction in the performance of these devices. In 0633. In another aspect, the medical device may be a certain cases, an implanted device may be combined with a drug-delivery pump (i.e., a medical device that includes a therapeutic agent (e.g., an anti-fibrotic agent) to minimize pump which is configured to deliver a biologically active US 2005/0281883 A1 Dec. 22, 2005 74 agent (e.g., a drug) at a regulated dose). These devices are ing (A) anthracyclines (e.g., doxorubicin and mitoxantrone), implanted within the body and may include an external (B) taxanes (e.g., paclitaxel, TAXOTERE and docetaxel), transmitter for programming the controlled release of drug, and (C) podophyllotoxins (e.g., etoposide); (D) immuno or alternatively, may include an implantable Sensor that modulators (e.g., Sirolimus, everolimus, tacrolimus); (E) provides the trigger for the drug delivery pump to release heat shock protein 90 antagonists (e.g., geldanamycin), (F) drug as physiologically required. Drug-delivery pumps may HMGCoA reductase inhibitors (e.g., simvastatin); (G) be used to deliver virtually any agent, but specific examples inosine monophosphate dehydrogenase inhibitors (e.g., include insulin for the treatment of diabetes, medication for mycophenolic acid, 1-alpha-25 dihydroxy Vitamin D); the relief of pain, chemotherapy for the treatment of cancer, (H)NF kappa B inhibitors (e.g., Bay 11-7082); (I) antimy anti-Spastic agents for the treatment of movement and mus cotic agents (e.g., Sulconizole), (J) p38 MAP kinase inhibi cular disorders, or antibiotics for the treatment of infections. tors (e.g., SB202190), and (K) angiogenesis inhibitors (e.g., Representative examples of drug delivery pumps for use in halofuginone bromide), as well as analogues and derivatives the practice of the invention include, without limitation, of the aforementioned. constant flow drug delivery pumps, programmable drug delivery pumps, intrathecal pumps, implantable insulin 0638 Regardless of the method of application of the drug delivery pumps, implantable OSmotic pumps, ocular drug to the device, the exemplary anti-fibrosing agents, used alone or in combination, should be administered under the delivery pumpS and implants, metering Systems, peristaltic following dosing guidelines. The total amount (dose) of (roller) pumps, electronically driven pumps, elastomeric anti-Scarring agent in or on the device may be in the range pumps, Spring-contraction pumps, gas-driven pumps (e.g., of about 0.01 lug-10 ug, or 10 ug-10 mg, or 10 mg-250 mg, induced by electrolytic cell or chemical reaction), hydraulic or 250 mg-1000 mg, or 1000 mg-2500 mg. The dose pumps, piston-dependent pumps and non-piston-dependent (amount) of anti-Scarring agent per unit area of device pumps, dispensing chambers, infusion pumps, passive Surface to which the agent is applied may be in the range of pumps, infusate pumpS and osmotically-driven fluid dis about 0.01 ug/mm-1 g/mm, or 1 ug/mm-10 ug/mm, or penSerS. 10 ug/mm-2501 g/mm, 250 ug/mm-1000 ug/mm, or 0634. In yet another aspect, the medical device may be a 1000 tug/mm-2500 ug/mm°. Soft tissue implant. Soft tissue implants are medical devices that may includes a volume replacement material for aug 0639 AS medical implants are made in a variety of mentation or reconstruction to replace a whole or part of a configurations and sizes, the exact dose administered will living structure. Soft tissue implants are used for the recon vary with device size, Surface area and design; however, Struction of Surgically or traumatically created tissue voids, certain principles can be applied in the application of this art. augmentation of tissueS or organs, contouring of tissues, the Drug dose can be calculated as a function of dose per unit restoration of bulk to aging tissues, and to correct Soft tissue area (of the portion of the device being coated), total drug folds or wrinkles (rhytides). Soft tissue implants may be dose administered, and appropriate Surface concentrations of used for the augmentation of tissue for cosmetic (aesthetic) active drug can be determined. Drugs are to be used at enhancement or in association with reconstructive Surgery concentrations that range from Several times more than to following disease or Surgical resection. Representative 10%, 5%, or even less than 1% of the concentration typically examples of Soft tissue implants include breast implants, used in a single chemotherapeutic Systemic dose application. chin implants, calf implants, cheek implants and other facial Preferably, the drug is released in effective concentrations implants, buttockS implants, and nasal implants. for a period ranging from 1-90 days. 0635 Soft tissue implants that release a therapeutic agent 0640 Provided below are exemplary dosage ranges for for reducing Scarring at the implant-tissue interface can be various anti-Scarring agents that can be used in conjunction used to enhance the appearance, increase the longevity, with medical implants in accordance with the invention. A) reduce the need for corrective Surgery or repeat procedures, Cell cycle inhibitors including doxorubicin and mitox decrease the incidence of pain and other Symptoms, and antrone. Doxorubicin analogues and derivatives thereof: improve the clinical function of implant. Accordingly, the total dose not to exceed 25 mg (range of 0.1 ug to 25 mg); present invention provides Soft tissue implants that are preferred 1 lug to 5 mg. The dose per unit area of 0.01 ug-100 coated or otherwise incorporate an anti-Scarring agent or a Aug per mm; preferred dose of 0.1 ug/mm-10 ug/mm. composition that includes an anti-Scarring agent. Mitoxantrone and analogues and derivatives thereof: total dose not to exceed 5 mg (range of 0.01 ug to 5 mg); preferred 0636. According to the present invention, any fibrosis 0.1 lug to 1 mg. The dose per unit area of the device of 0.01 inhibiting agent described above can be utilized in the ug-20 ug per mm ; preferred dose of 0.05 ug/mm-3 practice of this embodiment. Within one embodiment of the Aug/mm. B) Cell cycle inhibitors including paclitaxel and invention, medical implants may be adapted to release an analogues and derivatives (e.g., docetaxel) thereof: total agent that inhibits one or more of the four general compo dose not to exceed 10 mg (range of 0.1 ug to 10 mg); nents of the process of fibrosis (or Scarring), including: preferred 1 lug to 3 mg. The dose per unit area of the device formation of new blood vessels (angiogenesis), migration of 0.1 lug-0 ug per mm’; preferred dose of 0.25 ug/mm°-5 and proliferation of connective tissue cells (Such as fibro Aug/mm. (C) Cell cycle inhibitors such as podophyllotoxins blasts or Smooth muscle cells), deposition of extracellular (e.g., etoposide): total dose not to exceed 10 mg (range of matrix (ECM), and remodeling (maturation and organization 0.1 ug to 10 mg); preferred 1 g to 3 mg. The dose per unit of the fibrous tissue). By inhibiting one or more of the area of the device of 0.1 lug-10 ug per mm; preferred dose components of fibrosis (or Scarring), the overgrowth of of 0.25 ug/mm-5 ug/mm. (D) Immunomodulators includ granulation tissue may be inhibited or reduced. ing Sirolimus and everolimus. Sirolimus (i.e., rapamycin, 0637. Several examples of agents for use with medical RAPAMUNE): Total dose not to exceed 10 mg (range of 0.1 implants include the following: cell cycle inhibitors includ Aug to 10 mg); preferred 10 ug to 1 mg. The dose per unit area US 2005/0281883 A1 Dec. 22, 2005 75 of 0.1 ug-100 ug per mm; preferred dose of 0.5 lug/mm-10 to 10 mg); preferred 10 ug to 1 mg. The dose per unit area tag/mm. Everolimus and derivatives and analogues thereof: of 0.1 ug-100 ug per mm of Surface area; preferred dose of Total dose should not exceed 10 mg (range of 0.1 ug to 10 0.3 ug/mm-10 ug/mm. (F) Pimecrolimus and derivatives mg); preferred 10 ug to 1 mg. The dose per unit area of 0.1 and analogues thereof: Total dose should not exceed 10 mg Aug-100 ug per mm of Surface area; preferred dose of 0.3 (range of 0.1 ug to 10 mg); preferred 10 ug to 1 mg. The dose tag/mm-10 ug/mm. (E) Heat shock protein 90 antagonists per unit area of 0.1 ug-100 ug per mm of Surface area; (e.g., geldanamycin) and analogues and derivatives thereof: preferred dose of 0.3 ug/mm-10 ug/mm. and (G) ABT-578 total dose not to exceed 20 mg (range of 0.1 ug to 20 mg); and analogues and derivatives thereof: Total dose should not preferred 1 lug to 5 mg. The dose per unit area of the device exceed 10 mg (range of 0.1 ug to 10 mg); preferred 10 ug of 0.1 ug-10 ug per mm; preferred dose of 0.25 ug/mm-5 to 1 mg. The dose per unit area of 0.1 ug-100 ug per mm ug/mm. (F) HMGCoA reductase inhibitors (e.g., simvasta of Surface area; preferred dose of 0.3 ug/mm-10 ug/mm. tin) and analogues and derivatives thereof: total dose not to 0642. In a general method for coating a Surface of a exceed 2000 mg (range of 10.0 ug to 2000 mg); preferred 10 Synthetic implant, the composition is exposed to the aqueous lug to 300 mg. The dose per unit area of the device of 1.0 jug-1000 ug per mm; preferred dose of 2.5 lug/mm-500 environment, and a thin layer of the composition is then tag/mm. (G) Inosine monophosphate dehydrogenase inhibi applied to a Surface of the implant before Substantial inter tors (e.g., mycophenolic acid, 1-alpha-25 dihydroxy Vita reaction has occurred. In one embodiment, in order to min D) and analogues and derivatives thereof: total dose minimize cellular and fibrous reaction to the coated implant, not to exceed 2000 mg (range of 10.0 ug to 2000 mg); the components are Selected So as to result in a matrix that preferred 10 ug to 300 mg. The dose per unit area of the has a net neutral charge. Application of the composition to device of 1.0 ug-1000 ug per mm; preferred dose of 2.5 the implant Surface may be by extrusion, brushing, spraying tug/mm-500 ug/mm. (H)NF kappa B inhibitors (e.g., Bay (as described above), or by any other convenient means. 