USO09533056 B2

(12) United States Patent (10) Patent No.: US 9,533,056 B2 Rau et al. (45) Date of Patent: Jan. 3, 2017

(54) DIPEPTDE-BASED PRODRUG LINKERS 2012. O156259 A1 6, 2012 Rau et al. FOR ALPHATIC AMINE-CONTAINING 2012. O156260 A1 6, 2012 Rau et al. 2012,019 1039 A1 7/2012 Rau et al. DRUGS (71) Applicant: Ascendis Pharma A/S, Hellerup (DK) FOREIGN PATENT DOCUMENTS Inventors: Harald Rau, Heidelberg (DE); Torben WO WO 99,30727 6, 1999 (72) WO WO O2,38590 5, 2002 Le?mann, Neustadt an der Weinstrasse WO WO O2/O8318O 10, 2002 (DE) WO WO O2/O89789 11 2002 (73) Assignee: Ascendis Pharma AS, Hellerup (DK) WO WO 2004/O19993 3, 2004 WO WO 2004/043493 5, 2004 WO WO 2004/108070 12, 2004 (*) Notice: Subject to any disclaimer, the term of this WO WO 2009.095479 8, 2009 patent is extended or adjusted under 35 WO WO 2009.099763 8, 2009 U.S.C. 154(b) by 0 days. WO WO 2009 123768 A2 * 10, 2009 ...... A61K 9,06 (21) Appl. No.: 14/674,928 OTHER PUBLICATIONS Santos et al., “Cyclization-activated prodrugs. Synthesis, reactivity (22) Filed: Mar. 31, 2015 and toxicity of dipeptide esters of paracetamol. Bioorganic & Medicinal Chemistry Letters, 2005, 1595-1598, 15, Elsevier. (65) Prior Publication Data Product List of Jenkem Technology, USA (accessed and down loaded from www.jenkemusa.com on Jul. 28, 2009). US 2015/02O2317 A1 Jul. 23, 2015 Suaifan et al., “Effects of steric bulk and stereochemistry on the rates of diketopiperazine formation from N-aminoacyl-2.2- dimethylthiazolidine-4-carboxamides (Dmt dipeptide amides)-a model for a new prodrug linker system”. Tetrahedron, Nov. 27. Related U.S. Application Data 2006, 11245-11266,-62, 48, Elsevier, Amsterdam NL. Brady et al., “Design and Synthesis of a Pro-Drug of Vinblastine (63) Continuation of application No. 13/574,092, filed as Targeted at Treatment of Prostate Cancer with Enhanced Efficacy application No. PCT/EP2011/050821 on Jan. 21, and Reduced Systemic Toxicity”. J. Med. Chem., 2002, 4706-4715, 45, American Chemical Society. 2011, now Pat. No. 9,062,094. Wipf, et al., “Synthesis of Chemoreversible Prodrugs of ara-C with Variable Time-Release Profiles. Biological Evaluation, of Their (30) Foreign Application Priority Data Apoptotic Activity”. Bioorganic & Medicinal Chemistry, 1996, 1585-1596, 4, 10, Pergamon. Jan. 22, 2010 (EP) ...... 10151.465 Gomes et al., “Cyclization-activated Prodrugs'. Molecules.2007. 2484-2506, 12, MDPI. Hamel et al., “Cyclosporin A prodrugs: design, synthesis and (51) Int. Cl. biophysical properties”. J. Pept. Res., 2004, 147-154, 63, Blackwell A6 IK 38/00 (2006.01) Munksgaard. A6 IK 47/00 (2006.01) Greenwald et al., “A New Aliphatic Amino Prodrug System for the A6 IK 47/48 (2006.01) Delivery of Small Molecules and Proteins Utilizing Novel PEG A 6LX 3/553 (2006.01) Derivatives'. J. Med. Chem., 2004, 726-734, 47, American Chemi A6 IK 38/26 (2006.01) cal Society. Garman et al., “The preparation and properties of novel reversible A6M 5/9 (2006.01) polymer-protein conjugates', FEBS Letters, Nov. 1987, 361-365, (52) U.S. Cl. 223, 2, Elsevier Science Publishers BV. CPC ...... A61K 47/48338 (2013.01); A61 K3I/553 Lee et al., “Targeted Enzyme-Responsive Drug Carriers: Studies on (2013.01); A61K 38/26 (2013.01); A61 K the Delivery of a Combination of Drugs'. Angew. Chem., 2004, 1707-1710, 16, Wiley-VCH Verlag GmbH & Co. KGaA. Weinheim. 47/48215 (2013.01); A61K 47/48784 Shabat et al., "Chemical Adaptor Systems”. Chem. Eur, J., 2004, (2013.01); A61M 5/19 (2013.01) 2626-2634, 10, Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. (58) Field of Classification Search Antczak et al., “A New Acivicin Prodrug Designed for Tumor CPC ...... A61K 39/39558: A61K 47/48384; Targeted Delivery'. Bioorganic & Medicinal Chemistry, 2001, A61K 38.700 2843-2848, 9, Elsevier Science Ltd. See application file for complete search history. (Continued) Primary Examiner — Hasan Ahmed (56) References Cited Assistant Examiner — Kaipeen Yang U.S. PATENT DOCUMENTS (74) Attorney, Agent, or Firm — Frommer Lawrence & Haug LLP 5,952.294 A 9, 1999 Lazo et al. 6,613,879 B1 9, 2003 FireStone et al. (57) ABSTRACT 7,163,923 B2 1/2007 Sergeeva et al. The present invention relates to a prodrug or a pharmaceu 7,879,588 B2 2/2011 Vetter et al. tically acceptable salt thereof, comprising a drug linker 7,968,085 B2 6, 2011 Hersel et al. conjugate D-L, wherein D being a biologically active moiety 2008/0241102 A1 10, 2008 Hersel et al. containing an aliphatic amine group is conjugated to one or 2010/0291021 A1 11/2010 Vetter et al. more polymeric carriers via dipeptide-containing linkers L. 2011 0009315 A1 1/2011 Hersel et al. 2011/005.3848 A1 3/2011 Cleemann et al. Such carrier-linked prodrugs achieve drug releases with 2011/O112021 A1 5, 2011 Rau et al. therapeutically useful half-lives. The invention also relates 2011/0172390 A1 7, 2011 Vetter et al. to pharmaceutical compositions comprising said prodrugs 2011/022323.0 A1 9/2011 Hersel et al. and their use as medicaments. 2012fOO58084 A1 3/2012 Rau et al. 10 Claims, No Drawings US 9,533,056 B2 Page 2

