US 20120328702A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2012/0328702 A1 EDELSON et al. (43) Pub. Date: Dec. 27, 2012

(54) NANOPARTICLE COMPOSITIONS A6IP3/04 (2006.01) A6IP3 L/10 (2006.01) (75) Inventors: Jonathan EDELSON, Scarsdale, NY A6IP 7/2 (2006.01) (US); Timothy KOTYLA, Lowell, MA A6IP 7/02 (2006.01) (US): Klaus THEOBALD, Paoli, PA A61O 19/02 (2006.01) (US) A6IP35/00 (2006.01) A6IP 9/02 2006.O1 (73) Assignee: ANTERIOS, INC., New York, NY (US) A6IP 25/00 :08: A6IP27/02 2006.O1

A6IP 2L/00 (2006.01) (22) Filed: Jan. 23, 2012 A61O 19/08 (2006.01) Related U.S. Application Data St. iO :08: (60) Provisional application No. 61/435,780, filed on Jan. A6IPI3/08 (2006.01) 24, 2011. A6IP37/08 (2006.01) A6IPI3/10 (2006.01) Publication Classification A 6LX39/395 (2006.01) B82Y5/00 (2011.01) (51) Int. Cl. (52) U.S. Cl...... 424/490; 424/400; 424/130.1; 3. 6. 3:08: 424/239.1, 514/785; 424/65;977/773;977/906 A6 IK9/48 (2006.01) A6 IK 47/44 (2006.01) (57) ABSTRACT A6 IK 8/92 (2006.01) A61O 15/00 (2006.01) The present invention describes novel nanoparticle composi A6IP 7/10 (2006.01) tions, and systems and methods utilizing them for treating A6IP 7/00 (2006.01) disorders and/or conditions associated with the epidermal A6IP 7/08 (2006.01) and/or dermal level of the skin. Such disorders include acne, A6IP 7/4 (2006.01) hyperhidrosis, bromhidrosis, chromhidrosis, rosacea, hair A6IP 7/06 (2006.01) loss, dermal infection, actinic keratosis, facial wrinkles, A6IP3I/00 (2006.01) muscle contracture, and headache. Methods generally A6IP3L/2 (2006.01) involve administering nanoparticle compositions to the skin. US 2012/0328702 A1 Dec. 27, 2012

NANOPARTICLE COMPOSITIONS active agent), and systems and methods relating thereto, are also contemplated by the present invention. RELATED APPLICATIONS 0006 Nanoparticle compositions, such as those described in PCT patent application number PCT US06/46236, filed 0001. This application claims priority to and benefit of Dec. 1, 2006, published as WO 08/045,107 on Apr. 17, 2008, U.S. provisional application Ser. No. 61/435,780 filed Jan. and entitled “BOTULINUM NANOEMULSIONS: in PCT 24, 2011, the entire contents of which are incorporated herein patent application number PCT US07/86018, filed Nov.30, by reference. 2007, published as WO 08/070,538 on Jun. 12, 2008, and entitled “AMPHIPHILIC ENTITY NANOPARTICLES: BACKGROUND and/or in PCT patent application number PCT US09/48972, 0002 Nanoparticle compositions are useful in a variety of filed Jun. 26, 2009, published as WO 09/158,687 on Dec. 30, contexts. Nanoparticle compositions have proven to be par 2009, and entitled “DERMAL DELIVERY” (the contents of ticularly useful and/or effective in the context of medical all of which are incorporated herein by reference) have been applications, including administering therapeutic agents to Successfully used for topical administration of therapeutic patients in need thereof. Nanoparticle compositions have agents. The present inventors have undertaken extensive stud proven to be particularly useful and/or effective in the context ies of this class of composition, and, as described in the of topical administration of therapeutic agents (see, e.g., PCT Examples, have made important and Surprising findings that patent application number PCT US06/46236, filed Dec. 1, define certain embodiments and/or classes of Such composi 2006, published as WO 08/045,107 on Apr. 17, 2008, and tions as particularly and unexpectedly useful and/or advanta entitled “BOTULINUMNANOEMULSIONS: in PCT patent geous. application number PCT US07/86018, filed Nov. 30, 2007, 0007. In some embodiments, the present invention pro published as WO 08/070,538 on Jun. 12, 2008, and entitled vides particular nanoparticle compositions as described “AMPHIPHILIC ENTITY NANOPARTICLES: and/or in herein. In some embodiments, nanoparticle compositions PCT patent application number PCT US09/48972, filed Jun. comprise an oil, and aqueous dispersion medium, a surfactant 26, 2009, published as WO 09/158,687 on Dec. 30, 2009, and and, optionally, an independently biologically active agent entitled “DERMAL DELIVERY”: the contents of all of and/or a known therapeutic agent. which are incorporated herein by reference). 0008. In some embodiments, provided nanoparticle com 0003 Conditions or disorders associated with skin (in positions comprise 1349 oil. In some embodiments, provided cluding the surface of the skin, sweat glands, sebaceous nanoparticle compositions comprise polysorbate 80. In some glands, etc.) can cause a great deal of unhappiness and psy embodiments, provided nanoparticle compositions comprise chological debilitation for those who suffer from them, and propylparaben. In some embodiments, provided nanoparticle current treatments are not very successful and often have compositions comprise methylparaben. In some embodi undesirable side effects. For example, according to studies, ments, provided nanoparticle compositions comprise iso acne often leads to reduced self esteem, and sometimes even tonic sodium chloride Solution. In some embodiments, pro to depression or Suicide (see, e.g., Goodman, 2006, Aust. vided nanoparticle compositions comprise purified water. In Fam. Physician 35:503, 2006: Purvis et al., 2006, J. Paediatr. Some embodiments, provided nanoparticle compositions Child. Health 42:793; both of which are incorporated herein comprise gelatin. In some embodiments, provided nanopar by reference). Similar challenges are observed with hyper ticle compositions comprise sodium phosphate dibasic. In hidrosis (excessive Sweating), bromhidrosis (body odor), Some embodiments, provided nanoparticle compositions chromhidrosis (colored Sweat), psoriasis, dermal infection comprise concentrated hydrochloric acid. In some embodi (e.g., herpes simplex virus infection, human papillomavirus ments, provided nanoparticle compositions do not comprise infection, fungal infection, etc.), hair loss, actinic keratosis, any parabens. In some embodiments, provided nanoparticle rosacea, facial wrinkles, neck lines, platysma bands, neuro compositions do not comprise methylparaben. In some muscular disorders and conditions involving muscular spasm embodiments, provided nanoparticle compositions do not and/or contracture, and other afflictions of the skin. Nanopar comprise propylparaben. ticle compositions have been described which may be useful 0009. In some embodiments, nanoparticle compositions and/or effective for topical administration for treatment of comprise a nanoparticle composition, including, but not lim Such disorders, but there is a need for improved nanoparticle ited to, a nanoemulsion. In some embodiments, a nanoemul compositions for topical administration for more effective sion comprises 1349 oil, polysorbate 80, propylparaben, iso treatment of disorders such as those associated with the skin. tonic sodium chloride solution, methylparaben, a buffer Solution (comprising gelatin, Sodium phosphate dibasic, puri SUMMARY OF THE INVENTION fied water, and hydrochloric acid) and, optionally, a known therapeutic agent and/or independently active biologically 0004. The present invention provides particular composi active agent. In some embodiments, a nanoemulsion com tions as described herein. prises 1349 oil, polysorbate 80, isotonic sodium chloride 0005. In some embodiments, the present invention encom Solution, a buffer solution (comprising gelatin, Sodium phos passes the recognition that nanoparticle compositions (e.g., phate dibasic, purified water, and hydrochloric acid) and, nanoemulsions) are useful as therapeutic agents. Thus, nano optionally, a known therapeutic agent and/or independently particle compositions containing a known therapeutic agent active biologically active agent. In some embodiments, the and/or independently active biologically active agent, and nanoemulsion comprises oil and Surfactant at a ratio of 0.67: systems and methods relating thereto, are contemplated by 1. In some embodiments, a nanoemulsion comprises oil and the present invention. Empty nanoparticle compositions (e.g., an isotonic sodium chloride solution at a ratio of 1:10. In nanoparticle compositions that do not contain any known Some embodiments, a nanoemulsion comprises Surfactant therapeutic agent and/or independently active biologically and an isotonic sodium chloride solution at a ratio of 1:1.67. US 2012/0328702 A1 Dec. 27, 2012