11-7082) and analogues and derivatives thereof: total dose Following application of the composition to the implant not to exceed 200 mg (range of 1.0 ug to 200 mg); preferred Surface, inter-reaction is allowed to continue until complete 1 tug to 50 mg. The dose per unit area of the device of 1.0 and the three-dimensional matrix is formed. tug-100 ug per mm; preferred dose of 2.5 lug/mm-50 0643 Although this method can be used to coat the Aug/mm. (I) Antimycotic agents (e.g., Sulconizole) and ana Surface of any type of Synthetic implant, it is particularly logues and derivatives thereof: total dose not to exceed 2000 useful for implants where reduced thrombogenicity is an mg (range of 10.0 ug to 2000 mg); preferred 10 ug to 300 important consideration, Such as artificial blood vessels and mg. The dose per unit area of the device of 1.0 lug-1000 ug heart Valves, vascular grafts, vascular Stents, and Stent/graft per mm; preferred dose of 2.5 ug/mm-500 ug/mm. (J) p38 combinations. The method may also be used to coat implant MAPKinase Inhibitors (e.g., SB202190) and analogues and able Surgical membranes (e.g., monofilament polypropy derivatives thereof: total dose not to exceed 2000 mg (range lene) or meshes (e.g., for use in hernia repair). Breast of 10.0 ug to 2000 mg); preferred 10 ug to 300 mg. The dose implants may also be coated using the above method in order per unit area of the device of 1.0 ug-1000 ug per mm; to minimize capsular contracture. preferred dose of 2.5 ug/mm-500 ug/mm. (K) antiangio 0644. The compositions of the invention can also be genic agents (e.g., halofuginone bromide) and analogues and coated on a Suitable fibrous material, which can then be derivatives thereof: total dose not to exceed 10 mg (range of wrapped around a bone to provide Structural integrity to the 0.1 ug to 10 mg); preferred 1 lug to 3 mg. The dose per unit bone. The term "suitable fibrous material” as used herein, area of the device of 0.1 ug-10 ug per mm; preferred dose refers to a fibrous material which is substantially insoluble of 0.25 ug/mm-5 ug/mm. in water, non-immunogenic, biocompatible, and immiscible 0641. In addition to those described above (e.g., siroli with the crosslinkable compositions of the invention. The mus, everolimus, and tacrolimus), Several other examples of fibrous material may comprise any of a variety of materials immunomodulators and appropriate dosages ranges for use having these characteristics and may be combined with with medical implants include the following: (A) Biolimus crosslinkable compositions herein in order to form and/or and derivatives and analogues thereof: Total dose should not provide Structural integrity to various implants or devices exceed 10 mg (range of 0.1 ug to 10 mg); preferred 10 ug used in connection with medical and pharmaceutical uses. to 1 mg. The dose per unit area of 0.1 ug-100 ug per mm 0.645. The compositions of the present invention may also of Surface area; preferred dose of 0.3 ug/mm-10 ug/mm. be used to coat lenticules, which are made from either (B) Tresperimus and derivatives and analogues thereof: naturally occurring or Synthetic polymers. Total dose should not exceed 10 mg (range of 0.1 ug to 10 mg); preferred 10 ug to 1 mg. The dose per unit area of 0.1 0646 85. Treatment of Aneurysm tug-100 ug per mm of Surface area; preferred dose of 0.3 g/mm-10 ug/mm. (C) Auranofin and derivatives and ana 0647. The compositions can be extruded or molded in the logues thereof: Total dose should not exceed 10 mg (range shape of a String or coil, then dehydrated. The resulting of 0.1 ug to 10 mg); preferred 110 g to 1 mg. The dose per dehydrated String or coil can be delivered via catheter to the unit area of 0.1 ug-10g per mm of Surface area; preferred Site of a vascular malformation, Such as an aneurysm, for the dose of 0.3 ug/mm°-10 ug/mm . (D) 27-0-Demethylrapa purpose of vascular occlusion and, ultimately, repair of the mycin and derivatives and analogues thereof: Total dose malformation. The dehydrated string or coil can be delivered should not exceed 10 mg (range of 0.1 ug to 10 mg); in a compact size and will rehydrate inside the blood vessel, preferred 10 ug to 1 mg. The dose per unit area of 0.1 ug-100 Swelling Several times in size compared to its dehydrated Aug per mm of Surface area; preferred dose of 0.3 ug/mm-10 State, while maintaining its original shape. tag/mm. (E) Gusperimus and derivatives and analogues 0648. Thus, another embodiment of the invention is a thereof: Total dose should not exceed 10 mg (range of 0.1 ug method for treating an aneurysm, where steps (a) and (b) are US 2005/0281883 A1 Dec. 22, 2005 76 as described for the method of Sealing tissue, and step (c) A 40% (w/v) solution of that PEG powder was then prepared involves allowing a three-dimensional matrix to form in the by dissolving the powder in diluted HCl. The obtained desired shape, delivering it to the Site of interest, and Solution (pH =2.1) was then cosprayed with an equal volume allowing the matrix to rehydrate in Situ. of a 300 mM sodium phosphate/sodium carbonate buffer (pH 9.6). Gelation occurred almost immediately (<3 sec) 0649) 86. Other Uses and the gel obtained its firm, rubbery Solid properties in leSS 0650) As discussed in U.S. Pat. No. 5,752,974 to Rhee et than a minute. al., the compositions can be used to block or fill various lumens and Voids in the body of a mammalian Subject. The Example 2 compositions can also be used as bioSealants to Seal fissures or crevices within a tissue or structure (Such as a vessel), or Crosslinking Network from a Tetra Functional junctures between adjacent tissueS or structures, to prevent NHS-PEG and MC Bearing Amino Groups leakage of blood or other biological fluids. The compositions 0655 10% w/v of tetra functional NHS-PEG was mixed may also be used to Seal or close a fistula, where a Scar with a methylated collagen (“MC) bearing amino groups promoting agent or Sclerosing agent, e.g., Silk, may be and having a concentration of 22 mg/ml and pH 3-4 to form included in the composition to promote tissue closure. a homogeneous Stable acidic Solution. This Solution spon 0651. The compositions can also be used as a large taneously formed a three-dimensional matrix when mixed Space-filling device for organ displacement in a body cavity with 0.3 M phosphate/carbonate pH 9.6 solution. This basic during Surgical or radiation procedures, for example, to Solution activated the amines from MC which then reacted protect the intestines during a planned course of radiation to with the SG groups to form the amide bonds. The gelation the pelvis. occurred within Seconds to form a strong biomaterial that adhered well to the tissue and did not Swell after 24 hours of 0652 The compositions can also be coated onto the immersion in Saline Solution. interior Surface of a physiological lumen, Such as a blood vessel or Fallopian tube, thereby Serving as a Sealant to Example 3 prevent restenosis of the lumen following medical treatment, Such as, for example, balloon catheterization to remove Phase Separated Hydrogels of TA-PEG 912 with arterial plaque deposits from the interior Surface of a blood 3-SH vessel, or removal of Scar tissue or endometrial tissue from 0656 Phase separated hydrogels having 40 wt % water the interior of a Fallopian tube. A thin layer of the compo were prepared from of tri-functional acrylate-PEG (“TA Sition is preferably applied to the interior Surface of the PEG 912, 912 g/mol, Aldrich Chemicals, Milwaukee, vessel (for example, via catheter) immediately following Wis.)) and TP703-mercaptopropionate ("3-SH,” 711 g/mol, exposure to the aqueous environment. Because the compo Perstorp Polyols, Perstorp, Sweden). TA-PEG 912 (0.64 g) Sitions of the invention are not readily degradable in Vivo, was mixed with 3-SH (0.50g), to a 1:1 mole ratio in regards the potential for restenosis due to degradation of the coating to functional groups. A diluted acidic Solution (0.60 g) was is minimized. The use of a three-dimensional matrix having added to the mixture using a Vortex mixer, which formed a net neutral charge further minimizes the potential for latex. A basic aqueous buffer (0.16 g) (pH 9.6) was then restenosis. added, which caused the mixture to Solidify within a couple of seconds through the formation thiol-ether bonds. The EXPERIMENTAL formed hydrogel became quite firm in one minute or less. 0653. The following examples are put forth so as to provide those of ordinary skill in the art with a complete Example 4 disclosure and description of how to make and use the compounds of the invention, and are not intended to limit the Premixed Gelling Formulation (Premix) I Scope of what the inventors regard as their invention. Efforts 0657. A 1 ml syringe (syringe 1) equipped with a luer have been made to ensure accuracy with respect to numbers lock mixing connector was filled with a mixture of tetra (e.g., amounts, temperature, etc.) but Some errors and devia functional poly (ethylene glycol) Succinimidyl glutarate tions should be accounted for. Unless indicated otherwise, PEG-SG4 (50 mg) (Sunbio, Inc) and PEG-SH4 (tetra func parts are parts by weight, temperature is in C. and pressure tional poly (ethylene glycol) thiol) (50 mg) (Sunbio, Inc.) is at or near atmospheric. (referred to as “premix”). A 1 ml capped Syringe (Syringe 2) was filled with 0.25 ml of 6.3 mM HCl solution (pH 2.1). A Example 1 1 ml capped syringe (syringe 3) was filled with 0.25 ml 0.12 M monobasic sodium phosphate and 0.2 M sodium carbon Gelation of tetra Functional NHS-PEG, 10,000 ate (pH 9.7) buffer. The solid contents of syringe 1 and the mol. Wit. (NHS-PEG) with Tetra-Sulfhydryl PEG, acidic Solution of Syringe 2 were mixed through a mixing connector by repeatedly transferring the contents from one 10,000 mol. Wt (HS-PEG) Syringe to the other. After complete mixing, the entire 0654) A homogeneous mixture of pentaerythritol tetrakis mixture was pushed into one of the Syringes. The Syringe 1-(1'-oxo-5-Succimidylpentanoate)-2-poly(oxyethylene) containing the mixture then was attached to one inlet of an ether (“NHS-PEG,” 10,000 mol. wt., Aldrich Chemical Co., applicator (MICROMEDICS air assisted spray-applicator (Milwaukee, Wis.)) with (pentaerythritol tetrakis mercap (Model SA-6105)). Syringe 3 containing the pH 9.7 solution toethyl poly(oxyethylene) ether (“HS-PEG,” Aldrich was attached onto the other inlet of the applicator. The Chemical (Milwaukee, Wis.)) was obtained by mixing formulation was applied to a tissue Surface as Specified by approximately equal amounts of the two powders by weight. the applicator manufacturer. US 2005/0281883 A1 Dec. 22, 2005 77