(56) References Cited Luo et al., “A Hyaluronic Acid-Taxol Antitumor Bioconjugate Targeted to Cancer Cells'. Biomacromolecules, 200, 208-218, 1. American Chemical Society. OTHER PUBLICATIONS Greenwald et al., “Drug Delivery Systems Employing 1,4- or Greenwald et al., “Drug Delivery Systems Based on Trimethyl Lock 1,6-Elimination: Poly(ethylene glycol) Prodrugs of Amine-Contain ing Compounds”. J. Med. Chem., 1999, 3657-3667, 42, American Lactonization: Poly(ethylene glycol) Prodrugs of Amino-Contain Chemical Society. ing Compounds”. J. Med. Chem., 2000, 475-487. 43, American Testa et al., Metabolic Hydrolysis and Prodrug Design, 2003, p. 4. Chemical Society. Wiley-VCH. Testa et al., Metabolic Hydrolysis and Prodrug Design, 2003, p. 5, Peleg-Shulman et al., “Reversible PEGylation: A Novel Technology Wiley-VCH. to Release Native Interferon O2 over a Prolonged Time Period”. J. Cavallaro et al., “Polymeric Prodrug for Release of an Antitumoral Med. Chem., 2004, 4897-4904, 47. American Chemical Society. Agent by Specific Enzymes'. Bioconjugate Chem., 2001, 143-151, Caliceti et al., “Pharmacokinetic and biodistribution properties of 12, American Chemical Society. poly(ethylene glycol)-protein conjugates'. Advanced Drug Deliv ery, Reviews, 2003, 1261-1277, 55, Elsevier B.V. Testa et al., Metabolic Hydrolysis and Prodrug Design (Chapter 8: Duncan et al., “The Drawing Era of Polymer Therapeutics', Nature “The Hydrolysis of Carboxylic Acid Ester Prodrugs”), 419-534, Reviews Drug Discovery, 2003, 347-360, 2, Nature Publishing Wiley-VCH. Group. Bhatt et al., “Synthesis and in Vivo Antitumor Activity of Poly(L- Na et al., “Monitoring of peptide acylation inside degrading glutamic acid) Conjugates of 20 (S)-Camptothecin”. J. Med. Chem. PLGAmicrospheres by capillary electrophoresis and MALDLI 2003, 190-193, 46, American Chemical Society. TOF mass spectromary”, Journal of Controlled Release, 2003, Cheng et al., “Sythesis of Linear, B-Cyclodextrin-Based Polymers 291-299,92, Elsevier B.V. and Their Camptothecin Conjugates'. Bioconjugate Chem, 2003, 1007-1017, 14, American Chemical Society. * cited by examiner US 9,533,056 B2 1. 2 DIPEPTDE-BASED PRODRUG LINKERS Alternatively, the drugs may be conjugated to a carrier FOR ALPHATIC AMINE-CONTAINING through permanent covalent bonds. This approach is applied DRUGS to various classes of molecules, from So-called Small mol ecules, through natural products up to larger proteins. The present application is a continuation of U.S. patent Liraglutide is an example of a peptide drug that achieves application Ser. No. 13/574,092 filed on Jul. 19, 2012, which an extended half-life by permanent covalent modification claims priority from PCT Patent Application No. PCT/ with a palmitoyl moiety. The fatty acid alkyl chain serves to EP2011/050821 filed on Jan. 21, 2011, which claims priority provide albumin binding in vivo and the palmitoylated from European Patent Application No. EP 10 151 65.1 filed peptide forms an albumin complex that acts as a drug 10 reservoir in the blood stream. on Jan. 22, 2010, the disclosures of which are incorporated Albuferon is an example of a protein drug that achieves an herein by reference in their entirety. extended half-life by permanent covalent modification with another protein that in itself has a long half-life. The FIELD OF THE INVENTION corresponding fusion protein of albumin and interferon 15 alpha, Albuferon, exhibits a significantly extended half-life The present invention is directed to carrier-linked prod as compared to interferon alpha. rugs having temporary amide linkages between Substituted Many Small molecule medicinal agents, like alkaloids and dipeptide moieties and aliphatic amine groups of biologi anti-tumor agents, show low solubility in aqueous fluids. cally active entities such as peptides, proteins, natural prod One way to solubilize these small molecule compounds is to ucts or synthetic chemical compounds. Such carrier-linked conjugate the Small molecule compounds to hydrophilic prodrugs are characterized by slow release of unmodified (water-soluble) polymers. A variety of water-soluble poly biologically active entity. mers, such as human serum albumin, dextran, lectins, poly It is noted that citation or identification of any document (ethylene glycol) (PEG), poly(styrene-co-maleic anhy in this application is not an admission that Such document is dride), poly(N-hydroxypropylmethacrylamide), poly available as prior art to the present invention. 25 (divinyl ether-co-maleic anhydride), hyaluronic acid have Typically, carriers employed for extended time-action been described for this purpose (R. Duncan, Nature Rev. engineering in drug delivery are either used in a non Drug Disc., 2003, 2, 347-360). covalent fashion, with the drug physicochemically formu Covalent modification of biological molecules with poly lated into a solvent-carrier mixture, or by permanent cova (ethylene glycol) has been extensively studied since the late lent attachment of a carrier reagent to one of the drugs 30 1970s. So-called PEGylated proteins have shown improved functional groups. therapeutic efficacy by increasing Solubility, reducing immu Non-covalent drug encapsulation into polymeric carriers nogenicity, and increasing circulation half-live in vivo due to has been applied to depot formulations for long-acting reduced renal clearance and proteolysis by enzymes (see, for release profiles. Typically, the drug is mixed with carrier example, Caliceti P. Veronese F. M., Adv. Drug Deliv. Rev. material and processed in Such fashion, that the drug 35 2003, 55, 1261-1277). becomes distributed inside the bulk carrier. For instance However, many biological molecules such as IFN alfa 2. polymer-drug aggregates may be shaped as microparticles saquinavir or Somatostatin are inactive or show decreased which are administered as an injectable Suspension or the biological activity when a carrier is covalently conjugated to polymer-drug aggregates are formulated as gels which are the drug (T. Peleg-Shulman et al., J. Med. Chem., 2004, 47. administered in a single bolus injection. Known in the art are 40 4897-4904). also liposomal formulations, where the carrier may be a In order to avoid shortcomings imposed by either the polymeric or non-polymeric entity capable of Solubilizing non-covalent polymer mixtures or the permanent covalent the drug. Drug release occurs when the carrier Swells or attachment, it may be preferable to employ a prodrug physically deteriorates or chemical degradation allows dif approach for chemical conjugation of the drug to the poly fusion of the drug to the exterior and subsequently into the 45 mer carrier. In Such polymeric prodrugs, the biologically biological environment. Such chemical degradation pro active moieties (drugs, therapeutic, biological molecule, cesses may be autohydrolytic or enzyme-catalyzed. An etc.) are typically linked to the polymeric carrier moiety by example for a marketed drug based on bolus administration a temporary bond formed between the carrier moiety and a of a drug-polymer gel is Lupron Depot. An example for a hydroxy, amino or carboxy group of the drug molecule. marketed drug based on Suspended microparticles is Nutro 50 Prodrugs are therapeutic agents that are almost inactive pin Depot. An example for a marketed drug based on a per se but are predictably transformed into active molecular liposomal formulation is Doxil. entities (see B.Testa, J. M: Mayer in Hydrolysis in Drug and A disadvantage of the non-covalent approach is that in Prodrug Metabolism, Wiley-VCH, 2003, page 4). The car order to prevent uncontrolled, burst-type release of the drug, rier prodrug approach may be applied in Such a fashion that encapsulation of the drug has to be highly efficient by 55 the drug is released in vivo from the polymer in order to creating a sterically highly crowded environment. Restrain regain its biological activity. The reduced biological activity ing the diffusion of an unbound, water soluble drug molecule of the prodrug as compared to the released drug is of requires strong van der Waals contacts, frequently mediated advantage if a slow or controlled release of the drug is through hydrophobic moieties. Many conformationally sen desired. In this case, a relatively large amount of prodrug sitive drugs, such as proteins or peptides, are rendered 60 may be administered without concomitant side effects and dysfunctional during the encapsulation process and/or dur the risk of overdosing. Release of the drug occurs over time, ing Subsequent storage of the encapsulated drug. In addition, thereby reducing the necessity of repeated and frequent Such amino-containing drugs readily undergo side reactions administration of the drug. with carrier degradation products (see, for example, D. H. Prodrug activation may occur by enzymatic or non Lee et al., J. Contr. Rel., 2003, 92,291-299). Furthermore, 65 enzymatic cleavage of the temporary bond between the dependence of the release mechanism of the drug upon carrier and the drug molecule, or a sequential combination biodegradation may cause interpatient variability. of both, i.e. an enzymatic step followed by a non-enzymatic US 9,533,056 B2 3 4 rearrangement. In an enzyme-free in-vitro environment Such Amide bonds are usually much more stable against hydro as an aqueous buffer Solution, a temporary bond such as an lysis than ester bonds, and the rate of clevage of the amide ester or amide may undergo hydrolysis, but the correspond bond would be too slow for therapeutic utility in a carrier ing rate of hydrolysis may be much too slow and thus linked prodrug. Therefore it is advantageous to add struc outside the therapeutically useful range. In an in vivo 5 tural chemical components such as neighbouring groups in environment, esterases or amidases are typically present and order to exert control over the cleavability of the prodrug the esterases and amidases may cause significant catalytic amide bond. Such additional cleavage-controlling chemical acceleration of the kinetics of hydrolysis from twofold up to structures that are provided neither by the carrier entity nor several orders of magnitude (see, for example, R. B. Green by the drug are termed “linkers'. Prodrug linkers can have wald et al. J. Med. Chem. 1999, 42 (18), 3857-3867). 10 a strong effect on the rate of hydrolysis of a given temporary Prodrugs fall in two classes, bioprecursors and carrier bond. Variation of the chemical nature of these linkers linked prodrugs. Bioprecursors do not contain a carrier allows the engineering of the properties of the linker to a group and are activated by the metabolic creation of a great extent. functional group. In carrier-linked prodrugs the active Sub Several examples have been published of the prodrug stance is linked to a carrier moiety by a temporary linkage. 15 activation of amine-containing biologically active moieties The carrier may be biologically inert (for instance PEG) or by specific enzymes for targeted release. A prerequisite for may have targeting properties (for instance antibodies). This enzymatic dependence is that the structure of the linker invention is concerned with polymeric carrier-linked or displays a structural motif that is recognized as a substrate macromolecular prodrugs, where the carrier itself is a mac by a corresponding endogenous enzyme. In these cases, the romolecule Such as a carrier protein or polysaccharide or 20 cleavage of the temporary bond occurs in a one-step process poly(ethylene glycol). which is catalyzed by the enzyme. G. Cavallaro et al. Cleavage of a carrier prodrug generates a molecular entity (Bioconjugate Chem. 2001, 12, 143-151) describe the enzy (drug) of increased bioactivity and at least one side product, matic release of an antitumoral agent by the protease plas the carrier. After cleavage, the bioactive entity will reveal at min. Cytarabin is coupled via the tripeptide sequence D-Val least one previously conjugated and thereby protected func- 25 Leu-Lys to the polymer alpha, beta-poly(N-hydroxyethyl)- tional group, and the presence of this group typically con DL-aspartamide (PHEA). Enzymatic release of cytarabin is tributes to the drug's bioactivity. effected by the protease plasmin which concentration is In order to implement a prodrug strategy, at least one relatively high in various kinds of tumor mass. selected functional group in the drug molecule is employed Enzyme-catalyzed acceleration of prodrug cleavage is a for attachment of the carrier polymer. Preferred functional 30 desirable feature for organ or cellular targeting applications. groups are hydroxyl or amino groups. Consequently, both Targeted release of the bioactive entity is effected, only if an the attachment chemistry and hydrolysis conditions depend enzyme, that selectively cleaves the linkage, is specifically on the type of functional group employed. present in the organ or cell-type chosen for treatment. Numerous macromolecular prodrugs are described in the A major drawback of predominantly enzymatic cleavage literature where the temporary linkage is a labile ester bond. 35 is interpatient variability. Enzyme levels may differ signifi In theses cases, the functional group provided by the bio cantly between individuals resulting in biological variation active entity is either a hydroxyl group or a carboxylic acid of prodrug activation by the enzymatic cleavage. The (e.g. Y. Luo, M R Ziebell, G D Prestwich, “A Hyaluronic enzyme levels may also vary depending on the site of Acid Taxol Antitumor Bioconjugate Targeted to Cancer administration. For instance it is known that in the case of Cells’, Biomacromolecules 2000, 1, 208-218, J Cheng et al., 40 Subcutaneous injection, certain areas of the body yield more Synthesis of Linear, beta-Cyclodextrin Based Polymers and predictable therapeutic effects than others. To reduce this Their Camptothecin Conjugates, Bioconjugate Chem. 2003, unpredictable effect, non-enzymatic cleavage or intramo 14, 1007-1017, R. Bhatt et al, Synthesis and in Vivo Anti lecular catalysis is of particular interest (see, for example, B. tumor Activity of Poly(L-glutamic acid) Conjugates of Testa, J. M: Mayer in Hydrolysis in Drug and Prodrug 20(S)-Campthothecin, J. Med. Chem. 2003, 46, 190-193; R. 45 Metabolism, Wiley-VCH, 2003, page 5). B. Greenwald, A. Pendri, C. D. Conover, H. Zhao, Y. H. Furthermore, it is difficult to establish an in vivo-in vitro Choe, A. Martinez, K. Shum, S. Guan, J. Med. Chem., 1999, correlation of the pharmacokinetic properties for enzyme 42,3657-3667; B. Testa, J. M: Mayer in Hydrolysis in Drug dependent carrier-linked prodrugs. In the absence of a and Prodrug Metabolism, Wiley-VCH, 2003, Chapter 8). reliable in vivo-in vitro correlation optimization of a release Especially for therapeutic biomacromolecules but also for 50 profile becomes a cumbersome task. certain Small molecule drugs, it may be desirable to link the Other carrier prodrugs employing temporary linkages to carrier to amino groups of the bioactive entity (i.e. N-ter amino groups present in the drug molecule are based on a minus or lysine amino groups of proteins). This will be the cascade mechanism. Cascade cleavage is enabled by linker case if masking the drug's bioactivity requires conjugation compounds that are composed of a structural combination of of a certain amino group of the bioactive entity, for instance 55 a masking group and an activating group. The masking an amino group located in an active center or a region or group is attached to the activating group by means of a first epitope involved in receptor binding. Also, during prepara temporary linkage such as an ester or a carbamate. The tion of the prodrug, the amino groups may be more che activating group is attached to an amino-group of the drug moselectively addressed and serve as a better handle for molecule through a second temporary linkage, for instance conjugating the carrier and the drug because of their greater 60 a carbamate. The stability or susceptibility to hydrolysis of nucleophilicity as compared to hydroxylic or phenolic the second temporary linkage (e.g. carbamate) is dependent groups. This is particularly true for proteins and peptides on the presence or absence of the masking group. In the which may contain a great variety of different reactive presence of the masking group, the second temporary link functionalities, where non-selective conjugation reactions age is highly stable and unlikely to release the drug with lead to undesired product mixtures which require extensive 65 therapeutically useful kinetics. In the absence of the mask characterization or purification and may decrease reaction ing group, this linkage becomes highly labile, causing rapid yield and therapeutic efficiency of the product. cleavage and drug release. US 9,533,056 B2 5 6 The cleavage of the first temporary linkage is the rate released. The activating group is still connected to the limiting step in the cascade mechanism. This first step may polyacrylamide polymer after drug release. induce a molecular rearrangement of the activating group M.-R. Lee et al. describe (Angew. Chem. 2004, 116, Such as a 1.6-elimination. The rearrangement renders the 1707-1710) a similar prodrug system based on a mandelic second temporary linkage so much more labile that its acid activating group and an enzymatically cleavable ester cleavage is induced. Ideally, the cleavage rate of the first linked masking group. temporary linkage is identical to the desired release rate for Nevertheless, in these linkers the 16-elimination step still the drug molecule in a given therapeutic scenario. Further generates a highly reactive aromatic intermediate. Even if more, it is desirable that the cleavage of the second tempo the aromatic moiety remains permanently attached to the 10 polymeric carrier, side reactions with potentially toxic prod rary linkage is substantially instantaneous after its lability ucts or immunogenic effects may be caused. has been induced by cleavage of the first temporary bond. For these reasons, there is a need to provide novel linker Examples of polymeric prodrugs based on 1.6-elimination technologies for forming polymeric prodrugs of amine con have been described by R. B. Greenwald et al. J. Med. taining active agents using aliphatic prodrug linkers that are Chem., 1999, 42, 3657-3667 & PCT Patent Application 15 not enzyme-dependent and do not generate reactive aromatic WO-A-99/30727, F. M. H. DeGroot et al. (WO-A 02/83 180 intermediates during cleavage. and WO-A 04/43493A1), and D. Shabat et al. (WO-A A. J. Garman et al. (A. J. Garman, S. B. Kalindjan, FEBS 04/19993). Lett. 1987, 223 (2), 361-365, 1987) use PEG5000-maleic Examples of polymeric amino-containing prodrugs based anhydride for the reversible modification of amino groups in on trimethyl lock lactonization were described by R. B. tissue-type plasminogen activator and urokinase. Regenera Greenwald et al. J. Med. Chem. 2000, 43(3), 457-487: PCT tion of functional enzyme from PEG-uPA conjugate upon Patent Application No. WO-A-02/089789). In this prodrug incubation at pH 7.4 buffer by cleavage of the maleamic acid system, Substituted o-hydroxyphenyl-dimethylpropionic linkeage follows first order kinetics with a half-life of 6.1 h. acid is linked to PEG by an ester, carbonate, or carbamate A disadvantage of the maleamic acid linkage is the lack of group as a first temporary linkage and to amino groups of 25 stability of the conjugate at lower pH values. This limits the drug molecules by means of an amide bond as second applicability of the maleamic acid linkage to active agents temporary linkage. The rate-determining step in drug release which are stable at basic (high) pH values, as purification of is the enzymatic cleavage of the first linkage. This step is the active agent polymer conjugate has to be performed followed by fast amide cleavage by lactonization, liberating under basic (high pH) conditions to prevent premature an aromatic lactone side product. 30 prodrug cleavage. The disadvantage in the abovementioned prodrug systems More recently, R. B. Greenwald et al. (Greenwald et al. J. described by Greenwald, DeGroot and Shabat is the release Med. Chem. 2004, 47, 726-734 and WO-A 2004/108070) of highly reactive and potentially toxic aromatic Small described a PEG cascade prodrug system based on N,N-bis molecule side products like quinone methides or aromatic (2-hydroxyethyl)glycine amide (bicine) linker. In this sys lactones after cleavage of the temporary linkage. The poten 35 tem two PEG carrier molecules are linked via temporary tially toxic entities are released in a 1:1 stoichiometry with bonds to a bicine molecule coupled to an amino group of the the drug and can assume high in vivo concentrations. drug molecule. The first two steps in prodrug activation is A different group of cascade produgs with aromatic the enzymatic cleavage of the first temporary linkages activating groups based on 1.6-elimination structurally sepa connecting both PEG carrier molecules with the hydroxy rates the masking group and the carrier. This may be 40 groups of the bicine activating group. Different linkages achieved by employing a permanent bond between the between PEG and bicine are described resulting in different polymer carrier and the activating group. This stable bond prodrug activation kinetics. The second step in prodrug does not participate in the cascade cleavage mechanism. If activation is the cleavage of the second temporary linkage the carrier is not serving as a masking group and the connecting the bicine activating group to the amino group of activating group is coupled to the carrier by means of a 45 the drug molecule. The main disadvantage of this system is stable bond, release of potentially toxic side products such the connection of the polymer to the bicine linker via as the activating group is avoided. The stable attachment of temporary bonds and the slow hydrolysis rate of this second the activating group and the polymer also Suppresses the temporary bicine amide linkage (t/2>3 h in phosphate release of drug-linker intermediates with undefined pharma buffer) which results in the release of a bicine-modified cology. 50 prodrug intermediate that may show different pharmacoki Antczak et al. (Bioorg Med Chem 9 (2001) 2843-48) netic, immunogenic, toxicity and pharmacodynamic prop describe a reagent which forms the basis for a macromo erties as compared to the parent native drug molecule. lecular cascade prodrug system for amine-containing drug Dipeptides are frequently utilized for prodrug develop molecules. In this approach an antibody serves as the carrier, ment for targeting or targeted transport as they are Substrates a stable bond connects the antibody to an activating group, 55 for enzymes or biotransport systems. Less studied is the carrying an enzymatically cleavable masking group. Upon non-enzymatic route for dipeptide prodrug formation, enzymatic removal of the ester-linked masking group, a namely the ability to undergo intramolecular cyclization to second temporary bond cleaves and releases the drug com form the corresponding diketopiperazine (DKP) and release pound. the active drug. D. Shabat et al. (Chem. Eur. J. 2004, 10, 2626-2634) 60 Such dipeptides may be attached to a drug via ester bonds describe a polymeric prodrug system based on a mandelic as was described for dipeptide esters of the drug paracetamol acid activating group. In this system the masking group is (Santos, Gomes et al Bioorganic & Medicinal Chemistry linked to the activating group by a carbamate bond. The Letters, 2005). In this case, the cyclization reaction consists activating group is conjugated permanently to a polyacryl of a nucleophilic attack of the N-terminal amine of the amide polymer via an amide bond. After enzymatic activa 65 peptide on the ester carbon atom to form a tetrahedral tion of the masking group by a catalytic antibody, the intermediate. This is followed by a proton transfer from the masking group is cleaved by cyclization and the drug is amine to the leaving group oxyanion with simultaneous US 9,533,056 B2 7 8 formation of a peptide bond to give the cyclic DKP product attributed to it in U.S. Patent law; e.g., they can mean and free drug. The reaction has been described for ester “includes”, “included, “including, and the like; and that prodrugs for example for cyclosporin A (Hamel, A R; terms such as "consisting essentially of and “consists Hubler, F: Carrupt, A: Wenger, R M: Mutter, M. J. Pept. essentially of have the meaning ascribed to them in U.S. Res., Vol. 63, num. 2 (2004), p. 147-154). This method is Patent law, e.g., they allow for elements not explicitly applicable to hydroxyl-containing drugs in vitro but has recited, but exclude elements that are found in the prior art been found to compete with enzymatic hydrolysis of the or that affect a basic or novel characteristic of the invention. It is further noted that the invention does not intend to ester bond in vivo, as corresponding dipeptide esters encompass within the scope of the invention any previously released paracetamol at a much faster rate than in buffer disclosed product, process of making the product or method (Gomes et al, Molecules 12 (2007) 2484-2506). 10 of using the product, which meets the written description The problem of susceptibility of dipeptide-based prodrugs and enablement requirements of the USPTO (35 U.S.C. 112, to peptidases may be addressed by incorporating at least one first paragraph) or the EPO (Article 83 of the EPC), such that non-natural amino acid in the dipeptide motif. Correspond applicant(s) reserve the right to disclaim, and hereby dis ing prodrugs of cytarabine (Wipfet al. Bioorg. Med. Chem. close a disclaimer of any previously described product, 4 (1996) 1585-1596) and cyclosporine A (Hamel et al., J. 15 method of making the product, or process of using the Peptide Res. 63 (2004) 147-154) were synthesized and product. tested. Still, endogenous enzymes capable of cleaving ester bonds are not limited to peptidases, and the enzyme-depen SUMMARY OF THE INVENTION dence of Such prodrug cleavage still gives rise to unpredict able in vivo performance. Therefore, an object of the present invention is to provide Enzyme-dependence by design was engineered into DKP carrier-linked prodrug linkers suitable for drugs containing prodrugs as described in U.S. Pat. No. 7,163,923, where aliphatic amine groups from which free drug is released with dipeptide ester prodrugs were formylated at the amino therapeutically useful half-lives. terminus of the dipeptide, and enzymatic deformylation was This object is achieved by a polymeric prodrug or phar used as a trigger to set off diketopiperazine formation and 25 maceutically acceptable salt thereof comprising a drug subsequent cleavage of the ester-dipeptide bond followed by linker conjugate D-L, wherein drug release. Similarly, vinblastine conjugates bearing an D is an aliphatic amine containing biologically active moi oligopeptide were described (Brady et al., J. Med. Chem. 45 ety; and (2002) 4706-4715). Here, an octapeptide was attached by an L is a non-biologically active linker containing ester linkage to the 4-hydroxyl group of vinblastine and 30 i) a moiety L' represented by formula (I), found to undergo ester bond cleavage by DKP formation after specific enzymatic removal of the N-terminal hexapep tide. (I) Recently the scope of the DKP formation reaction was extended to amide prodrugs. U.S. Pat. No. 5,952.294 details 35 prodrug activation using diketopiperazine formation for dipeptidyl amide prodrugs of cytarabine. In this case, the temporary linkage was formed between the carbonyl of a dipeptide and the aromatic amino group of cytarabine. In another study, the utility of diketopiperazine activation was 40 demonstrated for even more stable aliphatic amide prodrugs (G. A. R. Y Suaifan et al., Tetrahedron 62 (2006) 11245 11266). Neither of these studies teaches how a slow-release effect can be achieved for Such conjugates as there is no carrier or other half-life extending moiety or functionality 45 wherein the dashed line indicates the attachment of L' to present in the compounds disclosed. an aliphatic amino group of D by forming an amide WO-A 2009/99763 describes dipeptide prodrugs of bio bond; active peptides such as GLP-1 capable of releasing the X is selected from 0, S or CH-R'". peptide through diketopiperazine formation of the dipeptidic R" and R'' are independently selected from H, OH, CH extension. In this case, the bioactive peptide moiety may 50 R. R. R. and R“ are independently selected from H carry an additional PEG chain on one of its amino acid side and C alkyl, chain residues to achieve extended circulation of the bioac R. Rare independently selected from H, C, alkyl, and tive peptide. A significant disadvantage of this approach is R5 that the PEG chain has to be linked to the peptide without R is selected from compromising its bioactivity, and it is well known that this 55 is difficult to achieve for many peptide-based bioactives. Furthermore, as the PEGylated peptide is bioactive, it may be expected that the dipeptidic promoiety has an effect on the peptide's bioactivity and may negatively affect its recep tor binding properties. As it is well known, that many 60 peptides may interact with more than one receptor and that sequence extensions may affect the balance of Such multiple co receptor binding, unpredictable in vivo performance and even side effects may occur. It is noted that in this disclosure and particularly in the 65 claims and/or paragraphs, terms such as "comprises”, “com -(o- prised', 'comprising and the like can have the meaning US 9,533,056 B2 10 -continued is used as a prodrug principle. In particular detail, it was Surprisingly found that diketopiperazine formation can be used for carrier-linked amide prodrugs. Specifically, the linkers used in these carrier-linked amide prodrugs are designed such that they consist of a carrier permanently NH attached to a dipeptide motif in Such a fashion that dike topiperazine-formation can still be employed as a self activation principle. Suprisingly, in these linker structures, the presence of the carrier entity still allows for therapeuti 10 cally useful autohydrolysis rates, an essential prerequisite for prodrug applications. In the present application the following terms are used as described below. “Prodrug': A prodrug is any compound that undergoes 15 biotransformation before exhibiting its pharmacological NH2 effects. Prodrugs can thus be viewed as drugs containing specialized non-toxic protective groups used in a transient manner to alter or to eliminate undesirable properties in the NH parent molecule. “Promoiety' refers to the part of the prodrug which is not \-- the biologically active moiety. Promoiety thus refers to the linker and the carrier, if a carrier is present. Preferably, one of the pair R/R is H and the other one “Carrier-linked prodrug or “carrier prodrug': A carrier is selected from R. linked prodrug is a prodrug that contains a temporary 25 linkage of a given active Substance with a transient carrier Preferably, one of R/R is H. group that produces improved physicochemical or pharma Optionally, one or more of the pairs R/R, R/R", cokinetic properties and that can be easily removed in Vivo, R/R may independently form one or more cyclic usually by a hydrolytic cleavage. fragments selected from C, cycloalkyl, 4 to 7 mem "Cascade prodrug': A cascade prodrug is a carrier prod bered heterocyclyl, or 9 to 11 membered heterobicy 30 rug for which the cleavage of the carrier group becomes clyl. effective only after unmasking an activating group. Optionally, R, R, R and R* are further substituted; "Polymeric cascade prodrug: A polymeric cascade pro Suitable Substituents are alkyl (such as C. alkyl). drug is a carrier prodrug that contains a temporary linkage alkenyl (such as C. alkenyl), alkynyl (such as C of a given active Substance with a transient polymeric carrier alkynyl), aryl (Such as phenyl), heteroalkyl, heteroalk 35 group for which the cleavage of the carrier becomes effec enyl, heteroalkynyl, heteroaryl (such as aromatic 4 to 7 tive only after unmasking an activating group. membered heterocycle) or halogen moieties. “Bioprecursor prodrug': A bioprecursor prodrug is a ii) a moiety L, which is a chemical bond or a spacer, and Lif prodrug that does not imply the linkage to a carrier group, is bound to a carrier group Z, but results from a molecular modification of the active wherein L' is substituted with one to four (preferably one) 40 principle itself. This modification generates a new com Li moieties, pound, able to be transformed metabolically or chemically, Z is PEG or a hydrogel, more preferably Z is a hydrogel, the resulting compound being the active principle. even more preferably Z is a PEG-based hydrogel; “Biotransformation': Biotransformation is the chemical optionally, L is further substituted. conversion of Substances by living organisms or enzyme Suitable Substituents are alkyl (Such as C. alkyl), alkenyl 45 preparations. (such as C- alkenyl), alkynyl (Such as C- alkynyl), aryl The previous definitions are based on IUPAC, as given (such as phenyl), heteroalkyl, heteroalkenyl, heteroalkynyl, under http://www.chem.qmul.ac.uk/iupac/medchem/(ac heteroaryl (such as aromatic 4 to 7 membered heterocycle) cessed on 8 Mar. 2004) or halogen moieties. "Linker: Cleavage-controlling chemical structures or The present invention addresses the disadvantages 50 groups present in carrier prodrugs that are not provided by described above. The invention provides for carrier-linked either the carrier entity or by the drug. prodrugs characterized by connecting a carrier via a dipep “Sustained release' or “substained release rate” means tide linker to a primary or secondary amino group of an that the administration intervals of the respective prodrug aliphatic amine-containing drug molecule. The carrier is are expanded. Drugs with a daily dosage may for example linked to the dipeptide linker via a permanent linkage and 55 be turned into a Sustained release form with a week-long or the bond between the dipeptide promoiety and the amine even longer interval between two administrations. containing drug molecule is a temporary amide linkage that A strong in vivo/in vitro correlation is observed, if the exhibits extended autohydrolysis at a therapeutically useful release kinetics exhibited by a hydrogel prodrug conjugate rate at pH 7.4 and 37°C., i.e. under physiological conditions. according to the present invention has a half-life in vivo that Due to the presence of a permanent bond between the 60 is not smaller than half the value exhibited by the same carrier and the DKP linker, the prodrugs according to the hydrogel prodrug conjugate in aqueous buffer of pH 7.4 at present invention ensure release of unmodified native drug 370 C. molecules from a stable conjugate comprising carrier and “Cis-amide conformation inducer refers to a moiety that linker moiety. stabilizes the preceeding cis-amide bond. Suitable cis-amide It was now Surprisingly found, that aliphatic amide bonds 65 conformation inducers are, for example, pseudoprolines. can undergo autohydrolysis at a rate that is useful for “Aliphatic amine containing biologically active moiety carrier-linked prodrug applications if cyclization-activation D” means the part, e.g. the moiety or fragment, of the drug US 9,533,056 B2 11 12 linker conjugate D-L, which results after cleavage in the More preferably, Z is a biodegradable polyethylene glycol drug D-H, the active agent, of known biological activity. In based water-insoluble hydrogel. addition, the Subterm “aliphatic amine containing' means The term “water-insoluble” refers to a swellable three dimensionally crosslinked molecular network forming the that the respective moiety D and analogously the corre hydrogel. The hydrogel if suspended in a large Surplus of sponding drug D-H contains at least one aliphatic fragment, water or aqueous buffer of physiological osmolality may and which at least one aliphatic fragment is substituted with take up a Substantial amount of water, e.g. up to 10-fold on at least one amino group. a weight per weight basis, and is therefore swellable but “Non-biologically active linker” means a linker which after removing excess water still retains the physical stabil does not show pharmacological effects. ity of a gel and a shape. Such shape may be of any geometry “Biologically active moiety D' means the part of the drug 10 and it is understood that such an individual hydrogel object linker conjugate, which results after cleavage in a drug D-H is to be considered as a single molecule consisting of of known biological activity. components wherein each component is connected to each Suitable carriers are polymers and can either be directly other component through chemical bonds. conjugated to the linker or via a non-cleavable spacer. The The term “PEG' or “pegylation residue' is used herein 15 exemplary for suitable water-soluble polymers characterized term “prodrug according to the invention” refers to carrier by repeating units. Suitable polymers may be selected from linked prodrugs of biologically active agents, wherein the the group consisting of polyalkyloxy polymers, hyaluronic carrier is PEG or a hydrogel, preferably a PEG-based acid and derivatives thereof, polyvinyl alcohols, polyoxazo hydrogel. The terms “PEG prodrug”, “PEG-linked prodrug. lines, polyanhydrides, poly(ortho esters), polycarbonates, “hydrogel prodrug and “hydrogel-linked prodrug” refer to polyurethanes, polyacrylic acids, polyacrylamides, poly prodrugs of biologically active agents transiently linked to a acry-lates, polymethacrylates, polyorganophosphaZenes, PEG or to a hydrogel, respectively, and are used synony polysiloxanes, polyvinylpyrrolidone, polycyanoacrylates, mously. and polyesters. Preferred are polyalkyloxy polymers, espe The term “polyethylene glycol based” or “PEG based” as cially polyethylene glycol polymers containing at least 10% understood herein means that the mass proportion of PEG 25 by weight ethylene oxide units, more preferably at least 25% chains or in the hydrogel is at least 10% by weight, prefer by weight, even more preferably at least 50% by weight ably at least 25%, based on the total weight of the hydrogel. A “hydrogel may be defined as a three-dimensional, The remainder can be made up of other polymers. hydrophilic or amphiphilic polymeric network capable of Such other polymers are preferably selected from the taking up large quantities of water. The networks are com group consisting of for example, 2-methacryloyl-oxyethyl 30 posed of homopolymers or copolymers, are insoluble due to phosphoyl cholins, hydrogels, PEG-based hydrogels, poly the presence of covalent chemical or physical (ionic, hydro (acrylic acids), poly(acrylates), poly(acrylamides), poly phobic interactions, entanglements) crosslinks. The cross (alkyloxy) polymers, poly(amides), poly(amidoamines), links provide the network structure and physical integrity. poly(amino acids), poly(anhydrides), poly(aspartamides), Hydrogels exhibit a thermodynamic compatibility with 35 water which allows them to swell in aqueous media. The poly(butyric acids), poly(glycolic acids), polybutylene chains of the network are connected in Such a fashion that terephthalates, poly(caprolactones), poly(carbonates), poly pores exist and that a Substantial fraction of these pores are (cyanoacrylates), poly(dimethylacrylamides), poly(esters), of dimensions between 1 nm and 1000 nm. poly(ethylenes), poly(ethyleneglycols), poly(ethylene “Free form” of a drug refers to the drug in its unmodified, oxides), poly(ethyl phosphates), poly(ethyloxazolines), poly 40 pharmacologically active form, Such as after being released (glycolic acids), poly(hydroxyethyl acrylates), poly(hy from a polymer conjugate. droxyethyloxazolines), poly(hydroxymethacrylates), poly The terms “drug”, “biologically active molecule”, “bio (hydroxypropylmethacrylamides), poly(hydroxypropyl logically active moiety”, “biologically active agent”, “active methacrylates), poly(hydroxypropyloxazolines), poly(imi agent, and the like mean any Substance which can affect nocarbonates), poly(lactic acids), poly(lactic-co-glycolic 45 any physical or biochemical properties of a biological organ acids), poly(methacrylamides), poly(methacrylates), poly ism, including but not limited to viruses, bacteria, fungi, (methyloxazolines), poly(organophosphaZenes), poly(ortho plants, animals, and humans. In particular, as used herein, esters), poly(oxazolines), poly(propylene glycols), poly(si biologically active molecules include any Substance loxanes), poly(urethanes), poly(vinyl alcohols), poly(vinyl intended for diagnosis, cure, mitigation, treatment, or pre amines), poly(vinylmethylethers), poly(vinylpyrrolidones), 50 vention of disease in humans or other animals, or to other silicones, celluloses, carbomethyl celluloses, hydroxypropyl wise enhance physical or mental well-being of humans or methylcelluloses, chitins, chitosans, dextrans, dextrins, gela animals. tins, hyaluronic acids and derivatives, mannans, pectins, The terms “spacer” or “spacer moieties’ refer to any rhamnogalacturonans, starches, hydroxyalkyl starches, moiety suitable for connecting two moieties, such as Clso 55 alkyl, C-so alkenyl or Clso alkinyl, which fragment is hydroxyethyl starches and other carbohydrate-based poly optionally interrupted by one or more groups selected from mers, Xylans, and copolymers thereof. —NH-, -N (C. alkyl)-, —O— —S , —C(O)—, Suitable carriers can either be directly conjugated to the —C(O)NH , —C(O)N(C. alkyl)-, —O C(O)—, linker or via a non-cleavable spacer. The term “polymer —S(O)— —S(O). , 4 to 7 membered heterocyclyl phe prodrug” refers to carrier-linked prodrugs of a biologically 60 nyl or naphthyl. active agent, wherein the carrier is a polymer. "Functional groups' mean groups of atoms within mol The term polymer describes a molecule comprised of ecules that exhibit a specific chemical activity. Examples are repeating structural units connected by chemical bonds in a amides, amines, alcohols, carbonyls, carboxylic acids, linear, circular, branched, crosslinked or dendrimeric way or thiols. a combination thereof, which can be of synthetic or biologi 65 “Protective groups' refers to a moiety which temporarily cal origin or a combination of both. Typically, a polymer has protects a functional group of a molecule during synthesis to a molecular weight of at least 1 kDa. obtain chemoselectivity in Subsequent chemical reactions. US 9,533,056 B2 13 14 Protective groups for alcohols are, for example, benzyl and cycloalkyl ring also includes bridged bicycles like norbo trityl, protective groups for amines are, for example, tert nane (norbonanyl) or norbonene (norbonenyl). Accordingly, butyloxycarbonyl, 9-fluorenylmethyloxycarbonyl and ben "Cs cycloalkyl means a cycloalkyl having 3 to 5 carbon Zyl and for thiols examples of protective groups are 2.4.6- atOmS. trimethoxybenzyl, phenylthiomethyl, acetamidomethyl, "Halogen' means fluoro, chloro, bromo or iodo. It is p-methoxybenzyloxycarbonyl, tert-butylthio, triphenylm generally preferred that halogen is fluoro or chloro. ethyl, 3-nitro-2-pyridylthio. 