In some embodiments, a nanoemulsion comprises the com cream and/or lotion formulations comprise purified water, ponents set forth in any of Examples 1-20. mineral oil, isopropyl myristate, white petrolatum, and emul 0010. In some embodiments, provided nanoparticle com Sifying wax. In some embodiments, provided cream and/or positions comprise a saline Solution. In some embodiments, lotion formulations comprise the components set forth in the saline solution comprises isotonic sodium chloride solu Examples 1-20. tion. In some embodiments, the saline solution comprises 0017. In some embodiments, the present invention pro isotonic sodium chloride solution and/or water. In some vides particular cream and/or lotion formulations as embodiments, the Saline Solution comprises isotonic sodium described herein. In some embodiments, provided cream and/ chloride Solution and methylparaben. In some embodiments, orlotion formulations comprise water. In some embodiments, the saline Solution comprises the components set forth in any provided cream and/or lotion formulations comprise meth of Examples 1-20. ylparaben. In some embodiments, provided cream and/or 0011. The present invention encompasses the recognition lotion formulations comprise mineral oil. In some embodi that provided nanoparticle compositions can be formulated ments, provided cream and/or lotion formulations comprise for delivery to a subject in need thereof via topical and/or isopropyl myristate. In some embodiments, provided cream transdermal administration (e.g., by lotions, creams, pow and/or lotion formulations comprise white petrolatum. In ders, ointments, liniments, gels, drops, etc.). In some embodi Some embodiments, provided cream and/or lotion formula ments, provided nanoparticle compositions are administered tions comprise emulsifying wax. In some embodiments, pro to a Subject in need thereof via topical and/or transdermal vided cream and/or lotion formulations comprise propylpa (e.g., by lotions, creams, powders, ointments, liniments, gels, raben. In some embodiments, provided cream and/or lotion drops, etc.) administration. formulations do not comprise any parabens. In some embodi 0012. The present invention encompasses the recognition ments, provided cream and/or lotion formulations do not that provided nanoparticle compositions can be particularly comprise methylparaben. In some embodiments, provided useful for topical and/or transdermal administration. The cream and/or lotion formulations do not comprise propylpa present invention encompasses the recognition that provided raben. nanoparticle compositions can be particularly useful for 0018. The present invention encompasses the recognition delivery of agents to the dermal level of the skin. In some that provided cream and/or lotion formulations can be par embodiments, provided nanoparticle compositions are for ticularly useful for formulating nanoparticle compositions, mulated for topical and/or transdermal delivery to a subject in Such as those described herein, for administration to a Subject. need thereof. In some embodiments, provided nanoparticle The present invention encompasses the recognition that pro compositions are administered to a Subject in need thereof via vided cream and/or lotion formulations can be particularly topical and/or transdermal delivery. useful for formulating nanoemulsions, such as those 0013 The present invention provides the surprising find described herein, for administration to a subject. ing of particular cream and/or lotion formulations with unex 0019. The present invention provides that surprising and/ pected and/or beneficial properties. or unexpected results can be achieved when cream and/or 0014. The present invention encompasses the recognition lotion formulations are formulated with particular provided that provided cream and/or lotion formulations can be used nanoparticle compositions as described herein. for delivery of agents to a subject in need thereof via topical 0020. In some embodiments, provided compositions com and/or transdermal (e.g., by lotions, creams, powders, oint prise a mixture of a provided nanoparticle composition and ments, liniments, gels, drops, etc.) administration. In some one or more pharmaceutically acceptable excipients. In some embodiments, provided cream and/or lotion formulations are embodiments, provided compositions comprise a mixture of administered to a subject in need thereof via topical and/or a provided nanoparticle composition, a saline solution, and a transdermal (e.g., by lotions, creams, powders, ointments, provided cream and/or lotion formulation, as described liniments, gels, drops, etc.) administration. In some embodi herein. ments, provided cream and/or lotion formulations are formu lated with provided nanoparticle compositions. In some 0021. In some embodiments, provided compositions com embodiments, provided cream and/or lotion formulations prise provided nanoparticle compositions. In some embodi that are formulated with provided nanoparticle compositions ments, provided compositions comprise provided cream and/ are useful and/or effective for topical administration to a or lotion formulations. In some embodiments, provided Subject. compositions comprise both provided nanoparticle composi 0015 The present invention encompasses the recognition tions and provided cream and/or lotion formulations. In some that provided cream and/or lotion formulations can be par embodiments, provided compositions comprise provided ticularly useful for topical and/or transdermal administration. nanoparticle compositions but do not comprise provided The present invention encompasses the recognition that pro cream and/or lotion formulations. In some embodiments, pro vided cream and/or lotion formulations can be particularly vided compositions comprise provided cream and/or lotion useful for delivery of agents to the dermal level of the skin. In formulations but do not comprise provided nanoparticle com Some embodiments, provided cream and/or lotion formula positions. tions are formulated for topical and/or transdermal delivery to 0022. In some embodiments, provided compositions com a Subject in need thereof. In some embodiments, provided prise a mixture of a provided nanoparticle composition and cream and/or lotion formulations are administered to a Sub one or more pharmaceutically acceptable excipients, e.g., for ject in need thereof via topical and/or transdermal delivery. topical and/or transdermal (e.g., by lotions, creams, powders, 0016. In some embodiments, provided cream and/or ointments, liniments, gels, drops, etc.) administration. lotion formulations comprise purified water, methylparaben, 0023 The present invention provides methods of treating mineral oil, isopropyl myristate, white petrolatum, emulsify conditions or disorders using any of the provided composi ing wax, and propylparaben. In some embodiments, provided tions (e.g., provided nanoparticle composition; cream and/or US 2012/0328702 A1 Dec. 27, 2012 lotion formulation; combination of provided nanoparticle 0027. The present invention provides methods of treating composition and cream and/or lotion formulation, etc.) as conditions or disorders by administering to a patient a pro described herein. vided composition as described herein (e.g., provided nano particle composition; cream and/or lotion formulation; com 0024. In some embodiments, such methods involve bination of provided nanoparticle composition and cream administration of a provided composition to a patient Suffer and/or lotion formulation, etc.). In some embodiments, the ing from and/or Susceptible to a disease, condition, or disor present invention provides methods of treating conditions or der. In some embodiments, such methods