Example 5 monobasic Sodium phosphate and 0.2 M Sodium carbonate (pH 9.7) buffer. The components were mixed and applied to Mycophenolic Acid in Premix a tissue Surface using the procedure described in Example 4. 0658) A 1 ml syringe (syringe 1) equipped with a luer Example 9 lock mixing connector was filled with a mixture of PEG SH4 (50 mg), PEG-SG4 (50 mg), and MPA (100 mg, Sifted.<100 micron). A 1 ml capped Syringe (syringe 2) was CPZ Loaded Microspheres in Premix filled with 0.25 ml of 6.3 mM HCl solution (pH 2.1). A 1 ml 0662. A 1 ml Syringe (Syringe 1) equipped with a luer capped syringe (syringe 3) was filled with 0.35 ml 0.24 M lock mixing connector was filled with a mixture of PEG monobasic Sodium phosphate and 0.4 M Sodium carbonate SG4 (50 mg), PEG-SH4 (50 mg), and 10% CPZ loaded (pH 10.0) buffer. The components were mixed and applied MePEG5000-PDLLA (65:35) microspheres prepared by to a tissue Surface using the procedure described in Example spray drying (50 or 100 mg) (prepared using the procedure 4. described in Example 11). A1 ml capped Syringe (Syringe 2) was filled with 0.25 ml of 6.3 mM HCl solution (pH 2.1). A Example 6 1 ml capped syringe (syringe 3) was filled 0.25 ml 0.12 M monobasic Sodium phosphate and 0.2 M Sodium carbonate Mycophenolic Acid and Disodium Salt of MPA (pH 9.7) buffer. The components were mixed and applied to (NA, MPA) in Premix a tissue Surface using the procedure described in Example 4. 0659 A 1 ml syringe (syringe 1) equipped with a luer Example 10 lock mixing connector was filled with a mixture of PEG SG4 (50mg) and PEG-SH4 (50 mg). A 1 ml capped syringe (syringe 2) was filled with 0.25 ml of 6.3 mM HCl solution MPA Loaded Microspheres in Premix (pH 2.1). A 1 ml capped Syringe (Syringe 3) was filled with 0663 A 1 ml syringe (syringe 1) equipped with a luer 0.25 ml 0.12 M monobasic sodium phosphate and 0.2 M lock mixing connector was filled with a mixture of PEG sodium carbonate (pH 9.7) buffer. A 1 ml syringe (syringe 4) SG4 (50 mg), PEG-SH4 (50 mg), and 10% MPA loaded equipped with luer-lock mixing connector was filled with MePEG5000-PDLLA65:35 microspheres prepared by spray MPA (5 mg) and Na MPA (95 mg), both sifted.<100 micron. drying (25 or 75 mg) (prepared using the procedure The contents of Syringe 4 and Syringe 2 were mixed through described in Example 11). A1 ml capped Syringe (Syringe 2) a mixing connector by repeatedly transferring the contents was filled with 0.25 ml 6.3 mM HCl solution (pH 2.1). A 1 from one Syringe to the other. This Solution was then used to ml capped syringe (syringe 3) was filled 0.35 ml 0.24 M reconstitute the Solids in Syringe 1. After complete mixing, monobasic Sodium phosphate and 0.4 M Sodium carbonate all of the formulation was pushed into one of the Syringes (pH 10.0) buffer. The components were mixed and applied which was then attached to one inlet of an applicator to a tissue Surface using the procedure described in Example (MICROMEDICS air assisted spray-applicator (Model 4. SA-6105)). Syringe 3 containing the pH 9.7 solution was attached onto the other inlet of the applicator. The formu Example 11 lation was applied to a tissue Surface as Specified by the applicator manufacturer. Preparation of Drug Loaded Microspheres by Spray Drying Example 7 0664 3.6 grams of methoxy poly(ethylene glycol 5000))- block-(poly (DL-lactide). (65:35 MePEG:PDLLA weight Chlorpromazine in Premix ratio) was dissolved in 200 ml methylene chloride. 400 mg 0660 A 1 ml syringe (syringe 1) equipped with a luer of a drug (mycophenolic acid (MPA), chlorpromazine (CPZ) lock mixing connector was filled with a mixture of PEG or paclitaxel (PTX)) was added and the resulting solution SG4 (50 mg), PEG-SH4 (50 mg), and CPZ (5 or 10 mg). A was spray dried (Buchi spray drier model B 191). Inlet 1 ml capped syringe (syringe 2) was filled with 0.25 ml of temperature 50 C., outlet temperature.<39 C., aspirator 6.3 mM HCl solution (pH 2.1). A 1 ml capped syringe 100%, flow rate 700 l/hr. The collected microspheres were (syringe 3) was filled 0.25 ml 0.12 M monobasic sodium dried under vacuum at room temperature overnight to pro phosphate and 0.2 M sodium carbonate (pH 9.7) buffer. The duce uniform, Spherical particles having Size ranges of leSS components were mixed and applied to a tissue Surface than about 10 microns (typically about 0.5 to about 2 using the procedure described in Example 4. microns). Example 8 Example 12 Paclitaxel Loaded Microspheres in Premix MPA Loaded Microspheres (<10 Micron) by the 0661. A 1 ml Syringe (Syringe 1) equipped with a luer W/O/W Emulsion Process lock mixing connector was filled with a mixture of PEG 0665 100 ml of freshly prepared 10% polyvinyl alcohol SG4 (50 mg), PEG-SH4 (50 mg), and 10% PTX loaded (PVA) solution and 10 ml of pH 3 acetic acid solution MePEG5000-PDLLA (65:35) microspheres prepared by Saturated with MPA was added into a 600 ml beaker. The spray drying (0.5 or 2 mg) (prepared using the procedure acidified PVA Solution was stirred at 2000 rpm for 30 described in Example 11. A 1 ml capped Syringe (Syringe 2) minutes. Meanwhile, a solution of 400 mg MPA and 800 mg was filled with 0.25 ml of 6.3 mM HCl solution (pH 2.1). A MePEG5000-PDLLA(65:35) in 20 ml dichloromethane was 1 ml capped syringe (syringe 3) was filled 0.25 ml 0.12 M prepared. The polymer/dichloromethane Solution was added US 2005/0281883 A1 Dec. 22, 2005 78 dropwise to the PVA solution while stirring at 2000 rpm with contents from one Syringe to the other. After complete a Fisher DYNA-MIX stirrer. After addition was complete, mixing, the entire mixture was pushed into one of the the Solution was allowed to stir for an additional 45 minutes. Syringes, which was then attached to one inlet of an appli The microSphere Solution was transferred to Several dispos cator (MICROMEDICS air assisted spray-applicator (Model able graduated polypropylene conical centrifuge tubes, SA-6105)). Syringe 3 containing the pH 9.7 solution was washed with pH 3 acetic acid solution saturated with MPA, attached onto the other inlet of the applicator. The formu and centrifuged at 2600 rpm for 10 minutes. The aqueous lation was applied to a tissue Surface as Specified by the layer was decanted and the Washing, centrifuging and applicator manufacturer. decanting was repeated 3 times. The combined, washed microSpheres were freeze-dried and vacuum dried to remove Example 17 any excess water. MPA Loaded Microspheres in Premix Example 13 0670) A 1 ml syringe (syringe 1) equipped with a luer lock mixing connector was filled with a mixture of PEG MPA Containing Microspheres (50-100 Micron) by SG4 (50 mg), PEG-SH4 (50 mg), and 10% PTX loaded the W/O/W Emulsion Process MePEG5000-PDLLA (65:35) microspheres (0.5 or 2 mg) 0666 Microspheres having an average size of about (prepared using the procedure described in Example 12 and 50-100 microns were prepared using a 1% PVA Solution and 13). A 1 ml capped syringe (syringe 2) was filled with 0.25 500 rpm Stirring rate using the same procedure described in ml of 6.3 mM HCl solution (pH 2.1). A 1 ml capped syringe Example 12. (syringe 3) was filled 0.25 ml 0.12 M monobasic sodium phosphate and 0.2 M sodium carbonate (pH 9.7) buffer. The Example 14 components were mixed and applied to a tissue Surface using the procedure described in Example 4. CPZ and PTX Containing Microspheres by the W/O/W Emulsion Process Example 18 0667 Paclitaxel (PTX) and chlorpromazine (CPZ) con taining microSpheres were prepared using the procedure Gelling Formulation (Premix) II described in Example 12 with the exception that the PVA 0671 A3 ml syringe (syringe 1) equipped with a luer Solution and the washing Solution does not have to be lock mixing connector was filled with a mixture of PEG acidified and Saturated with the drug. PTX and CPZ loaded SG4 (200 mg) and PEG-SH4 (200 mg). A 3 ml capped microSpheres were incorporated in premix as described in syringe (syringe 2) was filled with 1.0 ml of 6.3 mM HCl Example 17. Solution (pH 2.1). A 3 ml capped Syringe (Syringe 3) was filled 1 ml 0.12 M monobasic sodium phosphate and 0.2 M Example 15 sodium carbonate (pH 9.7) buffer. The components were mixed and applied to a tissue Surface using the procedure Paclitaxel Containing Micelles described in Example 4. 0668 MePEG2000 (41 g) and MePEG2000-PDLLA (60:40) (410 g) were combined in a vessel and heated to 75 Example 19 C. with Stirring. After the polymers were completely melted and mixed, the temperature was decreased to 55° C. Mean MPA Loaded Premix while, a PTX solution in tetrahydrofuran (46 g/200 ml) was 0672 A3 ml syringe (syringe 1) equipped with a luer prepared and poured into the polymer Solution under con lock mixing connector was filled with a mixture of PEG Stant Stirring. Stirring was continued for and additional hour. SG4 (200 mg), PEG-SH4 (200 mg), and MPA (200 mg or The PTX containing micelles were dried at 50 C. under 400 mg). A3 ml capped Syringe (Syringe 2) was filled with Vacuum to remove Solvent and were ground on a 2 mm mesh 1 ml of 6.3 mM HCl solution (pH 2.1). A 3 ml capped Screen after cooling. syringe (syringe 3) was filled 1.5 ml 0.24 M monobasic sodium phosphate and 0.4 M sodium carbonate (pH 10) Example 16 buffer. The components were mixed and applied to a tissue Surface using the procedure described in Example 4. Incorporation of PTX Loaded Micelles into Premix 0669 A 1 ml syringe (syringe 1) equipped with a luer Example 20 lock mixing connector was filled with a mixture of PEG Surgical Adhesions Model to Assess Fibrosis SG4 (50mg) and PEG-SH4 (50 mg). A 2 ml serum vial was Inhibiting Agents in Rats filled with 1.5 ml of 6.3 mM HCl solution (pH 2.1). A 1 ml capped syringe (syringe 2) was filled with 0.25 ml 0.12 M 0673. The rat caecal sidewall model is used to as to assess monobasic Sodium phosphate and 0.2 M Sodium carbonate the anti-fibrotic capacity of formulations in Vivo. Sprague (pH 9.7) buffer. A 2 ml serum vial was filled with 10% PTX Dawley rats are anesthetized with halothane. Using aseptic loaded micelles (2 mg or 8 mg) (prepared as in Example 15) precautions, the abdomen is opened via a midline incision. and reconstituted with 1 ml of the pH 2.1 solution. 0.25 ml The caecum is exposed and lifted out of the abdominal of the micelle Solution was removed with a 1 ml Syringe; the cavity. Dorsal and Ventral aspects of the caecum are Suc Syringe was attached to Syringe 1 containing the Solids cessively scraped a total of 45 times over the terminal 1.5 cm PEG-SG4 and PEG-SH4; and the components were mixed using a #10 Scalpel blade. Blade angle and pressure are through the mixing connector by repeatedly transferring the controlled to produce punctate bleeding while avoiding US 2005/0281883 A1 Dec. 22, 2005 79