4-methyltrity1. “4 to 7 membered heterocyclyl” or “4 to 7 membered “Protected functional groups' means a functional group heterocycle” means a ring with 4, 5, 6 or 7 ring atoms that protected by a protective group. may contain up to the maximum number of double bonds “Acylating agent’ means a moiety of the structure 10 (aromatic or non-aromatic ring which is fully, partially or R—(C=O)—, providing the acyl group in an acylation un-saturated) wherein at least one ring atom up to 4 ring reaction, optionally connected to a leaving group, Such as atoms are replaced by a heteroatom selected from the group acid chloride, N-hydroxy Succinimide, pentafluorphenol and consisting of Sulfur (including —S(O)— —S(O) ), oxy para-nitrophenol. gen and nitrogen (including=N(O)—) and wherein the ring 15 is linked to the rest of the molecule via a carbon or nitrogen “Alkyl means a straight-chain or branched carbon chain atom (unsubstituted 4 to 7 membered heterocyclyl). (unsubstituted alkyl). Optionally, each hydrogen of an alkyl Examples for a 4 to 7 membered heterocycles are azeti carbon may be replaced by a Substituent. dine, oxetane, thietane, furan, thiophene, pyrrole, pyrroline, “Heteroalkyl refers to analogs of alkyls in which one or imidazole, imidazoline, pyrazole, pyrazoline, oxazole, more than one methylene group is replaced by a heteroatom, oxazoline, isoxazole, isoxazoline, thiazole, thiazoline, iso Such as nitrogen, oxygen, Sulfur, phosphorus, or boron. If the thiazole, isothiazoline, thiadiazole, thiadiazoline, tetrahy methylene group is replaced by nitrogen, phosphorous or drofuran, tetrahydrothiophene, pyrrolidine, imidazolidine, boron, these heteroatoms may be further substituted. Suit pyrazolidine, oxazolidine, isoxazolidine, thiazolidine, iso able Substituents are alkyl, alkenyl, alkynyl, aryl, het thiazolidine, thiadiazolidine, Sulfolane, pyran, dihydropy eroalkyl, heteroalkenyl, heteroalkynyl, heteroaryl or halo 25 ran, tetrahydropyran, imidazolidine, pyridine, pyridazine, gen moieties (such as those described above). The terms pyrazine, pyrimidine, piperazine, piperidine, morpholine, heteroalkenyl and heteroalkynyl are defined accordingly. tetrazole, triazole, triazolidine, tetrazolidine, diazepane, "C. alkyl means an alkyl chain having 1 to 4 carbon azepine or homopiperazine. Optionally, each hydrogen of a atoms (unsubstituted C. alkyl), e.g. if present at the end of 4 to 7 membered heterocyclyl may be replaced by a sub a molecule: methyl, ethyl, n-propyl, isopropyl. n-butyl, 30 stituent. isobutyl, sec-butyl tert-butyl, or e.g. —CH2—, —CH2— “9 to 11 membered heterobicyclyl” or “9 to 11 membered CH , —CH(CH)—, CH2—CH2—CH2—, CH heterobicycle” means a heterocyclic system of two rings (CHs)— —C(CH) , when two moieties of a molecule with 9 to 11 ring atoms, where at least one ring atom is are linked by the alkyl group. Optionally, each hydrogen of shared by both rings and that may contain up to the maxi a C alkyl carbon may be replaced by a substituent. 35 mum number of double bonds (aromatic or non-aromatic Accordingly, "Clso alkyl means an alkyl chain having 1 to ring which is fully, partially or un-saturated) wherein at least 50 carbon atoms. The term C is defined accordingly. one ring atom up to 6 ring atoms are replaced by a heteroa "C-so alkenyl' means a branched or unbranched alkenyl tom selected from the group consisting of Sulfur (including chain having 2 to 50 carbon atoms (unsubstituted C-so —S(O)— —S(O) ), oxygen and nitrogen (including alkenyl), e.g. if present at the end of a molecule: 40 —N(O)—) and wherein the ring is linked to the rest of the -CH=CH, -CH=CH-CH, CH-CH=CH, molecule via a carbon or nitrogen atom (unsubstituted 9 to -CH=CH-CH CH-CH=CH-CH=CH, or e.g. 11 membered heterobicyclyl). —CH=CH , when two moieties of a molecule are linked Examples for a 9 to 11 membered heterobicycle are by the alkenyl group. Optionally, each hydrogen of a C-so indole, indoline, benzofuran, benzothiophene, benzoxazole, alkenyl carbon may be replaced by a substituent as further 45 benzisoxazole, benzothiazole, benzisothiazole, benzimida specified. Accordingly, the term “alkenyl relates to a carbon Zole, benzimidazoline, quinoline, quinazoline, dihydroqui chain with at least one carbon double bond. Optionally, one nazoline, quinoline, dihydroquinoline, tetrahydroquinoline, or more triple bonds may occur. The term C- alkenyl is decahydroquinoline, isoquinoline, decahydroisoquinoline, defined accordingly. tetrahydroisoquinoline, dihydroisoquinoline, benzazepine, "C-so alkynyl' means a branched or unbranched alkynyl 50 purine or pteridine. The term 9 to 11 membered heterobi chain having 2 to 50 carbon atoms (unsubstituted C-so cycle also includes spiro structures of two rings like 1,4- alkynyl), e.g. if present at the end of a molecule: —C=CH, dioxa-8-azaspiro4.5 decane or bridged heterocycles like —CH C=CH, CH, CH, C=CH, CH-C=C- 8-aza-bicyclo3.2.1]octane. Optionally, each hydrogen of a CH, or e.g. —C=C- when two moieties of a molecule are 9 to 11 membered heterobicyclyl may be replaced by a linked by the alkynyl group. Optionally, each hydrogen of a 55 substituent. Clso alkynyl carbon may be replaced by a substituent as In case the prodrugs according to the present invention further specified. Accordingly, the term “alkynyl relates to contain one or more acidic or basic groups, the invention a carbon chain with at lest one carbon carbon triple bond. also comprises their corresponding pharmaceutically or Optionally, one or more double bonds may occur. The term toxicologically acceptable salts, in particular their pharma C. alkynyl is defined accordingly. 60 ceutically utilizable salts. Thus, the prodrugs which contain “C., cycloalkyl or “C., cycloalkyl ring” means a cyclic acidic groups can be used according to the invention, for alkyl chain having 3 to 7 carbon atoms, which may have example, as alkali metal salts, alkaline earth metal salts or as carbon-carbon double bonds being at least partially saturated ammonium salts. More precise examples of Such salts (unsubstituted C-7 cycloalkyl), e.g. cyclopropyl, cyclobutyl, include Sodium salts, potassium salts, calcium salts, mag cyclopentyl, cyclohexyl, cyclohexenyl, cycloheptyl. Option 65 nesium salts or salts with ammonia or organic amines Such ally, each hydrogen of a cycloalkyl carbon may be replaced as, for example, ethylamine, ethanolamine, triethanolamine by a substituent. The term "C., cycloalkyl or “C., or amino acids. Prodrugs which contain one or more basic US 9,533,056 B2 15 16 groups, i.e. groups which can be protonated, can be present “Excipients’ refers to compounds administered together and can be used according to the invention in the form of with the therapeutic agent, for example, buffering agents, their addition salts with inorganic or organic acids. isotonicity modifiers, preservatives, stabilizers, anti-adsorp Examples for suitable acids include hydrogen chloride, tion agents, oxidation protection agents, or other auxiliary hydrogen bromide, phosphoric acid, Sulfuric acid, nitric agents. However, in some cases, one excipient may have acid, methanesulfonic acid, p-toluenesulfonic acid, naphtha dual or triple functions. lenedisulfonic acids, oxalic acid, acetic acid, tartaric acid, “Dry composition” means that the prodrug composition is lactic acid, salicylic acid, benzoic acid, formic acid, propi provided in a dry form in a container. Suitable methods for onic acid, pivalic acid, diethylacetic acid, malonic acid, drying are spray-drying and lyophilization (freeze-drying). 10 Such dry composition of prodrug has a residual water Succinic acid, pimelic acid, fumaric acid, maleic acid, malic content of a maximum of 10%, preferably less than 5% and acid, Sulfaminic acid, phenylpropionic acid, gluconic acid, more preferably less than 2% (determined according to Karl ascorbic acid, isonicotinic acid, citric acid, adipic acid, and Fischer). The preferred method of drying is lyophilization. other acids known to the person skilled in the art. If the “Lyophilized composition” means that the prodrug com prodrugs simultaneously contain acidic and basic groups in 15 position was first frozen and Subsequently Subjected to water the molecule, the invention also includes, in addition to the reduction by means of reduced pressure. This terminology salt forms mentioned, inner salts or betaines (Zwitterions). does not exclude additional drying steps which occur in the The respective salts of the prodrugs of the present invention manufacturing process prior to filling the composition into can be obtained by customary methods which are known to the final container. the person skilled in the art like, for example by contacting “Lyophilization' (freeze-drying) is a dehydration process, these with an organic or inorganic acid or base in a solvent characterized by freezing a composition and then reducing or dispersant, or by anion exchange or cation exchange with the Surrounding pressure and, optionally, adding heat to other salts. The present invention also includes all salts of allow the frozen water in the composition to sublime directly the prodrugs which, owing to low physiological compatibil from the Solid phase to gas. Typically, the Sublimed water is ity, are not directly suitable for use in pharmaceuticals but 25 collected by desublimation. which can be used, for example, as intermediates for chemi “Reconstitution' means the addition of a liquid to bring cal reactions or for the preparation of pharmaceutically back the original form of a composition. acceptable salts. “Reconstitution solution” refers to the liquid used to The term “pharmaceutically acceptable” means approved reconstitute the dry composition of a prodrug prior to by a regulatory agency, such as the EMEA (Europe) and/or 30 administration to a patient in need thereof. the FDA (US) and/or any other national regulatory agency “Container” means any container in which the prodrug for use in animals, preferably in humans. composition is comprised and can be stored until reconsti “Pharmaceutical composition” or “composition” means a tution. composition containing one or more active ingredients, for “Buffer or “buffering agent” refers to chemical com example a drug or a prodrug, and one or more inert ingre 35 pounds that maintain the pH in a desired range. Physiologi dients, as well as any product which results, directly or cally tolerated buffers are, for example, Sodium phosphate, indirectly, from combination, complexation or aggregation Succinate, histidine, bicarbonate, citrate and acetate, Sul of any two or more of the ingredients, or from dissociation phate, nitrate, chloride, pyruvate. Antacids such as Mg(OH)2 of one or more of the ingredients, or from other types of or ZnCO may be also used. Buffering capacity may be reactions or interactions of one or more of the ingredients. 40 adjusted to match the conditions most sensitive to pH Accordingly, the pharmaceutical compositions of the present stability. invention encompass any composition made by admixing a A “lyoprotectant” is a molecule which, when combined prodrug of the present invention and a pharmaceutically with a protein of interest, significantly prevents or reduces acceptable excipient. chemical and/or physical instability of the protein upon "Stable' and “stability” means that within the indicated 45 drying in general and especially during lyophilization and storage time the polymer conjugates remain conjugated and Subsequent storage. Exemplary lyoprotectants include Sug do not hydrolyze to a substantial extent and exhibit an ars. Such as Sucrose or trehalose; amino acids such as acceptable impurity profile relating to the biologically active monosodium glutamate or histidine; methylamines Such as agent. To be considered stable, the composition contains less betaine; lyotropic salts such as magnesium Sulfate; polyols than 10%, preferably less than 5% of the drug in its free 50 Such as trihydric or higher Sugar alcohols, e.g. glycerin, form. erythritol, glycerol, arabitol. Xylitol, Sorbitol, and mannitol; “Therapeutically effective amount’ means an amount ethylene glycol; propylene glycol, polyethylene glycol, sufficient to cure, alleviate or partially arrest the clinical pluronics; hydroxyalkyl starches, e.g. hydroxyethyl starch manifestations of a given disease and its complications. An (HES), and combinations thereof. amount adequate to accomplish this is defined as “therapeu 55 "Surfactant” refers to wetting agents that lower the sur tically effective amount. Effective amounts for each pur face tension of a liquid. pose will depend on the severity of the disease or injury as “Isotonicity modifiers' refer to compounds which mini well as the weight and general state of the subject. It will be mize pain that can result from cell damage due to osmotic understood that determining an appropriate dosage may be pressure differences at the injection depot. achieved using routine experimentation, by constructing a 60 The term “stabilizers' refers to compounds used to sta matrix of values and testing different points in the matrix, bilize the polymer prodrug. Stabilisation is achieved by which is all within the ordinary skills of a trained physician. strengthening of the protein-stabilising forces, by destabili Within the scope of this invention, therapeutically effective sation of the denatured state, or by direct binding of excipi amount relates to dosages that aim to achieve therapeutic ents to the protein. effect for an extended period of time, i.e. for 12 hours, or 24 65 "Anti-adsorption agents' refers to mainly ionic or non hours, or three days or longer, for instance one week or two ionic Surfactants or other proteins or soluble polymers used weeks. to coat or adsorb competitively to the inner surface of the US 9,533,056 B2 17 18 composition’s container. Chosen concentration and type of In case of diketopiperazine-activated prodrug cleavage, excipient depends on the effect to be avoided but typically the amine-containing nucleophile serves to attack the pro a monolayer of Surfactant is formed at the interface just drug amide carbonyl group and consequently induces above the CMC value. transamidation, and the permanent amide bond serves to “Oxidation protection agents’ refers to antioxidants such form a stabilized six-membered ring structure. as ascorbic acid, ectoine, glutathione, methionine, monoth The formation of the stabilized six-membered ring struc ioglycerol, morin, polyethylenimine (PEI), propyl gallate, ture is facilitated through a cis-amide conformation inducing Vitamin E, chelating agents such aus citric acid, EDTA, pseudoproline. Pseudoprolines are artificially created dipep hexaphosphate, thioglycolic acid. tides, which contain an oxazolidine or thiazolidine ring. In "Antimicrobial refers to a chemical substance that kills 10 peptide synthesis, pseudoprolines are used to increase sol or inhibits the growth of microorganisms, such as bacteria, vation and solubility. Due to the preference for a cis-amide fungi, yeasts, protozoans and/or destroys viruses. bond with the preceding residue of C2-substituted pseudo “PEG based as understood herein means that the mass prolines, their incorporation results in a kink conformation proportion of PEG chains in the hydrogel is at least 10% by of the peptide backbone which decreases aggregation, self weight, preferably at least 25%, based on the total weight of 15 association and B-structure formation. the hydrogel. The remainder can be made up of other spacers Preferred linker structures are composed of a dipeptide and/or oligomers or polymers, such as oligo- or polylysines. promoiety conjugated through a permanent linkage to a The term “hydrolytically degradable' or “biodegradable' polymer carrier. Corresponding prodrugs are composed of a refers within the context of the present invention to linkages dipeptide containing a permanent linkage to a polymer which are non-enzymatically hydrolytically degradable carrier and a temporary amide bond to an aliphatic amino under physiological conditions (aqueous buffer at pH 7.4. group-containing drug. 37° C.) with half-lives ranging from one hour to three Preferably, linkers of the present invention have a hydro months, include, but are not limited to, aconityls, acetals, lysis rate between 1 h and 2 years at pH 7.4 and 37° C. and carboxylic anhydrides, esters, imines, hydrazones, maleamic hydrolysis rates in buffer and plasma are essentially identi acid amides, ortho esters, phosphamides, phosphoesters, 25 cal, i.e. the hydrolysis rates exhibit a strong in vivo/in vitro phosphosilyl esters, silyl esters, Sulfonic esters, aromatic correlation. carbamates, combinations thereof, and the like. Preferred Preferably, D-H is a small molecule bioactive agent or a biodegradable linkages are esters, carbonates, phosphoesters biopolymer. and Sulfonic acid esters and most preferred are esters or Preferably, D-H is a biopolymer selected from the group carbonates. It is understood that for in vitro studies accel 30 of biopolymers consisting of proteins, polypeptides, oligo erated conditions like, for example, pH 9, 37° C., aqueous nucleotides, and peptide nucleic acids. buffer, may be used for practical purposes. “Oligonucleotides” means either DNA, RNA, single stranded or double-stranded, siRNA, miRNA, aptamers, and DETAILED DESCRIPTION OF EMBODIMENTS any chemical modifications thereof with preferably 2 to 35 1000 nucleotides. Modifications include, but are not limited It is to be understood that the descriptions of the present to, those which provide other chemical groups that incor invention have been simplified to illustrate elements that are porate additional charge, polarizability, hydrogen bonding, relevant for a clear understanding of the present invention, electrostatic interaction, and fluxionality to the nucleic acid while eliminating, for purposes of clarity, many other ele ligand bases or to the nucleic acid ligand as a whole. Such ments which are conventional in this art. Those of ordinary 40 modifications include, but are not limited to. 2'-position skill in the art will recognize that other elements are desir Sugar modifications, 5-position pyrimidine modifications, able for implementing the present invention. However, 8-position purine modifications, modifications at exocyclic because Such elements are well known in the art, and amines, substitution of 4-thiouridine, substitution of because they do not facilitate a better understanding of the 5-bromo or 5-iodo-uracil; backbone modifications, methyl present invention, a discussion of Such elements is not 45 ations, unusual base-pairing combinations such as the iso provided herein. bases isocytidine and isoguanidine and the like. Modifica The present invention will now be described in detail on tions can also include 3' and 5' modifications such as capping the basis of exemplary embodiments. and change of stereochemistry. In the present invention, the hydrolytic lability required Preferably, D-H is a polypeptide selected from the group for a temporary linkage may be introduced into the prodrug 50 of polypeptides consisting of ACTH, adenosine deaminase, amide bond by selecting the structural properties of the agallsidase, alfa-1 antitrypsin (AAT), alfa-1 proteinase linker for cyclization activation. In cyclization-activated inhibitor (API), alteplase, amylins (amylin, symlin), anis amide bond cleavage, the cleavage products are a free amine treplase, ancrod serine protease, antibodies (monoclonal or as part of the biologically active moiety and a cyclized polyclonal, and fragments or fusions), antithrombin III, residue. The linker structures of the present invention are 55 antitrypsins, aprotinin, asparaginases, atosiban, biphalin, designed Such that highly stable rings are formed as cleav bivalirudin, bone-morphogenic proteins, bovine pancreatic age products and the hydrolysis of the prodrug amide bond trypsin inhibitor (BPTI), cadherin fragments, calcitonin facilitates hydrolysis in a time range useful for drug delivery (salmon), collagenase, complement C1 esterase inhibitor, under physiological conditions. Preferred cyclic cleavage conotoxins, cytokine receptor fragments, DNase, dynor products are diketopiperazine rings. Prerequisite for Such 60 phine A, endorphins, enfuvirtide, enkephalins, erythropoi cyclization activation is the presence of an amine-containing etins, exendins, factor VII, factor VIIa, factor VIII, factor nucleophile in the linker structure and another amide bond VIIIa, factor IX, fibrinolysin, fibroblast growth factor (FGF), which is not the temporary amide prodrug bond but a growth hormone releasing peptide 2 (GHRP2), fusion pro permanent amide bond. Preferably, such linker structures teins, follicle-stimulating hormones, gramicidin, ghrelin, contain a cis-amide conformation inducer. Alternatively, the 65 desacyl-ghrelin, granulocyte colony stimulating factor cleavage might occur through intramolecular catalysis (G-CSF), galactosidase, glucagon, glucagon-like peptides, caused by neighbouring group effects. glucocerebrosidase, granulocyte macrophage colony stimu US 9,533,056 B2 19 20 lating factor (GM-CSF), human heat shock proteins (HSP), ride, 13-Deoxydoxorubicin hydrochloride, 17-Amino-17 phospholipase-activating protein (PLAP), gonadotropin demethoxygeldanamycin, 17-Aminogeldanamycin, 19-O- chorionic (hCG), hemoglobins, hepatitis B vaccines, hiru Methylgeldanamycin, 1-Methyl-D-tryptophan, din, human serine protease inhibitor, hyaluronidases, idurn 21-Aminoepothilone B, 2-Aminoaristeromycin, 2-Aminon onidase, immune globulins, influenza vaccines, interleukins eplanocin A, 3-Chloroprocainamide, 3-Deazaadenosine, (1 alfa, 1 beta, 2, 3, 4, 6, 10, 11, 12, 13, 21), IL-1 receptor 3-MATIDA, 447-480 Human alpha-fetoprotein, 4-Amin antagonist (rhIL-1 ra), insulins, insulin like growth factors, opyridine, 4-Aminosalicylic acid, 4-Chlorophenylthio insulin-like growth factor binding protein (rhIGFBP), inter DADMe-immucillin-A, 4-epi-Adriamycin, 4'-epi-Doxoru ferons (alfa 2a, alfa 2b, alfa 2c, beta 1a, beta 1b, gamma 1a, bicin, 5,4'-Diepiarbekacin, 5-Aminosalicylic acid, 5-Aza-2'- gamma 1b), intracellular adhesion molecule, keratinocyte 10 deoxycytidine, 5-azacitidine, 5'-Homoneplanocin A, growth factor (KGF), P-selectin glycoprotein ligand 6'-Homoneplanocin A, 8(R)-Fluoroidarubicin hydrochlo (PSGL), transforming growth factors, lactase, leptin, leu ride, 99mTc-c(RGDfK*)2HYNIC, 9-Aminocamptothecin, prolide, levothyroXine, luteinizing hormone, lyme vaccine, A-42867 pseudoaglycone, Abacavir Succinate, Abacavir Sul natriuretic peptides (ANP BNP. CNP and fragments), neu fate, Abanoquil mesilate, Abarelix, Acadesine, Acetyldina ropeptide Y, pancrelipase, pancreatic polypeptide, papain, 15 line, Acetylsalicylic acid lysine salt, Aciclovir, Acriflavine, parathyroid hormone, PDGF, pepsin, peptide YY. platelet Actinomycin D, Acycloguanosine, Acyclovir, Acyclovir activating factor acetylhydrolase (PAF-AH), prolactin, pro elaidate, Acyclovir oleate, Acyline, AD Peptide, Adaman tein C, thymalfasin, octreotide, secretin, sermorelin, Soluble tamine hydrochloride, Adamplatin-IV, Adefovir, Adefovir tumor necrosis factor receptor (TNFR), superoxide dis dipivoxil, Ademetionine tosylate sulfate, Adenallene, mutase (SOD), Somatropins (growth hormone), Somatoprim, Adenophostin A, Adenophostin B, Adenosine, Aerothricin 1, Somatostatin, streptokinase. Sucrase, terlipressin, tetanus Aerothricin 16, Aerothricin 41, Aerothricin 45, Aerothricin toxin fragment, tilactase, thrombins, thymosin, thyroid 5, Aerothricin 50, Aerothricin 55, Afamelanotide, Afloqua stimulating hormone, thyrotropin, tumor necrosis factor lone, Ageliferin diacetate, Ageliferin dihydrochloride, Aica (TNF). TNF receptor-IgG Fc, tissue plasminogen activator riboside, ALA hexyl ester, ALA Me ester, Aladapcin, Alami (tPA), TSH, urodilatin, urate oxidase, urokinase, vaccines, 25 fovir, Alatrofloxacin mesilate, Albolabrin, Alendronate vascular endothelial growth factor (VEGF), vasoactive Sodium, Alendronic acid sodium salt, Alestramustine, Alfu intestinal peptide, Vasopressin, Ziconotide, lectin and ricin. Zosin hydrochloride, Aliskiren fumarate. Alloferon-1, Alo Preferably, D-H is a protein prepared by recombinant gliptin benzoate, alpha-Difluoromethylornithine hydrochlo DNA technologies. ride, alpha-Human atrial natriuretic polypeptide, alpha Preferably, D-H is a protein selected from the group of 30 Methylnorepinephrine, alpha-Methyltryptophan, proteins consisting of antibodies, antibody fragments, single Altemicidin, Alvespimycin hydrochloride, Amantadine chain antigen binding proteins, catalytic antibodies and hydrochloride, Ambasilide, Ambazone, Ambroxol nitrate, fusion proteins. Amdoxovir, Ameltolide, Amelubant, Amezinium methylsul More preferably, D-H is a protein selected from the group fate, Amfenac sodium, Amidox, Amifostine hydrate, Ami of proteins consisting of antibody fragments, single chain 35 kacin, Amiloride hydrochloride, Aminocandin, Aminocap antigen binding proteins, catalytic antibodies and fusion roic acid, Aminoglutethimide, Aminoguanidine, proteins. Aminolevulinic acid hexyl ester, Aminolevulinic acid Preferably, D-H is a small molecule bioactive agent hydrochloride, Aminolevulinic acid methyl ester, Aminoqui selected from the group of agents consisting of central nuride, Aminosidine, Amisulpride, Amlexanox, Amlodipine, nervous system-active agents, anti-infective, anti-allergic, 40 Amlodipine besylate. AmoxanoX, Amoxicillin, Amoxicillin immunomodulating, anti-obesity, anticoagulants, antidi trihydrate. Amoxycillin trihydrate, Amphotericin, Ampho abetic, anti-neoplastic, antibacterial, anti-fungal, analgesic, tericin B, Ampicillin sodium, Amprenavir, Ampydin, Amri contraceptive, anti-inflammatory, steroidal, vasodilating, none, Amrubicin hydrochloride, Amselamine hydrobro vasoconstricting, and cardiovascular agents with at least one mide, Amthamine, Anakinra, Anamorelin hydrochloride, primary or secondary amino group. 45 Anatibant mesilate, Anginex, Angiopeptin acetate, Angio Preferably, D-H is a small molecule bioactive agent tensin II (human), Anisperimus, Antagonist-G, Antide, selected from the group of agents containing at least one Antide-1, Antide-2, Antide-3, Antiflammin-1, Antiflammin aliphatic primary amine group: (-)-Draflazine, (-)-Indocar 10, Antiflammin-2, Antiflammin-3, Antiflammin-4, Anti bazostatin B, (+)-23.24-Dihydrodiscodermolide, (+)-Disco flammin-5, Antiflammin-6, Antiflammin-7, Antiflammin-8, dermolide, (+)-R-Pramipexole, (R)-(+)-Amlodipine, (R)- 50 Antiflammin-9, Antileukinate, Antocin II. Apadenoson, (+)-TeraZosin, (R)-Ganciclovir cyclic phosphonate, (R)- Apcitide technetium (99mTc), Aphidicolin glycinate, Apixa Sulfinosine, (R)-Zacopride, (S)-(+)-Ketoprofen trometamol, ban, Aplonidine hydrochloride, Apoptozole 1, Apoptozole 1, (S)-Norfluoxetine, (S)-Oxiracetam, (S)-Sulfinosine, (S)-Za Apoptozole 2, Apoptozole 3, Apraclonidine hydrochloride, copride hydrochloride, 111 In-DTPA-Pro1, Tyrabombesin, Apricitabine, Arbekacin, Arbekacin Sulfate, Arborcandin A, 90Y-DOTAGA-substance P, 99TcDemobesin 3, 99Tc 55 Arborcandin B, Arborcandin C, Arborcandin D, Arborcan Demobesin 4, Ala11, D-Leu15Orexin B, Arg(Me)9] din E, Arborcandin F. Arenicin, Arenicin-1, Arenicin-2, MS-10, D11G.K26R,Y40YR-Plecta sin, D11G.M13K, Argatroban monohydrate, Argimesna, Arginine butyrate, K26R.Y40YR-Plecta sin, D9N.M13L.Q14R-Plectasin, Argiopine, Argiotoxin-636, Argipidine, Arotinolol hydro D9SQ14K.V36L-Plectasin, D-Tyr1Arg(Me)9 MS-10, chloride, Arterolane maleate, Asp(B14)-relaxin, Aspoxicil D-Tyr1AzaGly7. Arg(Me)9 MS-10, D-Tyr1 MS-10, 60 lin, Astromicin Sulfate, Atenolol, Atosiban, Atreleuton, G1n30-Pancreatic polypeptide (2-36), Glu10.Nle17, Atrial natriuretic factor (99-126), Avizafone, Avorelin, Nle30-Pancreatic polypeptide (2-36), Glut O-Pancreatic AZacitidine, AZacytidine, AZalanstat, AZaromycin SC, AZel polypeptide(2-36), L17K.K30R GLP-2 (1-33), Leu13 nidipine, AZetirelin, AZodicarbonamide, AZOxybacilin, Motilin, N5R,M13Y,N17R-Plectasin, Niel 7.Nle30-Pan Aztreonam, Aztreonam L-lysine, AZtreonam lysinate, AZu creatic polypeptide(2-36), psiCH2NHITpg4Vancomycin 65 mamide A, Baclofen, Bactobolin, Balapiravir hydrochloride, aglycon, Ser12-Humanin, Trp 19 MS-10, Tyr24-Huma Balhimycin, Baogongteng A, Barusiban, Batracylin, nin, 111 In-Pentetreotide, 13-Deoxyadriamycin hydrochlo Batroxostatin, Belactin A, Belactosin A, Belactosin C, US 9,533,056 B2 21 22 Benanomicin B, Benexate cyclodextrin, Benzocaine, Besi fosol tetrasodium, Deoxymethylspergualin, Deoxynegamy floxacin hydrochloride, Binodenoson, Bivalirudin, Bleomy cin, Deoxyspergualin hydrochloride, Deoxy variolin B, cin A2 sulfate, Boceprevir, Body protection compound-15, Desacetylvinblastinehydrazide/folate conjugate. Desferriox Bogorol A, Boholmycin, Brain natriuretic peptide, Brasili amine, des-F-sitagliptin, Desglugastrin tromethamine, cardin A, Bremelanotide, Brivanibalaninate, Brivaracetam, Deslorelin, Desmopressin acetate. Desulfated hirudin (54 Brodimoprim, Bromfenac sodium, Bromhexine hydrochlo 65), Desulfated hirugen, Detiviciclovir diacetate, Dexamfe ride, Brostallicin hydrochloride, B-Type natriuretic peptide, tamine sulfate, Dexamphetamine sulfate, DexelVucitabine, Bunazosin hydrochloride, Buserelin acetate. Butabindide, Dexibuprofen lysine, Dexketoprofen D.L-lysine, Dexketo Butamidine, Buteranol, Cabin 1, Caerulein diethylamine, profen imidazole salt, Dexketoprofen lysine, Dexketoprofen Calcium folinate, Calcium-like peptide 1, Calcium-like pep 10 trometamol, Dexormaplatin, Dextroamphetamine Sulfate, tide 2, Cambrescidin 800, Cambrescidin 816, Cambrescidin Dextronatrin, Dezinamide, Dezocitidine, Diadenosine tetra 830, Cambrescidin 844, Camostat mesilate, Camostat mesy phosphate, Diaveridine, Dichlorobenzoprim, Dicloguamine late, Canfosfamide hydrochloride, Capadenoson, Capeserod maleate, Didemnin X, Didemnin Y. Dideoxycytidine, hydrochloride, Capimorelin, Capravirine, CapraZamycin A, Difurazone, Dilevalol, Dilevalol hydrochloride, Dirucotide, CapraZamycin B, CapraZamycin C, CapraZamycin E, 15 Disagregin, Discodermolide, Disermolide, Disitertide, Diso CapraZamycin F. Capromorelin, Cap Savanil, Carafiban dium pamidronate, Disopyramide phosphate, di-Val-L-dC. maleate, Carbachol, Carbamazepine, Carbetocin, Carbovir, Docosylcidofovir, Dolastatin 14, Dolastatin C, Donitriptan Carboxyamidotriazole, Cariporide mesilate, Carisbamate, hydrochloride, Donitriptan mesilate, Doripenem, Dovitinib Carnosine Zinc complex (1:1), Carperitide, Carpipramine, Lactate, Doxazosin mesylate, Doxorubicin hydrochloride, Carumonam Sodium, Caspofungin acetate, Cavtratin, Cecro Doxycycline hyclate, Doxycycline hydrochloride ethanol pin A(1-11) D(12-37), Cecropin D, Cefaclor, Cefalexin hydrate, D-Penicillamine, Draflazine, Droxidopa, DTPA monohydrate, Cefcamate pivoxil hydrochloride, Cefcanel adenosylcobalamin, d-trans-Tetraplatin, Dumorelin, daloxate hydrochloride, Cefcapene pivoxil hydrochloride, Duramycin, Dyofin-1, Dyofin-2, Dyofin-9, Ebrotidine, Cefdaloxime, Cefdaloxime Pentexil Tosilate, Cefdinir, Ecenofloxacin hydrochloride, Echistatin, Edotreotide Cefditoren pivoxil, Cefepime, Cefetamet pivoxil, Cefetecol, 25 yttrium, Efegatran sulfate hydrate, Eflornithine hydrochlo Cefixime, Cefluprenam, Cefnmatilen hydrochloride hydrate, ride, Eglumegad hydrate, Eglumetad hydrate, Eicosylcido Cefnmenoxime hydrochloride, Cefninox sodium, Cefodiz fovir, Elacytarabine, Elaidic acid-Cytarabine, Elastatinal B, ime, Cefodizime sodium, Cefoselis sulfate, Cefotaxime Elastatinal C, Elpetrigine, Eltrombopag olamine, Elvucit sodium, Cefotetan disodium, Cefotiam cilexetil, Cefotiam abine, Emoxyl, Emtricitabine, Enalkiren, Endothelin, cilexetil hydrochloride, Cefotiam hexetil, Cefotiam hexetil 30 Endothelin 1, Enfluvirtide, Enigmol, Eniporide mesilate, hydrochloride, Cefotiam hydrochloride, Cefoxitin, Cefozo Entecavir, Enteric neural peptide, Entinostat, Epidoxorubi pran, CefoZopran hydrochloride, Cefoirome, Cefpodoxime cin, Epinastine hydrochloride, Epiroprim. Epirubicin hydro proxetil, Cefprenam, Cefprozil, Cefprozil monohydrate, chloride, Epithalon, Epofolate, Epostatin, Epsilon amin Cefauinome, Cefsulodin sodium, Ceftaroline, Ceftazidime, ocaproic acid, Eptaplatin, Eptifibatide, Eremomycin, Cefiteram pivoxil, Ceftibuten, Ceftizoxime alapivoxil, 35 Eribulin mesilate, Eribulin mesylate, Erucamide, Esafloxa Ceftobiprole, Ceftobiprole mediocaril, CeftraZonal bopentil, cin hydrochloride, Eslicarbazepine acetate, Etaquine, Etha CeftraZonal sodium, Ceftriaxone sodium, Cefuroxime, nolamine, Ethanolamine oleate, Ethiofos (former USAN), Cefuroxime axetil, Cefuroxime pivoxetil, Centanamycin, Ethyl aminobenzoate, Ethylthio-DADMe-immucillin-A, Cephalexin monohydrate, Ceranapril, Ceronapril, Cerulein, Ethynylcytidine, Etiracetam levo-isomer, Etravirine, Ceruletide diethylamine, Cetefloxacin, Cetrorelix acetate, 40 Etriciguat, Eurocin, Exalamide. Examorelin, EXatecan mesi Chlorofusin, Chloroorienticin A, Chloroorienticin B, Chlo late, Exenatide, Exenatide LAR, Exendin-4, Ezatiostat rotetain, Cibrostatin 1, Ciclopiroxolamine, Cidofovir, Cilas hydrochloride, Famciclovir, Famotidine, Famotidine bis tatin Sodium, Cilastatino, Cilengitide, Cimaterol, Cinitapride muth citrate, Fampridine, Favipiravir, Feglymycin, Fegly hygrogen tartrate, Cinnamycin, Cipamfylline, Circinamide, mycine, Felbamate, Felbinac lysine salt, Fenleuton, Fidar Cisapride hydrate, Cispentacin, Citicoline, Citrullimycine A, 45 estat, Fidexaban, Filaminast, Filarizone, Fingolimod Cladribine, Clavanin A(K), Clavanin E(3-23), Clitocine, hydrochloride, Fish amunine, Flucytosine, Fludarabine Clofarabine, Clopidogrel sulfate, Colivelin, Conantokin-R, phosphate, Fluorobenzyltriamterene, Fluorocytosine, Fluo Contulakin G, Cortagine, Coumamidine gammal, Couma rominoxidil, Fluoroneplanocin A. Flupirtine maleate, Flu midine gamma2, Cromoglycate lisetil hydrochloride, cyclic virucin B2, Fluvoxamine maleate, Folinic acid, Folinic acid Cidofovir, Cycloplatam, Cycloserine, Cyclotheonamide A, 50 calcium salt, Fortimicin A, Fosamprenavir calcium, Fosam Cyclothialidine, Cycloviolin A, Cycloviolin B, Cycloviolin prenavir Sodium, Fosaprepitant dimeglumine, Fosfomycin C, Cycloviolin D, Cygalovir, Cypemycin, CySmethynil. trometamol, Fosfomycin tromethamine, Fosteabine sodium Cystamidin A, Cystamine, CystaZosin, Cystocin, Cytallene, hydrate, Fradafiban, Freselestat, Frog neuromedin U. Fro Cytarabine, Cytarabine ocfosfate, Cytaramycin, Cytochlor, Vatriptan, Fudosteine, Furamidine, G1 peptide, Gabadur, Cytomodulin, Dabigatran, Dabigatran etexilate, DACH-Pt 55 Gabapentin, Gabexate mesilate, Galarubicin hydrochloride, (II)-bis-ascorbate, Dacopafant, Dactimicin, Dactinomycin, Gallinacin 1, Gallinacin 1alpha, Gallinacin 2, Galmic, Gal Dactylocycline A, Dactylocycline B, DADMe-Immucillin non, Galparan, Gammaphos, Ganciclovir, Ganciclovir G. Dalargin, D-allo-Ileu3 PYY(3-36), Danegaptide hydro elaidic acid, Ganciclovir monophosphate, Ganciclovir chloride, Daniquidone, Dapropterin dihydrochloride, Dap Sodium, Ganirelix, Ganirelix acetate, Garomefrine hydro sone, Darbufelone mesilate, Darifenacin hydrobromide, 60 chloride, Gemcitabine, Gemcitabine elaidate, Gemifloxacin Darinaparsin, Darunavir, Daunomycin, Daunorubicin, mesilate, Gibbosin, Gilatide, Giracodazole, Girodazole, Davasaicin, Davunetide, D-Cycloserine, Debrisoquin sul Girolline, Glaspimod, Glucagon-like peptide I (7-37), Glu fate, Debrisoquine Sulfate, Decahydromoenomycin A, Deca cosamine Sulfate, Gludopa, Glufanide, Glutathione mono planin, Decitabine, Declopramide, Deferoxamine, Degarelix ethyl ester, Glutathione monoisopropyl ester, Glycine-pro acetate, Dekafin 1, Dekafin 10, Delafloxacin, delta-Amin 65 line-Melphalan, Glycopin, Glycothiohexide alpha, olevulinic acid hydrochloride, Deltibant, Deltorphin E, Golotimod, Goralatide, Goserelin, Growth factor antago Denagliptin hydrochloride, Denibulin hydrochloride, Denu nist-116. Growth hormone releasing peptide 2, Growth US 9,533,056 B2 23 24 Inhibitory Peptide, Guanabenz, acetate, Guanadrel sulfate, Mexiletine hydrochloride, Micafungin sodium, Micronomi Guanethidine monosulfate, Guanfacine hydrochloride, Gus cin sulfate, Midalcipran hydrochloride, Midaxifylline, perimus hydrochloride, Gusperimus trihydrochloride, Mideplanin, Midoriamin, Milacainide tartrate, Milacemide Habekacin, Habekacin sulfate, Halovir A, Halovir B, Halo 2H), Milnacipran hydrochloride, Minamestane, Minocy Vir C. Halovir D. Halovir E. Hayumicin B, Hayumicin C1, cline hydrochloride, Minoxidil, Mirabegron, Miriplatin Hayumicin C2, Hayumicin D, Helvecardin A, Helvecardin hydrate, Mitomycin, Mitomycin C, MivaZerol, Mivobulin B, Hepavir B, Heptaminol AMP amidate, Heptaplatin, isethionate, Mizoribine, Mocetinostat dihydrobromide, Hexa-D-Arginine, Hexadecyl cidofovir, Hexadecyloxypro Modafinil, Modafinil sulfone, Moenomycin A chloride bis pyl-cidofovir, Hexaminolevulinate, Hexyl aminolevulinate, muth salt, Mofegiline, Mofegiline hydrochloride, Monami Hirudin desulfated, Hirulog-1. Histamine dihydrochloride, 10 docin, Monodansyl cadaverine, Monoethanolamine oleate, Histaprodifen. Histrelin, Histrelin acetate, Human adrenom Montirelin tetrahydrate, Mosapride citrate, Moxilubant, edulin, Human adrenomedullin (22-52), Human angio Moxilubant maleate, Mozenavir mesilate, m-Phenylene tensin II, Human corticotropin-releasing hormone, Human ethynylene, mu-Conotoxin IIIA, Multiple sclerosis vaccine, lactoferrin (1-11), Human proislet peptide, Human Secretin, muO-Conotoxin MrVIB, Muraminomicin A, Muraminomi Hydrostatin A, Hydroxyakalone, Hydroxycarbamide, 15 cin B, Muraminomicin C, Muraminomicin D, Muraminomi Hydroxyurea, Hypeptin, Ibutamoren mesilate, Icatibant cin E1, Muraminomicin E2, Muraminomicin F. Muramin acetate, Iclaprim, Icofungipen, Idarubicin hydrochloride, omicin G. Muraminomicin H. Muraminomicin I. Ilatreotide, Ilonidap, Imetit, Imidafenacin, Imidazenil, Imi