involve administra disorders by administering to a patient a composition con tion of a provided nanoparticle composition to a patient taining a provided nanoparticle composition (e.g., a Suffering from and/or susceptible to a disease, condition, or nanoemulsion) as described herein. In some embodiments, disorder associated with the dermal layer of the skin. In some administration is local administration. In some embodiments, embodiments, such methods involve administration of an empty nanoparticle composition (e.g., a nanoparticle compo local administration is topical administration. sition not containing a known therapeutic agent and/or inde 0028. In general, for nanoparticle compositions compris pendently active biologically active agent) to a patient Suffer ing a known therapeutic agent and/or independently active ing from and/or susceptible to a disease, condition, or biologically active agent, such nanoparticle compositions are disorder. In some embodiments, such methods involve arranged and constructed Such that an amount of therapeutic administration of a nanoparticle composition comprising at agent is delivered to a desired target site (e.g., to epidermal least one known therapeutic agent and/or independently and/or dermal structures) that is sufficient to treat the condi active biologically active agent to a patient Suffering from tion or disorder. In some embodiments, provided nanoparticle and/or Susceptible to a disease, condition, or disorder. In some compositions are arranged and constructed (e.g., through embodiments. Such methods involve administration of a selection and/or combination of agents, structure of compo nanoparticle composition and/or at least one known therapeu sition, etc.) such that they achieve the desired therapeutic tic agent and/or independently active biologically active effect upon administration to the skin. In some embodiments, agent formulated with a provided cream and/or lotion formu provided nanoparticle compositions are arranged and con lation to a patient Suffering from and/or Susceptible to a structed such that they do not induce unwanted clinical effects disease, condition, or disorder. In some embodiments, such inside and/or outside of the desired site of action (e.g., Surface methods involve administration of provided compositions via of skin, dermis, etc.). In some embodiments, provided nano topical and/or transdermal (e.g., by lotions, creams, powders, particle compositions are arranged and constructed Such that ointments, liniments, gels, drops, etc.) administration. they have systemic effects. 0029. In some embodiments, provided compositions may 0025. In some embodiments, the present invention pro be formulated and/or delivered so that systemic delivery is vides methods of treating any conditions or disorders. In achieved; in Some embodiments, provided compositions may Some embodiments, the present invention demonstrates that beformulated and/or delivered so that local, but not systemic, certain compositions as described herein can achieve con delivery is achieved. trolled delivery of active agents efficiently and specifically to 0030 The present disclosure specifically demonstrates biologically relevant target sites (e.g., particular tissues, loca effective and efficient delivery of a therapeutic agent (and, in tions within the skin, cells, etc.). In some embodiments, the particular, a large biologic agent, such as botulinum toxin) to present invention demonstrates controlled delivery and/or the dermis using provided compositions. For example, in achievement of therapeutic effect in a certain biologically Some embodiments, the present invention provides methods relevant target site without significant side effects associated comprising administration of a composition as described with delivery to other areas. herein without clinically significant side effects. To give but 0026. In some embodiments, the present invention pro one example, when topical delivery is contemplated, clini vides methods of treating conditions or disorders associated cally significant side effects include, but are not limited to, with epidermal and/or dermal structures (e.g., Sweat glands, unwanted systemic side effects, damage to nervous tissue sebaceous glands, hair follicles, etc.). In some embodiments, underlying the dermis (e.g., neuronal paralysis), unwanted the present invention demonstrates that provided composi effects on muscles (e.g., muscle paralysis), and/or undesir tions as described herein (e.g., provided nanoparticle compo able blood levels of therapeutic agent, etc. The present inven sition; cream and/or lotion formulation; combination of pro tion provides the Surprising benefits and/or capabilities of vided nanoparticle composition and cream and/or lotion particular provided compositions (e.g., provided nanoparticle formulation, etc.) can deliver active agents efficiently and compositions and/or provided cream and/or lotion formula specifically to the dermis, and that provided compositions as tions) as compared with nanoparticle compositions in gen described herein can have therapeutic effects upon adminis eral. tration to the skin of a Subject. In some embodiments, the 0031. The present invention further provides technologies present invention demonstrates dermal delivery and/or for identifying the component or components present in the achievement of therapeutic effect without significant side provided compositions that are responsible for the observed effects associated with delivery to other areas (e.g., to Sub activity of nanoparticle compositions and/or cream and/or dermal or extradermal structures and/or to tissues other than lotion formulations. To the extent that such technologies iden dermis). In some embodiments, provided compositions as tify component(s) that can achieve the observed results inde described herein (e.g., provided nanoparticle composition; pendent of a nanoparticle structure, the present invention also cream and/or lotion formulation; combination of provided provides use in medicine, and in particular in the treatment of nanoparticle composition and cream and/or lotion formula conditions or disorders associated with dermal structures tion; etc.) can transdermally deliver active agents, such as (e.g., Sweat glands, sebaceous glands, hair follicles, etc.), of therapeutic agents (e.g., botulinum toxins, monoclonal anti compositions containing one or more individual components bodies, etc.). of provided compositions. As used herein, a “provided com US 2012/0328702 A1 Dec. 27, 2012

position” may contain one or more individual components of of a peptide. It will be apparent from the context in which the nanoparticle compositions and/or cream and/or lotion formu term is used whether it refers to a free amino acid or a residue lations. of a peptide. 0032. This application refers to various patent and non 0036 Animal: As used herein, the term “animal' refers to patent publications, all of which are incorporated herein by any member of the animal kingdom. In some embodiments, reference. "animal' refers to humans, at any stage of development. In Some embodiments, “animal' refers to non-human animals, DEFINITIONS at any stage of development. In certain embodiments, the 0033. Abrasion: The term “abrasion,” as used herein, non-human animal is a mammal (e.g., a rodent, a mouse, a rat, refers to any means of altering, disrupting, removing, or a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, destroying the top layer of the skin. In some embodiments, and/or a pig). In some embodiments, animals include, but are abrasion refers to a mechanical means of altering, disrupting, not limited to, mammals, birds, reptiles, amphibians, fish, removing, or destroying the top layer of the skin. In some and/or worms. In some embodiments, an animal may be a embodiments, abrasion refers to a chemical means of alter transgenic animal, genetically-engineered animal, and/or a ing, disrupting, removing, or destroying the top layer of skin. clone. To give but a few examples, agents such as exfoliants, fine 0037 Approximately: As