(12) Patent Application Publication (10) Pub. No.: US 2014/0234271 A1 MILLER Et Al

US 20140234271 A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2014/0234271 A1 MILLER et al. (43) Pub. Date: Aug. 21, 2014

(54) COMPOSITIONS FOR PROLIFERATION OF A61O 19/00 (2006.01) CELLS AND RELATED METHODS A61O 19/08 (2006.01) A68/98 (2006.01) (71) Applicant: The Hospital for Sick Children, A618/365 (2006.01) Toronto (CA) A618/49 (2006.01) A61K 8/64 (2006.01) (72) Inventors: Freda D. MILLER, Toronto (CA); A613 L/4 (2006.01) David KAPLAN, Toronto (CA); A618/4I (2006.01) Kristen SMITH, San Clemente, CA GOIN33/50 (2006.01) (US); Maryline PARIS, Toronto (CA); A6135/36 (2006.01) Sibel NASKA, Toronto (CA) (52) U.S. Cl. (73) Assignee: THE HOSPITAL FOR SICK crimitate E938th, CHILDREN, Toronto (CA) 38/1808 (2013.01); A61 K3I/353 (2013.01): A6 IK3I/5375 (2013.01); A61O 700 (21) Appl. No.: 14/270,021 (2013.01); A61O 19/00 ESO 1-1. (2013.01); A61K 8/981 (2013.01); A61K 8/365 (22) Filed: May 5, 2014 (2013.01); A61K 8/498 (2013.01); A61K 8/49 Related U.S. Application Data (2013.01); A61K 8/64 (2013.01); A61 K3I/14 (2013.01); A61K 8/416 (2013.01); G0IN (63) Continuation of application No. 13/075,647, filed on 33/5044 (2013.01) Mar. 30, 2011, now Pat. No. 8,748,177, which is a USPC ...... 424/93.7: 514/573; 514/456; 514/239.2: continuation-in-part of application No. PCT/US2009/ 435/375; 435/29; 506/10; 435/6.12: 435/6.13; 058723, filed on Sep. 29, 2009. 435/721 (60) Provisional application No. 61/101443, filed on Sep. 30, 2008, provisional application No. 61/426,160, (57) ABSTRACT filed on Dec. 22, 2010, provisional application No. We have discovered that pó3 inhibition results in increased 61/367,780, filed on Jul 26, 2010. cellular proliferation. We have also performed a screen for agents capable of increasing cellular proliferation, (e.g., of Publication Classification stem cells such as skin-derived precursors (SKPs)). The invention therefore invention provides compositions, meth (51) Int. Cl. ods, and kits for increasing proliferation of cells, using com A 6LX3/55.75 (2006.01) pounds that decrease p63 expression or activity or using the A6 IK38/8 (2006.01) compounds described herein. The invention also features A6 IK3I/353 (2006.01) methods of using these compounds for increasing hair A 6LX3/5.375 (2006.01) growth, improving skin health, or promoting skin repair in a A61O 700 (2006.01) Subject. Patent Application Publication Aug. 21, 2014 Sheet 1 of 28 US 2014/0234271 A1