Severe tissue damage. The left Side of the abdomen is Formulations with various concentrations of antifibrotic retracted and everted to expose a Section of the peritoneal drugs are tested against control animals to assess inhibition wall that lies proximal to the caecum. The Superficial layer of adhesion formation. of muscle (transverses abdominis) is excised over an area of 1x2 cm, leaving behind torn fibers from the second layer of 0678 Rabbits are anesthetized with an IM injection of muscle (internal oblique muscle). Abraded Surfaces are ketamine/Zylazine. An endotracheal tube is inserted for tamponaded until bleeding Stops. The abraded caecum is maintenance of anesthesia with halothane. The animal is then positioned over the sidewall wound and attached by placed prone on the operating table on top of a heating pad two sutures. The formulation is applied over both sides of and the skin over the lower half of the back is shaved and the abraded caecum and over the abraded peritoneal Side prepared for Sterile Surgery. A longitudinal midline skin wall. A further two Sutures are placed to attach the caecum incision is made from L-1 to L-5 and down the lumbosacral to the injured sidewall by a total of 4 sutures and the fascia. The fascia is incised to expose the tips of the Spinous abdominal incision is closed in two layers. After 7 dayS, processes. The paraspinous muscles are dissected and animals are evaluated post mortem with the extent and retracted from the Spinous process and lamina of L-4. A Severity of adhesions being Scored both quantitatively and laminectomy is performed at L-4 by removal of the Spinal qualitatively. process with careful bilateral excision of the laminae, thus creating a Small 5x10 mm laminectomy defect. Hemostasis Example 21 is obtained with Gelfoam. The test formulations are applied to the injury Site and the wound is closed in layers with Surgical Adhesions Model to Assess Fibrosis Vicryl Sutures. The animals are placed in an incubator until Inhibiting Agents in Rabbits recovery from anesthesia and then returned to their cage. 0674) The rabbit uterine horn model is used to assess the 0679. Two weeks after Surgery, the animals are anesthe anti-fibrotic capacity of formulations in vivo. Mature New tized using procedures Similar to those described above. The Zealand White (NZW) female rabbits are placed under animals are euthanized with Euthanyl. After a skin incision, general anesthetic. Using aseptic precautions, the abdomen the laminectomy site is analyzed by dissection and the is opened in two layers at the midline to expose the uterus. amount of adhesion is Scored using Scoring Systems pub Both uterine horns are lifted out of the abdominal cavity and lished in the scientific literature for this type of injury. assessed for size on the French Scale of catheters. Horns between #8 and #14 on the French Scale (2.5-4.5 mm Example 23 diameter) are deemed suitable for this model. Both uterine horns and the opposing peritoneal wall are abraded with a Tendon Surgical Adhesions Model to Assess #10 scalpel blade at a 45° angle over an area 2.5 cm in length Fibrosis Inhibiting Agents in Rabbits and 0.4 cm in width until punctuate bleeding is observed. Abraded Surfaces are tamponaded until bleeding Stops. The 0680 This model is used to investigate whether adhesion individual horns are then opposed to the peritoneal wall and of the tendons can be prevented by local slow release of Secured by two Sutures placed 2 mm beyond the edges of the drugs known to inhibit fibrosis. Polymeric formulations are abraded area. The formulation is applied and the abdomen is loaded with drugs and implanted around injured tendons in closed in three layers. After 14 days, animals are evaluated rabbits. In animals without fibrosis-inhibiting formulations, post mortem with the extent and Severity of adhesions being adhesions develop within 3 weeks of flexor tendon injury if Scored both quantitatively and qualitatively. immobilization of the tendon is maintained during that period. An advantage of rabbits is that their tendon anatomy Example 22 and cellular behaviour during tendon healing are similar to those in man except for the rate of healing that is much faster Spinal Surgical Adhesions Model to Assess Fibrosis in rabbits. Inhibiting Agents in Rabbits 0681 Rabbits are anesthetized and the skin over the right 0675 Extensive scar formation and adhesions often hindlimb is shaved and prepared for Sterile Surgery. Sterile occur after lumbar Spine Surgery involving the vertebrae. Surgery is performed aided by an operating microScope. A The dense and thick fibrous tissue adherent to the Spine and longitudinal midline skin incision is made on the Volvar adjacent muscles must be removed by Surgery. Unfortu aspect of the proximal phalange in digits 2 and 4. The nately, fibrous adhesions usually reform after the Secondary Synovial sheath of the tendons is carefully exposed and Surgery. Adhesions are formed by proliferation and migra incised transversally to access the flexor digitorum profun tion of fibroblasts from the surrounding tissue at the site of duS distal to the flexor digitorum Superficialis bifurcation. Surgery. These cells are responsible for the healing response Tendon injury is performed by gently lifting the flexor after tissue injury. Once they have migrated to the wound digitorum profundus with curved forceps and incising trans they lay down proteins Such as collagen to repair the injured versally through half of its substance. The formulation tissue. Overproliferation and Secretion by these cells induce containing the test drug formulation is applied around the local obstruction, compression and contraction of the Sur tendons in the sheath of one of the two digits randomly rounding tissues with accompanying Side effects. Selected. The other digit is left untreated and is used as a control. The sheath is then repaired with 6-0 nylon Suture. 0676 The rabbit laminectomy spinal adhesion model An immobilizing 6-0 nylon Suture is inserted through the described herein is used to investigate Spinal adhesion transverse metacarpal ligament into the tendon/sheath com prevention by local slow release of antifibrotic drugs. plex to immobilize the tendon and the Sheath as a Single unit 0677 Five to six animals are included in each experi to encourage adhesion formation. The wound is closed with mental group to allow for meaningful Statistical analysis. 4-0 interrupted Sutures. A bandage is applied around the US 2005/0281883 A1 Dec. 22, 2005

hindpaw to further augment immobilization of the digits and 6. The composition of claim 5, wherein the amine ensure comfort and ambulation of the animals. The animals reactive groups contain an electrophilically reactive carbo are recovered and returned to their cage. nyl group Susceptible to nucleophilic attack by a primary or 0682. Three weeks after surgery, the animals are anes Secondary amine. thetized. After a skin incision, the tissue plane around the 7. The composition of claim 5, wherein the amine Synovial sheath is dissected and the tendon-sheath complex reactive groups are Selected from carboxylic acid esters, acid harvested en block and transferred in 10% phosphate buff chloride groups, anhydrides, ketones, aldehydes, halo, iso ered formaldehyde for histopathology analysis. The animals cyanato, thioisocyanato, epoxides, activated hydroxyl are then euthanized. After paraffin embedding, Serial 5-um groups, olefins, carboxyl, Succinimidyl ester, SulfoSuccinim thin cross-sections are cut every 2 mm through the sheath idyl ester, maleimido, epoxy, and etheneSulfonyl. and tendon complex. Sections are Stained with H&E and 8. The composition of claim 1, wherein the nucleophilic Movat's Stains to evaluate adhesion growth. Each Slide is groups are Sulfhydryl groupS and the electrophilic groups are digitized using a computer connected to a digital microscope Sulfhydryl-reactive groups. camera (Nikon Micropublisher cooled camera). Morphom 9. The composition of claim 8, wherein the sulfhydryl etry analysis is then performed using image analysis Soft reactive groups are Selected So as to form a thioester, ware (ImagePro). Thickness and area of adhesion defined as imido-thioester, thioether, or disulfide linkage upon reaction the Substance obliterating the Synovial Space are measured with the sulfhydryl groups. and compared between formulation-treated and control ani 10. The composition of claim 9, wherein the sulfhydryl mals. reactive groups form a disulfide linkage and have the structure -S-S-Ar where Ar is a Substituted or unsub 0683. It is to be understood that while the invention has Stituted nitrogen-containing heteroaromatic moiety or a non been described in conjunction with the preferred specific heterocyclic aromatic group Substituted with an electron embodiments thereof, the foregoing description, as well as withdrawing moiety. the examples that are presented above, are intended to illustrate and not limit the scope of the invention. Other 11. The composition of claim 9, wherein the sulfhydryl aspects, advantages and modifications will be apparent to reactive groups form a thioether linkage and are Selected those skilled in the art to which the invention pertains. All from maleimido, Substituted maleimido, haloalkyl, epoxy, patents, patent applications, journal articles, and other ref imino, aziridino, olefins, and C.f3-unsaturated aldehydes and erences cited herein are incorporated by reference in their ketones. entireties. 12. The composition of claim 8, wherein the sulfhydryl reactive groups are Selected from mixed anhydrides, ester We claim: derivatives of phosphorus, ester derivatives of p-nitrophe 1. A dry powder composition comprised of nol, p-nitrothiophenol and pentafluorophenol; esters of Sub Stituted hydroxylamines, including N-hydroxyphthalimide a first component having a core Substituted with m esters, N-hydroxySuccinimide esters, N-hydroxySulfoSuc nucleophilic groups, where me2; and cinimide esters, and N-hydroxyglutarimide esters, esters of a Second component having a core Substituted with n 1-hydroxybenzotriazole, 3-hydroxy-3,4-dihydro-benzotri electrophilic groups, where ne2 and m+n>4, aZin-4-one, 3-hydroxy-3,4-dihydro-quinazoline-4-One, car bonylimidazole derivatives, acid chlorides, ketenes, and wherein the nucleophilic and electrophilic groups are isocyanates. non-reactive in a dry environment but are rendered 13. The composition of claim 1, wherein the number of reactive upon exposure to an aqueous environment nucleophilic groups in the mixture is approximately equal to Such that the components inter-react in the aqueous the number of electrophilic groups in the mixture. environment to form a three-dimensional composition. 2. The composition of claim 1, wherein the nucleophilic 14. The composition of claim 13, wherein the ratio of and electrophilic groupS undergo a nucleophilic Substitution moles of nucleophilic groups to moles of electrophilic reaction, a nucleophilic addition reaction, or both. groups is about 2:1 to 1:2. 3. The composition of claim 1, wherein the nucleophilic 15. The composition of claim 14, wherein the ratio is 1:1. groups are selected from -NH, -NHR', -N(R'), -SH, 16. The composition of claim 1, wherein the core is -OH, -COOH, -CH-OH, -H, -PH, -PHR', Selected from hydrophilic polymers, hydrophobic polymers, P(R), -NH-NH,-CO-NH-NH2, and -CHN, amphiphilic polymers, C-1 hydrocarbyls, and heteroatom where R is a hydrocarbyl group, and each R' may be the containing C2 hydrocarbyls. Same or different. 17. The composition of claim 16, wherein the core is a 4. The composition of claim 1, wherein the electrophilic Synthetic or naturally occurring hydrophilic polymer. groups are selected from -CO-Cl, -(CO)-O- 18. The composition of claim 17, wherein the hydrophilic (CO)-R (where R is an alkyl group), -CH=CH-CH=O polymer is a linear, branched, dendrimeric, hyperbranched, and -CH=CH-C(CH)=O, halo, -N=C=O, or Star polymer. -N=C=S, -SOCH=CH, -O(CO)-C=CH, 19. The composition d of claim 17, wherein the hydro -O(CO)C(CH)=CH, -S-S-(CHN), -O(CO)- philic polymer is Selected from polyalkylene oxides, poly C(CH2CH)=CH, CH=CH-C=NH, COOH, ols; poly(oxyalkylene)-Substituted diols and polyols; poly -(CO)O-N(COCH), -CHO, -(CO)O- oxyethylated Sorbitol; polyoxyethylated glucose, N(COCH-)-S(O).OH, and -NCCOCH). poly(acrylic acids) and analogs and copolymers thereof; 5. The composition of claim 1, wherein the nucleophilic polymaleic acids; polyacrylamides; poly(olefinic alcohols); groups are amino groups and the electrophilic groups are poly(N-vinyl lactams); polyoxazolines; polyvinylamines; amine-reactive groups. and copolymers thereof. US 2005/0281883 A1 Dec. 22, 2005