used herein, the terms “approxi particles (e.g., magnesium or aluminum particles), acids mately” or “about in reference to a number are generally (e.g., alpha-hydroxy acids or beta-hydroxy acids), and/or taken to include numbers that fall within a range of 5%, 10%, alcohols may cause abrasion. In general, permeation enhanc 15%, or 20% in either direction (greater than or less than) of ers such as those described, for example, by Donovan (see, the number unless otherwise stated or otherwise evident from e.g., U.S. Patent Publications 2004/009180 and 2005/ the context (except where such number would be less than 0% 175636; and PCT Publication WO 04/06954; all of which are or exceed 100% of a possible value). incorporated herein by reference), and Graham (see, e.g., 0038 Biologically active agent: As used herein, the phrase U.S. Pat. No. 6,939,852 and U.S. Patent Publication 2006/ “biologically active agent” refers to any Substance that has 093624; both of which are incorporated herein by reference), activity in a biological system and/or organism. For instance, etc., are expected to cause abrasion. Of course, those of ordi a Substance that, when administered to an organism, has a nary skill in the art will appreciate that a particular agent may biological effect on that organism is considered to be biologi cause abrasion when present at one concentration, or in asso cally active. In some embodiments, where a Substance (e.g., a ciation with one or more other agents, but may not cause polypeptide, nucleic acid, antibody, etc.) is biologically abrasion under different circumstances. Thus, whether or not active, a portion of that Substance that shares at least one a particular material is an "abrasive agent' depends on con biological activity of the whole substance is typically referred text. Abrasion can readily be assessed by those of ordinary to as a “biologically active' portion. skill in the art, for example by observation of redness or 0039. Botulinum nanoparticle composition: The term irritation of the skin and/or histologic examination of skin “botulinum nanoparticle composition, as used herein, refers showing alteration, disruption, removal, or erosion of the to any nanoparticle composition described herein in which at Stratum COrneum. least one nanoparticle includes botulinum toxin. The botuli 0034 Administration: The term “administration,” as used num toxin may be present within the nanoparticle, on the herein refers to the delivery and/or administration of a pro nanoparticle Surface and/or within a micellar membrane vided composition to a subject. For example, the present defining the nanoparticle. invention contemplates routes of delivering or administering 0040. Botulinum toxin: The term “botulinum toxin,” as that include, but are not limited to, topical and/or transdermal used herein, refers to any neurotoxin produced by (e.g., by lotions, creams, liniments, ointments, powders, gels, Clostridium botulinum. Except as otherwise indicated, the drops, deodorants, antiperspirants, Sunscreens, etc.). term encompasses fragments orportions (e.g., the light chain 0035 Amino acid: As used herein, term “amino acid.” in and/or the heavy chain) of Such neurotoxin that retain appro its broadest sense, refers to any compound and/or Substance priate activity (e.g., muscle relaxant activity). The phrase that can be incorporated into a polypeptide chain. In some “botulinum toxin, as used herein, encompasses the botuli embodiments, an amino acid has the general structure H.N- num toxin serotypes type A, type Ab, type Af, type B, type Bf C(H)(R)—COOH. In some embodiments, an amino acid is a type C1, type C2, type D, type E, type F, and type G. mutants naturally-occurring amino acid. In some embodiments, an thereof; variants thereof; fragments thereof; characteristic amino acid is a synthetic amino acid; in some embodiments, portions thereof; and/or fusions thereof. In some embodi an amino acid is a D-amino acid; in Some embodiments, an ments, botulinum toxin is present as any of the Subtypes amino acid is an L-amino acid. “Standard amino acid refers described in Sakaguchi, 1982, Pharmacol. Ther:, 19:165; to any of the twenty standard L-amino acids commonly found and/or Smith et al., 2005, Infect. Immun., 73:5450; both of in naturally occurring peptides. "Nonstandard amino acid which are incorporated herein by reference. Botulinum toxin, refers to any amino acid, other than the standard amino acids, as used herein, also encompasses both a botulinum toxin regardless of whether it is prepared synthetically or obtained complex (i.e., for example, the 300, 600, and 900 kD com from a natural Source Amino acids, including carboxy- and/or plexes) as well as the purified (i.e., for example, isolated) amino-terminal amino acids in peptides, can be modified by botulinum toxin (i.e., for example, about 150 kD). “Purified methylation, amidation, acetylation, and/or Substitution with botulinum toxin' is defined as a botulinum toxin that is iso other chemical groups that can change the peptide's circulat lated, or Substantially isolated, from other proteins, including ing half-life without adversely affecting their activity. Amino proteins that form a botulinum toxin complex. A purified acids may participate in a disulfide bond. The term "amino toxin may be greater than 80% pure, greater than 85% pure, acid is used interchangeably with “amino acid residue.” and greater than 90% pure, greater than 95% pure, greater than may refer to a free amino acid and/or to an amino acid residue 98% pure, and/or greater than 99% pure. Those of ordinary US 2012/0328702 A1 Dec. 27, 2012

skill in the art will appreciate that the present invention is not tide molecules. In some embodiments, polymeric molecules limited to any particular source of botulinum toxin. For are considered to be “homologous' to one another if their example, botulinum toxin for use in accordance with the sequences are at least 25%, at least 30%, at least 35%, at least present invention may be isolated from Clostridium botuli 40%, at least 45%, at least 50%, at least 55%, at least 60%, at num, may be chemically synthesized, may be produced least 65%, at least 70%, at least 75%, at least 80%, at least recombinantly (i.e., in a host cell or organism other than 85%, at least 90%, at least 95%, or at least 99% identical. In Clostridium botulinum), etc. Some embodiments, polymeric molecules are considered to 0041 Cosmetic formulation: The term "cosmetic formu be "homologous to one another if their sequences are at least lation' is used herein to refer to a topically applied composi 25%, at least 30%, at least 35%, at least 40%, at least 45%, at tion that contains one or more agents having cosmetic prop least 50%, at least 55%, at least 60%, at least 65%, at least erties. To give but a few examples, a cosmetic formulation 70%, at least 75%, at least 80%, at least 85%, at least 90%, at may be a skin softener, nutrition lotion type emulsion, cleans least 95%, or at least 99% similar. ing lotion, cleansing cream, skin milk, emollient lotion, mas 0047. Identity: As used herein, the term “identity” refers to sage cream, emollient cream, make-up base, lipstick, facial the overall relatedness between polymeric molecules, e.g., pack or facial gel, cleaner formulation Such as shampoos, between polynucleotide molecules (e.g., DNA molecules rinses, body cleanser, hair-tonics, or soaps, and/or a derma and/or RNA molecules) and/or between polypeptide mol tological composition Such as a lotion, ointment, gel, cream, ecules. Calculation of the percent identity of two nucleic acid patch, deodorant, antiperspirant, and/or spray. sequences, for example, can be performed by aligning the two 0042 Cream: The term “cream” refers to a spreadable sequences for optimal comparison purposes (e.g., gaps can be composition, typically formulated for application to the skin. introduced in one or both of a first and a second nucleic acid Creams typically contain an oil and/or fatty acid based-ma sequences for optimal alignment and non-identical sequences trix. Creams formulated