Muraminomicin Z1, Muraminomicin Z2, Muraminomicin quimod, Immunosine, Impentamine, Incyclinide, Indano Z3, Muraminomicin Z4, Muramyl dipeptide C. Mureidomy cine, Indantadol hydrochloride, Indium In 111 pentetreotide, cin A, Mureidomycin B. Mureidomycin C, Mureidomycin Indolicidin-11, Indolicidin-4, Indolicidin-8, Indomethacin D, Muroctasin, Mycestericin E. Myriocin, Nafamostat mesi trometamol, Indomethacin tromethamine, Indoxam, Inoga late, Nafamo.stat mesylate, Nafarelin acetate, Naglivan, tran, Insulin chain B (9-23) peptide, Intrifiban, Iobenguane Nagrestipen, Namitecan, Naproxen piperazine (2:1), Napsa 131I. Iodorubidazone (p), Iotriside, Irsogladine maleate, gatran, Neboglamine, Nebostinel, Nebracetam fumarate, Isatoribine, Iseganan hydrochloride, Isepamicin Sulfate, Iso 25 Nelarabine, Neldazosin, Nelzarabine, Nemifitide ditriflutate, batzelline A, Isobatzelline B, Isobatzelline C, Isobatzelline Nemonoxacin, Neo-acridine, Neomycin B-arginine conju D, Isobutyramide, Isodoxorubicin, Isopropamide iodide gate, Neomycin B-hexaarginine conjugate, Neomycin-acri Neramexane Iturelix, Janthinomycin A, Janthinomycin B, Janthinomycin hydrochloride, Neridronate, Neridronic acid, Nesiritide, C, Jaspine B, K9-Retrocyclin-1, Kahalalide F, Kaitocepha 30 Netamiftide trifluoroacetate, Netilmicin sulfate, Neurome lin, Kanamycin, Kanamycin A. kappa-Conotoxin P VIIA, din U-25, Neuropeptide S. Neutrophil-activating factor, Karnamicin B1, Kassinatuerin-1, Katanosin A, Katanosin B, Niacinamide, Nicotinamide, Niduline, Nisin, Nitrovin, Katanosin B. Ketoprofen lysine, Ketorolac trometamol. Nocathiacin I, Nocathiacin II, Nocathiacin III, Nocathiacin Ketorolac tromethamine, Kistamicin A, L-4-Oxalysine, IV, NO-Gabapentin, Nolatrexed dihydrochloride, NO-Me Labetalol hydrochloride, Labradimil, Ladakamycin (for 35 salamine, Noraristeromycin, Nuvanil, O6-Benzylguanine, merly), Lagatide, Lamifiban, Lamivudine, Lamotrigine, Ocimumoside A, Octacosamicin A, Octacosamicin B, Oct Lanicemine 2CS)-hydroxySuccinate, Lanicemine hydrochlo reother, Octreotide acetate, Octreotide LAR, Oglufanide ride, Lanomycin, Lanreotide acetate, Lanthiopeptin, Lara disodium, Olamufloxacin, Olamufloxacin mesilate, Zotide acetate, Lazabemide hydrochloride, L-DOPA 3-O- Olcegepant, Olradipine hydrochloride, Omaciclovir, glucoside, L-DOPA 4-O-glucoside, L-Dopa methyl ester 40 Ombrabulin, Ombrabulin hydrochloride, omega-Conotoxin hydrochloride, L-Dopamide, Lecirelin, Leconotide, CVID, omega-Conotoxin MVIIA, Omiganan pentahydro Lenalidomide, Lenampicillin hydrochloride, Leucettamine chloride, Onnamide A, Opiorphin, Orbofiban acetate. Ori A, Leucovorin calcium, Leuprolide acetate, Leuprorelin enticin A, Orienticin B, Orienticin C, Orienticin D. Orita acetate, Leurubicin, Leustroducsin A, Leustroducsin B, vancin, Oseltamivir carboxylate, Oseltamivir phosphate, Leustroducsin C. Leustroducsin H. Levetiracetam, 45 Otamixaban, Otenabant hydrochloride, Ovothiol A, Oxali Levodopa, Levodopa 3-O-glucoside, Levodopa 4-O-gluco platin, Oxalysine-L, Oxazofurin, Oxcarbazepine, Oxigluta side, Levodopa methyl ester hydrochloride, Levofolinate tione sodium, Oxiracetam, OXolide, Oxynor, Oxyphenars calcium, Levoleucovorin calcium, Levonadifloxacin argi ine, Oystrisin, Ozarelix, Pachastrissamine, Pachymedusa nine salt, L-Histidinol, L-Homothiocitruline, Liblomycin, dacnicolor Tryptophyllin-1, Paecilaminol, Pafuramidine Linagliptin, Lingual antimicrobial peptide, Linifanib, Lin 50 maleate, Palau'amine, Paldimycin B, Pamidronate sodium, topride, Liraglutide, Lirexapride, Lirimilast, Lisdexamfet p-Aminoclonidine hydrochloride, Pancopride, Papuamide amine mesilate, Lisinopril, L-Lysine-d-amphetamine dime A, Papuamide B, Papuamide C, Papuamide D, Parasin I, Sylate, Lobaplatin, Lobophorin A, Lobradimil, Lobucavir, Parathyroid hormone (human recombinant), Paromomycin, Lobucavir, Lodenosine, Loloatin B, Lomeguatrib, Lom Pasireotide, Paulomycin, Paulomycin A, Paulomycin A2, etrexol, Lonafarnib, Loracarbef hydrate, Loviride, Loxorib 55 Paulomycin B, Paulomycin C, Paulomycin D. Paulomycin ine, L-Simexonyl homocysteine, L-Thiocitrulline, Lym E. Paulomycin F. PaZufloxacin, PaZufloxacin mesilate, PEG phostin, Lysine acetylsalicylate, Lysobactin, Mabuterol Vancomycin, Pelagiomicin C, Peldesine, Pelitrexol, Pem hydrochloride, Magainin II, Makaluvamine A, Makalu etrexed disodium, Penciclovir, Penicillamine, Penicillin G vamine A, Makaluvamine B. Makaluvamine C. Managlinat procaine, Pentafuside, Pentamidine gluconate, Pentamidine dialanetil, Matristatin A2, Maxadilan, Melagatran, Melano 60 isethionate, Pentamidine lactate, Peplomycin, Peptide Leu tan, Melanotan I, Melanotan II, Melevodopa hydrochloride, cine Arginine, Peramivir, Perphenazine 4-aminobutyrate, Memantine hydrochloride, Memno-peptide A, Meprobam Pexiganan acetate, Phakellistatin 5, Phe-Arg-beta-naphthyl ate, Meriolin-3, Mersacidin, Mesalamine, Mesalazine, Met amide, Phentermine, Phortress, Phospholine, Pibutidine araminol, Metazosin, Meterelin, Metformin hydrochloride, hydrochloride, Piceasin, Picumeterol fumarate, Pidorubicin, Methotrexate, Methyl aminolevulinate, Methyl bestatin, 65 Pimagedine, Pimeloylanilide o-aminoanilide, Piproxen, Methyldopa, Methylthio-DADMe-immucillin-A, Piracetam, Pirarubicin, Piscidin 1, Piscidin 2, Piscidin 3, Metirosine, Metoclopramide hydrochloride, Metyrosine, Pivampicillin, Pixantrone maleate, Pluraflavin A. Plurafla US 9,533,056 B2 25 26 vin B, Plusbacin A1, Plusbacin A2, Plusbacin A3, Plusbacin Teicoplanin-A2-1, Teicoplanin-A2-2, Teicoplanin-A2-3, A4, Plusbacin B1, Plusbacin B2, Plusbacin B3, Plusbacin Teicoplanin-A2-3, Teicoplanin-A2-5, Telavancin hydro B4, PMEO-5-Me-DAPy, Pneumocandin A0, Pneumocandin chloride, Telinavir, Temozolomide, Temurtide, Tenidap, B0, Pneumocandin B0 2-phosphate, Pneumocandin D0, Tenidap sodium, Tenofovir, Tenofovir DF, Tenofovir diso Polaprezinc, Polydiscamide A, Polymer bound human leu proxil fumarate, Terazosin hydrochloride, Teriparatide, Ter kocyte elastase inhibitor, Poststatin, PPI17-24, Pradefovir lipressin, Tertomotide, Tetracosyl cidofovir, Tetracycline mesylate, Pradimicin C, Pradimicin E, Pradimicin FA-2, hydrochloride, Tetrafibricin, Tetrahydrobiopterin, Texeno Pralatrexate, Pralmorelin, Pramipexole hydrochloride, mycin A, Textilinin-1, Tezacitabine, TGP. Thanatin, Thep Pramlintide acetate, Pranedipine tartrate, Prazosin hydro rubicin, Thermozymocidin, Thioacet, Thiothio. Thr10-Con chloride, Prefolic A. Pregabalin, Preladenant, Prezatide cop 10 tulakin G, Thrazarine, Thymalfasin, Thymic humoral factor per acetate, Primaquine phosphate, Prinomide trometh gamma-2, Thymoctonan, Thymopentin, Thymosin alpha 1. amine, Probestin, Procainamide hydrochloride, Procaine Tiamdipine, Tifluvirtide, Tigecycline, Tigilcycline, Tilargi hydrochloride, Procaine Penicillin, Pro-diazepam, Propep nine hydrochloride, Timirdine diethanesulfonate, tin, Propeptin T. Prostatin, Protegrin IB-367, Prucalopride, Timodepres sin, Timogen, Tipifarnib, Tiplimotide, TNF Prucalopride hydrochloride, Prucalopride succinate, 15 alpha protease enzyme inhibitor, Tobramycin, Tocainide Pseudomycin A", Pseudomycin B, Pyloricidin B, Pymadin, hydrochloride, Tokaramide A, Tomopenem, Topostatin, Tor Pyrazinamide, Pyrazinoylguanidine, Pyridazomycin, Pyrif citabine, ToSufloxacin, ToSufloxacin tosilate, Tranexamic erone, Pyrimethamine, Pyrodach-2, Quinelorane hydrochlo acid, Trantinterol hydrochloride, Tranylcypromine sulfate, ride, R-(+)-Aminoindane, R9K-Retrocyclin, Ragaglitazar Trelanserin, Tresperimus triflutate, Tribavirin, Trichomycin L-arginine salt, Ralfinamide, Ramoplanin A1, Ramoplanin A, Triciribine, Triciribine phosphate, Triciribine-5'-mono A2, Ramoplanin A3. Ramorelix, Rat adrenomedullin, phosphate, Trientine hydrochloride, Trimazosin hydrochlo Ravidomycin N-Oxide, Razaxaban hydrochloride, Reblasta ride, Trimetrexate glucuronate, Trimexautide, Trimidox, Tri tin, Receptor mediated permeabilizer, Recombinant human platin tetranitrate, Triproamylin acetate, Trovafloxacin, parathyroid hormone (1-84), Recombinant Jerdostatin, Trovafloxacin hydrate, Trovafloxacin hydrochloride mesy Regadenoson, Relaxin-3/INSL5 chimeric peptide, Relco 25 late, Trovafloxacin mesilate, Trovafloxacin mesylate, Trox vaptan, Remacemide hydrochloride, Remofovir mesylate, acitabine, Trybizine hydrochloride, Tubastrine ((+)- Residuimod, Restricticin, Retaspimycin hydrochloride, enantiomer), Tuftsin, Tumor necrosis factor-alpha protease Retigabine hydrochloride, Rhodopeptin C1, Rhodopeptin inhibitor, Tyroservaltide, Tyroservatide, Tyrphostin 47, Tyr C2, Rhodopeptin C3, Rhodopeptin C4, Rhodostreptomycin phostin AG-213, Ubenimex, Ubenimex methyl ester, A, Rhodostreptomycin B. Ribamidine hydrochloride, Riba 30 Ubestatin, Ubidine, Uroguanylin, Valaciclovir, Valacyclovir, Virin, Ribavirin eicosenate cis, Ribavirin eicosenate trans, ValboroPro, Valganciclovir hydrochloride, Valnemulin, Ribavirin elaidate, Ribavirin oleate, Rilmazafone hydro Valomaciclovir Stearate, Valonomycin A, Valonomycin B, chloride dihydrate, Riluzole, Rimacalib hydrochloride, Valopicitabine, Valpromide, Valrocemide, Vamicamide, Rimeporide hydrochloride, Riociguat, Ritipenem acoxil, Vancomycin hydrochloride, Vancoresmycin, Vapitadine r-Jerdostatin, RobalZotan hydrochloride, Robalzotan tartrate 35 hydrochloride, Varespladib, Varespladib methyl, Varesp hydrate, Rociclovir, Romurtide, Rotigaptide, Roxifiban ladib mofetil, Vasonatrin peptide, Velnacrine maleate, acetate, Ruboxyl, Rufinamide, Rumycin 1, Rumycin 2, Venorphin, Vesiculin, Vigabatrin, Vilazodone hydrochlo S(-)-Norketamine, Sabarubicin hydrochloride, Sabiporide ride, Vindesine, Viramidine hydrochloride, Viranamycin-B, mesilate, Safinamide mesilate, Safingol, Sagamacin, Sam Virgisin-1, Virgisin-2, Vitamin B3, W Peptide, Xemilofiban, patrilat, Sampirtine, Saprisartan, Sapropterin dihydrochlo 40 Xenoxin-1, Xenoxin-2, Xenoxin-3, Xylocydine. Yttrium-90 ride, Saquinavir, Saquinavir mesilate, Sardomozide, Sardo edotreotide, Zalcitabine, Zanamivir, Ziconotide, Zileuton, mozide hydrochloride, Satoribine, Satraplatin, Zofenoprilat arginine, Zoniporide hydrochloride, Zorubicin Saussureamine C, Saxagliptin, Secobatzelline A, Secobat hydrochloride, Zelline B, Seglitide, Selanc, Selank, Seletracetam, Semapi Preferably, D-H is a small molecule bioactive agent mod hydrochloride, Semparatide, Senicapoc, Sepimostat 45 selected from the group of agents containing at least one mesilate, Seproxetine, Seraspenide, Sermorelin, Sevelamer aliphatic secondary amine group: (-)-2-(2-Bromohexade carbonate, Sevelamer hydrochloride. Shepherdin, Shiga canoyl)paclitaxel, ()-3-O-Acetylspectaline hydrochloride, ( vaccine, Siamycin I, Siamycin II, Sibrafiban, Sifuvirtide, )-3-O-tert-Boc-spectaline hydrochloride, (-)-Bemoradan, Silodosin, Silver sulfadiazine, Sipatrigine, Sitafloxacin (-)-Bupivacaine hydrochloride, (-)-Calicheamicinone, (-)- hydrate, Sitagliptin phosphate monohydrate, S-Nitrosoglu 50 Cicloprolol, (-)-Conophylline, (-)-Draflazine, (-)-Efar tathione, Sofigatran, Sonedenoson, Sotirimod, Sparfloxacin, oxan, (-)-Halofenate, (-)-Indocarbazostatin B, (-)-Nebiv Sperabillin A, Sperabillin B. Sperabillin C, Sperabillin D, olol, (-)-Norchloro-18F fluoro-homoepibatidine, (-)- Sphingofungin F. Spifloxacin hydrate, Spinorphin, Spisu Salbutamol hydrochloride, (-)-Salmeterol, (-)-Ternatin, losine, Spisulosine 285, Squalamine lactate, Stearyl-norleu (+)-(S)-Hydroxychloroquine, (+)-AZacalanolide A, (+)- cine-VIP Streptomycin, Stressin1-A, Styloguanidine, Sub 55 Efaroxan, (+)-Indocarbazostatin, (+)-Isamoltan, (+)-Pemed stance P(8-11), Sulcephalosporin, Sulfinosine, Sulfircin, olac, (+)-R-Pramipexole, (+)-Scyphostatin, (+)-SNAP Sulfostin, Sulphazocine, Sulphostin, Sultamicillin tosylate, 7180, (+)-Sotalol, (+)-threo-Methylphenidate Sunflower trypsin inhibitor-1, Surfen, Synadenol, Syngua hydrochloride, (R)-(+)-Amlodipine, (R)-Albuterol hydro nol, Synthetic human secretin, Synthetic neutrophil inhibitor chloride, (R)-Bicalutamide, (R)-Clevidipine, (R)-Ganciclo peptide, Synthetic porcine secretin, Tabilautide, Tabimo 60 vir cyclic phosphonate, (R)-Niguldipine hydrochloride, (R)- relin, Tacedinaline, Tacrine hydrochloride, Tafenoquine Suc NSP-307, (R)-Teludipine, (R)-Thionisoxetine, (R)- cinate, Tageflar, Talabostat, Talaglumetad hydrochloride, Tulobuterol, (R)-Zacopride, (R,R)-Formoterol tartrate, (S)- Talampanel, Talipexole dihydrochloride, Tallimustine Acetorphan, (S)-Clevidipine, (S)-N- hydrochloride, Talopterin, Talotrexin, Taltirelin, Tanespimy Desmethyltrimebutine, (S)-Noremopamil, (S)- cin, Tanogitran, Targinine, Targinine hydrochloride, Tariba 65 Rivoglitazone, (S)-Sotalol, (S)-Zacopride hydrochloride, virin hydrochloride, Technetium (99mTc) apcitide, Techne 111 In-DTPA-Pro1,Tyr4 bombesin, 90Y-DOTAGA-sub tium (99mTc) depreotide, Technetium Tc 99m depreotide, stance P, 99TcDemobesin 3, 99TcDemobesin 4, Ala 1 1, US 9,533,056 B2 27 28 D-Leu15IOrexin B, Arg(Me)9 MS-10, D11G.K26R, Amethocaine hydrochloride, Amfebutamone hydrochloride, Y40YR-Plecta sin, D11G.M13K.K26R,Y40YR-Plecta Amibegron hydrochloride, Amifostine hydrate, Amiglu sin, D9N.M13L.Q14R-Plectasin, D9SQ14K.V36L mide, Amikacin, Amiloride hydrochloride, Amineptine, Plectasin, D-Tyr1Arg(Me)9 MS-10, D-Tyr1AzaGly7, Aminocandin, Aminochinol, Aminoglutethimide, Amin Arg(Me)9 MS-10, D-Tyr1 MS-10, Gln20-Pancreatic oguanidine, Aminoquinol, Aminoquinuride, Amisulpride, polypeptide (2-36), Glu10.Nle17.Nle30-Pancreatic poly Amitivir, Amlodipine, Amlodipine besylate. Amobarbital, peptide (2-36), Glut O-Pancreatic polypeptide(2-36), Amocarzine, Amodiaquine, Amosulalol hydrochloride, L17K.K30R GLP-2 (1-33), Leu13-Motilin, N5R.M13Y. Amoxapine, Amoxicillin, Amoxicillin trihydrate. Amoxycil N17R-Plectasin, Nle17.Nle30-Pancreatic polypeptide lin trihydrate, Ampicillin sodium, AmpiroXicam, Amprena (2-36), IN-Melle4-cyclosporin, psiCH2NHITpg4Vanco 10 Vir, Amrinone, Amsacrine, Amsilarotene, Amsulosin hydro mycin aglycon, Ser12-Humanin, Trp19 MS-10, Tyr24 chloride, Amtolimetin guacil, Amylobarbitone, Anabasine Humanin, 111 In-Pentetreotide, 12-Methylthiovinblastine hydrochloride, Anagrelide hydrochloride, Anakinra, dihydrochloride, 14beta-Hydroxydocetaxel, 14beta-Hy Anamorelin hydrochloride, Anandamide, Anatibant mesi droxydocetaxel-1,14-acetonide, 14beta-Hydroxytaxotere, late, Andolast, Androxolutamide, Anginex, Angiopeptin 15bbeta-Methoxyardeemin, 16-Aza-epothilone B, 16-Meth 15 acetate, Angiotensin II (human), Anidulafungin, Anisperi yloxazolomycin, 17-Amino-17-demethoxygeldanamycin, mus, Ansamycin, Antagonist-G, Antide, Antide-1, Antide-2, 17-Aminogeldanamycin, 18-Hydroxycoronaridine, Antide-3, Antiflammin-1, Antiflammin-10, Antiflammin-2, 18-Methoxycoronaridine, 19-O-Methylgeldanamycin, Antiflammin-3, Antiflammin-4, Antiflammin-5, Antiflam 1-Deoxynoirimycin, 2,7-Dibromocryptolepine, 2-Bromo min-6, Antiflammin-7, Antiflammin-8, Antiflammin-9, Anti 7-nitrocryptolepine, 2'-Palmitoylpaclitaxel, 3-Bromometh leukinate, Antimycin A11, Antimycin A12, Antimycin A13, cathinone, 3-Chloroprocainamide, 3-Fluorothalidomide, Antimycin A14, Antimycin A15, Antimycin A16, Antocin II, 3-Indole, 4,5-Dianilinophthalimide, 4,6-diene-Cer, 447-480 Apadenoson, Apadoline, Apalcillin Sodium, Apaxifylline, Human alpha-fetoprotein, 4-Chlorophenylthio-DADMe-im Apcitide technetium (99mTc). Apicularen A, Apicularen B. mucillin-A, 4-Ethynylstavudine, 4-Hydroxyatomoxetine, Apilimod, Apilimod mesylate, Apiodionene, Aplaviroc 5,4'-Diepiarbekacin, 5-Fluorodeoxyuridine, 5-Iodofrederi 25 hydrochloride, Aplidine, Aplindore fumarate, Aplonidine camycin A, 5-Methylurapidil, 5-Phenylthioacyclouridine, hydrochloride, Apoptozole 1, Apoptozole 2, Apraclonidine 6,7-Dichloroisatin 3-oxime, 6alpha-7-Epipaclitaxel, 6-Mer hydrochloride, Apremilast, Aprepitant, Aprikalim, Aprosu captopurine, 7-Bromo-2-chlorocryptolepine, 7-Deoxytaxol. late Sodium, Aptiganel, Aranidipine, Aranorosin, Aranorosi 7-Oxostaurosporine, 9.9-Dihydrotaxol. 99mTc-c(RGDfK*) nol A, Aranorosinol B, Aranose, Aranoza, Araprofen, Arba 2HYNIC, 9-Nitropaullone, A-42867 pseudoaglycone, Aba 30 clofen placarbil sodium, Arbekacin, Arbekacin sulfate, cavir Succinate, Abacavir Sulfate, Abafungin, Abarelix, Abe Arborcandin A, Arborcandin B, Arborcandin C, Arborcandin carnil, Abitesartan, Acamprosate calcium, Acarbose, D. Arborcandin E. Arborcandin F. Arbutamine hydrochlo Acebutolol hydrochloride, Aceclofenac, Acemannan, Ace ride, Archazolid A, Archazolid B, Archazolide A, Arcyri neuramic acid sodium salt, Acetamidoxolutamide, Acet acyanin A, Ardeemin, Arenicin, Arenicin-1, Arenicin-2, aminophen, Acetazolamide, Acetohexamide, Acetorphan, 35 Arformoterol tartrate, Argatroban monohydrate, Argimesna, Acetylcysteine, Acetyldinaline, Aciclovir, AcitaZanolast, Arginine butyrate, Argiopine, Argiotoxin-636, Argipidine, Acomustine, Acotiamide hydrochloride hydrate, AcreoZast, Argyrin A, Argyrin B, Arhalofenate, Aripiprazole, Arisosta Actarit, Actinomycin D, Actinoplanone B. Aculeacin tin A, Arofylline, Arotinolol hydrochloride, Arterolane Agamma, Acycloguanosine, Acyclovir, Acyclovir elaidate, maleate, Artilide fumarate, Ascosalipyrrolidinone A, Acyclovir oleate, Acyline, AD Peptide, Adamantyl globot 40 Ascosalipyrrolidinone B, Asobamast, Asp(B14)-relaxin, riaosylceramide, Adaphostin, Adaprolol maleate, Adaprolol Asperlicin B, Asperlicin C, Asperlicin D, Asperlicin E, oxalate, Adatanserin hydrochloride, Adecypenol, Aspoxicillin, Astemizole, Ataciguat, Atalaphillidine, Ata Adelmidrol, Adenopeptin, Aderbasib, Adjudin, Adosopine, laphillinine, Ataquimast, Ataquimast hydrochloride, Ataza Adozelesin, Adrafinil, Adrogolide hydrochloride, Aerothri navir Sulfate, Atenolol, Atevirdine mesylate, Atipamezole, cin 1, Aerothricin 16, Aerothricin 41, Aerothricin 45, Aero 45 Atizoram, Atomoxetine hydrochloride, Atorvastatin, AtorV thricin 5, Aerothricin 50, Aerothricin 55, Aeruginosamide, astatin calcium, Atosiban, Atrial natriuretic factor (99-126), Afamelanotide, Afeletecan hydrochloride, Afobazol, Afoba Aureobasidin A, Auristatin E, Auristatin PE, Avasimibe, Zole, Agelasphin 517. Agelasphin 564, Ageliferin diacetate, Avicin D, Avicin G. Avitriptan, Avizafone, Avorelin, Ageliferin dihydrochloride, Aglaiastatin C, Agomelatine, AVosentan, AVrainvillamide, Axitinib, Axitirome, Alacepril, Aladapcin, Aladotril, Alatriopril, Alatrofloxacin 50 AZ-36041, Azaromycin SC, AZasetron, AZasetron hydro mesilate, Albendazole, Albifylline, Albolabrin, Albuterol chloride, AZathioprine, Anatoxin, AZelnidipine, AZepinosta nitrate, Albuterol sulfate, Alchemix, Alfuzosin hydrochlo tin, AZetirelin, AZidothymidine, AZidothymidine phospho ride, Alinidine, Alisamycin, Aliskiren fumarate, Alizapride, nate, AZilsartan, AZilsartan medoxomil, Aztreonam, Alkasar-18, Allantoxamic acid, Alloferon-1, Allopurinol, Aztreonam L-lysine, AZtreonam lysinate, AZumamide A, Alminoprofen, Almitrine bismesylate, Almitrine dimeSylate, 55 AZumamide E. Bactobolin, Bafetinib, Balaglitazone, Bala Almorexant, Almotriptan, Alniditan, Aloracetam, Alosetron mapimod, Balanol, Balaperidone, Balhimycin, Balicatib, hydrochloride, Alosetron maleate, Aloxistatin, alpha-C-Ga Balofloxacin, Balofloxacin dihydrate, Balsalazide disodium, lactosylceramide, alpha-Galactosylceramide, alpha-Galac Bamaquimast, Bambuterol, Bamirastine hydrate, Banoxan tosylceramide-BODIPY, alpha-Human atrial natriuretic trone, Baogongteng A, Barixibat, Barnidipine hydrochlo polypeptide, alpha-Lactosylceramide, alpha-Methylepi 60 ride, Barusiban, Basifungin, Batimastat, Batoprazine, nephrine, alpha-Methyltryptophan, alpha-Nornicotine, Batroxostatin, Batzelline A, Batzelline B, Batzelline C, alpha-Sialosylcholesterol sodium salt, Alprafenone hydro Beauveriolide I, Beauveriolide III, Becampanel, Becate chloride, Alprenolol hydrochloride, Alprenoxime hydro carin, Bederocin, Bedoradrine sulfate. Befol. Befunolol chloride. Alsterpaullone. Altemicidin, Altromycin A, Altro hydrochloride, Begacestat, Belactin A, Belactin B, Belacto mycin C, Alvespimycin hydrochloride, Alvimopan hydrate, 65 sin A, Belactosin C, Belaperidone, Belinostat, Belotecan Amacetam hydrochloride, Amamistatin A, Amamistatin B, hydrochloride, Bemoradan, Benadrostin, Benafentrine Ambazone, AmbroXol nitrate, Ameltolide, Amesergide, dimaleate, Benanomicin A, Benanomicin B, Benatoprazole, US 9,533,056 B2 29 30 Benazepril hydrochloride, Bendrofluazide, Bendroflumethi Cefprozil monohydrate, Cefiguinome, Cefsulodin Sodium, azide, Benexate cyclodextrin, Benidipine hydrochloride, Ceftaroline, Ceftaroline fosamil acetate, Ceftazidime, Ceft Benperidol, Benzimidavir, Benzylpenicillin sodium, Ber eram pivoxil, Ceftibuten, Ceftizoxime alapivoxil, Ceftizox lafenone hydrochloride, Besonprodil, beta-CCM, beta ime sodium, Ceftobiprole, Ceftobiprole mediocaril, Ceftra Methyl-6-chloromelatonin, Betamipron, beta-Sialosylcho Zonal bopentil, CeftraZonal sodium, Ceftriaxone sodium, lesterol sodium salt, beta-Tethymustine, Betaxolol Cefuroxime, Cefuroxime axetil, Cefuroxime pivoxetil, hydrochloride, Bevantol hydrochloride, Bevantolol hydro Celiprolol hydrochloride, Cemadotin hydrochloride, Cen chloride, Bezafibrate, Bicalutamide, Biemnidin, Bifemelane tanamycin, Cephalexin monohydrate, Cephazolin Sodium, hydrochloride, Bifeprunox mesilate, Bimatoprost, Binode Ceratamine A, Ceratamine B, Cerebrocrast, Cerebroside A, noson, BinoSpirone mesylate, Biotinylated idraparinux, 10 Cerebroside B, Cerebroside C, Cerebroside D, Cerulein, Bioxalomycin alpha 1, Bipranol hydrochloride, Bis(7)-cog Ceruletide diethylamine, Cethromycin, Cetrorelix acetate, nitin, Bis(7)-tacrine, Bisantrene hydrochloride, Bisnafide Cevipabulin, Chackol, Chaetocin, Chetocin, Chinoin-169, mesilate, Bisnafide mesylate, Bisoprolol fumarate, Bitolt Chloptosin, Chlorazicomycin, Chlordiazepoxide hydrochlo erol mesylate, Bivalirudin, Bizelesin, Bleomycin A2 sulfate, ride, Chlorofusin, Chloroorienticin A, Chloroorienticin B, Boc-Belactosin A, Boceprevir, Boc-Lysinated betulonic 15 Chloropeptin II, Chlorotetain, Chlorothiazide, Chlorprop acid, Body protection compound-15, Bogorol A, Bohemine, amide, Chlorpropham, Chlortalidone, Chlortenoxicam, Boholmycin, Bopindolol, Bortezomib, Bosentan, Bosutinib, Chlorthalidone, Chondramide A, Chondramide B, Chondr Bradyzide, Brain natriuretic peptide, Brasilicardin A, Breca amide C, Chondramide D, Cibenzoline succinate, navir, Bremelanotide, Brimonidine tartrate, Brinazarone, Ciclosporin, Cifenline Succinate, Cilastatin Sodium, Cilas Brinzolamide, Brivanib, Brivanibalaninate, Brivudine, Bro tatino, Cilazapril, Cilengitide, Cilnidipine, Cilostazol, Cilu mantan, Bromantane, Bromazepam, Bromocriptine mesi previr, Cimadronate sodium, Cimaterol, Cimetidine, Cime late, Bromotopsentin, Bromovinyldeoxyuridine, Brostalli tidine bismuth citrate, Cimetidine bismuth L-tartrate, cin hydrochloride, Brovavir, B-Type natriuretic peptide, Cimetidine nitrate, Cinacalcet hydrochloride, Cinaldipine, Bucillamine, Bucladesine sodium, Buflomedil pyridoxal Cinalukast, Cinitapride hygrogen tartrate, Cinnabaramide A, phosphate, Bulaquine, Bumetanide, Bupivacaine hydrochlo 25 Cinnamycin, Cinnoxicam, Cipamfylline, Cipemastat, Cip ride, Bupropion hydrochloride, Buserelin acetate. Butabin ralisant, Ciprofloxacin hydrochloride, Ciprofloxacin silver dide, Buteranol, Butobarbitone, Butoctamide salt, Ciprokiren, Ciproxifan, Circinamide, Cisapride hemisuccinate. Butofilolol, Butyl flufenamate. ButyZamide, hydrate, Citropeptin, Citrullimycine A, Clamikalant, Clause Cabazitaxel, Cabergoline, Cabin 1, Cadralazine, Cadro namine A, Clavanin A(K), Clavanin E(3-23), Clazosentan, floxacin hydrochloride, Caerulein diethylamine, Calcium 30 Clazosentan sodium, Clevidipine butyrate, Clevudine, Clin folinate, Calcium-like peptide 1, Calcium-like peptide 2, damycin hydrochloride, Clitocine, Clobenpropit, Clonaze Caldaret hydrate, Caldiamide sodium, Calicheamicin pam, Clonidine, Clonidine hydrochloride (hydrochloride), gammal aglycone, Calindol Dihydrochloride, Calothrixin Clopamide, Clopenphendioxan, Clopidogrel Sulfate, Clora A, Cambrescidin 800, Cambrescidin 816, Cambrescidin nolol hydrochloride, CloraZepate dipotassium, Clospip 830, Cambrescidin 844, Camostat mesilate, Camostat mesy 35 ramine hydrochloride, Cloturin, Clozapine, Coenzyme late, Camprofen, Canadensol, Candesartan, Candesartan PQQ, Collabomycin A, Coleneuramide, Colivelin, Colurac cilexetil, Candesartan hexetil, Candoxatril, Candoxatrilat, etam, Complestatin, Conagenin, Conantokin-R, Coniosetin, Canertinib dihydrochloride, Canfosfamide hydrochloride, Conivaptain hydrochloride, Conophylline, Contulakin G, Cangrelor tetrasodium, Capecitabine, Capimorelin, Cortagine, Coumamidine gammal, Coumamidine gamma2. CapraZamycin A, CapraZamycin B, CapraZamycin C, 40 Covidarabine, Creatine phosphate, Crilvastatin, Crisinatol CapraZamycin E. CapraZamycin F. Capridine beta, Capro mesilate, Cronidipine, Cryptophycin 52, Cyclamenol, Cyclo lactin A, Caprolactin B, Capromorelin, Capsavanil, Cap His-Pro, Cyclocreatine, Cyclolinopeptide A, Cyclolino Sazepine, Carabersat, Caracasanamide, Caracasandiamide, peptide B, Cyclomarin A, Cyclopenthiazide, Cyclophosph Carafiban maleate, Carbazomadurin A, Carbazomadurin B, amide, Cycloserine, Cyclosporin, Cyclosporin A, Carbazomycin G. Carbazomycin H. Carbetocin, Carbidopa, 45 Cyclosporin J. Cyclosporine, Cyclosporine A, Cyclotheona Carbovir, Carfilzomib, Cariprazine hydrochloride, Carme mide A, Cyclothialidine, Cycloviolin A, Cycloviolin B, thizole, Carmofur, Carmoterol hydrochloride, Carmoxirole Cycloviolin C, Cycloviolin D, Cygalovir, Cymserine, Cype hydrochloride, Carmustine, Carnosine Zinc complex (1:1), mycin, Cystamidin A, Cystemustine, Cystocin, Cytoblastin, Carperitide, Carprofen, Carquinostatin A, Carsatirin, Car Cytochalasin B, Cytomodulin, Cytotrienin A, Cytotrienin I, teolol hydrochloride, Carteramine A, Carumonam Sodium, 50 Cytotrienin II, Cytotrienin III, Cytotrienin IV, Cytoxazone, Carvedilol, Carvotroline hydrochloride, Carzelesin, Caspo Dabelotine mesilate, Dabigatran, Dabigatran etexilate, fungin acetate, Catramilast, Cavtratin, Cebaracetam, Cecro DabuZalgron hydrochloride, Dacinostat, Dactimicin, Dac pin A(1-11) D(12-37), Cecropin D, Cediranib, Cefaclor, tinomycin, Dactylocycline A, DADMe-Immucillin-G, Cefalexin monohydrate, Cefazolin sodium, Cefbuperazone DADMe-Immucillin-H, Daglutril, Dalargin, Dalbavancin, sodium, Cefcamate pivoxil hydrochloride, Cefcanel, Cef 55 Dalcetrapib, Dalcotidine, Dalfopristin mesilate, D-allo-Ileu3 canel daloxate hydrochloride, Cefcapene pivoxilhydrochlo PYY(3-36), DANA, Danegaptide hydrochloride, Danu ride, Cefdaloxime, Cefdaloxime Pentexil Tosilate, Cefdinir, sertib, Dapivirine, Daporinad, Dapropterin dihydrochloride, Cefditoren pivoxil, Cefepime, Cefetamet pivoxil, Cefetecol, Darbufelone, DarglitaZone, Darinaparsin, Darunavir, Das Cefixime, Cefluprenam, Cefnmatilen hydrochloride hydrate, antafil. Dasatinib, Davasaicin, Davunetide, Daxalipram, Cefnmenoxime hydrochloride, Cefninox sodium, Cefodiz 60 D-Cycloserine, Debromoshermilamine, Decahydromoeno ime, Cefodizime sodium, Cefonicid sodium, Cefoperazone mycin A, Decaplanin, Decatromicin A, Decatromicin B, sodium, Cefoselis sulfate, Cefotaxime sodium, Cefotetan Declopramide, Deferobiotin, Deferoxamine, Degarelix disodium, Cefotiam cilexetil, Cefotiam cilexetil hydrochlo acetate, Degrasyn, Dehydrodidemnin B, Dehydrodolastatin ride, Cefotiam hexetil, Cefotiam hexetil hydrochloride, 13, Dekafin 1, Dekafin 10, Delaminomycin A, Delamino Cefotiam hydrochloride, Cefoxitin, Cefozopran, Cefozopran 65 mycin B, Delaminomycin C, Delapril hydrochloride, Dela hydrochloride, Ce?pimizole sodium, Ce?piramide sodium, Virdine mesilate, Delfaprazine hydrochloride, Delimotecan Cefpirome, Cefpodoxime proxetil, Ce?prenam, Cefprozil, sodium, Deltibant, Deltorphin E. Delucemine hydrochlo US 9,533,056 B2 31 32 ride, Demethylallosamidin, Demethylasterriquinone B-1, Emicerfont, Emivirine, Emonapride, Emricasan, Enalapril Demetomidine, Demexiptiline hydrochloride, Denibulin maleate, Enalapril nitrate, Enalaprilat, Enalkiren, Enaza hydrochloride, Denopamine, Denufosol tetrasodium, drem, Endothelin, Endothelin 1, Enfluvirtide, Eniluracil, Deoxycoformycin, Deoxymethylspergualin, Deoxymulun Enkastin (D), Enkastin AD, Enkastin AE, Enkastin ID, docandin, Deoxynegamycin, Deoxynoirimycin, Deox Enkastin IE, Enkastin VD, Enkastin VE, Enocitabine, yspergualin hydrochloride, Depsipeptide, Deriglidole, Enoxacin, Enoximone, Entecavir, Enteric neural peptide, Desacetylvinblastinehydrazide, Desacetylvinblastinehy Entinostat, Enzastaurin hydrochloride, Eperezolid, Eper drazide/folate conjugate, Desbutyl benflumetol, Desbutyl eZolid N-oxide, Epervudine, Epibatidine, Epidepride-(125I), halofantrine hydrochloride, Desferri-danoxamine, Desferri Epiderstatin, Epipachysamine E. Epithalon, Epocarbazolin nordanoxamine, Desferrioxamine, Desferri-salmycin A, 10 A, Epocarbazolin B, Epofolate, Epolactaene, Eponemycin, Desferri-salmycin B. Desferri-salmycin C, Desferri-salmy Epostatin, Epoxomicin, Epristeride, Eprobemide, Epro cin D. Desglugastrin tromethamine, Desipramine hydro tirome, Eptastigmine tartrate, Eptifibatide, Erbulozole, chloride, Desloratadine, Deslorelin, Desmin-370, Desmo Erdosteine, Eremomycin, Ergotamine tartrate, Eribaxaban, pressin acetate. Desulfated hirudin (54-65), Desulfated Eritoran tetrasodium, Erlosamide, Erlotinib hydrochloride, hirugen, Detomidine hydrochloride, Devazepide, Dexeca 15 Ersentilide, Ersentilide hydrochloride, Ertapenem sodium, dotril, Dexefaroxan, Dexfenfluramine hydrochloride, Erythromycin Salnacedin, Erythromycin stinoprate, Escu Dexketoprofen imidazole salt, Dexketoprofen meglumine, leogenin A, Esculeoside A, Eserine salicylate, Esmolol Dexlansoprazole, Dexloxiglumide, Dexmedetomidine hydrochloride, Esperamicin A1. Esperatrucin, Etacizin, hydrochloride, Dexmethylphenidate hydrochloride, Dex Etamsylate, Etaquine, Ethacizin, Ethamsylate, Ethimizol. niguldipine hydrochloride, Dexpemedolac, DexraZoxane Ethiofos, Ethosuximide, Ethoxy-idazoxan, Ethyl hydrochloride, Dexsotalol, Dextronatrin, Dexylosylbenano loflazepate, Ethylthio-DADMe-immucillin-A, Etidocaine mycin A, d-Fenfluramine hydrochloride, D-Fluviabactin, hydrochloride, Etodolac, Etodolic acid, Etravirine, Eugeno DHA-paclitaxel, Diaplasinin, Diazepinomicin, DiaZoxide, dilol, Eurocin, Eurystatin A, Eurystatin B. Examorelin, Dichlorobenzoprim, Diclofenac potassium, Diclofenac Exenatide, Exenatide LAR, Exendin-4, Ezatiostat hydro Sodium, Diclofenac Zinc salt, Didanosine, Didemnin X. 25 chloride, Ezlopitant, Fabesetron, Fabesetron hydrochloride, Didemnin Y. Dideoxyinosine, Diethyl-lactam, Diethylnor Fadolmidine hydrochloride, Falnidamol, Famotidine bis spermine. Difurazone, Diheteropeptin, Dihydrexidine, muth citrate, Fanapanel, Fargilitazar, Fasidotril, Fasobegron Dihydro-alpha-ergokryptine meSylate, Dihydroavenanthr hydrochloride, Fasoracetam, Fasudil hydrochloride, Fegly amide D. Dihydroeponemycin, Dihydroergotamine mesy mycin, Feglymycine, Felodipine, Fenoldopam mesilate, late, Dilevalol, Dilevalol hydrochloride, Dimelamol, Dim 30 Fenoterol hydrobromide, Fenoximone, Fepradinol, Ferro ethynur, Dimiracetam, di-mPEG5-Atazanavir, Dinapsoline, chloroquine, Ferroquine, Ferulinolol, Fidarestat, Fiduxosin Dinoxyline, Dioxolane T. Dioxolane thymine nucleoside, hydrochloride, Filarizone, Fimasartan, Fimbrigal P. Fina Diperamycin, Dipivefrine hydrochloride, Dipranol hydro floxacin hydrochloride, Finasteride, Fipamezole hydrochlo chloride, Diduafosol tetrasodium, Dirithromycin, Dirlo ride, Fish amunine, Flecainide acetate, Flibanserin, tapide, Dirucotide, Disagregin, Disalazine, Discodermide, 35 Flindokalner, Flomoxef sodium, Flopristin, Flopristine, Discodermide acetate, Discorhabdin D, Discorhabdin P. Florbetaben, Flovagatran sodium, Floxuridine, Flucytosine, Discorhabdin S, Discorhabdin T. Discorhabdin U, Disiter Flufenoxine, Flumezapine, Fluodipine, Fluorocytosine, tide, Dithiosteine, d-Methamphetamine hydrochloride, Fluorofur, Fluoroindolocarbazole A, Fluoroindolocarbazole Dobutamine hydrochloride, Dobutamine phosphate, Docar B, Fluoroindolocarbazole C, Fluorouracil, Fluoxetine pamine, Docetaxel, Docetaxol. Dofetilide, Dolasetron, 40 hydrochloride, Fluparoxan, Flupirtine maleate, Fluspirilene, Dolasetron mesilate, Dolastatin 10, Dolastatin 13, Dolastatin Flutamide, Fluvirucin B2, Foetidine 1, Foetidine 2, Folinic 14, Dolastatin 15, Dolastatin C, Dolastatin D, Domitroban acid, Folinic acid calcium salt, Fomepizole, Fondaparin calcium hydrate, Domperidone, Donitriptan hydrochloride, Sodium, Fondaparinux sodium, Fonsartan potassium, Donitriptan mesilate, Dopexamine, Dopexamine hydrochlo Forasartan, Foretinib, Formobactin, Formoterol fumarate, ride, Doramapimod, Doripenem, Dormitroban, Dorrigocin 45 Forodesine hydrochloride, Fosalvudine tidoxil, Fosampre A, Dorrigocin B. Dorzolamide hydrochloride, Dovitinib navir calcium, Fosamprenavir Sodium, Fosaprepitant, Fos Lactate, Doxifluridine, DoxoTam 12, d-Pseudoephedrine aprepitant dimeglumine, Fosopamine, Fosphenytoin hydrochloride, Draflazine, Dronedarone hydrochloride, Sodium, Fostamatinib, Fostamatinib disodium, Fotemustine, Droperidol, Droximavir, d-threo-Methylphenidate hydro Fozivudine tidoxil, Fradafiban, Franidipine hydrochloride, chloride, DTPA-adenosylcobalamin, Duloxetine hydrochlo 50 Freselestat, Frog neuromedin U. Frovatriptan, Frusemide, ride, Dumorelin, Duocarmycin A, Duocarmycin B1, Duo Ftorafur. Furaldipine. Furavir, Furnidipine, Furosemide, G1 carmycin B2, Duocarmycin C1, Duocarmycin C2, peptide, Gabadur, Gabapentin enacarbil, Gabexate mesilate, Duocarmycin SA, Duramycin, Dutasteride, Dynemicin A, Gaboxadol, Gadobenate dimeglumine, Gadobenic acid Dynemicin C, Dyofin-1, Dyofin-2, Dyofin-9, EbalZotan, dimeglumine salt, Gadocoletic acid trisodium salt, Gado Ebanicline, Ebrotidine, Ecadotril, Echistatin, Ecomustine, 55 denterate, Gadodiamide, Gadodiamide injection, Gado Ecteinascidin 1560, Ecteinascidin 722, Ecteinascidin 729, linium DTPA, Gadolinium DTPA-BMA, Gadomelitol, Ecteinascidin 736, Ecteinascidin 743, Ecteinascidin 745, Gadopentetate dimeglumine, Gadoterate meglumine, Gado Ecteinascidin 770, Ecteinascidin 875, Edaglitazone, Edo versetamide, Galactomycin I, Galactomycin II, Galdanse nentan hydrate, Edotecarin, Edotreotide yttrium, Edoxaban tron, Gallinacin 1, Gallinacin 1alpha, Gallinacin 2, Galmic, tosilate, Efaproxiral sodium, Efaroxan, Efavirenz, Efegatran 60 Galnon, Galocitabine, Galparan, Gammaphos, Ganciclovir, sulfate hydrate, Efepristin, Eflucimibe, Eflumast, Efo Ganciclovir elaidic acid, Ganciclovir monophosphate, nidipine hydrochloride ethanol, Eformoterol fumarate, Elac Ganefromycin alpha, Ganefromycin beta, Ganglioside ridar, Elagolix sodium, Elarofiban, Elastatinal B, Elastatinal GM1, Ganirelix, Ganirelix acetate, Ganstigmine hydrochlo C, Elbanizine, Eldacimibe, Elesclomol, Eletriptan, Elgo ride, Gantofiban, Garenoxacin mesilate, Garomefrine hydro dipine hydrochloride, Elinafide mesilate, Elinogrel potas 65 chloride, Gastrazole, Gastrophenzine, Gatifloxacin, Gavesti sium, Elliptinium acetate, Elliptoside A, Elliptoside E. Elna nel sodium, Gedocarnil, Gefitinib, Gefitinib hydrochloride, dipine, Elopiprazole, Eltrombopag olamine, Embusartan, Gemopatrilat, Gibbosin, Gidazepam, Gilatide, Gilvusmycin, US 9,533,056 B2 33 34 Giracodazole, Giripladib, Girodazole, Girolline, Givinostat, ride, Ketanserin, Kifunensine, Kinetin, Kistamicin A, Glaspimod, Glibenclamide, Gliclazide, Glidobactin PF-1, Kopsinine, Korupensamine A, Korupensamine B, Korupen Glimepiride, Glipizide, Gliquidone, Glucagon-like peptide I samine C, Kosinostatin, Kurasoin B. Kynostatin-227. (7-37), Glucarolactam potassium, Glucolanomycin, Glu Kynostatin-272, Labedipinedilol A, Labedipinedilol B, dopa, Glufanide, Glufosfamide, Glutapyrone, Glutathione Labetalol hydrochloride, Labradimil, Lacidipine, Lacos monoethyl ester, Glutathione monoisopropyl ester, Glute amide, Lactosylphenyl trolox, Ladasten, Ladostigil tartrate, thimide, Glyburide, Glycine-proline-Melphalan, Glycopin, Laflunimus, Lafutidine, Lagatide, Lamectacin, Lamifiban, Glycopril, Glycothiohexide alpha, Gold talaporfin sodium, Landiolol, Lanepitant, Lanreotide acetate, Lansoprazole, Golotimod, Gomisin A glycinosuccinate sodium salt, Gora Lanthiopeptin, Lapatinib ditosylate, LaraZotide acetate, latide, Goserelin, Gosogliptin hydrochloride, Granisetron 10 Laromustine, Larotaxel dihydrate, Lasinavir, Latamoxef hydrochloride, Grepafloxacin hydrochloride, Growth factor Sodium, Latrunculin S. Lavanduquinocin, LaZabemide antagonist-116. Growth hormone releasing peptide 2. hydrochloride, Lecimibide, Lecirelin, Leconotide, Ledaz Growth Inhibitory Peptide, Guanabenz acetate, Guanadrel erol, Leflunomide, Lefradafiban, Leinamycin, Leminopra sulfate, Guanethidine monosulfate, Guanfacine hydrochlo Zole, Lemuteporfin, Lemuteporphin, Lenalidomide, Lenam ride, Gusperimus hydrochloride, Gusperimus trihydrochlo 15 picillin hydrochloride, Lenapenem hydrochloride, ride, Gyp setin, Habekacin, Habekacin sulfate, Halimide, Lenapenem hydrochloride hydrate, Lercanidipine hydro Halofuginone hydrobromide, Halovir A. Halovir B. Halovir chloride, Lerisetron, Lestaurtinib, Leteprinim potassium, C, Halovir D, Halovir E. Harkoseride, Helvecardin A, LetraZuril, Leualacin, Leucovorin calcium, Leuprolide Helvecardin B, Heptaminol AMP amidate, Heptylstigmine acetate, Leuprorelin acetate, Leurubicin, Levalbuterol tartrate, Herquline B, Hesperadin, Hexa-D-Arginine, hydrochloride, Levobetaxolol hydrochloride, Levobunolol Himastatin, Hirudin desulfated, Hirulog-1. Hispidospermi hydrochloride, Levobupivacaine hydrochloride, Levofoli din, Histamine dihydrochloride. Histaprodifen, Histrelin, nate calcium, Levoleucovorin calcium, Levonadifloxacin Histrelin acetate, Homoepibatidine, Homoindanomycin, arginine salt, Levonebivolol, Levosalbutamol hydrochlo Hormaomycin, Human adrenomedulin, Human adrenom ride, Levosimendan, Levosulpiride, L-Fluviabactin, L-Flu edulin (22-52), Human angiotensin II, Human corticotro 25 vibactin, L-Fluvibactine, L-Histidinol, L-Homothiocitrul pin-releasing hormone, Human lactoferrin (1-11), Human line, Liarozole, Liarozole hydrochloride, Liblomycin, proislet peptide, Human Secretin, Hyaluronan, Hyaluronate Licostinel, Lidamidine hydrochloride, Lidanserin, Lido sodium, Hydralazine hydrochloride, Hydrochlorothiazide, caine hydrochloride, Lifarizine, Lifarizine hydrochloride, Hydroflumethiazide, Hydrostatin A, Hydroxyakalone, Lignocaine, Limazocic, Linaprazan, Linarotene, Linazolast, Hydroxycarbamide, Hydroxychloroquine sulfate, Hydroxy 30 Linetastine, Linezolid, Linezolid oxide, Lingual antimicro mycotrienin A, Hydroxymycotrienin B. Hydroxyurea, bial peptide, Linifanib, Linopristin, Linopristine, Lino Hymenistatin 1, Hypeptin, Ibipinabant, Ibodutant, Ibo troban, Lintitript, Lintopride, Lipohexin, Lipoxazolidinone pamine, Ibrolipim, Ibutamoren mesilate, Ibutilide fumarate, A. Lipoxazolidinone B, Lipoxazolidinone C. Liraglutide, Icatibant acetate, Icopezil maleate, IdaZoxan hydrochloride, Lirexapride, Lirimilast, Lisdexamfetamine mesilate, Lisino Idrabiotaparinux sodium, Idrapril, Ifetroban, Ifosfamide, 35 pril, Lisuride maleate, Lisuride TTS, Litoxetine, Lixivaptan, Iganidipine hydrochloride, Iguratimod, Ilaprazole, Ilat L-Lysine-d-amphetamine dimesylate, 1-Nebivolol, Lobat reotide, Ilepatril, Ilomastat, Imatinib mesylate, Imetit, amide C. Lobatamide F, Lobeglitazone sulfate, Lobenzarit Imexon, Imidacrine, Imidapril, Imidapril hydrochloride, sodium, Lobophorin A, Lobophorin B, Lobradimil, Lobu Imidazoacridinone, Imidazole 24b. Imipemide, Imipenem, cavir, Lodamin, Lodenafil. Lofexidine hydrochloride, Imirestat, Imisopasem manganese, Immepip, Immepyr, 40 Loloatin B, Lomefloxacin hydrochloride, Lomeguatrib, Immethridine, Immucillin-H. Immunosine, Imoproxifan, Lometrexol, Lopinavir, Loprinone hydrochloride, Loracar Impentamine, Implitapide, Improgan, Incadronate, Incad bef hydrate, Lorazepam, Lorcaserin, Lorglumide, Lornoxi ronic acid sodium salt, Indacaterol, Indanomycin, Indant cam, Losartan, Lotrafiban, Loviride, Loxiglumide, LOXista adol hydrochloride, Indapamide, Indeloxazine hydrochlo tin, Loxoribine, L-Simexonyl homocysteine, ride, Indibulin, Indinavir sulfate, Indisetron hydrochloride, 45 L-Thiocitrulline, L-threitol ceramide, L-threo-C6-pyri Indisulam, Indium In 111 pentetreotide, Indole-3-propionic dinium-ceramide-bromide, Lubazodone hydrochloride, acid, Indolicidin-11, Indolicidin-4, Indolicidin-8, Indolmy Lufironil, Lumiracoxib, Lurosetron, Luzindole, Lycopersi cin, Indomethacin phenethylamide, Indomethacin-Simvas cin, Lymphostin, Lysinated-betulonic acid, Lysobactin, tatin, Indoramin hydrochloride, Inogatran, Inosine pra Lysuride maleate, Mabuterol