20. The composition of claim 19, wherein the hydrophilic electrophilic group; L and L are linking groups; and polymer is a polyalkylene oxide or polyols Selected from p and q are integers from 0-1. polyethylene glycol and poly(ethylene oxide)-poly(propy 35. The composition of claim 1, wherein the components lene oxide) copolymers. inter-react to form covalent bonds, noncovalent bonds, or 21. The composition of claim 19, wherein the hydrophilic both. polymer is a polyol Selected from glycerol, polyglycerol and 36. The composition of claim 35, wherein the noncovalent propylene glycol. bonds are ionic bonds, hydrogen bonds, or the association of 22. The composition of claim 19, wherein the hydrophilic hydrophobic molecular Segments. polymer is a poly(oxyalkylene)-substituted polyol selected 37. The composition of claim 36, wherein all of the from mono-, di- and tri-polyoxyethylated glycerol, mono molecular segments are the same. and di-polyoxyethylated propylene glycol, and mono- and 38. The composition of claim 1, further comprising a di-polyoxyethylated trimethylene glycol. biologically active agent. 23. The composition of claim 19, wherein the hydrophilic 39. The composition of claim 38, further comprising a polymer is a poly(acrylic acid), analog or copolymer thereof pharmaceutically acceptable carrier. Selected from poly(acrylic acid), poly(methacrylic acid), 40. The composition of claim 39, wherein the pharma poly(hydroxyethylmethacrylate), poly(hydroxyethylacry ceutically acceptable carrier is a micelle. late), poly(methylalkylsulfoxide acrylates), and poly(methy 41. The composition of claim 39, wherein the pharma lalkylsulfoxide methacrylates). ceutically acceptable carrier is a microSphere. 24. The composition of claim 19, wherein the hydrophilic 42. The composition of claim 41, wherein the micro polymer is a polyacrylamide Selected from polyacrylamide, Sphere comprises a degradable polymer. poly(methacrylamide), poly(dimethylacrylamide), poly(N- 43. The composition of claim 42, wherein the degradable isopropylacrylamide), and copolymers thereof. polymer is a polyester. 25. The composition of claim 19, wherein the hydrophilic 44. The composition of claim 43, wherein the polyester is polymer is a poly(olefinic alcohol) selected from poly(Vinyl a glycolide/lactide copolymer. alcohols) and copolymers thereof. 45. The composition of claim 42, wherein the degradable 26. The composition of claim 19, wherein the hydrophilic polymer comprises residues of one or more monomers Selected from the group consisting of lactide, lactic acid, polymer is a poly(N-vinyl lactam) Selected from poly(Vinyl glycolide, glycolic acid, e-caprolactone, gamma-caprolac pyrrolidones), poly(Vinyl caprolactams), and copolymers tone, hydroxyvaleric acid, hydroxybutyric acid, beta-buty thereof. rolactone, gamma-butyrolactone, gamma-Valerolactone, 27. The composition of claim 19, wherein the hydrophilic y-decanolactone, Ödecanolactone, trimethylene carbonate, polymer is a polyoxazoline Selected from poly(methylox 1,4-dioxane-2-one or 1,5-dioxepan-2one.). azoline) and poly(ethyloxazoline). 46. The composition of claim 39, wherein the pharma 28. The composition of claim 17, wherein the hydrophilic ceutically acceptable carrier is a nanosphere. polymer is Selected from proteins, carboxylated polysaccha 47. The composition of claim 46, wherein the nanosphere rides, aminated polysaccharides, and activated polysaccha comprises a degradable polymer. rides. 48. The composition of claim 47, wherein the degradable 29. The composition of claim 28, wherein the hydrophilic polymer is a polyester. polymer is Selected from collagen and glycosaminoglycans. 49. The composition of claim 48, wherein the polyester is 30. The composition of claim 16, wherein the core is a a glycolide/lactide copolymer. hydrophobic polymer Selected from polylactic acid and 50. The composition of claim 47, wherein the degradable polyglycolic acid. polymer comprises residues of one or more monomers 31. The composition of claim 16, wherein the core is an Selected from the group consisting of lactide, lactic acid, amphiphilic polymer. glycolide, glycolic acid, e-caprolactone, gamma-caprolac 32. The composition of claim 16, wherein the core is a tone, hydroxyvaleric acid, hydroxybutyric acid, beta-buty

C.2-14 hydrocarbyl Selected from alkanes, diols, polyols, and rolactone, gamma-butyrolactone, gamma-Valerolactone, polyacids. y-decanolactone, Ödecanolactone, trimethylene carbonate, 33. The composition of claim 16, wherein the core is a 1,4-dioxane-2-one or 1,5-dioxepan-2one.). heteroatom-containing C-1 hydrocarbyl selected from di 51. The composition of claim 38, wherein the biologically and poly-electrophiles. active agent is an anti-fibrotic agent. 34. The composition of claim 1, 52. The composition of claim 51, wherein the anti-fibrotic agent inhibits cell regeneration. wherein the first component has the Structure of formula 53. The composition of claim 51, wherein the anti-fibrotic (I) agent inhibits angiogenesis. X(L)l-R (I) 54. The composition of claim 51, wherein the anti-fibrotic and the Second component has the Structure of formula agent inhibits fibroblast migration. 55. The composition of claim 51, wherein the anti-fibrotic (II) agent inhibits fibroblast proliferation. Y-(L)-R', (II) 56. The composition of claim 51, wherein the anti-fibrotic wherein m and n are integers from 2-12 and m+n>4, R agent inhibits deposition of extracellular matrix. and R' are independently selected from hydrophilic 57. The composition of claim 51, wherein the anti-fibrotic polymers, hydrophobic polymers, amphiphilic poly agent inhibits tissue remodeling. mers, C. hydrocarbyls, and heteroatom-containing 58. The composition of claim 51, wherein the anti-fibrotic C. hydrocarbyls, X is a nucleophilic group; Y is an agent is an angiogenesis inhibitor. US 2005/0281883 A1 Dec. 22, 2005 82

59. The composition of claim 51, wherein the anti-fibrotic 90. The composition of claim 51, wherein the anti-fibrotic agent is a 5-lipoxygenase inhibitor or antagonist. agent is a topoisomerase inhibitor. 60. The composition of claim 51, wherein the anti-fibrotic 91. The composition of claim 51, wherein the anti-fibrotic agent is a chemokine receptor antagonist. agent is a DNA cleaving agent. 61. The composition of claim 51, wherein the anti-fibrotic 92. The composition of claim 51, wherein the anti-fibrotic agent is a cell cycle inhibitor. agent is an antimetabolite. 62. The composition of claim 51, wherein the anti-fibrotic 93. The composition of claim 51, wherein the anti-fibrotic agent is a taxane. agent inhibits adenosine deaminase. 63. The composition of claim 51, wherein the anti-fibrotic 94. The composition of claim 51, wherein the anti-fibrotic agent is an anti-microtubule agent. agent inhibits purine ring Synthesis. 64. The composition of claim 51, wherein the anti-fibrotic 95. The composition of claim 51, wherein the anti-fibrotic agent is paclitaxel. agent is a nucleotide interconversion inhibitor. 65. The composition of claim 51, wherein the anti-fibrotic 96. The composition of claim 51, wherein the anti-fibrotic agent is an analogue or derivative of paclitaxel. agent inhibits dihydrofolate reduction. 66. The composition of claim 51, wherein the anti-fibrotic 97. The composition of claim 51, wherein the anti-fibrotic agent is a Vinca alkaloid. agent blocks thymidine monophosphate. 67. The composition of claim 51, wherein the anti-fibrotic 98. The composition of claim 51, wherein the anti-fibrotic agent is camptothecin or an analogue or derivative thereof. agent causes DNA damage. 68. The composition of claim 51, wherein the anti-fibrotic agent is a podophyllotoxin. 99. The composition of claim 51, wherein the anti-fibrotic 69. The composition of claim 68, wherein the podophyl agent is a DNA intercalation agent. lotoxin is etoposide or an analogue or derivative thereof. 100. The composition of claim 51, wherein the anti 70. The composition of claim 51, wherein the anti-fibrotic fibrotic agent is a RNA synthesis inhibitor. agent is an anthracycline. 101. The composition of claim 51, wherein the anti 71. The composition of claim 70, wherein the anthracy fibrotic agent is a pyrimidine Synthesis inhibitor. cline is doxorubicin or an analogue or derivative thereof. 102. The composition of claim 51, wherein the anti 72. The composition of claim 70, wherein the anthracy fibrotic agent inhibits ribonucleotide Synthesis or function. cline is mitoxantrone or an analogue or derivative thereof. 103. The composition of claim 51, wherein the anti 73. The composition of claim 51, wherein the anti-fibrotic fibrotic agent inhibits thymidine monophosphate Synthesis agent is a platinum compound. or function. 74. The composition of claim 51, wherein the anti-fibrotic 104. The composition of claim 51, wherein the anti agent is a nitroSourea. fibrotic agent inhibits DNA synthesis. 75. The composition of claim 51, wherein the anti-fibrotic 105. The composition of claim 51, wherein the anti agent is a nitroimidazole. fibrotic agent causes DNA adduct formation. 76. The composition of claim 51, wherein the anti-fibrotic 106. The composition of claim 51, wherein the anti agent is a folic acid antagonist. fibrotic agent inhibits protein Synthesis. 77. The composition of claim 51, wherein the anti-fibrotic 107. The composition of claim 51, wherein the anti agent is a cytidine analogue. fibrotic agent inhibits microtubule function. 78. The composition of claim 51, wherein the anti-fibrotic 108. The composition of claim 51, wherein the anti agent is a pyrimidine analogue. fibrotic agent is a cyclin dependent protein kinase inhibitor. 79. The composition of claim 51, wherein the anti-fibrotic 109. The composition of claim 51, wherein the anti agent is a fluoropyrimidine analogue. fibrotic agent is an epidermal growth factor kinase inhibitor. 80. The composition of claim 51, wherein the anti-fibrotic 110. The composition of claim 51, wherein the anti agent is a purine analogue. fibrotic agent is an elastase inhibitor. 81. The composition of claim 51, wherein the anti-fibrotic 111. The composition of claim 51, wherein the anti agent is a nitrogen mustard or an analogue or derivative fibrotic agent is a factor Xa inhibitor. thereof. 112. The composition of claim 51, wherein the anti 82. The composition of claim 51, wherein the anti-fibrotic fibrotic agent is a farnesyltransferase inhibitor. agent is a hydroxyurea. 113. The composition of claim 51, wherein the anti 83. The composition of claim 51, wherein the anti-fibrotic fibrotic agent is a fibrinogen antagonist. agent is a mytomicin or an analogue or derivative thereof. 114. The composition of claim 51, wherein the anti 84. The composition of claim 51, wherein the anti-fibrotic fibrotic agent is a guanylate cyclase Stimulant. agent is an alkyl Sulfonate. 115. The composition of claim 51, wherein the anti 85. The composition of claim 51, wherein the anti-fibrotic fibrotic agent is a heat shock protein 90 antagonist. agent is a benzamide or an analogue or derivative thereof. 116. The composition of claim 115, wherein the heat 86. The composition of claim 51, wherein the anti-fibrotic Shock protein 90 antagonist is geldanamycin or an analogue agent is a nicotinamide or an analogue or derivative thereof. or derivative thereof. 87. The composition of claim 51, wherein the anti-fibrotic 117. The composition of claim 51, wherein the anti agent is a halogenated Sugar or an analogue or derivative fibrotic agent is a guanylate cyclase Stimulant. thereof. 118. The composition of claim 51, wherein the anti 88. The composition of claim 51, wherein the anti-fibrotic fibrotic agent is a HMGCoA reductase inhibitor. agent is a DNA alkylating agent. 119. The composition of claim 118, wherein the HMG 89. The composition of claim 51, wherein the anti-fibrotic CoA reductase inhibitor is simvastatin or an analogue or agent is an anti-microtubule agent. derivative thereof. US 2005/0281883 A1 Dec. 22, 2005