according to the present invention can be disregarded for comparison purposes). In certain may contain nanoparticles and may be capable of Substan embodiments, the length of a sequence aligned for compari tially complete penetration (e.g., of Such nanoparticles) son purposes is at least 30%, at least 40%, at least 50%, at through the skin upon topical administration. Such a cream least 60%, at least 70%, at least 80%, at least 90%, at least could also act as a carrier for incorporated materials (e.g., for 95% or 100% of the length of the reference sequence. The example, for one or more known therapeutic agents and/or nucleotides at corresponding nucleotide positions are then independently active biologically active agents). compared. When a position in the first sequence is occupied 0043. Dispersion medium: The term "dispersion medium' by the same nucleotide as the corresponding position in the as used herein, refers to a liquid medium in which particles second sequence, then the molecules are identical at that (e.g., empty nanoparticles and/or nanoparticles containing position. The percent identity between the two sequences is a one or more known therapeutic agents and/or independently function of the number of identical positions shared by the active biologically active agents) are dispersed. In general, a sequences, taking into account the number of gaps, and the dispersion is formed when at least two immiscible materials length of each gap, which needs to be introduced for optimal are combined. An “oil-in-water dispersion is one in which alignment of the two sequences. The comparison of oily particles are dispersed within an aqueous dispersion sequences and determination of percent identity between two medium. A “water-in-oil dispersion is one in which aqueous sequences can be accomplished using a mathematical algo particles are dispersed within an oily dispersion medium. rithm. For example, the percent identity between two nucle Those of ordinary skill in the art will appreciate that a disper otide sequences can be determined using the algorithm of sion can beformed from any two immiscible media and is not Meyers and Miller (CABIOS, 1989, 4: 11-17), which has limited strictly to combinations of aqueous and oily media. been incorporated into the ALIGN program (version 2.0) The term “dispersion medium' therefore applies broadly to using a PAM120 weight residue table, a gap length penalty of any dispersion medium notwithstanding that it is common to 12 and a gap penalty of 4. The percent identity between two refer to “aqueous” and "oily” categories. nucleotide sequences can, alternatively, be determined using 0044) Encapsulated: The term “encapsulated (also the GAP program in the GCG Software package using an “encapsulate' or “encapsulating) is used hereinto mean that NWSgapdna. CMP matrix. the encapsulated entity is completely Surrounded by another 0048. In conjunction with: As used herein, the phrase material. To give but one example, a biologically active agent “delivered in conjunction with and/or “administered in con (e.g., botulinum toxin) may be encapsulated within a nano junction with refers to the co-delivery and/or co-administra particle in an emulsion. Such encapsulation may beachieved, tion of two or more substances or agents. In some embodi for example, during formation of a nanoparticle composition ments, according to the present invention, the phrase is used (e.g., a nanoemulsion), for example during microfluidization. herein in reference to delivery of a biologically active agent To give but another example, known therapeutic agents and/ with nanoparticles and/or provided nanoparticle composi or independently active biologically active agents are not tions. A Substance or agent is delivered in conjunction with encapsulated within empty nanoparticles in an emulsion. nanoparticles when the Substance or agent is combined with 0045 Empty nanoparticle composition: The term “empty nanoparticles and/or nanoparticle compositions; is encapsu nanoparticle composition, as used herein, refers to a nano lated or completely surrounded by nanoparticles; is embed particle composition which does not include a known thera ded within a nanoparticle micellar membrane; and/or is asso peutic agent and/or an independently active biologically ciated with the outer surface of a nanoparticle micellar active agent. membrane. A substance or agent to be delivered in conjunc 0046) Homology: As used herein, the term “homology’ tion with nanoparticles and/or nanoparticle compositions refers to the overall relatedness between polymeric mol may or may not be covalently linked to the nanoparticles ecules, e.g., between polynucleotide molecules (e.g., DNA and/or nanoparticle compositions. A substance or agent to be molecules and/or RNA molecules) and/or between polypep delivered in conjunction with nanoparticles and/or nanopar US 2012/0328702 A1 Dec. 27, 2012 ticle compositions may or may not be attached to the nano butynin, glycopyrrolate, propantheline bromide, benz particles and/or nanoparticle compositions by adsorption tropine, etc.), beta-blockers, antidepressants, anxiolytics, forces. In some embodiments, according to the present inven talc, baby powder, and/or combinations thereof. Exemplary tion, the phrase is used herein in reference to simultaneous known therapeutic agents known to have a particular biologi administration of a composition comprising empty nanopar cal effect on Sebaceous glands include botulinum toxin, ticles with another composition comprising a known thera cleansers or soaps, a topical bactericidal (e.g., benzoyl per peutic agent and/or independently active biologically active oxide, triclosan, and/or chlorhexidine gluconate), topical agent. In Such embodiments, a known therapeutic agent and/ antibiotics (e.g., externally-applied erythromycin, clindamy or independently active biologically active agent is not part of cin, tetracycline, etc.), oral antibiotics (e.g., erythromycin, the empty nanoparticle composition, but instead, is adminis tetracycline, oxytetracycline, doxycycline, minocycline, tered separately to the Subject (e.g., either as a separate com lymecycline, trimethoprim, etc.), hormonal treatments (e.g., position, or having been admixed and/or formulated together estrogen/progestogen oral contraceptives, low dose spirono with the empty nanoparticle composition. In some embodi lactone, cortisone, etc.), a keratolytic (i.e., a substance that ments, a known therapeutic and/or independently active bio dissolves keratin plugging pores), benzoyl peroxide, a topical logically active agent is not incorporated nanoparticles of a retinoid (e.g., tretinoin RETIN-AR, adapalene DIF nanoparticle composition; in some embodiments, a known FERINR, and tazarotene TAZORACR, retinol, isotretin therapeutic and/or independently active biologically active oin, etc.), oral retinoids (e.g., isotretinoin ACCUTANER, agent is not encapsulated within nanoparticles of a nanopar AMNESTEEMTM, SOTRETTM, CLARAVISTM), retinoic ticle composition; in some embodiments, a known therapeu acids, a natural product with anti-acne activity (e.g., aloe Vera, tic and/or independently active biologically active agent is not aruna, haldi i.e., turmeric, papaya, etc.), azelaic acid (brand otherwise in association with nanoparticles of the nanopar names AZELEXTM, FINACEAR, FINEVINR, SKINOREN, ticle composition). etc.), anti-inflammatory agents (e.g., naproxen, ibuprofen, 0049 Independently active biologically active agent: The rofecoxib, etc.), nicotinamide (i.e., vitamin B3), tea tree oil term “independently active biologically active agent” refers (melaleuca oil), aminolevulinic acid, azithromycin, methy to an agent that shows biological activity whether or not the laminolevuninate, nadifloxacine, PRK124, talarozole, zileu agent is present in a nanoparticle composition as described ton, rofecoxib, zinc, an agent described in Krowchuk (2000, herein. In some embodiments, one or more particular biologi Pediatric Dermatology, 47:841-857; incorporated herein by cal activities of the agent is/are