hydrochloride, Macitentan, nobex, Inosiplex, Insulin chain B (9-23) peptide, Intaxel 50 Magainin II, Makaluvamine C. Makaluvamine D. Makalu (from Himalayan Yew), Integramycin, Intoplicine, Intri vamine E. Makaluvamine F, Makaluvone, Managlinat fiban, Iobenguane 131I]. Iobitridol, Iodixanol, Iodoproxy dialanetil, Manidipine hydrochloride, Manifaxine hydro fan, Iodorubidazone (p), Iofetamine hydrochloride 1-123, chloride, Manitimus, Mannopeptimycin alpha, Mannopep Iofratol, Iohexol, Iolopride (123I), Iomeprol, Iopamidol, timycin beta, Mannopeptimycin delta, Mannopeptimycin Iopentol, Iopromide, Iotriside, Iotrol, Iotrolan, Ioversol, 55 epsilon, Mannopeptimycin gamma, Manumycin A, Ioxilan, Ioxipride, Ipazilide fumarate. Iptakalim hydrochlo Manumycin B. Manumycin C, Manumycin E. Manumycin ride, Irbesartan, Irciniastatin A, Irciniastatin B. IroXanadine, F. Manumycin G. Manzamine A. Manzamine B. Manzamine Irtemazole, Isaglidole, IsaglitaZone, Isalsteine, Isatoribine, C. Manzamine D. Manzamine E. Manzamine F. Maprotiline Isavuconazonium chloride hydrochloride, Iseganan hydro hydrochloride, Maraviroc, Maribavir, Marimastat, Maropi chloride, Isepamicin Sulfate, Isocarboxazid, Isofagomine 60 tant, Masilukast, Masitinib mesylate, Masnidipine hydro tartrate, Isoniazid, Isoquine, Isosegoline A, Isovanihuper chloride, MASTPROM, Matlystatin A, Matlystatin B, Zine A, Ispronicline, Isradipine, Itasetron, Itopride, Itopride Matlystatin D. Matlystatin E. Matly statin F. Matristatin A1, hydrochloride, Itriglumide, Iturelix, Ixabepilone, Janthino Matristatin A2, Matristatin B1, Matristatin D1, Matristatin mycin A, Janthinomycin B. Janthinomycin C, Jaspamide, E1, Matristatin F1, Maxadilan, Mazokalim, Mebrofenin, Jasplakinolide, K9-Retrocyclin-1, Kahalalide F, Kaito 65 Mecamylamine hydrochloride, Meclinertant, Meclofena cephalin, Kanglemycin A, kappa-Conotoxin PVIIA, Kassi mate sodium, Medetomidine, Mefenamic acid, Mefloquine natuerin-1, Katanosin A, Katanosin B. Ketamine hydrochlo hydrochloride, Megovalicin A, Megovalicin B. Megovalicin US 9,533,056 B2 35 36 C, Megovalicin D. Megovalicin G. Megovalicin H, Mela Nepadutant, Nepaprazole, Nepicastat hydrochloride, Nera gatran, Melanotan, Melanotan I, Melanotan II, Meldonium, tinib, Nerfilin I, Nesbuvir, Nesiritide, Netamiftide trifluoro Melogliptin, Meloxicam, Meluadrine, Meluadrine tartrate, acetate, Netilmicin sulfate, Netivudine, Netoglitazone, Memno-peptide A, Memoquin, Mephenytoin, Mephobarbi Neu5Ac2en, Neuromedin U-25, Neuropeptide S. Neutro tal, Mepindolol sulfate, Mepindolol transdermal patch, phil-activating factor, Nevirapine, Ngercheumicin A, Mepirodipine hydrochloride, Mepivacaine hydrochloride, Ngercheumicin B. Ngercheumicin C, Ngercheumicin D, Mercaptopurine, Merimepodlib, Meriolin-3, Meropenem, Ngercheumicin E, Nibentan, Nicardipine hydrochloride, Mersacidin, Mesopram, Metaglidasen, Me-Talnetant, Nicavir, Nicorandil, Nicotredole, Niduline, Nifedipine, Metanicotine, Metaproterenol sulfate, Meterelin, Metergo Nifekalant hydrochloride, Nifurzide, Niguldipine hydro line, Metesind glucuronate, Metformin hydrochloride, 10 chloride, Nilotinib hydrochloride monohydrate, Nilutamide, Methamphetamine hydrochloride, Methanobactin, Methicil Nilvadipine, Nimesulide, Nimodipine, Nipradolol, Nisin, lin sodium, Methimepip, Methoctramine, Methoin, Metho Nisoldipine, Nitazoxanide, Nitracrine dihydrochloride trexate, Methoxatin, Methyclothiazide, Methyl bestatin, hydrate, Nitrazepam, Nitrendipine, Nitrofenac, Nitroparac Methylhistaprodifen, Methylhomoindanomycin, Methyl etamol, Nitroso-nifedipine, Nitrosopine, Nitrovin, Nivad phenidate hydrochloride, Methylphenobarbital, Methylphe 15 ipine, Nizatidine, Noberastine, Noberastine citrate, nobarbitone, Methylthio-DADMe-immucillin-A, Methypra Nocathiacin I, Nocathiacin II, Nocathiacin III, Nocathiacin nolol, Methysergide maleate, Meticillin sodium, IV, NO-ciprofloxacin, N-Octyl-beta-valienamine, NO-ibu Metipranolol, Metoclopramide hydrochloride, Metolazone, profen, Nolatrexed dihydrochloride, Nolomirole hydrochlo Metoprolol fumarate, Metoprolol succinate, Metoprolol tar ride, NO-Nifedipine, Nooglutil, Nooglutyl, NO-Paraceta trate, Mezacopride, Mibefradil, Mibefradil hydrochloride, mol, Norastemizole, Nordazepam, Norfloxacin, Micafungin sodium, Michellamine B, Microcin 25, Micro Nornicotine, Norsegoline, Nortopixantrone hydrochloride, cin J25, Microcolin A, Microcolin B, Micronomicin sulfate, Nortopsentin A, Nortopsentin B, Nortopsentin C, Nor Midafotel, Midaxifylline, Mideplanin, Midesteine, top sentin D, Nortriptyline hydrochloride, Nostocyclopep Midostaurin, Milacemide2H. Milataxel, Milbemycin tide M1, N-Retinoyl-D-glucosamine, N-tert butyl isoquine, alpha-9, Milfasartan, Milrinone, Milrinone lactate, Mimo 25 Nubiotic 2, Nutlin-3, Nutlin-3A, Nutlin-3-enantiomer A, pezil, Minalrestat, Minaprine hydrochloride, Minopafant, Nuvanil, O6-Benzylguanine, Obatoclax mesylate, Mipragoside, Mirabegron, Mirisetron, Mirodenafil hydro Oberadillol, Oberadilol monoethyl maleate, Octacosamicin chloride, Mitiromycin, Mitomycin, Mitomycin C. Mitoxan A, Octacosamicin B, Octreother, Octreotide acetate, Oct trone hydrochloride, Mitoxantrone hydrochloride, Mivaz reotide LAR, Odanacatib, O-Demethylmurrayafoline A, erol, Mivobulin isethionate, Mivotilate, Mixanpril, 30 Oglufanide disodium, Olanexidine hydrochloride, Olanzap Mizolastine, Mobazol, Mocetinostat dihydrobromide, ine, Olanzapine pamoate, Olaparib, Olcegepant, Olmesar Moclobemide, Modecainide, Modipafant, Moenomycin A tan, Olmesartan medoxomil, Olprinone hydrochloride, Olra chloride bismuth salt, Moexipril hydrochloride, Moexiprilat, dipine hydrochloride, Omaciclovir, Omapatrilat, Monamidocin, Monodansyl cadaverine, Mono-L-asparty1 Ombrabulin, Ombrabulin hydrochloride, omega-Conotoxin chlorin e6, Monophosphoryl lipid A. Montirelin tetrahy 35 CVID, omega-Conotoxin MVIIA, Omeprazole, Omiganan drate, Moracizine hydrochloride, Moranolin, Moricizine pentahydrochloride, Onnamide A, Ontazolast, Opaviraline, hydrochloride, Morniflumate, Mosapramine hydrochloride, OPC-17083, Opiorphin, Orbifloxacin, Orbofiban acetate, Mosapride citrate, Motesanib diphosphate, Motretinide, Orciprenaline sulphate. Orienticin A, Orienticin B. Orienti Moxalactam disodium, Moxifetin hydrogen maleate, Moxi cin C, Orienticin D. Oritavancin, Orlipastat, Orlistat, floxacin hydrochloride, Moxonidine hydrochloride hydrate, 40 Ortataxel, Oseltamivir carboxylate, Oseltamivir phosphate, Mozavaptan hydrochloride, mu-Conotoxin IIIA, Multiple Osemozotan hydrochloride, Osutidine, Otamixaban, Otastat sclerosis vaccine, muO-Conotoxin MrVIB, Muraminomicin potassium, Otenabant hydrochloride, Oteracil potassium, A, Muraminomicin B, Muraminomicin C, Muraminomicin Ovalicin A, Ovothiol B, Oxamflatin, Oxatomide, Oxazepam, D, Muraminomicin E1, Muraminomicin E2, Muraminomi Oxeclosporin, Oxiglutatione sodium, Oximidine III, Oxonic cin F. Muraminomicin G, Muraminomicin H, Muraminomi 45 acid, Oxprenolol hydrochloride, Oxymetazoline hydrochlo cin I, Muraminomicin Z1, Muraminomicin Z2, Muramin ride, Oxymethacyl, Oxymorphazole dihydrochloride, omicin Z3, Muraminomicin Z4, Muramyl dipeptide C. Oxynor, Oxypertine, Oystrisin, Ozarelix, Ozenoxacin, Mureidomycin A, Mureidomycin B. Mureidomycin C, Pachymedusa dacnicolor Tryptophyllin-1, Pachysamine E. Mureidomycin D. Mureidomycin E, Mureidomycin F, Paclitaxel, Paclitaxel ceribate, Pactimibe, Padeliporfin Mureidomycins, Muroctasin, Mycalamide A, Mycalamide 50 potassium, Pafenolol, Palau'amine, Paldimycin B, Palinavir, B. Myxovirescin A1, Myxovirescin B, N1,N8-Bisnorcym Palindore fumarate, Palmidrol, Palmitoylethanolamide, serine, N1-Phenethylnorcymserine, N4-Hexadecyl-dC Palosuran Sulfate, Pamapimod, p-Aminoclonidine hydro AZT, Naamidine A, N-Acetylcolchinol, N-Acetylcysteine, chloride, Pancopride, Pancratistatin disodium phosphate, N-Acetylesperamycin A1, N-Acetylesperamycin Alb, Pancratistatin-3,4-cyclic phosphate sodium salt, Panobinos N-Acetylesperamycin A2, N-Acetyl-L-cysteine, Nadolol. 55 tat, Pantethine, Pantoprazole, Papuamide A, Papuamide B, Nafadotride, Nafamostat mesilate, Nafamostat mesylate, Papuamide C. Papuamide D, Paracetamol, Paraherquamide Nafarelin acetate, Naglivan, NagreStipen, Nagstatin, Nal E. Paraherquamide F, Paraherquamide G, Parasin I, Para trindole, Naluzotan hydrochloride, Naminidil, Naproxen thyroid hormone (human recombinant), Parcetasal, Pardo piperazine (2:1), Napsagatran, Napsamycin A, Napsamycin prunox hydrochloride, Parodilol hemifumarate, Parogrelil B, Napsamycin C, Napsamycin D. Naratriptan hydrochlo 60 hydrochloride, Paroxetine, Paroxetine ascorbate, Paroxetine ride, Nardeterol, Nateglinide, Navelbine, Navuridine, Naxi camsilate, Paroxetine hydrochloride, Paroxetine mesilate, fylline, Nazasetron, Nazasetron hydrochloride, N-demeth Pasireotide, Pazelliptine trihydrochloride, Pazelliptine trihy ylated sildenafil. N-Desmethylmilameline, Nebivolol. drochloride monohydrate, Pazopanib hydrochloride, Neboglamine, Nebostinel, Necrostatin-1, Nefiracetam, Nei PEG40000-Paclitaxel, PEG5000-Paclitaxel, PEG-vancomy humicin, Nelfinavir mesilate, Nemifitide ditriflutate, Nem 65 cin, Peldesine, Pelitinib, Pelitrexol, Pemetrexed disodium, onapride, Neo-acridine, Neomycin B-arginine conjugate, Pemirolast, Pemirolast potassium, Pemoline, Penbutolol sul Neomycin B-hexaarginine conjugate, Neomycin-acridine, fate, Penciclovir, Penicillin G procaine, Penicillin G sodium, US 9,533,056 B2 37 38 Pentafuside, Pentobarbital sodium, Pentobarbitone sodium, Rafabegron, Ragaglitazar L-arginine salt, Ralfinamide, Pentostatin, Peplomycin, Pepticinnamin E. Peptide Leucine Raltegravir potassium, Raltitrexed, Raluridine, Ramatroban, Arginine, Peramivir, Perfosfamide, Pergolide mesylate, Per Ramelteon, Ramipril, Ramoplanin A1, Ramoplanin A2, indopril, Perzinfotel, Pexiganan acetate, PG-camptothecin, Ramoplanin A3. Ramorelix, Ramosetron hydrochloride, Phakellistatin 5, Phakellistatin 7, Phakellistatin 8, Phakel Ranbezolid hydrochloride, Ranimustine, Ranirestat, Raniti listatin 9, Phe-Arg-beta-naphthylamide, Phendioxan, dine bismuth citrate, Ranitidine Bismutrex, Ranitidine Phenelfamycin F. Phenelzine sulfate, Phenobarbital, Pheno hydrochloride, Ranitidine nitrate, Ranitidine zinc chloride barbitone, Phenochalasin A, Phenochalasin B, Phenoxazole, complex, Ranitidine Zinc citrate, Ranolazine, Rasagiline Phenserine tartrate, Phentolamine mesilate, Phenylisohy mesilate, Rat adrenomedullin, Razaxaban hydrochloride, dantoin, Phenylpseudohydantoin, Phenytoin sodium, Phe 10 RaZupenem, Rebamipide, Rebamipide bismuth citrate valin, Phomopsichalasin, Phortress, Phosphazid, Phosphe tetramethyledamine, Rebamipide bismuth L-tartrate tetram nytoin sodium, Photofrin II, Physostigmine salicylate, ethyledamine, Rebimastat, Reblastatin, Reboxetine mesi Piboserod hydrochloride, Pibrozelesin hydrochloride, Pibu late, Receptor mediated permeabilizer, Recombinant human tidine hydrochloride, Piceasin, Piclamilast, Picotamide, parathyroid hormone (1-84), Recombinant Jerdostatin, Picumeterol fumarate, Pidobenzone, Pidolacetamol, Pidol 15 Regadenoson, Reglitazar, Relacatib, Relaxin-3/INSL5 chi ate magnesium, Pidotimod, Pifatidine, Pikamilone, Piketo meric peptide, Remacemide hydrochloride, Remikiren mesi profen, Pilsicainide hydrochloride, Pimagedine, Pimavan late, Reminertant, Remoxipride, Remoxipride hydrochloride serin tartrate, Pimeloylanilide o-aminoanilide, Pimobendan, monohydrate, Repaglinide, Reparixin, Repertaxin, Repino Pimozide, Pinacidil, Pindolol, Pioglitazone, Pioglitazone tan, Repinotan hydrochloride, Repirinast, Reproterol hydro hydrochloride, Pipalamycin, Piperacillin sodium, Piper chloride, Resatorvid, Resequinil, Reserpine, Retaspimycin afizine A, Piperafizine B, Piproxen, Piragliatin, Pirbuterol hydrochloride, Retigabine hydrochloride, Retosiban, hydrochloride, Pirenzepine hydrochloride, Piroxicam, Revaprazan hydrochloride, Reversine, Revizinone, Rhodo Piroxicam cinnamate, Piroxicam pivalate, Piscidin 1, Pisci peptin C1, Rhodopeptin C2, Rhodopeptin C3, Rhodopeptin din 2, Piscidin 3, Pittsburgh Compound B, Pivagabine, C4, Rhodostreptomycin A, Rhodostreptomycin B. Ricase Pivampicillin, Pixantrone maleate, Platencin, Platensimy 25 tron, Rifabutin, Rifalazil, Rifamexil, Rifampicin, Rifampin, cin, Plerixafor hydrochloride, Plevitrexed, Plinabulin, Pliti Rifapentine, Rifaximin, Rilmazafone hydrochloride dihy dep sin, Plusbacin A1, Plusbacin A2, Plusbacin A3, Plusba drate, Rilmenidine dihydrogen phosphate, Rilpivirine, cin A4, Plusbacin B1, Plusbacin B2, Plusbacin B3, Rimonabant, Rimoterol hydrobromide, Ripisartan, Riso Plusbacin B4, Pneumocandin A0, Pneumocandin B0, Pneu tilide, Rispenzepine hydrochloride, Ritonavir, Ritonavir mocandin B0 2-phosphate, Pneumocandin D0, Polaprezinc, 30 oxymethylphosphate, Rivanicline, RivaroXaban, Rivoglita Polydiscamide A, Polyglutamate camptothecin, Polymer Zone, Rizatriptan benzoate, Rizatriptan Sulfate, r-Jerdostatin, bound human leukocyte elastase inhibitor, Polythiazide, Robenacoxib, Robotnikinin, Rocepafant, Roflumilast, Porfimer sodium, Poststatin, Pozanicline hydrochloride, Rolapitant hydrochloride, Rolipram, Rolofylline, Romergo PPI17-24, Pradimicin A, Pradimicin B, Pradimicin C, line, Romidepsin, Romurtide, Ropinirole hydrochloride, Pradimicin D, Pradimicin E, Pradimicin FA-1, Pradimicin 35 Ropivacaine hydrochloride, Rosabulin, Roscovitine, FA-2, Pradimicin FL, Pradimicin FS ((+)-enantiomer), RosiglitaZone maleate, Rosonabant, Rotigaptide, Roxatidine Pradimicin L, Pradimicin S, Pradimicin T1, Pradimicin T2, acetate hydrochloride, Roxatidine bismuth citrate, Roxifiban Pradofloxacin, Pralatrexate, Pralmorelin, Pralnacasan, acetate, Roxindole mesilate, Ruboxistaurin hydrochloride, Pramipexole hydrochloride, Pramipide, Pramiracetam RubOxistaurin mesilate hydrate, Rumycin 1, Rumycin 2, hydrochloride, Pramlintide acetate, Pranazepide, Prane 40 Rupintrivir, Ruprintrivir, Sabiporide mesilate, Safinamide dipine tartrate, Pranidipine, Pranlukast hydrate, Pratosartan, mesilate, Safironil, Sagamacin, Sagandipine, Salazodine, Prefolic A. Premafloxacin, Premafloxacin hydrochloride, Salazopyridazin, Salazosulfapyridine, Salbostatin, Salbuta Premafloxacin magnesium, Prezatide copper acetate, Prima mol nitrate, Salbutamol Sulfate, Salcaprozic acid sodium quine phosphate, Primidone, Prinomastat, Prinomide salt, Salicylihalamide A, Salicylihalamide B, Salinamide A, tromethamine, Prisotinol, Pristinamycin IA, Pristinamycin 45 Salinosporamide A, Saliphenylhalamide, Salmaterol, Sal IB, Pristinamycin IIA, Pristinamycin IIB, Proamipide, meterol, Salmeterol Xinafoate, Salmisteine, Salubrinal, Probestin, Procainamide hydrochloride, Procaine Penicillin, Samixogrel, Sampatrilat, Sampidine, Sanglifehrin A, San Procaterol hydrochloride hemihydrate, Procysteine, Pro-di glifehrin B, Sanglifehrin C, Sanglifehrin D. Sanilvudine, azepam, Proglumide, Propacetamol hydrochloride, Sapacitabine, Saprisartan, Sapropterin dihydrochloride, Propafenone hydrochloride, Propeptin, Propeptin T. Propra 50 Saquinavir, Saquinavir mesilate, Sardomozide, SardomoZ nolol hydrochloride, Prostatin, Protaxel, Protegrin IB-367, ide hydrochloride, Saredutant, Sarizotan hydrochloride, Proterguride, Protriptyline hydrochloride, Proxodolol, Pru Satoribine, Saussureamine C, Saviprazole, Sazetidine-A, calopride, Prucalopride hydrochloride, Prucalopride succi Schizandrin glycinosuccinate sodium salt, Scyphostatin, nate, Pruvanserin hydrochloride, Pseudoephedrine hydro SecinH-3, Secobarbital sodium, Secobatzelline A, Secobat chloride, Pseudomycin A", Pseudomycin B', Psymberin, 55 Zelline B, Seglitide, Segoline A, Segoline B, Selanc, Selank, Ptidepsin, Pumaprazole, Pumosetrag hydrochloride, Purva Selodenoson, Semagacestat, Semapimod hydrochloride, lanol A, Pyloricidin B, Pymeprazole, Pyrazinoylguanidine, Semaxanib, Semaxinib, Semoxind, Semparatide, Sepimo Pyrazoloacridine, Pyridinostatin, Pyridone-6, Pyriferone, stat mesilate, Seraspenide, Sermorelin, Sertindole, Sertra Pyrindamycin A. Pyrindamycin B, Pyroxamide, Pyrrocidine line, Sertraline hydrochloride, Setamycin, Setastine hydro A. Pyrrocidine B, Pyrroloquinoline quinone, Pyrrolosporin 60 chloride, Setazindol, Setipafant, Sezolamide hydrochloride, A. Pyrrophenone, Quarfloxin, Quinagolide hydrochloride, Shepherdin, Shermilamine D, Shiga vaccine, Shishijimicin Quinalbarbitone sodium, Quinapril hydrochloride, Quinet A, Shishijimicin B, Shishijimicin C, Sialosylcholesterol haZone, Quinotolast sodium, Quinoxapeptin C. Quinupristin alpha sodium salt, Sialosylcholesterol-beta sodium salt, Sia mesilate, R9K-Retrocyclin, Rabeximod, rac-Debromoflus mycin I, Siamycin II, Sibanomicin, Sibenadet hydrochlo tramine E. rac-Deoxypseudophrynaminol, Racecadotril, rac 65 ride, Sibrafiban, Sifuvirtide, Sildenafil citrate, Sildenafil Ptilocaulin nitrate, rac-threo-Methylphenidate hydrochlo mesilate, Sildenafil nitrate, Sildenafil N-oxide, Silodosin, ride, Radequinil, Radezolid, Radolmidine hydrochloride, SiltenZepine, Simendan, Simotaxel, Sinorphan, Sitamaquine US 9,533,056 B2 39 40 hydrochloride, Sivelestat sodium hydrate, Sivifene, SNAP Contulakin G, Threitol ceramide, Thymalfasin, Thymallene, 7292, SNAP-7493, S-Nitrosoglutathione, Soblidotin, Socor Thymectacin, Thymic humoral factor gamma-2, Thymocto romycin, Sofigatran, Sofinicline, Solabegron hydrochloride, nan, Thymodolic acid, Thymopentin, Thymosin alpha 1. Solimastat, Solpecainol hydrochloride. Somocystinamide A, Tiamdipine, Tiamenidine, Tianeptine Sodium, Tiapafant, Sorafenib, Sorafenib tosylate, Soraprazan, Sorbicillactone Tiapride hydrochloride, Tidembersat, Tienartan, Tienoxolol A, Soretolide, Sorivudine, Sotalol hydrochloride, Sotrastau hydrochloride, Tifenazoxide, Tifluvirtide, Tigecycline, Tigil rin, Sparfloxacin, Sparoxomycin A1, Sparoxomycin A2, cycline, Tilarginine hydrochloride, Tilisolol hydrochloride, Sperabillin A, Sperabillin B. Sperabillin C, Sperabillin D, Timodepressin, Timodolic acid, Timogen, Timolol hemihy Spermidine/lysine3/CTS6, Spermine dialdehyde, Spermine/ drate, Timolol maleate, Tinazoline hydrohloride, Tiobutarit, lysine4/CTS8, Spinorphin, Spiralizone B, Spirapril, 10 Tiocol 33, Tiocol 54, Tiplimotide, Tipranavir, Tiracizine Spiriprostil, Spiroglumide, Spiroquinazoline, Spiruchostatin hydrochloride, Tirofiban hydrochloride, Tiropramide, Tisar A, Spiruchostatin B, Sprodiamide, Squalamine lactate, tan, Tivirapine, Tizanidine hydrochloride, TNF-alpha pro Stampidine, Stavudine, Stearyl-norleucine-VIP Sterenin C, tease enzyme inhibitor, Toborinone, Tocainide hydrochlo Streptomycin, Stressin1-A, Styloguanidine, Suberanilohy ride, Tokaramide A, Tolafentrine, Tolbutamide, Tolfenamic droxamic acid, Suberoylanilide hydroxamic acid, Substance 15 acid, Tolvaptan, Tomatine, Tomeglovir, Tomopenem, P(8-11), Sufotidine bismuth citrate, Sufugolix, SulaZuril, Tomoxetine hydrochloride, Tonabersat, Topixantrone hydro Sulcephalosporin, Sulfasalazine, Sulfostin, Sulofenur, Sul chloride, Topostatin, TopSentin, Top sentine B1, Torasemide, phasalazine, Sulphostin, Sulpiride, Sulpiride L-(-), Sul Torsemide, Tosedostat, Tozasertib, Trabectedin, Tramazo prostone, Sultamicillin tosylate, Sultopride, Sulukast, line, Trandolapril, Trandolaprilat, Tranilast, trans,trans Sumanirole maleate, Sumatriptan Succinate, Sunflower Ceratospongamide, Transdihydrolisuride, Trantinterol trypsin inhibitor-1, Sunitinib malate, Super-Leu-Dox, hydrochloride, Trapoxin A, Trapoxin B, Trecetilide fumar Suplatast tosilate, Suprofenac 1, Suprofenac 2, Suprofenac ate, Treprostinil diethanolamine, Tresperimus triflutate, 3, Suradista, Suramin sodium, Surfen, Surinabant, Surona Trewiasine, Triacetyl dynemicin C, Trichostatin D, Tricif crine maleate, Susalimod, Symplostatin 1, Synguanol, Syn erol, Trientine hydrochloride, Trifluproxim, Trifluridine, thadotin, Synthetic human secretin, Synthetic neutrophil 25 Trimetazidine, Trimetrexate glucuronate, Trimexautide, inhibitor peptide, Synthetic porcine secretin, Tabilautide, Triproamylin acetate, TroglitaZone, Trombodipine, Tropise Tabimorelin, Tacapenem pivoxil, Tacedinaline, Tadalafil. tron, Troquidazole, Trovirdine hydrochloride, Troxipide, Tafenoquine Succinate, Tageflar, Talaglumetad hydrochlo Trunkamide A, Tryptamide, Tubastrine ((+)-enantiomer), ride, Talaporfin gold Sodium, Talaporfin Sodium, Talarozole, Tubingensin B, Tubulysin A, Tubulysin B, Tubulysin C, Talibegron, Talibegron hydrochloride, Tallimustine hydro 30 Tuftsin, Tulathromycin A, Tulathromycin B, Tulobuterol chloride, Talnetant, Talniflumate, Talopterin, Talotrexin, hydrochloride, Tulobuterol-(R), Tulopafant, Tumor necrosis Taltirelin, Taltobulin, Taludipine hydrochloride, Talviraline, factor-alpha protease inhibitor, Turbostatin 1, Turbostatin 2, Tamandarin A, Tamandarin B, Tamatinib fosdium, Turbostatin 3, Turbostatin 4, Turofexorate isopropyl, Turo Tamibarotene, Tamsulosin hydrochloride, Tanaproget, Tan steride ((-)-enantiomer), Tyropeptin A10, Tyropeptin A2, dutinib, Tanespimycin, Tanogitran, TAP-doxorubicin, 35 Tyropeptin A6, Tyropeptin A7, Tyropeptin A9, Tyroser Taranabant, Tarazepide, Targinine, Targinine hydrochloride, valtide, Tyroservatide, Ubenimex, Ubenimex methyl ester, Tariquidar, Tasidotin hydrochloride, Tasimelteon, Tasosar Ubestatin, Ubidine, Udenafil. Ufenamate, Ukrain, Ulicycla tan, Taurohyodeoxycholic acid, Tauroiodeoxycholic acid, mide, Ulifloxacin, Ulithiacyclamide A, Ulvenzepine hydro Taurolidine, Tauropyrone, Taurosteine, Taxuyunnanine, chloride, Uncialamycin, Uniroyal JR, Unnarmicin A, Unnar Tazanolast, Tazofelone, Tazopsine, Tebanicline, Tecadeno 40 micin C, Upidosin, Upidosin hydrochloride hydrate, son, Tecalcet hydrochloride, Tecastemizole, Technetium Urapidil, Uroguanylin, Urukthapelstatin A, Utibapril, (99mTc) apcitide, Technetium (99mTc) bicisate, Technetium Utibaprilat, V. Vabicaserin hydrochloride, Valaciclovir, (99mTc) depreotide, Technetium Tc 99m depreotide, Tech Valacyclovir, Valganciclovir hydrochloride, Valnemulin, netium Tc99m bicisate, Tecovirimat, Tegafur, Tegaserod Valomaciclovir Stearate, Valrocemide, Valrubicin, Valsartan, maleate, Teglicar, Teicoplanin-A2-1, Teicoplanin-A2-2, Tei 45 Valsartan axetil, Valsartan cilexetil, Valsartan pivoxil, Val coplanin-A2-3, Teicoplanin-A2-5, Telaprevir, Telatinib, spodar, Vancomycin hydrochloride, Vandetanib, Vanidipin Telavancin hydrochloride, Telbivudine, Telcagepant, Telina edilol, Vaninolol, Vapitadine hydrochloride, Vardenafil. vir, Teludipine hydrochloride, Temacrazine mesilate, Tema Vardenafil dihydrochloride, Vardenafil hydrochloride floxacin hydrochloride, Temocapril hydrochloride, Temocil hydrate, Varenicline tartrate, Variapeptin, Varlitinib, lin Sodium, Temoporfin, Temurtide, Tenatoprazole, Tenovin 50 Vasonatrin peptide, Vatalanib Succinate, Vatanidipine, 6, Tenoxicam, Terbequinil, Terbogrel, Terbutaline sulfate, Vatanidipine hydrochloride, V-Echinocandin, Velneperit, Terestigmine tartrate, Terguride, Teriflunomide, Teri Venorphin, Veralipride, Verongamine, Verticillatine, Vesicu paratide, Terlakiren, Terlipressin, Ternatin, Terodiline lin, Vesnarinone, Vestipitant mesylate. V-Glycopeptide, hydrochloride, Tertatolol hydrochloride, Tertomotide, Ter Vicenistatin, Vilazodone hydrochloride, Vildagliptin, Vilox utroban Sodium, Tesetaxel, Tetrabromostyloguanidine, Tet 55 azine hydrochloride, Vincristine sulfate, Vindesine, Vinflu racaine hydrochloride, Tetrahydrobiopterin, Tetrahydroder nine, Vinfosiltine sulfate, Vinleucinol, Vinorelbine, Vinxal citin 1, Tetrahydroechinocandin B, Tetrahydrolipstatin, tine Sulfate, Vinylamycin, Virginamycin M2, Virginiamycin Tetrazolast meglumine, Tetrindol, Tetrindole, Texenomycin M1, Virgisin-1, Virgisin-2, Viscosin, Vistonuridine, Vitile A, Textilinin-1, Tezampanel, Tezosentan, TGP. Thalido Vuamide, Voclosporin, Vofopiitant hydrochloride, Voglibose, mide, Thanatin, Theopederin D. Theoperidin E. Theophyl 60 Volpristin, Voreloxin, Vorinostat, Voxergolide hydrochlo line. Thiacymserine. Thiamet-G, Thiamphenicol, Thiamylal, ride, W Peptide, Watanidipine, Watanidipine hydrochloride, Thiatolserine. Thiazinotrienomycin B. Thiazinotrienomycin Xamoterol fumarate, Xemilofiban, Xenomin 1, Xenomin 2, F. Thiazinotrienomycin G. Thiazohalostatin, Thiocoraline, Xenoxin-1, Xenoxin-2, Xenoxin-3, Ximelagatran, Xip Thiocoraline A, Thiocoraline NF. Thiofedrine. Thiomarinol, amide, Yatakemycin, Yohimbine. Yttrium-90 edotreotide, Thiomarinol B, Thiomarinol C, Thiomarinol D, Thiomarinol 65 Zabicipril hydrochloride, Zabiciprilat hydrochloride, Zabo E. Thiomarinol F. Thionisoxetine-(R). Thiopental sodium, floxacin hydrochloride, Zafirlukast, Zampanolide, Zanami Thiopentone sodium, Thioviridamide. Thioxamycin, Thr10 Vir, Zanapezil fumarate, Zankiren, Zardaverine, Zatosetron, US 9,533,056 B2 41 42 Zatosetron maleate, Zelandopam hydrochloride, Zibotentan, A preferred aspect of the present invention is a carrier Ziconotide, Zidampidine, Zidovudine, Zilpaterol, Zinc aceX linked prodrug comprising a biodegradable hydrogel as amate, Ziprasidone hydrochloride, Ziprasidone mesilate, carrier, wherein a number of permanent linkages of the Zofenoprilat arginine, Zolasartan, Zolmitriptan, Zonam backbone moieties exist with the linker L to which the panel, Zorubicin hydrochloride, biologically active moiety is covalently attached. D may contain further functional groups besides at least Ideally, the hydrogel-connected drug-linker conjugates one aliphatic amino group to which the promoiety is bound, are dispersed homogeneously throughout the hydrogel Such further functional group may be aliphatic or aromatic according to the invention, and may or may not be present amines, amides, alcohols, carbonyls, carboxylic acids, on the Surface of the hydrogel according to the invention. thiols. The term “aliphatic' (aliphatic fragment) means any 10 The functional groups may be attached to a linear chain. aliphatic fragment known to a person skilled in the art. In this case, the functional groups may be spaced regularly Preferably, the carrier group Z (PEG or hydrogel) is a or irregularly across the chain, or alternatively, the chain polymer with a molecular weight 500 g/mol. may be terminated by two dendritic moieties, providing for In one embodiment, the carrier Z may be a PEG moiety. the total of functional groups. Such PEG moiety may be attached to the biologically active 15 Remaining reactive functional groups which are not con agent through one or more linkages. In case of one linkage, nected to a transient prodrug linker or to a spacer connected the corresponding PEG in the PEG prodrug monoconjugate to a transient prodrug linker may be capped with Suitable may be linear or branched. In case of more than one linkage, blocking reagents. such as in a bisconjugate, the corresponding PEG in the PEG Preferably, the covalent attachment formed between the prodrug may be linear or branched. Bisconjugates may reactive functional groups provided by the backbone moi contain one or two transient linkages, and PEG may be eties and the prodrug linker are permanent bonds. Suitable linear or branched or may contain a mixture of one linear functional groups for attachment of the prodrug linker to the and one branched chain. In case the bisconjugate contains hydrogel according to the invention include but are not one transient linkage and one linear and one branched chain limited to carboxylic acid and derivatives, carbonate and the transient linkage may be on either chain. In case a 25 derivatives, hydroxyl, hydrazine, hydroxylamine, maleamic branched PEG chain is used, there may be one or more acid and derivatives, ketone, amino, aldehyde, thiol and branching units. disulfide. A branched PEG is a PEG molecule consisting of a Such biodegradable hydrogel may be composed of back branching point connecting two or more PEG chains, to bone moieties interconnected by hydrolytically degradable form a molecule with one anchoring point for attachment to 30 bonds. The backbone moiety is characterized by a number of the biologically active agent. This could be two 20 kDa PEG functional groups, comprising interconnected biodegradable chains joined together to form one branched 40 kDa PEG functional groups and hydrogel-connected drug-linker con molecule. In the case where the molecule contains two or jugates, and optionally capping groups. This means that a three branching points, the molecule is referred to 3- and backbone moiety is characterized by a number of hydrogel 4-armed PEG, respectively. 35 connected drug-linker conjugates; functional groups, com In Summary and within the restrictions mentioned above, prising biodegradable interconnected functional groups; and the PEG polymer is not limited to a particular structure and optionally capping groups. Preferably, the Sum of intercon can be linear, branched, or multi-branched. nected biodegradable functional groups and drug-linker con Preferably, Z is a hydrogel and more preferably a PEG jugates and capping groups is 16-128, preferred 20-100, based hydrogel. Preferably, the covalent attachment formed 40 more preferred 24-80 and most preferred 30-60. between the linker and the hydrogel Z is a permanent bond. Preferably, the sum of interconnected functional groups The term "PEG based as understood herein means that the and hydrogel-connected drug-linker conjugates and capping mass proportion of PEG chains in the hydrogel is at least groups of a backbone moiety is equally divided by the 10% by weight, preferably at least 25%, based on the total number of PEG-based polymeric chains extending from the weight of the hydrogel. The remainder can be made up of 45 branching core. For instance, if there are 32 interconnected spacers and/or oligomers or polymers, such as oligo- or functional groups and hydrogel-connected drug-linker con polylysines. jugates and capping groups, eight groups may be provided Moreover, the term “water-insoluble” refers to a swellable by each of the four PEG-based polymeric chains extending three-dimensionally crosslinked molecular network forming from the core, preferably by means of dendritic moieties the hydrogel. If Suspended in a large Surplus of water or 50 attached to the terminus of each PEG-based polymeric aqueous buffer of physiological osmolality the hydrogel may chain. Alternatively, four groups may be provided by each of take up a Substantial amount of water, e.g. up to 10-fold on eight PEG-based polymeric chains extending from the core a weight per weight basis, and is therefore swellable but or two groups by each of sixteen PEG-based polymeric after removing excess water still retains the physical stabil chains. If the number of PEG-based polymeric chains ity of a gel and a shape. Such shape may be of any geometry 55 extending from the branching core does not allow for an and it is understood that Such an individual hydrogel object equal distribution, it is preferred that the deviation from the is to be considered as a single molecule consisting of mean number of the Sum of interconnected functional components wherein each component is connected to each groups and hydrogel-connected drug-linker conjugates and other component through chemical bonds. capping groups per PEG-based polymeric chain is kept to a According to this invention, the hydrogel may be com 60 minimum. posed of backbone moieties interconnected by hydrolyti Preferably, a backbone moiety is further characterized by cally degradable bonds. having a branching core, from which at least three PEG Preferably, the backbone moiety has a molecular weight based polymeric chains extend. Accordingly, in a preferred in the range of from 1 kDa to 20 kDa, more preferably from aspect the backbone reagent comprises a branching core, 1 kDa to 15 kDa and even more preferably from 1 kDa to 10 65 from which at least three PEG-based polymeric chains kDa. The backbone moieties are preferably also PEG-based, extend. Such branching cores may be comprised of poly- or comprising one or more PEG chains. oligoalcohols in bound form, preferably suitably substituted US 9,533,056 B2 43 44 derivatives of pentaerythritol, tripentaerythritol, hexaglyc It is preferred that the sum of interconnected functional erine, Sucrose, Sorbitol, fructose, mannitol, glucose, cellu groups and reactive functional groups of a backbone moiety lose, amylases, starches, hydroxyalkyl starches, polyvinyl is equally divided by the number of PEG-based polymeric alcohols, dextranes, hyaluronans, or branching cores may be chains extending from the branching core. If the number of comprised of poly- or oligoamines such as ornithine, PEG-based polymeric chains extending from the branching diaminobutyric acid, trilysine, tetralysine, pentalysine, core does not allow for an equal distribution, it is preferred hexalysine, heptalysine or oligolysine, nonalysine, decaly that the deviation from the mean number of the sum of sine, undecalysine, dodecalysine, tridecalysine, tetradecaly interconnected and reactive functional groups per PEG sine, pentadecalysine or oligolysines, low-molecular weight based polymeric chain is kept to a minimum. PEI, polyvinylamines, hexaglycerine, tripentaerythritol, in 10 More preferably, the sum of interconnected and reactive bound form. functional groups of a backbone moiety is equally divided Preferably, the branching core extends three to sixteen by the number of PEG-based polymeric chains extending PEG-based polymeric chains, more preferably four to eight. from the branching core. For instance, if there are 32 Preferred branching cores may be comprised of pentaeryth interconnected functional groups and reactive functional ritol, ornithine, diaminobutyric acid, trilysine, tetralysine, 15 groups, eight groups may be provided by each of the four pentalysine, hexalysine, heptalysine or oligolysine, low PEG-based polymeric chains extending from the core, pref molecular weight PEI, hexaglycerine, tripentaerythritol in erably by means of dendritic moieties attached to the ter bound form. Preferably, the branching core extends three to minus of each PEG-based polymeric chain. Alternatively, sixteen PEG-based polymeric chains, more preferably four four groups may be provided by each of eight PEG-based to eight. Preferably, a PEG-based polymeric chain is a linear polymeric chains extending from the core or two groups by poly(ethylene glycol) chain, of which one end is connected each of sixteen PEG-based polymeric chains. to the branching core and the other to a hyperbranched Such multi-arm PEG derivatives may be connected to dendritic moiety. It is understood that a polymeric PEG dendritic moieties to obtain additional functional groups. based chain may be terminated or interrupted by alkyl or aryl Preferably, each dendritic moiety has a molecular weight in groups optionally Substituted with heteroatoms and chemi 25 the range of from 0.4 kDa to 4 kDa, more preferably 0.4kDa cal functional groups. to 2 kDa. Preferably, each dendritic moiety has at least 3 Preferentially, a backbone moiety is characterized by branchings and at least 4 reactive functional groups, and at having a branching core, from which at least three chains most 63 branchings and 64 reactive functional groups, extend. Such branching cores may be provided by suitably preferred at least 7 branchings and at least 8 reactive substituted derivatives of poly- or oligoalcohols, preferably 30 functional groups and at most 31 branchings and 32 reactive pentaerythritol, tripentaerythritol, hexaglycerine, Sucrose, functional groups. sorbitol, fructose, mannitol, glucose, cellulose, amyloses, Examples for such dendritic moieties are comprised of starches, hydroxyalkyl starches, polyvinylalcohols, dex trilysine, tetralysine, pentalysine, hexalysine, heptalysine, tranes, hyualuronans, or branching cores may be provided octadecalysine, nonadecalysine in bound form. Examples by suitably substituted derivatives of poly- or oligoamines 35 for such preferred dendritic moieties are comprised of Such as trilysine, tetralysine, pentalysine, hexalysine, hep trilysine, tetralysine, pentalysine, hexalysine, heptalysine in talysine, octalysine, nonalysine, decalysine, undecalysine, bound form, most preferred trilysine, pentalysine or hep dedecalysine, tridecalysine, tetradecalysine, pentadecaly talysine, ornithine, diaminobutyric acid in bound form. sine or oligolysines, polyethyleneimines, polyvinylamines. Preferably, such dendritic moieties are comprised of Preferably, the branching core extends three to sixteen 40 lysine, dilysine, trilysine, tetralysine, pentalysine, hexaly chains, more preferably four to eight. Preferably, such chain sine, heptalysine, most preferred trilysine, pentalysine or is a linear polyethylene glycol chain, of which one end is heptalyine, in bound form. connected to the branching core and the other to a hyper Most preferably, the hydrogel of the prodrugs of the branched dendritic moiety. present invention is characterized in that the backbone Preferably, a PEG-based polymeric chain is a suitably 45 moiety has a quarternary carbon of formula C(A-Hyp), substituted polyethylene glycol derivative (PEG based). wherein each A is independently a poly(ethylene glycol) Preferred structures for corresponding PEG-based poly based polymeric chain terminally attached to the quarternary meric chains extending from a branching core contained in carbon by a permanent covalent bond and the distal end of a backbone moiety are multi-arm PEG derivatives as, for the PEG-based polymeric chain is covalently bound to a instance, detailed in the products list of Jenkem Technology, 50 dendritic moiety Hyp, each dendritic moiety Hyp having at USA (accessed by download from www.jenkemusa.com on least four groups representing the interconnected functional Jul. 28, 2009), 4ARM-PEG Derivatives (pentaerythritol groups and biodegradable and permanent linkages. core), 8ARM-PEG Derivatives (hexaglycerin core) and Preferably, each A is independently selected from the 8ARM-PEG Derivatives (tripentaerythritol core). Most pre formula —(CH2)(OCHCH),X , wherein n1 is 1 or 2: ferred are 4arm PEG Amine (pentaerythritol core) and 4arm 55 n is an integer in the range of from 5 to 50; and X is a PEG Carboxyl (pentaerythritol core), 8arm PEG Amine functional group covalently linking A and Hyp. (hexaglycerin core), 8arm PEG Carboxyl (hexaglycerin Preferably, A and Hyp are covalently linked by an amide core), 8arm PEG Amine (tripentaerythritol core) and 8arm functional group. PEG Carboxyl (tripentaerythritol core). Preferred molecular Preferably, the dendritic moiety Hyp is a hyperbranched weights for such multi-arm PEG-derivatives in a backbone 60 polypeptide. Preferably, the hyperbranched polypeptide moiety are 1 kDa to 20 kDa, more preferably 2.5 kDa to 15 comprises lysine in bound form, most preferably Hyp is kDa and even more preferably 5 kDa to 10 kDa. It is heptalysinyl in bound form. Preferably, each dendritic moi understood that the terminal amine groups of the above ety Hyp has a molecular weight in the range of from 0.4kDa mentioned multi-arm molecules are present in bound form in to 4 kDa, more preferably in the range of from 0.4 kDa to the backbone moiety to provide further interconnected func 65 2 kDa. It is understood that a backbone moiety C(A-Hyp) tional groups and reactive functional groups of a backbone can consist of the same or different dendritic moieties Hyp moiety. and that each Hyp can be chosen independently. Each US 9,533,056 B2 45 46 moiety Hyp consists of between 5 and 32 lysines, preferably Preferred biodegradable linkages are esters, carbonates, of at least 7 lysines, i.e. each moiety Hyp is comprised of phosphoesters and Sulfonic acid esters and most preferred between 5 and 32 lysines in bound form, preferably of at are esters or carbonates. It is understood that for in vitro least 7 lysines in bound form. Most preferably Hyp is studies accelerated conditions like, for example, pH 9, 37° comprised of heptalysinyl. C., aqueous buffer, may be used for practical purposes. The reaction of polymerizable functional groups a back Permanent linkages are non-enzymatically hydrolytically bone reagent, more specifically of Hyp with the polymeriZ degradable under physiological conditions (aqueous buffer able functional groups of PEG based crosslinker reagents at pH 7.4, 37° C.) with half-lives of six months or longer, results in a permanent amide bond. Such as, for example, amides. One preferred backbone moiety is shown below, dashed 10 The degradation of the hydrogel is a multi-step reaction lines indicate interconnecting biodegradable linkages to where a multitude of degradable bonds is cleaved resulting crosslinker moieties and n is an integer of from 5 to 50: in degradation products which may be water-soluble or