120. The composition of claim 51, wherein the anti 145. The composition of claim 51, wherein the anti fibrotic agent is a hydroorotate dehydrogenase inhibitor. fibrotic agent is an NO antagonist. 121. The composition of claim 51, wherein the anti 146. The composition of claim 51, wherein the anti fibrotic agent is an IKK2 inhibitor. fibrotic agent is a p38 MAP kinase inhibitor. 122. The composition of claim 51, wherein the anti 147. The composition of claim 146, wherein the p38 MAP fibrotic agent is an IL-1 antagonist. kinase inhibitor is SB 202190. 123. The composition of claim 51, wherein the anti 148. The composition of claim 51, wherein the anti fibrotic agent is an ICE antagonist. fibrotic agent is a phosphodiesterase inhibitor. 124. The composition of claim 51, wherein the anti 149. The composition of claim 51, wherein the anti fibrotic agent is an IRAK antagonist. fibrotic agent is a TGFbeta inhibitor. 125. The composition of claim 51, wherein the anti 150. The composition of claim 51, wherein the anti fibrotic agent is an IL-4 agonist. fibrotic agent is a thromboxane A2 antagonist. 126. The composition of claim 51, wherein the anti 151. The composition of claim 51, wherein the anti fibrotic agent is an immunomodulatory agent. fibrotic agent is a TNF alpha antagonist. 127. The composition of claim 51, wherein the anti 152. The composition of claim 51, wherein the anti fibrotic agent is Sirolimus or an analogue or derivative fibrotic agent is a TACE inhibitor. thereof. 153. The composition of claim 51, wherein the anti 128. The composition of claim 51, wherein the anti fibrotic agent is a tyrosine kinase inhibitor. fibrotic agent is everolimus or an analogue or derivative 154. The composition of claim 51, wherein the anti thereof. fibrotic agent is a vitronectin inhibitor. 129. The composition of claim 51, wherein the anti 155. The composition of claim 51, wherein the anti fibrotic agent is tacrolimus or an analogue or derivative fibrotic agent is a fibroblast growth factor inhibitor. thereof. 156. The composition of claim 51, wherein the anti 130. The composition of claim 51, wherein the anti fibrotic agent is a protein kinase inhibitor. fibrotic agent is biolmus or an analogue or derivative 157. The composition of claim 51, wherein the anti thereof. fibrotic agent is a PDGF receptor kinase inhibitor. 131. The composition of claim 51, wherein the anti 158. The composition of claim 51, wherein the anti fibrotic agent is tresperimus or an analogue or derivative fibrotic agent is an endothelial growth factor receptor kinase thereof. inhibitor. 132. The composition of claim 51, wherein the anti 159. The composition of claim 51, wherein the anti fibrotic agent is auranofin or an analogue or derivative fibrotic agent is a retinoic acid receptor antagonist. thereof. 160. The composition of claim 51, wherein the anti 133. The composition of claim 51, wherein the anti fibrotic agent is a platelet derived growth factor receptor fibrotic agent is 270demethylrapamycin or an analogue or kinase inhibitor. derivative thereof. 161. The composition of claim 51, wherein the anti 134. The composition of claim 51, wherein the anti fibrotic agent is a fibrinogen antagonist. fibrotic agent is gusperimus or an analogue or derivative 162. The composition of claim 51, wherein the anti thereof. fibrotic agent is an antimycotic agent. 135. The composition of claim 51, wherein the anti 163. The composition of claim 162, wherein the the fibrotic agent is pimecrolimus or an analogue or derivative antimycotic agent is Sulconizole. thereof. 164. The composition of claim 51, wherein the anti 136. The composition of claim 51, wherein the anti fibrotic agent is a bisphosphonate. fibrotic agent is ABT-578 or an analogue or derivative 165. The composition of claim 51, wherein the anti thereof. fibrotic agent is a phospholipase A1 inhibitor. 137. The composition of claim 51, wherein the anti 166. The composition of claim 51, wherein the anti fibrotic agent is an inosine monophosphate dehydrogenase fibrotic agent is a histamine H1/H2/H3 receptor antagonist. (IMPDH) inhibitor. 167. The composition of claim 51, wherein the anti 138. The composition of claim 137, wherein the IMPDH fibrotic agent is a macrollide antibiotic. inhibitor is mycophenolic acid or an analogue or derivative 168. The composition of claim 51, wherein the anti thereof. fibrotic agent is a GPIb/IIIa receptor antagonist. 139. The composition of claim 137, wherein the IMPDH 169. The composition of claim 51, wherein the anti inhibitor is 1alpha25 dihydroxy Vitamin D or an analogue fibrotic agent is an endothelin receptor antagonist. or derivative thereof. 170. The composition of claim 51, wherein the anti 140. The composition of claim 51, wherein the anti fibrotic agent is a peroxisome proliferator-activated receptor fibrotic agent is a inhibitor. agonist. 141. The composition of claim 51, wherein the anti 171. The composition of claim 51, wherein the anti fibrotic agent is a MCP-1 antagonist. fibrotic agent is an estrogen receptor agent. 142. The composition of claim 51, wherein the anti 172. The composition of claim 51, wherein the anti fibrotic agent is a MMP inhibitor. fibrotic agent is a Somastostatin analogue. 143. The composition of claim 51, wherein the anti 173. The composition of claim 51, wherein the anti fibrotic agent is an NF kappa B inhibitor. fibrotic agent is a neurokinin 1 antagonist. 144. The composition of claim 143, wherein NF kappa B 174. The composition of claim 51, wherein the anti inhibitor is Bay 11-7082. fibrotic agent is a neurokinin 3 antagonist. US 2005/0281883 A1 Dec. 22, 2005 84

175. The composition of claim 51, wherein the anti 205. The composition of claim 198, wherein the fibrosing fibrotic agent is a VLA-4 antagonist. agent is an arterial vessel wall irritant. 176. The composition of claim 51, wherein the anti 206. The composition of claim 198, wherein the fibrosing fibrotic agent is an osteoclast inhibitor. agent is or comprises Silk. 177. The composition of claim 51, wherein the anti 207. The composition of claim 198, wherein the fibrosing fibrotic agent is a DNA topoisomerase ATP hydrolyzing agent is or comprises Silkworm Silk. inhibitor. 208. The composition of claim 198, wherein the fibrosing 178. The composition of claim 51, wherein the anti agent is or comprises Spider Silk. fibrotic agent is an angiotensin I converting enzyme inhibi 209. The composition of claim 198, wherein the fibrosing tor. agent is or comprises recombinant Silk. 179. The composition of claim 51, wherein the anti 210. The composition of claim 198, wherein the fibrosing fibrotic agent is an angiotensin II antagonist. agent is or comprises raw silk. 180. The composition of claim 51, wherein the anti 211. The composition of claim 198, wherein the fibrosing fibrotic agent is an enkephalinase inhibitor. agent is or comprises hydrolyzed Silk. 181. The composition of claim 51, wherein the anti 212. The composition of claim 198, wherein the fibrosing fibrotic agent is a peroxisome proliferator-activated receptor agent is or comprises acid-treated Silk. gamma agonist insulin Sensitizer. 213. The composition of claim 198, wherein the fibrosing 182. The composition of claim 51, wherein the anti agent is or comprises acylated Silk. fibrotic agent is a protein kinase C inhibitor. 214. The composition of claim 198, wherein the fibrosing 183. The composition of claim 51, wherein the anti agent is or comprises mineral particles. fibrotic agent is a ROCK (rho-associated kinase) inhibitor. 184. The composition of claim 51, wherein the anti 215. The composition of claim 198, wherein the fibrosing fibrotic agent is a CXCR3 inhibitor. agent is or comprises talc. 185. The composition of claim 51, wherein the anti 216. The composition of claim 198, wherein the fibrosing fibrotic agent is an Itk inhibitor. agent is or comprises chitosan. 186. The composition of claim 51, wherein the anti 217. The composition of claim 198, wherein the fibrosing fibrotic agent is a cytosolic phospholipase A-alpha inhibi agent is or comprises polylysine. tor. 218. The composition of claim 198, wherein the fibrosing 187. The composition of claim 51, wherein the anti agent is or comprises fibronectin. fibrotic agent is a PPAR agonist. 219. The composition of claim 198, wherein the fibrosing 188. The composition of claim 51, wherein the anti agent is or comprises bleomycin. fibrotic agent is an immunosuppressant. 220. The composition of claim 198, wherein the fibrosing 189. The composition of claim 51, wherein the anti agent is or comprises CTGF. fibrotic agent is an Erb inhibitor. 221. The composition of claim 198, wherein the fibrosing 190. The composition of claim 51, wherein the anti agent is in the form of a particulate. fibrotic agent is an apoptosis agonist. 222. The composition of claim 221, wherein the particu 191. The composition of claim 51, wherein the anti late is a biodegradable particulate. fibrotic agent is a lipocortin agonist. 223. The composition of claim 222, wherein the biode 192. The composition of claim 51, wherein the anti gradable particulate comprises a material Selected from the fibrotic agent is a VCAM-1 antagonist. group consisting of polyester, polyanhydride, poly(anhy 193. The composition of claim 51, wherein the anti dride ester), poly(ester-amide), poly(ester-urea), polyorthoe fibrotic agent is a collagen antagonist. Ster, polyphosphoester, polyphosphazine, polycyanoacry 194. The composition of claim 51, wherein the anti late, collagen, chitosan, hyaluronic acid, chromic cat gut, fibrotic agent is an alpha 2 integrin antagonist. alginate, Starch, cellulose and cellulose ester. 