improved in a nanoparticle reference) and/or in Johnson et al. (2000, American Family composition; in some embodiments, one or more biological Physician, 62:1823-1830 and 1835-1836; incorporated activities of the agent is/are not improved in a nanoparticle herein by reference), and/or combinations thereof. Exem composition. plary known therapeutic agents known to have a particular 0050. Isolated: As used herein, the term “isolated refers biological effect on hair follicles include minoxidil (RO to a substance and/or entity that has been (1) separated from GAINE(R)/REGAINE(R), finasteride (PROPECIAR), dutas at least Some of the components with which it was associated teride (AVODARTR), an antiandrogen (e.g., ketoconazole, when initially produced (whether in nature and/or in an fluconazole, Spironolactone, etc.), saw palmetto, caffeine, experimental setting), and/or (2) produced, prepared, and/or copper peptides, nitroxide spin labels TEMPO and TEM manufactured by the hand of man. Isolated Substances and/or POL, unsaturated fatty acids (e.g., gamma linolenic acid), entities may be separated from at least about 10%, about 20%, hedgehog agonists, azelaic acid and Zinc in combination, about 30%, about 40%, about 50%, about 60%, about 70%, Chinese knotweed, pumpkin seed, tretinoin, Zinc, Stinging about 80%, about 90%, or more of the other components with nettle, Tempol alcohol-based gel (e.g., MTS-01, etc.), Aldara, which they were initially associated. In some embodiments, alefacept, AS101, bimatoprost, capsaicin, efalizumab, isolated substances and/or entities are more than 90%, 91%, FK506, GP11046, GP11511, hydroxychloroquine, latano 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% pure. prost, MK0906, roXithromycin, Targretin Gel 1%, tetrapep 0051 Known therapeutic agent: As used herein, the term tide aldehyde proteasome inhibitor (e.g., NEOSH 101, etc.), “known therapeutic agent” describes a biologically active and/or combinations thereof. Exemplary known therapeutic agent known, prior to its incorporation in a nanoparticle com agents known to have a particular biological effect on the position, to have a particular biological effect e.g., on a der treatment and/or prevention of psoriasis include, but are not mal structure (e.g., for example, on Sweat glands, sebaceous limited to, botulinum toxin; coal tar; dithranol (anthralin); a glands, hair follicles, etc). In some embodiments, a known corticosteroid such as desoximetasone (TOPICORTR); a therapeutic agent describes a biologically active agent known Vitamin D3 analog (e.g., calcipotriol); a retinoid; argan oil; prior to filing of the present application to have a particular topical administration of psoralen with exposure to ultravio biological effect, e.g., on a dermal structure (e.g., for let A light (PUVA); milk thistle; methotrexate; cyclosporine; example, on Sweat glands, sebaceous glands, hair follicles, the antimetabolite tioguanine; hydroxyurea; Sulfasalazine; etc). Exemplary known therapeutic agents known to have a mycophenolate mofetil, azathioprine; tacrolimus; and/or particular biological effect on Sweat glands include alumi antibody-based therapeutics (e.g., alefacept num chloride, aluminum chlorohydrate, aluminum chlorohy |AMEVIEVER, etanercept EMBREL(R), infliximab drex compounds, aluminum dichlorohydrate, aluminum REMICADE(R), efalizumab RAPTIVAR), etc). dichlorohydrex compounds, aluminum sesquichlorohydrate, 0052 Microfluidized: As used herein, the term “microflu aluminum sesquichlorohydrex compounds, aluminum Zirco idized' means exposed to high shear forces. In some embodi nium tetrachlorohydrex gly, aluminum Zirconium trichloro ments, such exposure to high shear forces is accomplished by hydrex gly, ammonium alum, aluminum Sulfate compounds, exposure to high pressure; in some embodiments such high aluminum zirconium compounds, botulinum toxin, oral pressure is within the range of about 15,000 psi to about medication (e.g., diphenhydramine hydrochloride, hydrox 26,000 psi. In some embodiments, such exposure to high yZine, glycopyrrolate, etc.), anticholinergic drugs (e.g., oxy shear forces is accomplished by cavitation. In some embodi US 2012/0328702 A1 Dec. 27, 2012

ments, such exposure to high shear forces is accomplished by nanoemulsion as described in PCT patent application number passing a sample through an instrument Such as, for example, PCT/US06/026918, filed Jul. 11, 2006, published as WO a Microfluidizer R (Microfluidics Corporation/MFIC Corpo 08/010,788 on Jan. 24, 2008, and entitled “COMPOSITIONS ration) or other like device that may be useful in creating a AND METHODS FOR MAKING AND USING uniform nanoparticle composition. In some embodiments, a NANOEMULSIONS''' (incorporated herein by reference). In sample is microfluidized through exposure to high shear Some embodiments, a nanoparticle composition comprises a forces for a period of time less than about 10 minutes. In some nanoemulsion as described in PCT patent application number embodiments, the period of time is less than about 9, about 8, PCT US06/46236, filed Dec. 1, 2006, published as WO about 7, about 6, about 5, about 4, about 3, about 2, or about 08/045,107 on Apr. 17, 2008, and entitled “BOTULINUM 1 minute(s). In some embodiments, the period of time is NANOEMULSIONS''' (incorporated herein by reference). In within the range of about 1-about 2 minutes. In some embodi Some embodiments, a nanoparticle composition comprises ments, the period of time is about 30 seconds. In some amphiphilic entity nanoparticles as described in PCT patent embodiments, a sample is “microfluidized' through a single application number PCT/US07/86018, filed Nov. 30, 2007, exposure to high shear forces; such embodiments are referred published as WO 08/070,538 on Jun. 12, 2008, and entitled to as “single pass' microfluidization. “AMPHIPHILIC ENTITY NANOPARTICLES (incorpo 0053 Nanoemulsion: An emulsion is traditionally defined rated herein by reference). In some embodiments, a nanopar in the art “as a system ... consisting of a liquid dispersed with ticle composition comprises particles as described in PCT or without an emulsifier in an immiscible liquid usually in application serial number PCT/US08/65329, filed May 30, droplets of larger than colloidal size' Medline Plus Online 2008, published as PCT publication WO 08/151,022 on Dec. Medical Dictionary, Merriam Webster (2005). The term 11, 2008, and entitled “NUCLEIC ACID NANOPAR "nanoemulsion, as used herein, refers to an emulsion in TICLES AND USES THEREFOR” (incorporated herein by which at least some of the droplets (or particles) have diam reference). In some embodiments, a nanoparticle composi eters in the nanometer size range. As will be understood by tion comprises particles as described in PCT patent applica those of ordinary skill in the art, a nanoemulsion is charac tion number PCT/US07/86040, filed Nov. 30, 2007, pub terized by droplets or particles one thousandfold smaller than lished as PCT publication WO 08/140,594 on Nov. 20, 2008, microemulsion droplets or particles. and entitled "PEPTIDE NANOPARTICLES AND USES THEREFOR” (incorporated herein by reference). In some 0054 Nanoparticle: As used herein, the term “nanopar embodiments, a nanoparticle composition comprises par ticle' refers to any particle having a diameterofless than 1000 ticles as described in PCT patent application number PCT nanometers (nm). In some embodiments, a nanoparticle has a US09/48972, filed Jun. 26, 2009, published as WO 09/158, diameter of less than 300 nm, as defined by the National 687 on Dec. 30, 2009, and entitled “DERMAL DELIVERY” Science Foundation. In some embodiments, a nanoparticle (incorporated herein by reference). In some embodiments, has a diameter of less than 100 nm as defined by the National nanoparticle compositions are stable. In some embodiments, Institutes of Health. In some embodiments, nanoparticles are nanoparticle compositions include one or more biologically micelles in that they comprise an enclosed compartment, active agents to be delivered in conjunction with the nanopar separated from the bulk solution by a micellar membrane. A ticles. In some embodiments, nanoparticle compositions are “micellar membrane' comprises amphiphilic entities which empty nanoparticle compositions (e.g., they do not contain have aggregated to Surround and enclose a space or compart any known therapeutic agents and/or independently active ment (e.g., to define a lumen). biologically active agents). 