40 Preferably, C(A-Hyp) has a molecular weight in the water-insoluble. However each water-insoluble degradation range of from 1 kDa to 20 kDa, more preferably from 1 kDa product further comprises degradable bonds so that it can be to 15 kDa, more preferably from 2.5 kDa to 15 kDa and even cleaved in that water-soluble degradation products are more preferably 5 kDa to 10 kDa. obtained. These water-soluble degradation products may Preferably, L is attached to Z through a thiosuccinimide 45 comprise one or more backbone moieties. It is understood group which in turn is attached to the hydrogel's backbone that released backbone moieties may, for instance, be per moiety through a spacer, such as an oligoethylene glycol manently linked to spacer or blocking groups and/or prod chain. Preferably, the linkage of this spacer chain to the rug-linker degradation products. backbone moiety is a permanent bond, preferably an amide In Such hydrogel-linked prodrugs according to the inven 50 tion, it is desirable that almost all release of biologically bond. active agent (>90%) has occurred before a significant Preferably, L is a chemical bond. amount of release of the backbone degradation products Biodegradability of the hydrogels according to the present (<10%) has taken place. This can be achieved by adjusting invention is achieved by introduction of hydrolytically the hydrogel-linked prodrugs half-life versus the hydrogel degradable bonds. 55 degradation kinetics. The terms “hydrolytically degradable”, “biodegradable' To introduce the hydrolytically cleavable bonds into the or “hydrolytically cleavable”, “auto-cleavable', or “self hydrogel carrier of the invention, the backbone moieties can cleavage”, “self-cleavable', “transient’ or “temporary” refer be directly linked to each other by means of biodegradable within the context of the present invention to bonds and bonds. linkages which are non-enzymatically hydrolytically 60 In one embodiment, the backbone moieties of the biode degradable under physiological conditions (aqueous buffer gradable hydrogel carrier may be linked together directly, at pH 7.4, 37° C.) with half-lives ranging from one hour to i.e. without crosslinker moieties. The hyperbranched den three months, include, but are not limited to, aconityls, dritic moieties of two backbone moieties of such biodegrad acetals, carboxylic anhydrides, esters, imines, hydrazones, able hydrogel may either be directly linked through an maleamic acid amides, ortho esters, phosphamides, phos 65 interconnected functional group that connects the two hyper phoesters, phosphosilyl esters, silyl esters, Sulfonic esters, branched dendritic moieties. Alternatively, two hyper aromatic carbamates, combinations thereof, and the like. branched dendritic moieties of two different backbone moi US 9,533,056 B2 47 48 eties may be interconnected through two spacer moieties Preferably, the poly(ethylene glycol) based crosslinkers connected to a backbone moiety and on the other side have a molecular weight in the range of from 0.5 kDa to 5 connected to a crosslinking moiety separated by an inter kDa. connected functional groups. Preferably, backbone moieties may be linked together 5 In a preferred embodiment of the present invention the through crosslinker moieties, each crosslinker moiety being crosslinker moiety consists of a PEG chain, which is sym terminated by at least two of the hydrolytically degradable metrically connected through ester bonds to two alpha, bonds. In addition to the terminating degradable bonds, the omega-aliphatic dicarboxylic spacers provided by backbone crosslinker moieties may contain further biodegradable moieties through permanent amide bonds. bonds. Thus, each end of the crosslinker moiety linked to a backbone moiety shows a hydrolytically degradable bond, 10 The dicarboxylic acids of the spacer moieties connected and additional biodegradable bonds may optionally be pres to the backbone moiety and on the other side connected to ent in the crosslinker moiety. a crosslinking moiety consists of 3 to 12 carbon atoms, most The hydrogel may contain one or more different types of crosslinker moieties, preferably one. The crosslinker moiety preferably between 5 and 8 carbon atoms and may be may be a linear or branched molecule and preferably is a 15 substituted at one or more carbon atom. Preferred substitu linear molecule. In a preferred embodiment of the invention, ents are alkyl groups, hydroxy groups or amido groups or the crosslinker moiety is connected to backbone moieties by at least two biodegradable bonds. Substituted amino groups. One or more of the aliphatic If present in a hydrogel used as carrier in the prodrugs dicarboxylic acids methylene groups may optionally be according to the invention as degradable interconnected substituted by O or NH or alkyl-substituted N. Preferred functional group, preferred biodegradable linkages are car alkyl is linear or branched alkyl with 1 to 6 carbon atoms. boxylic esters, carboxylic anhydrides, carbonates, phospho esters and sulfonic acid esters; more preferably carboxylic Preferably, there is a permanent amide bond between the esters, carbonates, phosphoesters and Sulfonic acid esters hyperbranched dendritic moiety and the spacer moiety con and most preferred are carboxylic esters or carbonates. 25 nected to a backbone moiety and on the other side is In one embodiment, a crosslinker moiety consists of a connected to a crosslinking moiety. polymer. Preferably, crosslinker moieties have a molecular One preferred crosslinker moiety is shown below: dashed weight in the range of from 0.5 kDa to 5 kDa, more lines indicate interconnecting biodegradable linkages to preferably, from 1 kDa to 4 kDa, even more preferably from backbone moieties: 1 kDa to 3 kDa. 30 In addition to oligomeric or polymeric crosslinking moi eties, low-molecular weight crosslinking moieties may be used, especially when hydrophilic high-molecular weight O ', s backbone moieties are used for the hydrogel formation. O q O , Preferably, the polyethyleneglycol based crosslinker moi 35 eties are hydrocarbon chains comprising ethylene glycol units, optionally comprising further functional groups, wherein the poly(ethylene glycol) based crosslinker moi eties comprise at least each methylene glycol units, wherein wherein q is an integer of from 5 to 50. m is an integer in the range of from 3 to 100, preferably from 40 Preferably, the hydrogel carrier is composed of backbone 10 to 70. Preferably, the poly(ethylene glycol) based cross moieties interconnected by hydrolytically degradable bonds. linker moieties have a molecular weight in the range of from 0.5 kDa to 5 kDa. More preferably, the backbone moieties comprise a If used in reference to a crosslinker moiety or a PEG branching core of the following formula: based polymeric chain connected to a branching core, the 45 term “PEG-based refers to a crosslinker moiety or PEG based polymeric chain comprising at least 20 weight 96 ethylene glycol moieties. In one embodiment, monomers constituting the polymeric crosslinker moieties are connected by biodegradable bonds. 50 Such polymeric crosslinkers may contain up to 100 biode gradable bonds or more, depending on the molecular weight wherein the dashed line indicates attachment to the remain of the crosslinker moiety and the molecular weight of the monomer units. Examples for Such crosslinkers are poly der of the backbone moiety. lactic acid or polyglycolic acid based. It is understood that 55 More preferably, the backbone moieties comprise a struc Such polylactic acid) or poly(glycolic acid) chain may be ture of the following formula: terminated or interrupted by alkyl or aryl groups and that they may optionally be substituted with heteroatoms and chemical functional groups. Preferably, the crosslinker moieties are PEG based, pref 60 erably represented by only one PEG based molecular chain. C O , Preferably, the poly(ethylene glycol) based crosslinkers are -ha 1N-"Nu-Npi N -(4 hydrocarbon chains comprising ethylene glycol units, optionally comprising further functional groups, wherein the poly(ethylene glycol) based crosslinker moieties comprise at 65 wherein n is an integer of from 5 to 50 and the dashed line least each methylene glycol units, wherein m is an integer indicates attachment to the remainder of the backbone in the range of from 3 to 100, preferably from 10 to 70. moiety. US 9,533,056 B2 49 50 Preferably, backbone moiety comprises a hyperbranched wherein one of the dashed lines indicates attachment to the moiety Hyp. hyperbranched moiety Hyp and the second dashed line More preferably, the backbone moiety comprises a hyper indicates attachment to the rest of the molecule; and wherein branched moiety Hyp of the following formula: m is an integer of from 2 to 4. Preferably, the backbone moieties are linked together through crosslinker moieties having the following structure