195. The composition of claim 51, wherein the anti 224. The composition of claim 221, wherein the particu fibrotic agent is a TNF alpha inhibitor. late is non-biodegradable. 196. The composition of claim 51, wherein the anti 225. The composition of claim 224, wherein the non fibrotic agent is a nitric oxide inhibitor. biodegradable particulate comprises a material Selected 197. The composition of claim 51, wherein the anti from the group consisting of polyester, polyurethane, Sili fibrotic agent is a cathepsin inhibitor. cone, polyethylene, polypropylene, polystyrene, polyacry 198. The composition of claim 38, wherein the biologi late, polymethacrylate, and Silk. cally active agent is a fibrosing agent. 226. The composition of claim 221, wherein the particu 199. The composition of claim 198, wherein the fibrosing late is a particulate form of a member Selected from the agent promotes regeneration. group consisting of Silk, talc, Starch, glass, Silicate, Silica, 200. The composition of claim 198, wherein the fibrosing calcium phosphate, calcium Sulfate, calcium carbonate, agent promotes angiogenesis. hydroxyapatite, Synthetic mineral, polymethylmethacrylate, 201. The composition of claim 198, wherein the fibrosing Silver nitrate, ceramic and other inorganic particles. agent promotes fibroblast migration. 227. The composition of claim 38, wherein the biologi 202. The composition of claim 198, wherein the fibrosing cally active agent promotes bone growth. agent promotes fibroblast proliferation. 228. The composition of claim 198, wherein the fibrosing 203. The composition of claim 198, wherein the fibrosing agent promotes bone growth. agent promotes deposition of extracellular matrix (ECM). 229. The composition of claim 228, wherein the fibrosing 204. The composition of claim 198, wherein the fibrosing agent that promotes bone growth is a bone morphogenic agent promotes tissue remodeling. protein. US 2005/0281883 A1 Dec. 22, 2005

230. The composition of claim 228, wherein the fibrosing 245. The method of claim 244, wherein the second buffer agent that promotes bone growth is an Osteogenic growth Solution neutralizes the effect of the first buffer Solution, so factor. that the nucleophilic groups of the first component regain 231. The composition of claim 230, wherein the osteo their nucleophilic character and inter-react with the electro genic growth factor is Selected from transforming growth philic groups of the Second component. factor, platelet-derived growth factor, and fibroblast growth 246. The method of claim 241, wherein the composition, factor. first buffer Solution and second buffer Solution are housed 232. The composition of claim 198, wherein the compo Separately in a multiple-compartment Syringe System having Sition further comprises a pharmaceutical agent that induces a multiple barrels, a mixing head, and an exit orifice, Step Sclerosis (a Sclerosant). (b)(i) comprises adding the first buffer solution to the barrel 233. The composition of claim 232, wherein the sclero housing the composition to dissolve the composition and Sant is Selected from the group consisting of ethanol, dim form a homogeneous Solution, and extruding the homoge ethyl Sulfoxide, Sucrose, Sodium chloride, dextrose, glyc neous Solution into the mixing head; step (b)(ii) comprises erin, minocycline, tetracycline, doxycycline, polidocanol, Simultaneously extruding the Second buffer Solution into the Sodium tetradecyl Sulfate, Sodium morrhuate, and Sotrade mixing head; and step (c) further comprises extruding the col. resulting composition through the orifice onto a Surface. 234. The composition of claim 232, wherein the sclero 247. A method of Sealing tissue of a patient comprising Sant is a Surfactant. the Steps of: 235. The composition of claim 198, wherein the compo (a) providing the dry powder composition of claim 1; Sition further comprises an inflammatory cytokine. 236. The composition of claim 235, wherein the inflam (b) rendering the nucleophilic and electrophilic groups matory cytokine is Selected from the group consisting of reactive by exposing the composition to an aqueous TGFB, PDGF, VEGF, bFGF, TNFC, NGF, GM-CSF, IGF-a, environment to effect inter-reaction; wherein Said expo IL-1, IL-1-B, IL-8, IL-6, and growth hormone. Sure comprises: 237. The composition of claim 198, wherein the compo (i) dissolving the composition in a buffer Solution Sition further comprises an agent that Stimulates cell prolif having a pH within the range of about 1.0 to 5.5 to eration. form a homogeneous Solution, and 238. The composition of claim 237, wherein the agent that Stimulates cell proliferation is selected from the group (ii) adding a buffer solution having a pH within the consisting of dexamethasone, isotretinoin (13-cis retinoic range of about 6.0 to 11.0 to the homogeneous acid), 17-B-estradiol, estradiol, 1-C-25 dihydroxyvitamin Solution to form a mixture, and D, diethylstibesterol, cyclosporine A, L-NAME, all-trans (c) placing the mixture into contact with tissue and retinoic acid (ATRA), and analogues and derivatives thereof. allowing a three-dimensional composition to form and 239. The composition of claim 38, where the biologically Seal the tissue. active agent is mixed with the first and Second components 248. A method of preventing adhesions between tissues of to form a mixture. a patient comprising the Steps of: 240. The composition of claim 38, where the biologically active agent is chemically coupled to the first component or (a) providing the dry powder composition of claim 1; to the Second component. (b) rendering the nucleophilic and electrophilic groups 241. A method of forming a three-dimensional matrix reactive by exposing the composition to an aqueous comprising the Steps of: environment to effect inter-reaction; wherein Said expo (a) providing the dry powder composition of claim 1; and Sure comprises: (i) dissolving the composition in a first buffer Solution (b) rendering the nucleophilic and electrophilic groups having a pH within the range of about 1.0 to 5.5 to reactive by exposing the composition to an aqueous form a homogeneous Solution, and environment to effect inter-reaction; wherein Said expo Sure comprises: (ii) adding a second buffer Solution having a pH within the range of about 6.0 to 11.0 to the homogeneous (i) dissolving the composition in a first buffer Solution Solution to form a mixture, and having a pH within the range of about 1.0 to 5.5 to form a homogeneous Solution, and (c) placing the mixture into contact with tissue and allowing a three-dimensional composition to form on (ii) adding a second buffer Solution having a pH within the tissue. the range of about 6.0 to 11.0 to the homogeneous 249. A method of forming a three-dimensional matrix on Solution; and a Surface of a device comprising the Steps of: (c) allowing a three-dimensional composition to form. (a) providing the dry powder composition of claim 1; and 242. The method of claim 241, wherein the composition (b) rendering the nucleophilic and electrophilic groups is formed without input of any external energy. reactive by exposing the composition to an aqueous 243. The method of claim 241, wherein the composition environment to effect inter-reaction; wherein Said expo is formed by polymerization. Sure comprises: 244. The method of claim 241, wherein the pH of the first buffer solution is selected to retard the reactivity of the (i) dissolving the composition in a first buffer Solution nucleophilic groups on the first component by rendering the having a pH within the range of about 1.0 to 5.5 to nucleophilic groups relatively non-nucleophilic. form a homogeneous Solution, and US 2005/0281883 A1 Dec. 22, 2005 86

(ii) adding a second buffer Solution having a pH within (c) a Second buffer Solution having a pH within the range the range of about 6.0 to 11.0 to the homogeneous of about 6.0 to 11.0, Solution; and wherein each component is packaged separately and (c) applying the homogeneous Solution to a Surface of a admixed immediately prior to use. device; and 255. The kit of claim 254, wherein prior to use, each (d) allowing the three-dimensional matrix to form. component is in a separate Sterile package. 250. A method of preventing scarring in the vicinity of a 256. The kit of claim 254, further comprising a delivery medical implant comprising the Steps of device. 257. The kit of claim 256, wherein the delivery device is (a) providing the dry powder composition of claim 51; a multi-component spray device. (b) rendering the nucleophilic and electrophilic groups 258. The kit of claim 257, wherein the multi-component reactive by exposing the composition to an aqueous Spray device is a multiple-compartment Syringe System environment to effect inter-reaction; wherein Said expo having multiple barrels, a mixing head, and an exit orifice. Sure comprises: 259. The kit of claim 258, wherein the dry powder composition, the first buffer solution, and the second buffer (i) dissolving the composition in a first buffer Solution Solution are housed Separately in the multiple-compartment having a pH within the range of about 1.0 to 5.5 to Syringe System. form a homogeneous Solution, and 260. The kit of claim 256, wherein the delivery device is (ii) adding a second buffer Solution having a pH within a pressurized delivery device. the range of about 6.0 to 11.0 to the homogeneous 261. The kit of claim 260, wherein the pressurized deliv Solution to form a mixture; and ery System comprises: (c) applying the mixture to a Surface of a medical implant a plurality of fluid component inlets each adapted to and allowing a three-dimensional matrix to form on the communicate with a Source of different fluid compo Surface of the medical implant; and nents, (d) placing the medical implant into an animal host, at least one carrier fluid inlet adapted to communicate wherein release of the anti-fibrotic agent from the with a Source of a preSSurized carrier fluid, matrix inhibits scarring in the animal host. a diffuser Surface located downstream from the plurality 251. The method of claim 250, wherein the anti-fibrotic of fluid component inlets and the at least one carrier agent is released into tissue in the vicinity of the implant fluid inlet; and after deployment of the implant. 