0055 Nanoparticle composition: As used herein, the term "nanoparticle composition” refers to any Substance that con 0056. Not contaminated with: The phrase “not contami tains at least one nanoparticle. In some embodiments, a nano nated with, when used herein to refer to a nanoparticle com particle composition is a uniform collection of nanoparticles. position, is synonymous with “substantially free of and In some embodiments, nanoparticle compositions are disper describes a nanoparticle composition containing no more sions or emulsions. In general, a dispersion or emulsion is than about 50% of the recited material. For example, if a formed when at least two immiscible materials are combined. nanoparticle composition is said to be “substantially free of An “oil-in-water dispersion is one in which oily particles (or particles whose diameter is outside of a stated range, then no hydrophobic or non-polar) are dispersed within an aqueous more than about 50% of the particles in that composition have dispersion medium. A “water-in-oil dispersion is one in diameters outside of the range. In some embodiments, no which aqueous (or hydrophilic or polar) particles are dis more than 25% of the particles are outside of the range. In persed within an oily dispersion medium. Those of ordinary some embodiments, no more than 20%, 19%, 18%, 17%, skill in the art will appreciate that a dispersion can be formed 16%, 15%, 14%, 13%, 12%, 10%, 9%, 8%, 7%, 6%. 5%, 4%, from any two immiscible media and is not limited strictly to 3%, 2%, 1%, 0.5% or less of the particles have diameters combinations of aqueous and oily media. The term "disper outside of the stated range. sion medium therefore applies broadly to any dispersion 0057. Nucleic acid: As used herein, the term “nucleic medium notwithstanding that it is common to refer to “aque acid, in its broadest sense, refers to any compound and/or ous and "oily categories. In some embodiments, nanopar Substance that is or can be incorporated into an oligonucle ticle compositions are nanoemulsions. In some embodi otide chain. In some embodiments, a nucleic acid is a com ments, nanoparticle compositions comprise micelles. In pound and/or substance that is or can be incorporated into an Some embodiments, a nanoparticle composition comprises oligonucleotide chain via a phosphodiester linkage. In some particles such as those described in U.S. Pat. No. 7,763,663, embodiments, “nucleic acid refers to individual nucleic acid issued on Jul. 27, 2010, and entitled “POLYSACCHARIDE residues (e.g., nucleotides and/or nucleosides). In some CONTAINING BLOCK COPOLYMER PARTICLES AND embodiments, “nucleic acid refers to an oligonucleotide USES THEREOF (incorporated herein by reference). In chain comprising individual nucleic acid residues. As used Some embodiments, a nanoparticle composition comprises a herein, the terms "oligonucleotide' and “polynucleotide' can US 2012/0328702 A1 Dec. 27, 2012

be used interchangeably. In some embodiments, “nucleic nanoparticles according to the present invention. In some acid encompasses RNA as well as single and/or double embodiments, a premix contains two or more immiscible stranded DNA and/or cDNA. Furthermore, the terms “nucleic Solvents. In some embodiments, a premix contains compo acid, “DNA “RNA and/or similar terms include nucleic nents that self-assemble into nanoparticles. In some embodi acid analogs, e.g., analogs having other than a phosphodiester ments, a premix contains components that self-assemble into backbone. For example, the so-called “peptide nucleic acids.” micelles. In some embodiments, a premix contains one or which are known in the art and have peptide bonds instead of more amphiphilic entities as described in co-pending PCT phosphodiester bonds in the backbone, are considered within application serial number PCT/US07/86018, filed Nov.30, the scope of the present invention. The term “nucleotide 2007, published as WO 08/070,538 on Jun. 12, 2008, and sequence encoding an amino acid sequence' includes all entitled AMPHIPHILICENTITY NANOPARTICLES. In nucleotide sequences that are degenerate versions of each Some embodiments, a premix contains one or more known other and/or encode the same amino acid sequence. Nucle therapeutic agents and/or independently active biologically otide sequences that encode proteins and/or RNA may active agents. In some embodiments, a premix does not con include introns. Nucleic acids can be purified from natural tain any known therapeutic agents and/or independently Sources, produced using recombinant expression systems and active biologically active agents. In some embodiments, a optionally purified, chemically synthesized, etc. Where premix is agitated, mixed, and/or stirred; in Some embodi appropriate, e.g., in the case of chemically synthesized mol ments, a premix is agitated, mixed, and/or stirred prior to ecules, nucleic acids can comprise nucleoside analogs such as being Subjected to high shear force. In some embodiments, a analogs having chemically modified bases or Sugars, back premix comprises at least one solubilized component (i.e., at bone modifications, etc. A nucleic acid sequence is presented least one component that is in solution); in Some such in the 5' to 3' direction unless otherwise indicated. The term embodiments, the premix is subjected to high shear force “nucleic acid segment' is used hereinto refer to a nucleic acid after such solubilization is achieved. sequence that is a portion of a longer nucleic acid sequence. In 0061 Pure: As used herein, a substance and/or entity is many embodiments, a nucleic acid segment comprises at least “pure' if it is substantially free of other components. For 3, 4, 5, 6,7,8,9, 10, or more residues. In some embodiments, example, a preparation that contains more than about 90% of a nucleic acid is or comprises natural nucleosides (e.g., a particular substance and/or entity is typically considered to adenosine, thymidine, guanosine, cytidine, uridine, deoxyad be a pure preparation. In some embodiments, a Substance enosine, deoxythymidine, deoxyguanosine, and deoxycyti and/or entity is at least 91%, at least 92%, at least 93%, at least dine); nucleoside analogs (e.g., 2-aminoadenosine, 2-thio 94%, at least 95%, at least 96%, at least 97%, at least 98%, or thymidine, inosine, pyrrolo-pyrimidine, 3-methyladenosine, at least 99% pure. 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uri 0062 Refractory: The term “refractory” as used herein, dine, 2-aminoadenosine, C5-bromouridine, C5-fluorouri refers to any subject that does not respond with an expected dine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cy clinical efficacy following the administration of provided tidine, C5-methylcytidine, 2-aminoadenosine, compositions as normally observed by practicing medical 7-deaZaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-OX personnel. oguanosine, O(6)-methylguanine, and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., 0063. Self-administration: The term “self-administra methylated bases); intercalated bases; modified Sugars (e.g., tion.” as used herein, refers to the situation where a subject 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hex has the ability to administer a composition to him or herself ose); and/or modified phosphate groups (e.g., phosphorothio without requiring medical Supervision. In some embodi ates and 5'-N-phosphoramidite linkages). In some embodi ments, self-administration may be performed outside of a ments, the present invention is specifically directed to clinical setting. To give but one example, in Some embodi “unmodified nucleic acids.” meaning nucleic acids (e.g., ments, a