Y& O1n-N-1s O >1*

wherein q is an integer from 3 to 100. More preferably, the backbone moieties of the hydrogel are linked together through moieties of the following for mula:

O O O O

y w O 1a-on O F

wherein the dashed lines indicate attachment to a backbone

30 moiety, respectively, and wherein n is 45. In an alternative preferred embodiment, the backbone moieties of the hydrogel are linked together through moi eties of the following formula:

wherein the dashed lines indicate attachment to the rest of wherein the dashed lines indicate attachment to a backbone the molecule and carbon atoms marked with asterisks indi moiety, respectively, and wherein n is 22. cate in a preferred embodiment S-configuration. 45 The hydrolysis rate of the biodegradable bonds between backbone and crosslinker moieties is influenced or deter mined by the number and type of connected atoms adjacent to the PEG-ester carboxy group. For instance, by selecting from succinic, adipic or glutaric acid for PEG ester forma 50 tion it is possible to vary the degradation half-lives of the However, it is understood that hyperbranched moieties biodegradable hydrogel carrier. Hyp as shown above may also be in R-confirmation or may The hydrogel-linked prodrug of the present invention can be racemic. be prepared starting from the hydrogel of the present inven tion by convenient methods known in the art. It is clear to 55 a practitioner in the art that several routes exist. For example, the prodrug linker mentioned above to which the biologically active moiety is covalently attached can be Preferably, the backbone moieties are attached to at least reacted with the reactive functional groups of the hydrogel one spacer of the following formula: of the present invention with or with the prodrug linker 60 already bearing the active moiety in part or as whole.

In a preferable method of preparation, the hydrogel is generated through chemical ligation reactions. The hydrogel may be formed from two macromolecular educts with complementary functionalities which undergo a reaction 65 Such as a condensation or addition. One of these starting materials is a crosslinker reagent with at least two identical functional groups and the other starting material is a homo US 9,533,056 B2 51 52 multifunctional backbone reagent. Suitable functional backbone reagent. It is understood that the lysines can be groups present on the crosslinker reagent include terminal partially or fully protected by protective groups during the amino, carboxylic acid and derivatives, maleimide and other coupling steps and that also the final backbone reagent may alpha,beta unsaturated Michael acceptors like vinylsulfone, contain protective groups. A preferred building block is thiol, hydroxyl groups. Suitable functional groups present in bis-boc lysine. Alternatively, instead of sequential additions the backbone reagent include but are not limited to amino, of lysine residues, a dendritic poly-lysine moiety may be carboxylic acid and derivatives, maleimide and other alpha, assembled first and subsequently coupled to the 4-arm PEG beta unsaturated Michael acceptors like vinylsulfone, thiol, tetra amine or 8-arm PEG octa amine. It is desirable to hydroxyl groups. obtain backbone reagent carrying 32 amino groups, conse 10 quently seven lysines would be attached to each arm of a If the crosslinker reagent's reactive functional groups are 4-arm PEG, or three lysines would be attached to each arm used substoichiometrically with respect to backbone reac of a 8-arm PEG. In another embodiment, the multi-arm PEG tive functional groups, the resulting hydrogel will be a derivative is a tetra- or octa carboxy PEG. In this case, the reactive hydrogel with free reactive functional groups dendritic moieties may be generated from glutaric or aspar attached to the backbone structure. 15 tic acid, and the resulting backbone reagent would carry 32 Optionally, the prodrug linker may be first conjugated to carboxy groups. It is understood that all or a fraction of the the biologically active agent and the resulting prodrug linker backbone reagents functional groups may be present in a conjugate may then react with the hydrogel's reactive func free form, as salts or conjugated to protecting groups. It is tional groups. Alternatively, after activation of one of the understood that due to practical reasons the backbone functional groups of the prodrug linker, the linker-hydrogel reagent's number of lysines per PEG-arm will be between conjugate may be contacted with biologically active agent in six and seven, more preferably approximately seven. the second reaction step and excess biologically active agent A preferred backbone reagent is shown below:

HN ~ NH2 O NH NH2

O

O HN H N s NH2H C O N N

O O

N H HN NH2

O

in N 28 NH2

may be removed by filtration after conjugation of the Synthesis of the crosslinker reagent starts from a linear biologically active agent to the hydrogel-bound prodrug PEG chain with a molecular weight ranging from 0.2 to 5 linker. kDa, more preferably from 0.6 to 2 kDa, which is esterified A preferred process for the preparation of a prodrug 60 with a half ester of a dicarboxylic acid, preferably adipic according to the present invention is as follows: acid or glutaric acid. A preferred protecting group for half A preferred starting material for the backbone reagent ester formation is the benzylic group. The resulting bis synthesis is a 4-arm PEG tetra amine or 8-arm PEG octa dicarboxylic acid PEG half esters are converted into more amine, with the PEG reagent having a molecular weight reactive carboxy compounds, such as acyl chlorides or ranging from 2000 to 10000 Dalton, most preferably fom 65 active esters, eg pentafluorophenyl or N-hydroxysuccinim 2000 to 5000 Da. To such multi-arm PEG-derivatives, lysine ide esters, most preferred N-hydroxysuccinimde esters, of residues are coupled sequentially to form the hyperbranched which a preferred selected structure is shown below. US 9,533,056 B2 53 54

O O O O O O

N - O ---iii. O1n-N-1N O ---iii. O-N

O O wherein each m independently is an integer ranging from 2 10 reagents, carrying a reactive thiol group on the linker moiety to 4, and q is an integer of from 3 to 100. to form hydrogel-linked prodrugs according to the present More preferred is the following structure: invention.