252. A method of promoting Scarring in the vicinity of a an outlet extending through the diffuser Surface, medical implant comprising the Steps of wherein the diffuser surface is adapted to receive fluid components thereon and has a shape effective to direct (a) providing the dry powder composition of claim 198; and maintain each received fluid component in a dif (b) rendering the nucleophilic and electrophilic groups ferent flow path toward the outlet for mixing and reactive by exposing the composition to an aqueous dispensing therethrough by the pressurized carrier fluid environment to effect inter-reaction; wherein Said expo from the at least one carrier fluid inlet. Sure comprises: 262. The kit of claim 261, wherein the pressurized carrier fluid is pressurized air. (i) dissolving the composition in a first buffer Solution 263. The kit of claim 261, wherein the fluid components having a pH within the range of about 1.0 to 5.5 to are the first buffer Solution and the second buffer Solution. form a homogeneous Solution, and 264. The kit of claim 254, wherein the kit further com (ii) adding a second buffer Solution having a pH within prises a biologically active agent and the medical application the range of about 6.0 to 11.0 to the homogeneous involves delivering the biologically active agent. Solution; and 265. The kit of claim 264, wherein the biologically active agent is packaged with the dry powder composition. (c) applying the mixture to a Surface of a medical implant 266. The kit of claim 265, further comprising a pharma and allowing a three-dimensional matrix to form on the ceutically acceptable carrier packaged with the biologically Surface of the medical implant; and active agent and the dry powder composition. (d) placing the medical implant into an animal host, 267. The kit of claim 264, wherein the biologically active wherein release of the fibrotic agent from the matrix agent is packaged as a Solution with the first buffer. inhibits Scarring in the animal host. 268. The kit of claim 264, wherein the biologically active 253. The method of claim 252, wherein the fibrotic agent agent is packaged as a Solution with the Second buffer. is released into tissue in the vicinity of the implant after 269. The kit of claim 264, further comprising a pharma deployment of the implant. ceutically acceptable carrier as a fourth component. 254. A kit for use in for use in medical applications, 270. The kit of claim 269, wherein the biologically active comprising: agent is packaged with the pharmaceutically acceptable (a) the dry powder composition of claim 1, carrier. 271. The kit of claims 254, wherein the kit further (b) a first buffer solution having a pH within the range of comprises living cells or genes, and the medical application about 1.0 to 5.5; and involves delivering the living cells or genes. US 2005/0281883 A1 Dec. 22, 2005 87

272. The kit of claim 254, wherein the medical application 289. The composition of claim 288, wherein mid-3. involves adhering or Sealing biological tissue. 290. The composition of claim 288, wherein m=3. 273. The kit of claims 254, wherein the medical applica 291. The composition of claim 288, wherein m=4. tion involves bioadhesion. 292. The composition of claim 288, wherein n=4. 274. The kit of claims 254, wherein the medical applica 293. The composition of claim 288, wherein the electro tion involves an ophthalmic application. philic groups are Succinimidyl moieties. 275. The kit of claims 254, wherein the medical applica 294. The composition of claim 288, wherein all n are tion involves tissue augmentation. identical, and all m are identical. 276. The kit of claims 254, wherein the medical applica 295. The composition of claim 281, wherein the amino tion involves adhesion prevention. acid residues as cysteine. 277. The kit of claims 254, wherein the medical applica 296. The composition of claim 295, wherein mid-3. tion involves forming a Synthetic implant or coating a 297. The composition of claim 295, wherein m=3. Synthetic implant. 298. The composition of claim 295, wherein m=4. 278. The kit of claims 254, wherein the medical applica 299. The composition of claim 295, wherein n=4. tion involves the treatment of aneurysms. 300. The composition of claim 295, wherein the electro 279. The kit of claims 254, wherein the medical applica philic groups are Succinimidyl moieties. tion is a laparoscopic procedure. 301. The composition of claim 295, wherein all n are 280. A kit for use in for use in medical applications, identical, and all m are identical. comprising: 302. A crosslinkable composition comprised of: (a) a first component having a core Substituted with m (a) a first crosslinkable component having m nucleophilic nucleophilic groups, where me2, groups, wherein me2; and (b) a Second component having a core Substituted with n (b) a Second crosslinkable component having n electro electrophilic groups, where ne2 and m+n>4, philic groups capable of reaction with the m nucleo (c) a first buffer solution having a pH within the range of philic groups to form covalent bonds, about 1.0 to 5.5; and wherein ne2 and m+ne5, the first component comprises two or more amino acid residues Selected from the (d) a second buffer Solution having a pH within the range group consisting of amino acids comprising primary of about 6.0 to 11.0, amine groupS and amino acids comprising thiol groups, wherein the nucleophilic and electrophilic groups are the Second component comprises a polyethylene glycol non-reactive in a dry environment but are rendered moiety, the electrophilic groups are Succinimidyl moi reactive upon exposure to an aqueous environment eties, and each of the first and Second crosslinkable Such that the components inter-react in the aqueous components is biocompatible, Synthetic, and nonimmu environment to form a three-dimensional composition, nogenic, and further whererin crosslinking of the com and further wherein each component is packaged sepa position results in a biocompatible, nonimmunogenic, rately and admixed immediately prior to use. crosslinked matrix. 281. A crosslinkable composition comprised of: 303. The composition of claim 302, wherein mid-3. 304. The composition of claim 302, wherein m=3. (a) a first crosslinkable component having m nucleophilic 305. The composition of claim 302, wherein m=4. groups, wherein me2; and 306. The composition of claim 302, wherein n=4. (b) a Second crosslinkable component having n electro 307. The composition of claim 302, wherein the electro philic groups capable of reaction with the m nucleo philic groups are Succinimidyl moieties. philic groups to form covalent bonds, 308. The composition of claim 302, wherein all n are identical, and all m are identical. wherein ne2 and m+ne5, the first component comprises 309. The composition of claim 302, wherein the selected two or more amino acid residues Selected from the amino acid residues as lysine. group consisting of amino acids comprising primary 310. The composition of claim 309, wherein mid-3. amine groups and amino acids comprising thiol groups, 311. The composition of claim 309, wherein m=3. the Second component comprises a polyethylene glycol 312. The composition of claim 309, wherein m=4. moiety, and each of the first and Second crosslinkable 313. The composition of claim 309, wherein n=4. components is biocompatible, Synthetic, and nonimmu 314. The composition of claim 309, wherein the electro nogenic, and further wherein crosslinking of the com philic groups are Succinimidyl moieties. position results in a biocompatible, nonimmunogenic, 315. The composition of claim 309, wherein all n are crosslinked matrix. identical, and all m are identical. 282. The composition of claim 281, wherein mid-3. 316. The composition of claim 302, wherein the selected 283. The composition of claim 281, wherein m=3. amino acid residues as cysteine. 284. The composition of claim 281, wherein m=4. 317. The composition of claim 316, wherein mid-3. 285. The composition of claim 281, wherein n=4. 318. The composition of claim 316, wherein m=3. 286. The composition of claim 281, wherein the electro 319. The composition of claim 316, wherein m=4. philic groups are Succinimidyl moieties. 320. The composition of claim 316, wherein n=4. 287. The composition of claim 281, wherein all n are 321. The composition of claim 316, wherein the electro identical, and all m are identical. philic groups are Succinimidyl moieties. 288. The composition of claim 281, wherein the amino 322. The composition of claim 316, wherein all n are acid residues as lysine. identical, and all m are identical. US 2005/0281883 A1 Dec. 22, 2005 88

323. A crosslinkable composition comprised of: 333. The composition of claim 332, wherein mid-3. (a) a first crosslinkable component having m nucleophilic 334. The composition of claim 332, wherein m=3. groups, wherein me2; and 335. The composition of claim 332, wherein m=4. 336. The composition of claim 332, wherein n=4. (b) a Second crosslinkable component having n electro 337. The composition of claim 332, wherein the electro philic groups capable of reaction with the m nucleo philic groups are Succinimidyl moieties. philic groups to form covalent bonds, 338. The composition of claim 332, wherein all n are wherein ne2 and m+ne5, the first component comprises identical, and all m are identical. two or more amino acid residues Selected from the 339. The composition of claim 332, wherein the multi group consisting of amino acids comprising primary functionally activated polyethylene glycol is tetrafunction amine groups and amino acids comprising thiol groups, ally activated polyethylene glycol. the Second component comprises a multifunctionally 340. The composition of claim 332, wherein the multi activated polyethylene glycol, and each of the first and functionally activated polyethylene glycol is a Star-branched Second crosslinkable components is biocompatible, polyethylene glycol. Synthetic, and nonimmunogenic, and further wherein 341. The composition of claim 323, wherein the amino crosslinking of the composition results in a biocompat acid residues are cysteine. ible, nonimmunogenic, crosslinked matrix. 342. The composition of claim 341, wherein mid-3. 324. The composition of claim 323, wherein mid-3. 343. The composition of claim 341, wherein m=3. 325. The composition of claim 323, wherein m=3. 326. The composition of claim 323, wherein m=4. 344. The composition of claim 341, wherein m=4. 327. The composition of claim 323, wherein n=4. 345. The composition of claim 341, wherein n=4. 328. The composition of claim 323, wherein the electro 346. The composition of claim 341, wherein the electro philic groups are Succinimidyl moieties. philic groups are Succinimidyl moieties. 329. The composition of claim 323, wherein all n are 347. The composition of claim 341, wherein all n are identical, and all m are identical. identical, and all m are identical. 330. The composition of claim 323, wherein the multi 348. The composition of claim 341, wherein the multi functionally activated polyethylene glycol is tetrafunction functionally activated polyethylene glycol is tetrafunction ally activated polyethylene glycol. ally activated polyethylene glycol. 331. The composition of claim 323, wherein the multi 349. The composition of claim 341, wherein the multi functionally activated polyethylene glycol is a Star-branched functionally activated polyethylene glycol is a Star-branched polyethylene glycol. polyethylene glycol. 332. The composition of claim 323, wherein the amino acid residues are lysine.