facial cosmetic cream may be administered by a polynucleotides and residues, including nucleotides and/or Subject in one’s own home. 0064. Shear force: As used herein, the term “shear force' nucleosides) that have not been chemically modified. refers to a force that is parallel or tangential to the face of a 0058 Patient: As used herein, the term “patient” or “sub material, as opposed to a force that is perpendicular to the face ject” refers to any organism to which a provided composition of a material. In some embodiments, a composition is may be administered, e.g., for experimental, diagnostic, pro exposed to high shear forces in order to produce a uniform phylactic, cosmetic, and/or therapeutic purposes. Typical nanoparticle composition. Any method known in the art can patients include animals (e.g., mammals such as mice, rats, be used to generate high shear forces. In some embodiments, rabbits, non-human primates, and humans). In some embodi cavitation is used to generate high shear forces. In some ments, a patient is a human. embodiments, high pressure homogenization is used to gen 0059 Pharmaceutically acceptable: The term “pharma erate high shear forces. Alternatively or additionally, high ceutically acceptable' as used herein, refers to agents that, shear force may be administered by exposure to high pres within the scope of Sound medical judgment, are Suitable for sure, for example about 15,000 psi. In some embodiments, use in contact with the tissues of human beings and animals such high pressure is within the range of about 18,000 psi to without excessive toxicity, irritation, allergic response, or about 26,000 psi; in some embodiments, it is within the range other problem or complication, commensurate with a reason of about 20,000 psi to about 25,000 psi. In some embodi able benefit/risk ratio. ments, and to give but one example, a Microfluidizer R. Pro 0060 Premix: As used herein, the term “premix' refers to cessor (Microfluidics Corporation/MFIC Corporation) or any combination of components that is Subsequently used to other like device is used to generate high shear force. Microf generate a nanoparticle composition according to the present luidizer R. Processors provide high pressure and a resultant invention. For example, a premix is any collection of ingre high shear rate by accelerating a composition through micro dients that, when Subjected to high shear forces, generates channels (typically having dimensions on the order of 75 US 2012/0328702 A1 Dec. 27, 2012

microns) at a high Velocity (typically in the range of 50 four (24) months, about two (2) weeks to about twelve (12) m/s-300 m/s) for size reduction to the nanoscale range. As the months, about two (2) months to about five (5) months, etc. fluid exits the microchannels it forms jets which collide with For example, if a population of nanoemulsion particles is jets from opposing microchannels. In the channels the fluid Subjected to prolonged storage, temperature changes, and/or experiences high shear (up to 10 1/s) which is orders of pH changes and a majority of the nanoparticles in the com magnitude higher than that of conventional technologies. Jet position maintain a diameter within a stated range (for collisions result in mixing at Submicron levels. Therefore, in example, between approximately 10 nm and approximately Such devices, high shear and/or impact can achieve particle 120 nm), the nanoparticle composition is stable. For some size reduction and mixing of multiphase. In some embodi Such populations, a majority is more than about 50%, about ments, a sample is exposed to high shear forces for a period of 60%, about 70%, about 80%, about 90%, about 95%, about time less than about 10 minutes. In some embodiments, the 96%, about 97%, about 98%, about 99%, about 99.5%, about period of time is less than about 9 minutes, about 8 minutes, 99.6%, about 99.7%, about 99.8%, about 99.9% or more. In about 7 minutes, about 6 minutes, about 5 minutes, about 4 Some embodiments of the invention, where a provided com minutes, about 3 minutes, about 2 minutes, or about 1 minute. position comprises a known therapeutic agent and/oran inde In some embodiments, the period of time is within the range pendently active biologically active agent, the provided com of about 1 minute to about 2 minutes; in some embodiments, position is considered stable if the concentration of the period of time is less than about 1 minute; in some therapeutic agent and/or an independently active biologically embodiments, the period of time is about 30 seconds. In some active agent (e.g., botulinum toxin) is maintained in the com embodiments, a sample is “microfluidized' through a single position over the designated period of time under a designated exposure to high shear forces; such embodiments are referred set of conditions. In some embodiments of the invention, to herein as “single pass' microfluidization. where a provided composition comprises at least one thera 0065. Similarity: As used herein, the term “similarity” peutic agent and/or an independently active biologically refers to the overall relatedness between polymeric mol active agent, the provided composition is considered stable if ecules, e.g., between polynucleotide molecules (e.g., DNA the concentration of therapeutic agent and/or an indepen molecules and/or RNA molecules) and/or between polypep dently active biologically active agent (e.g., botulinum toxin) tide molecules. Calculation of percent similarity of polymeric is maintained in the composition over the designated period molecules to one another can be performed in the same man of time under a designated set of conditions. In some embodi ner as a calculation of percent identity, except that calculation ments of the invention, where a provided composition com of percent similarity takes into account conservative substi prises a therapeutic agent and/or an independently active tutions as is understood in the art. biologically active agent, the provided composition is con 006.6 Small Molecule: In general, a “small molecule' is a sidered stable if the bioavailability of the active agent (e.g., molecule that is less than about 5 kilodaltons (kD) in size. In botulinum toxin) is maintained over the designated period of Some embodiments, the Small molecule is less than about 4 time under a designated set of conditions. As will be appre kD, 3 kD, about 2 kD, or about 1 kD. In some embodiments, ciated by those of skill in the art, bioavailability of a thera the small molecule is less than about 800 daltons (D), about peutic agent and/or an independently active biologically 600 D, about 500 D, about 400 D, about 300 D, about 200 D, active agent in a nanoemulsion, in Some embodiments, may or about 100 D. In some embodiments, a small molecule is reflect the amount or concentration of the agent in active form less than about 2000 g/mol, less than about 1500 g/mol, less (e.g., not degraded or otherwise inactivated); in some than about 1000 g/mol, less than about 800 g/mol, or less than embodiments, bioavailability may be independent of amount about 500 g/mol. In some embodiments, small molecules are or concentration of the active agent. That is, in some embodi non-polymeric. In some embodiments, in accordance with ments, bioavilability may be maintained although amount or the present invention, Small molecules are not proteins, concentration increases (e.g., up to X % of study level) or polypeptides, oligopeptides, peptides, polynucleotides, oli decreases (e.g., down to Y 96 of starting level). gonucleotides, polysaccharides, glycoproteins, proteogly 0068 Substantially: As used herein, the term “substan CanS, etc. tially” refers to the qualitative condition of exhibiting total or 0067 Stable: The term “stable,” when applied to provided near-total extent or degree of a characteristic or property of compositions herein, means that the compositions maintain interest. One of ordinary skill in the biological arts will under one or more aspects of their physical structure (e.g., size stand that biological and chemical phenomena