O O O O O O

N - O ---* O 1-S-1NO ---* O-N s

O O q ~ 45 wherein r is either 1 or 2, preferably 1. After loading the drug-linker conjugate to the function Alternatively, the bis-dicarboxylic acid PEG half esters alized maleimido group-containing hydrogel, all remaining may be activated in the presence of a coupling agent Such as 30 functional groups are capped with a Suitable blocking DCC or HOBt or PyBOP. reagent, Such as mercaptoethanol, to prevent undesired In an alternative embodiment, the backbone reagent car side-reactions. ries carboxy groups and the corresponding crosslinker A particularly preferred method for the preparation of a reagent would be selected from ester-containing amino prodrug of the present invention comprises the steps of terminated PEG-chains. 35 (a) reacting a compound of formula C(A-X1), wherein Backbone reagent and crosslinker reagent may be polym A-X1 represents. A before its binding to Hyp or a erized to form the hydrogel according to the invention using precursor of Hyp and X1 is a suitable functional group, inverse emulsion polymerization. After selecting the desired with a compound of formula Hyp'-X2, wherein Hyp'- stoichiometry between backbone and crosslinker functional X2 represents Hyp before its binding to A or a precursor groups, backbone and crosslinker are dissolved in DMSO 40 of Hyp and X2 is a suitable functional group to react and a suitable emulgator with an appropriately selected HLB with X1; value, preferably Arlacel P135, is employed to form an (b) optionally reacting the resulting compound from step inverse emulsion using a mechanical stirrer and controlling (a) in one or more further steps to yield a compound of the stirring speed. Polymerization is initiated by the addition formula C(A-Hyp) having at least four functional of a suitable base, preferably by N.N.N',N'-tetramethylene 45 diamine. After stirring for an appropriate amount of time, the groups: reaction is quenched by the addition of an acid, such as (c) reacting the at least four functional groups of the acetic acid and water. The beads are harvested, washed, and resulting compound from step (b) with a polyethyl fractionated according to particle size by mechanical siev eneglycol based crosslinker precursor, wherein the ing. Optionally, protecting groups may be removed at this 50 crosslinker precursor is used in a Sub-stoichiometric Stage. amount compared to the total number of functional In an alternative embodiment of this invention, multi groups of C(A-Hyp) to yield a hydrogel; functional moieties are coupled to the reactive functional (d) reacting remaining un-reacted functional groups (rep groups of the polymerized reactive hydrogel to increase the resenting the reactive functional groups of the back number of functional groups which allows increasing the 55 bone comprised in the hydrogel) in the hydrogel back drug load of the hydrogel. Such multi-functional moieties bone of step (c) with a covalent conjugate of may be provided by suitably substituted derivatives of biologically active moiety and transient prodrug linker lysine, dilysine, trilysine, tetralysine, pentalysine, hexaly or first reacting the un-reacted functional groups with sine, heptalysine, or oligolysine, low-molecular weight PEI. the transient prodrug linker and Subsequently with the Preferably, the multi-functional moiety is lysine. 60 biologically active moiety; Further, such hydrogel according to the invention may be (e) optionally capping remaining un-reacted functional functionalized with a spacer carrying a different reactive groups to yield a prodrug of the present invention. functional group than provided by the hydrogel. For instance Such hydrogel is preferably comminuted by mechanical maleimide reactive functional groups may be introduced processes Such as stirring, crushing, cutting pressing, or into the hydrogel by coupling a suitable heterobifunctional 65 milling, and optionally sieving. For emulsion polymeriza spacer such as Mal-PEG6-NHS to the hydrogel. Such func tion, the reaction mixture is comprised of the dispersed tionalized hydrogel can be further conjugated to drug-linker phase and the continuous phase. US 9,533,056 B2 55 56 For the dispersed phase, backbone reagent and crosslinker stent. Most preferably, the hydrogel is formed into micropar reagent are mixed in a ratio aminefactive ester of 5:1 to ticulate beads which can be administered as Subcutaneous or 1.05:1, preferably of 2:1 to 1.05:1 and are dissolved in intramuscular injectably by means of a standard Syringe. DMSO to give a to give a solution with a concentration of Such soft beads may have a diameter of between 1 and 500 5 to 50 g per 100 mL, preferably 7 to 30 g per 100 ml, more 5 micrometer. preferably 7.5 to 20 g per 100 ml and most preferably 10 to Preferably, such beaded carrier-linked hydrogel prodrugs 20 g per 100 ml. have a diameter of between 10 and 100 micrometer if The continuous phase is any solvent, that is not miscible Suspended in an isotonic aqueous formulation buffer, most with DMSO, not basic, aprotic and shows a viscosity lower preferably a diameter of between 20 and 100 micrometer, than 10 Pas. Preferably, the solvent is not miscible with 10 most preferably a diameter of between 25 and 80 microm DMSO, not basic, aprotic, shows a viscosity lower than 2 eter. Pas and is non-toxic. More preferably, the solvent is a Preferably, such beaded carrier-linked hydrogel prodrugs saturated linear or branched hydrocarbon with 5 to 10 can be administered by injection through a needle Smaller carbon atoms. Most preferably, the solvent is n-heptane. than 0.6 mm inner diameter, preferably through a needle To form an emulsion of the dispersed phase in the 15 smaller than 0.3 mm inner diameter, more preferably continuous phase, an emulsifier is added to the continuous through a needle small than 0.25 mm inner diameter, even phase before adding the dispersed phase. The amount of more preferably through a needle smaller than 0.2 mm inner emulsifier is 2 to 50 mg per mL dispersed phase, more diameter, and most preferably through a needle Small than preferably 5 to 20 mg per mL dispersed phase, most pref 0.16 mm inner diameter. erably 10 mg per mL dispersed phase. It is understood that the terms “can be administered by The emulsifier has an HLB-value of 3 to 8. Preferably, the injection”, “injectable' or “injectability” refer to a combi emulsifier is a triester of sorbitol and a fatty acid or an nation of factors such as a certain force applied to a plunger poly(hydroxyl fatty acid)-poly(ethylene glycol) conjugate. of a syringe containing the carrier-linked hydrogel prodrugs More preferably, the emulsifier is an poly(hydroxy-fatty according to the invention Swollen in a liquid at a certain acid)-polyethylene glycol conjugate, with a linear poly(eth 25 concentration (w/v) and at a certain temperature, a needle of ylene glycol) of a molecular weight in the range of from 0.5 a given inner diameter connected to the outlet of Such kDa to 5 kDa and poly(hydroxy-fatty acid) units of a Syringe, and the time required to extrude a certain volume of molecular weight in the range of from 0.5 kDa to 3 kDa on the carrier-linked hydrogel prodrugs according to the inven each end of the chain. Most preferably, the emulsifier is tion from the Syringe through the needle. poly(ethylene glycol) dipolyhydroxy stearate, Cithrol DPHS 30 In order to provide for injectability, a volume of 1 mL of (Cithrol DPHS, former Arlacel P135, Croda International the carrier-linked hydrogel prodrugs swollen in water to a Plc). concentration of at least 5% (w/v) and contained in a syringe Droplets of the dispersed phase are generated by stirring holding a plunger of a diameter of 4.7 mm can be extruded with an axial flow impeller with a geometry similar to at room temperature within 10 seconds by applying a force stirrers such as Isojet, Intermig, Propeller (EKATO Rühr 35 of less than 60 Newton, such as less than 50 Newton, and Mischtechnik GmbH, Germany), most preferably simi preferably by applying a force of less than 40 Newton. lar to Isojet or Propeller with a diameter of 50 to 90% of the Preferably injectability measurement is carried out for the reactor diameter. Preferably, stirring is initated before addi carrier-linked hydrogel prodrugs of the present invention tion of the dispersed phase. Stirrer speed is set to 0.6 to 2.4 swollen in water to a concentration of ca. 15% (w/v). m/s, such as 0.8 to 2.3 m/s, preferably to 0.6 to 1.7 m/s. The 40 By consequence, the prodrugs according to the present dispersed phase is added at room temperature, and the invention show the beneficial effect of a controlled release concentration of the disperse phase is 2% to 70%, preferably rate in respect of the released drug D-H. Preferably, a 5 to 50%, more preferably 10 to 40%, and most preferably Sustained release rate is obtained. Sustained release means 20 to 35% of the total reaction volume. The mixture of that the administration intervals of the respective prodrug dispersed phase, emulsifier and continuous phase is stirred 45 are expanded. For example, prodrugs according to the for 5 to 60 min before adding the base to the effect present invention which are based on drugs commonly polymerization. applied once or several times a day provide therapeutically 5 to 10 equivalents (referred to each amide bond to be effective levels for at least three days, more preferably for at formed) of a base are added to the mixture of dispersed and least one week and even more preferably for at least one continuous phase. The base is aprotic, non nucleophilic and 50 month. soluble in the disperse phase. Preferably, the base is aprotic, The prodrug according to the present invention show non nucleophilic, well soluble in both disperse phase and excellent in vivo/in vitro correlation of linker cleavage, a DMSO. More preferably, the base is aprotic, non nucleop high degree of enzyme independence and show a higher hilic, well soluble in both disperse phase and DMSO, an stability at lower pH, resulting in a pH dependent cleavage. amine base and non-toxic. Most preferably, the base is 55 A strong in vivo/in vitro correlation is observed, if the N.N.N',N'-tertramethylethylene diamine (TMEDA). Stirring release kinetics exhibited by a carrier-linked prodrug con in the presence of base is continued for 1 to 16 h. jugate according to the invention in vivo has a half-life that During stirring, droplets of dispersed phase are hardened is not smaller than half the value exhibited by the same to become crosslinked hydrogel beads according to the carrier-linked prodrug conjugate in aqueous buffer of pH 7.4 invention which can be collected and fractionation accord 60 at 37°C., wherein the release kinetics in vivo is measured as ing to size is performed on a vibrational continuous sieving plasma levels of free drug. It is understood that in the case machine with a 75 um and a 32 um deck to give hydrogel of soluble carriers, release kinetics may be recorded as microparticles according to the invention. hydrolysis kinetics. The hydrogel for the prodrug of the present invention can Another aspect of the present invention are pharmaceu be obtained from the preparation methods in form of micro 65 tical compositions of the carrier-linked prodrugs described particles. In a preferred embodiment of the invention, the before. Such pharmaceutical compositions contain one or reactive hydrogel is a shaped article Such as a mesh or a more excipients, selected from the groups consisting of US 9,533,056 B2 57 58 (i) Buffering agents: physiologically tolerated buffers to (viii) Viscosifiers or viscosity enhancers: retard settling of maintain pH in a desired range. Such as Sodium phosphate, the particles in the vial and Syringe and are used in order to bicarbonate. Succinate, histidine, citrate and acetate, Sul facilitate mixing and resuspension of the particles and to phate, nitrate, chloride, pyruvate. Antacids Such as Mg(OH)2 make the Suspension easier to inject (i.e., low force on the or ZnCO may be also used. Buffering capacity may be Syringe plunger). Suitable viscosifiers or viscosity enhancers adjusted to match the conditions most sensitive to pH are, for example, carbomer viscosifiers like Carbopol 940, stability. Carbopol Ultrez, 10, cellulose derivatives like hydroxypro (ii) Isotonicity modifiers: to minimize pain that can result pylmethylcellulose (hypromellose, HPMC) or diethylam from cell damage due to osmotic pressure differences at the inoethyl cellulose (DEAE or DEAE-C), colloidal magne injection depot. Glycerin and sodium chloride are examples. 10 Effective concentrations can be determined by osmometry sium silicate (Veegum) or sodium silicate, hydroxyapatite using an assumed osmolality of 285-315 mCsmol/kg for gel, tricalcium phosphate gel, Xanthans, carrageenans like S. Satia gum UTC 30, aliphatic poly(hydroxy acids), such as (iii) Preservatives and/or antimicrobials: multidose par poly(D.L- or L-lactic acid) (PLA) and poly(glycolic acid) enteral preparations may require the addition of preserva 15 (PGA) and their copolymers (PLGA), terpolymers of D.L- tives at a sufficient concentration to minimize the risk of lactide, glycolide and caprolactone, poloxamers, hydrophilic patients becoming infected upon injection and correspond poly(oxyethylene) blocks and hydrophobic poly(oxypropyl ing regulatory requirements have been established. Typical ene) blocks to make up a triblock of poly(oxyethylene)-poly preservatives include m-cresol, phenol, methylparaben, eth (oxypropylene)-poly(oxyethylene) (e.g. Pluronic(R), ylparaben, propylparaben, butylparaben, chlorobutanol, polyetherester copolymer, such as a polyethylene glycol benzyl alcohol, phenylmercuric nitrate, thimerosol, Sorbic terephthalate/polybutylene terephthalate copolymer, Sucrose acid, potassium Sorbate, benzoic acid, chlorocresol, and acetate isobutyrate (SAIB), dextran or derivatives thereof, benzalkonium chloride. combinations of dextrans and PEG, polydimethylsiloxane, (iv) Stabilizers: Stabilisation is achieved by strengthening collagen, chitosan, polyvinyl alcohol (PVA) and derivatives, of the protein-stabilising forces, by destabilisation of the 25 polyalkylimides, poly (acrylamide-co-diallyldimethyl denatured stater, or by direct binding of excipients to the ammonium (DADMA)), polyvinylpyrrolidone (PVP), gly protein. Stabilizers may be amino acids such as alanine, cosaminoglycans (GAGs) such as dermatan Sulfate, chon arginine, aspartic acid, glycine, histidine, lysine, proline, droitin Sulfate, keratan Sulfate, heparin, heparan Sulfate, Sugars such as glucose, Sucrose, trehalose, polyols such as hyaluronan, ABA triblock or AB block copolymers com glycerol, mannitol, Sorbitol, salts such as potassium phos 30 phate, sodium Sulphate, chelating agents such as EDTA, posed of hydrophobic A-blocks, such as polylactide (PLA) hexaphosphate, ligands such as divalent metal ions (zinc, or poly(lactide-co-glycolide) (PLGA), and hydrophilic calcium, etc.), other salts or organic molecules Such as B-blocks, such as polyethylene glycol (PEG) or polyvinyl phenolic derivatives. In addition, oligomers or polymers pyrrolidone. Such block copolymers as well as the above such as cyclodextrins, dextran, dendrimers, PEG or PVP or 35 mentioned poloxamers may exhibit reverse thermal gelation protamine or HSA may be used. behavior (fluid state at room temperature to facilitate admin (v) Anti-adsorption agents: Mainly ionic or inon-ionic istration and gel state above sol-gel transition temperature at Surfactants or other proteins or Soluble polymers are used to body temperature after injection). coat or adsorb competitively to the inner surface of the (ix) Spreading or diffusing agent: modifies the permeabil composition’s container, e.g. poloxamer (Pluronic F-68), 40 ity of connective tissue through the hydrolysis of compo PEG dodecyl ether (Brij35), polysorbate 20 and 80, dextran, nents of the extracellular matrix in the intrastitial space Such polyethylene glycol, PEG-polyhistidine, BSA and HSA and as, but not limited to, hyaluronic acid, a polysaccharide gelatines. Chosen concentration and type of excipient found in the intercellular space of connective tissue. A depends on the effect to be avoided but typically a mono spreading agent Such as, but not limited to, hyaluronidase layer of surfactant is formed at the interface just above the 45 temporarily decreases the viscosity of the extracellular CMC value. matrix and promotes diffusion of injected drugs. (vi) Lyo- and/or cryoprotectants: During freeze- or spray (X) Other auxiliary agents: Such as wetting agents, vis drying, excipients may counteract the destabilising effects cosity modifiers, antibiotics, hyaluronidase. Acids and bases caused by hydrogen bond breaking and water removal. For Such as hydrochloric acid and sodium hydroxide are auxil this purpose, Sugars and polyols may be used, but corre 50 iary agents necessary for pH adjustment during manufac sponding positive effects have also been observed for sur ture. factants, amino acids, non-aqueous solvents, and other pep The composition of a prodrug according to the invention tides. Trehalose is particulary efficient at reducing moisture may be provided as a liquid composition, a Suspension induced aggregation and also improves thermal stability composition or as a dry composition. potentially caused by exposure of protein hydrophobic 55 groups to water. Mannitol and Sucrose may also be used, In one embodiment, the pharmaceutical composition of a either as Solely of cryoprotectant or in combination with each prodrug according to the invention is a dry composition. other where higher ratios of mannitol:Sucrose are known to Suitable methods of drying are, for example, spray-drying enhance physical stability of a lyophilized cake. Mannitol and lyophilization (freeze-drying). Preferably, the pharma may also be combined with trehalose. Trehalose may also be 60 ceutical composition of prodrug is dried by lyophilization. combined with sorbitol or sorbitol may be used as the sole Preferably, the prodrug is sufficiently dosed in the com protectant. Starch or starch derivatives may also be used. position to provide therapeutically effective amounts of (vii) Oxidation protection agents: antioxidants such as biologically active agent for at least 12 hours in one appli ascorbic acid, ectoine, methionine, glutathione, monothio cation. More preferably, one application of prodrug is Suf glycerol, morin, polyethylenimine (PEI), propyl gallate, 65 ficient for at least one day, more preferably for at least 3 Vitamin E, chelating agents such aus citric acid, EDTA, days, more preferably for at least 1 week and most prefer hexaphosphate, thioglycolic acid. ably for at least 4 weeks. US 9,533,056 B2 59 60 In one embodiment of the present invention, the compo needle and a container comprising the carrier-linked prodrug sition of prodrug is provided as a single dose, meaning that composition for use with the Syringe. In case of a dry the container in which it is Supplied contains one pharma composition, the container may have one chamber contain ceutical dose. ing the dry carrier-linked prodrug composition, and a second In another embodiment, the composition is provided as a chamber comprising a reconstitution solution. In more pre multiple dose composition, meaning that it contains more ferred embodiments, the injection device is other than a than one therapeutic dose. Preferably, a multiple dose com simple hypodermic syringe and so the separate container position contains at least 2 doses. Such multiple dose with carrier-linked prodrug composition is adapted to composition of prodrug can either be used for different engage with the injection device Such that in use the liquid patients in need thereof or is intendend for use in one patient, 10 wherein the remaining doses are stored after the application or Suspension or reconstituted dry composition in the con of the first dose until needed. tainer is in fluid connection with the outlet of the injection In another aspect of the present invention the prodrug device. Examples of administration devices include but are composition is comprised in a container. For liquid or not limited to hypodermic syringes and pen injector devices. Suspension compositions, the container is preferably a single 15 Particularly preferred injection devices are the pen injectors, chamber Syringe. For dry compositions, preferably the con in which case the container is a cartridge, preferably a tainer is a dual-chamber Syringe. The dry composition disposable cartridge. according to the present invention is provided in a first A preferred kit of parts comprises a needle and a container chamber of the dual-chamber Syringe and reconstitution containing the dry carrier-linked prodrug composition solution is provided in a second chamber of the dual according to the present invention and optionally further chamber Syringe. containing a reconstitution solution, the container being Prior to applying the dry composition of prodrug to a adapted for use with the needle. Preferably, the container is patient in need thereof, the dry composition is reconstituted. a dual-chamber Syringe. Reconstitution can take place in the container in which the In another aspect, the invention provides a cartridge dry composition of prodrug is provided, such as in a vial, 25 containing a composition of carrier-linked prodrug as here Syringe, dual-chamber Syringe, ampoule, and cartridge. inbefore described for use with a pen injector device. The Reconstitution is done by adding a predefined amount of cartridge may contain a single dose or multiplicity of doses reconstitution solution to the dry composition. Reconstitu of carrier-linked prodrug. tion solutions are sterile liquids, such as water or buffer, In one embodiment of the present invention the Suspen which may contain further additives, such as preservatives 30 and/or antimicrobials, such as, for example, benzylalcohol sion composition of carrier-linked prodrug does not only and cresol. Preferably, the reconstitution solution is sterile comprise a carrier-linked prodrug and one or more excipi Water. ents, but also other biologically active agents, either in their A further aspect is a method of preparing a reconstituted free form or as prodrugs or carrier-linked prodrugs such as composition comprising a therapeutically effective amount 35 PEG prodrugs or hydrogel prodrugs. Preferably, such addi of prodrug, and optionally one or more pharmaceutically tional one or more biologically active agent is a prodrug, acceptable excipients, the method comprising the step of more preferably a PEG or hydrogel prodrug. contacting the composition of the present invention with a In an alternative embodiment, the carrier-linked prodrug reconstitution solution. composition according to the present invention is combined Another aspect is a reconstituted composition comprising 40 with a second biologically active compound in Such way that a therapeutically effective amount of a prodrug according to the carrier-linked prodrug composition according to the the invention, and optionally one or more pharmaceutically invention is administered to a patient in need thereof first, acceptable excipients. followed by the administration of the second compound. Another aspect of the present invention is the method of Alternatively, the carrier-linked prodrug composition is manufacturing a liquid or Suspension composition of carrier 45 administered to a patient in need thereof after another linked prodrug. In one embodiment, such composition is compound has been administered to the same patient. made by Yet another aspect of the present invention is a carrier (i) admixing the carrier-linked prodrug with one or more linked prodrug of the present invention or a pharmaceutical excipients, composition of the present invention for use as a medica (ii) transferring amounts of the liquid or Suspension 50 ment. composition equivalent to single or multiple doses into Yet another aspect of the present invention is a carrier Suitable containers, and (iii) sealing the containers. linked prodrug of the present invention or a pharmaceutical Another aspect of the present invention is the method of composition of the present invention for use in a method of manufacturing a dry composition of carrier-linked prodrug. 55 treating or preventing diseases or disorders which can be In one embodiment, such composition is made by treated by the biologically active moiety released from the (i) admixing the carrier-linked prodrug with one or more carrier-linked prodrug according to the present invention. excipients, Another subject of the present invention is a method for (ii) transferring amounts equivalent to single or multiple the synthesis of a carrier-linked prodrug or a pharmaceuti doses into Suitable containers, 60 cally acceptable salt thereofas defined above. Carrier-linked (iii) drying the composition in said containers, and prodrugs or precursors of carrier-linked prodrugs according (iv) sealing the containers. to the present invention may be prepared by known methods Suitable containers are vials, Syringes, dual-chamber or in accordance with the reaction sequences described Syringes, ampoules, and cartridges. below. The starting materials used in the preparation (Syn Another aspect is a kit of parts. For liquid and Suspension 65 thesis) of prodrugs of the invention or precursors thereofare compositions, and when the administration device is simply known or commercially available, or can be prepared by a hypodermic syringe, the kit may comprise the Syringe, a known methods or as described below. US 9,533,056 B2 61 62 All reactions for the synthesis of the carrier-linked prod EXAMPLES rugs according to the present invention including precursors Such as the moiety L'according to the formula (I) are perse Example 1 well-known to the skilled person and can be carried out under Standard conditions according to or analogously to Synthesis of Linker Reagent Intermediate (1) procedures described in the literature, for example in Hou ben-Weyl, Methoden der Organischen Chemie (Methods of Organic Chemistry). Thieme-Verlag, Stuttgart, or Organic Reactions, John Wiley & Sons, New York. Depending on the circumstances of the individual case, in order to avoid side 10 reactions during the synthesis of a carrier-linked prodrug or OH a precursor thereof, it can be necessary or advantageous to temporarily block functional groups by introducing protec tive groups and to deprotect them in a later stage of the 15 synthesis, or introduce functional groups in the form of precursor groups which in a later reaction step are converted w into the desired functional groups. Such synthesis strategies boc and protective groups and precursor groups which are Suit able in an individual case are known to the skilled person. If desired, the carrier-linked prodrugs or precursors can be purified by customary purification procedures, for example NH by recrystallization or chromatography. / The carrier-linked prodrugs according to the present fmoc invention or a pharmaceutically acceptable Salt thereof may 25 be prepared by a method comprising the step of reacting a Fmoc-5,5-dimethyl-L-proline (0.9 mmol) is loaded onto prodrug precursor L-Y with a biologically active drug D-H 2-chlorotrityl resin (0.6 mmol) according to manufacturers to obtain the drug linker conjugate D-L by forming an amide instructions. After fimoc removal, a solution of boc-Lys bond, wherein Y is a leaving group. (fmoc)-OH (3 mmol), HATU (3 mmol), and collidine (6 30 mmol) is added to the resin and incubated for 30 min. This In respect of the prodrug precursor L-Y, L has the same procedure is repeated once. Intermediate 1 is cleaved from meaning as indicated above in connection with the drug resin according to the following procedure: linker conjugate D-L. The same holds true for the analogous The resin is washed with DCM, dried in vacuo and treated employment of the prodrug precursor L'-Y in respect of the two times for 30 minutes with 6/4 (v/v) DCM/HFIP. Eluates moiety L' represented by formula (I). 35 are combined, volatiles are removed under a nitrogen stream Y is a leaving group. Such leaving groups are known to and product 1 is purified by RP-HPLC and analyzed by a person skilled in the art. Preferably, Y is chloride, bromide, RP-HPLC-MS. fluoride, nitrophenoxy, imidazolyl. N-hydroxySuccinimidyl, N-hydroxybenzotriazolyl, N-hydroxyazobenzotriazolyl, Example 2 pentafluorophenoxy, 2-thiooxo-thiazolidinyl, or N-hydroxy 40 SulfoSuccinimidyl. Synthesis of Exendin-4 Linker Intermediate (2) In case the synthesis of a carrier-linked prodrug according to the present invention is carried out by employing a precursor L'-Y, a drug linker intermediate (L-D) is obtained 45 by reacting L'-Y with the biologically active drug D-H by oHis forming an amide bond. In Such case, said drug linker N-Exendin intermediate L'-D is reacted further to obtain the drug linker H 50 conjugate D-L by adding the moiety Land the carrier group O Z to said drug linker intermediate L'-D. It has to be indicated that the addition of L and/or Z to L'-D may be performed HN" in several steps by preparing further intermediate com pounds prior to obtaining the drug linker conjugate D-L. 55 Alternatively, a prodrug precursor L*-Y may be employed instead of L'-Y, wherein L* is selected from a fragment of L', L' containing at least one protecting group or L' addi NH tionally containing precursors of L and/or Z. 60 Another subject of the present invention is the use of prodrugs or a pharmaceutically acceptable salt thereof com prising a drug linker conjugate D-L as pharmaceuticals or medicaments, respectively. With respect of the definitions of SH the drug linker conjugate D-L as well as further Substituents 65 Such as L' the same explanations as laid out above in the 1 (0.10 mmol), PyBOP (0.10 mmol), and DIEA (0.20 context of the prodrug as such apply. mmol) are dissolved in 2 ml of dry DME Mixture is added US 9,533,056 B2 63 64 to 250 mg side-chain protected Exendin-4 (J. Eng et al., J. Example 5 Biol.Chem. 1992, 267, 11, 7402-7405) on-resin (25 umol; synthesized by Fmoc-strategy on Rink amide resin) and agitated for 30 min at room temperature. Resin is washed Synthesis of Linker Reagent Intermediate (4) with DMF (10 times) and DCM (10 times). Fmoc-group is removed by agitating the resin with 2/2/96 (v/v/v) piperi dine/DBU/DMF (two times, 10 min each) and washing with DMF (ten times). Trt-mercaptopropionic acid (0.1 mmol), PyBOB (0.1 mol), and DIEA (0.2 mmol) in DMF are added to the resin and agitated for 30 min at room temperature. 2 10 is cleaved from resin according to the following procedure: The resin is washed with DCM, dried in vacuo and treated with 2 ml of TFA cleavage cocktail (TFA/TES/Water/DTT 95/2/2/1) per 100 mg resin for 60 min at room temperature. 15 Volatiles are removed under a nitrogen stream. Nonpolar side products and protecting groups are removed by pre cipitating peptide from diethyl ether. Precipitate is dried in WaCO. Crude 2 is dissolved in acetonitrile/water 1/1 and purified NH by RP-HPLC. fmoc Example 3 Fmoc-5,5-dimethyl-D-proline (0.9 mmol) was loaded onto 2-chlorotrityl resin (0.6 mmol) according to manufac Synthesis of PEG-Linker-Exendin-4 Conjugate (3) 25 turer's instructions. After finnoc removal, a solution of boc Lys(fmoc)-OH (3 mmol), HATU (3 mmol), and collidine (6 mmol) was added to the resin and incubated for 30 min. This 3 procedure was repeated once. Intermediate 4 was cleaved O 30 from resin according to the following procedure: ois The resin was washed with DCM, dried in vacuo and N-Exendin H treated two times for 30 minutes with 6/4 (v/v) DCM/HFIP. N Eluates were combined, volatiles were removed under a O nitrogen stream and product 4 was purified by RP-HPLC 35 and analyzed by RP-HPLC-MS. H.N." Example 6

40 Synthesis of Amoxapine Linker Intermediate (5)

NH

O 45 O

N PEG40kDa N 50 boc1 4,7. O 2 (12 mg) is dissolved in 500 ul of 1/1 acetonitrile/water C and 120 mg 40 kDa methoxy poly(ethylene glycol) maleimido-propionamide in 1 ml of 1/1 acetonitrile/water are added. 300 ul of 0.25M sodium phosphate buffer pH 7.0 are added and solution is acidified after 10 min with 300 ul 55 acetic acid. 3 is purified by cation exchange chromatogra NH2 O phy, desalted, and then lyophilized.

Example 4 60 Exendin-4 Release In Vitro 5 (26 umol), PyBOP (32 umol), and DIEA (63 umol) were dissolved in 250 uL of anhydrous DME Amoxapine (36 Release of exendin from 3 is effected by hydrolysis in 50 umol) was added, and the mixture was stirred at room mM sodium phosphate buffer at pH 7.4 and 37°C. Unmodi 65 temperature for 50 min. Piperidine (63 uL) was added and fied native exendin-4 is released as assessed by RP-HPLC/ stirring was continued for 20 minutes. The crude product MS. was purified by RP-HPLC. Yield 83%. US 9,533,056 B2 65 66 Example 7 PyBOP benzotriazole-1-yl-oxy-tris-pyrrolidino-phospho nium hexafluorophosphate Synthesis of PEG-Linker-Amoxapine Conjugate (6) RP-HPLC reversed-phase high performance liquid chroma tography 5 TES triethylsilane TFA trifluoroacetic acid

The invention claimed is: N 10 1. A prodrug or a pharmaceutically acceptable salt thereof N compr1S1ng:

H2N,A. O a drug linker conjugate D-L. C wherein D is a biologically active moiety containing an N aliphatic amine containing group; and 15 wherein L is a linker containing: % i) a moiety L' represented by formula (I):

O NH O

(I)

PEG-2OkDA-OMe

To a solution of 5 (1.4 mg) in 50 uL of DMSO were added 25 a solution of 63 mg 20 kDa methoxy poly(ethylene glycol) NHS ester in 500 uL ml of DMSO and 7 ul of DIPEA. After incubation at 22° C. for 30 min, the reaction mixture was frozen and lyophilized. Dry samples were treated with TFA/DCM 1:1(v/v) for 20 30 min, diluted with MeCN and purified by RP-HPLC. wherein the dashed line indicates the attachment of Example 8 L' to the aliphatic amine group of D by forming an amide bond; Amoxapine Release In Vitro 35 wherein X is selected from O, S, and CH-R'': wherein R' and R" are independently selected from Release of amoxapine from 6 was effected by hydrolysis H, OH, and CH: in 50 mM sodium phosphate buffer at pH 7.4 and 37° C. wherein R. R. R', and R“ are independently Unmodified amoxapine is released as assessed by RP Selected from H and C alkyl; HPLC/MS. 40 wherein R and R" are independently selected from t=1.75 h. H, C, alkyl, and R. and While this invention has been described in conjunction wherein R is selected from: with the specific embodiments outlined above, it is evident that many alternatives, modifications, and variations will be 45 apparent to those skilled in the art. Accordingly, the pre ferred embodiments of the invention as set forth above are intended to be illustrative, not limiting. Various changes may be made without departing from the spirit and scope of the inventions as defined in the following claims. 50 co ABBREVIATIONS Boc t-butyloxycarbonyl -(o- DBU 1,3-diazabicyclo[5.4.0]undecene 55 DCM dichloromethane DIEA diisopropylethylamine DMF N,N-dimethylformamide DMSO dimethylsulfoxide NH DTT dithiothreitol 60 Fmoc 9-fluorenylmethoxycarbonyl HATU O-(7-AZabenzotriazol-1-yl)-N,N,N',N'-tetramethyl uronium hexafluorophosphate HFIP hexafluoroisopropanol MeCN acetonitrile 65 MS mass spectrometry PEG poly(ethylene glycol) US 9,533,056 B2 67 68 -continued 3. The prodrug according to claim 1: wherein the carrier group Z is a polymer with a molecular weight a 500 g/mol. 4. A pharmaceutical composition comprising: a prodrug of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. 5. The pharmaceutical composition according to claim 4: wherein the pharmaceutical composition is dry. s 6. The pharmaceutical composition according to claim 4: 10 wherein the prodrug is sufficiently dosed in the compo sition to provide a therapeutically effective amount of and biologically active agent for at least 12 hours in one wherein, optionally, one or more of the pairs R/R', application. R/R“, and R/R may independently form one or 7. A kit of parts comprising: more cyclic fragments selected from C, 15 a needle; and cycloalkyl, 4 to 7 membered heterocyclyl, and 9 to a container containing: 11 membered heterobicyclyl; and reconstitution solution; and the dry composition according to claim 5 configured for wherein, optionally, R, RR', and R* are further use with the needle. Substituted; and 8. The kit of parts according to claim 7: ii) a moiety Lif, which is a chemical bond or a spacer, wherein the container is a dual-chamber Syringe; and and L is bound to a carrier group Z: wherein one of the two-chambers of the dual-chamber wherein L' is substituted with one to four L. moieties: Syringe contains the dry pharmaceutical composition wherein Z is a water-soluble polymer selected from the and the second chamber of said dual-chamber Syringe group consisting of polyalkyloxy polymers, 25 contains the reconstitution solution. hyaluronic acid and derivatives thereof, polyvinyl 9. The prodrug according to claim 1: alcohols, polyoxazolines, polyanhydrides, poly(or wherein L is a spacer. 10. A method for the synthesis of a prodrug or a phar tho esters), polycarbonates, polyurethanes, poly maceutically acceptable salt thereof according to claim 1, acrylic acids, polyacrylamides, polyacrylates, comprising: polymethacrylates, polyorganophosphaZenes, poly- 30 a step of reacting a prodrug precursor L-Y or L'-Y with siloxanes, polyvinylpyrrolidone, polycyanoacry a biologically active drug D-H, to obtain the drug linker lates, and polyesters; and conjugate D-L or a drug linker intermediate D-L' by wherein, optionally, L is further substituted. forming an amide bond; 2. The prodrug according to claim 1: wherein Y is a leaving group. wherein L is a chemical bond. k k k k k