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COMPOSITIONS FOR PROLIFERATION OF embodiments, the compound is a p63 antibody, oran antigen CELLS AND RELATED METHODS binding fragment thereof; an RNAi molecule that decreases p63 expression, or a nucleic acid encoding the RNAi mol CROSS-REFERENCE TO RELATED ecule; or a polypeptide Substantially identical to a dominant APPLICATIONS negative form of p63, a fragment thereof, or a nucleic acid 0001. This application is a continuation of U.S. applica encoding such a polypeptide, where the polypeptide or the tion Ser. No. 13/075,647, filed Mar. 30, 2011, which is a fragment has dominant negative p63 activity. In particular continuation-in-part of International Application No. PCT/ embodiments, the compounds are (a) alprostadil and US2009/058723, filed Sep. 29, 2009, which, in turn, claims kaempferol or (b) alprostadil and pramoxine. In embodi the benefit of U.S. Provisional Application No. 61/101,443, ments where the cell is cultured in vitro, the culture may filed Sep. 30, 2008. Application Ser. No. 13/075,647 also include at least one additional growth factor (e.g., FGF2, claims the benefit of U.S. Provisional Application Nos. EGF, NGF, or a combination thereof). The method may 61/367,780, filed Jul. 26, 2010, and 61/426,160, filed Dec. 22, involve no substantial change in the differentiation rate of the 2010. Each of these applications is hereby incorporated by cells. reference. 0008. In another aspect, the invention features a composi tion including (a) an isolated cell (e.g., a stem cell Such as a BACKGROUND OF THE INVENTION SKP, or any cell described herein) and (b) one or more com pounds selected from the group consisting of acyclovir, 0002 The invention relates to compositions and methods alprostadil, aristolochic acid, carbadox, chlorpyrifos, cyclo useful in the proliferation of cells, particularly stem cells such creatine, 7,4'-dimethoxyisoflavone, dorzolamide, S(-)eticlo as skin-derived precursors (SKPs). Also provided are meth pride, evoxine, guaifenesin, hydroxyprogesterone, ods for treating a subject having a disease or condition where kaempferol, linamarin, mexicanolide, MG 624, pramoxine, an increase in SKP proliferation is desired. tannic acid, targinine, alpha-methyllycacontine, mecamy 0003 Expensive growth factors are often required for cell lamine hexamethonium, alsterpaullone, and yohimbic acid, proliferation, and even then, expansion is often not optimal. or an analog thereof, where the compound is present in an Thus, molecules which replace or enhance the actions of amount sufficient to promote proliferation of the stem cell. In growth factors and allow increased expansion of cells in particular embodiments, the compounds are (a) alprostadil culture are desirable. and kaempferol or (b) alprostadil and pramoxine. 0004. In addition, there are inadequate methods for regen 0009. In another aspect, the invention features a composi erating skin or inducing hair growth in a Subject (e.g., for tion including (a) an isolated cell (e.g., a stem cell Such as a treatment of a disease or condition where regenerating skin or SKP, or any cell described herein) and (b) one or more com inducing hair growth is beneficial). pounds that decreases p63 expression or activity, where the 0005 Thus, there is a need for molecules that promote the compound is present in an amount Sufficient to promote pro proliferation and self-renewal of cells such as SKPs. These liferation of the cell. In particular embodiments, the com molecules may be highly advantageous for cosmetic and pounds are (a) alprostadil and kaempferol or (b) alprostadil medical purposes. and pramoxine. 0010. In either of the above two aspects, the composition SUMMARY OF THE INVENTION may further include a growth factor (e.g., FGF2, EGF. NGF, 0006 We have discovered that p63 inhibits proliferation or a combination thereof). The composition may be capable of skin-derived precursors (SKPs) and, further have screened of Supporting cell proliferation. for and identified compounds that increase proliferation of 0011. In another aspect, the invention features a kit includ SKPs and neuroblastoma cells. On the basis of these discov ing (a) one or more compounds that decreases p63 expression eries, the invention features methods and compositions useful or activity (e.g., any described herein) or one or more com for increasing cell proliferation, as well as methods for pounds selected from the group consisting of acyclovir, increasing SKP proliferation in Subject (e.g., for treating dis alprostadil, aristolochic acid, carbadox, chlorpyrifos, cyclo eases and conditions where increased SKP proliferation is creatine, 7,4'-dimethoxyisoflavone, dorzolamide, S(-)eticlo desired). pride, evoxine, guaifenesin, hydroxyprogesterone, 0007 Accordingly, in a first aspect, the invention features kaempferol, linamarin, mexicanolide, MG 624, pramoxine, a method of increasing proliferation of a cell (e.g., cultured in tannic acid, targinine, alpha-methyllycacontine, mecamy vitro). The method includes contacting the cell with a suffi lamine hexamethonium, alsterpaullone, and yohimbic acid, cient amount of one or more of a compound that decreases or an analog thereof, and (b) instructions for use of (a) to (e.g., selectively decreases) p63 expression or activity or a promote cell proliferation, hair growth, skin repair, or skin compound selected from the group consisting of acyclovir, health. In particular embodiments, the compounds are (a) alprostadil, aristolochic acid, carbadox, chlorpyrifos, cyclo alprostadil and kaempferol or (b) alprostadil and pramoxine. creatine, 7,4'-dimethoxyisoflavone, dorzolamide, S(-)eticlo 0012. In another aspect, the invention features a composi pride, evoxine, guaifenesin, hydroxyprogesterone, tion including (a) one or more compounds that decreases p63 kaempferol, linamarin, mexicanolide, MG 624, pramoxine, expression or activity (e.g., any described herein) or one or tannic acid, targinine, alpha-methyllycacontine, mecamy more compounds selected from the group consisting of acy lamine hexamethonium, alsterpaullone, and yohimbic acid, clovir, alprostadil, aristolochic acid, carbadox, chlorpyrifos, or an analog thereof, under conditions that Support cell pro cyclocreatine, 7,4'-dimethoxyisoflavone, dorzolamide, S(-) liferation. The cell may be from a tumor cell line, may be a eticlopride, evoxine, guaifenesin, hydroxyprogesterone, stem cell (e.g., a SKP), or any cell described hereincapable of kaempferol, linamarin, mexicanolide, MG 624, pramoxine, proliferation. The cell may be a human or non-human cell tannic acid, targinine, and yohimbic acid, or an analog (e.g., from a mammal Such as a mouse or rat). In certain thereof, and (b) a topically Suitable excipient. The composi US 2014/0234271 A1 Aug. 21, 2014

tion may be in the form of a cream or a lotion (e.g., a mois may be administered topically, systemically, or by any route turizing lotion). In particular embodiments, the compounds described herein. In certain embodiments, the compound that are (a) alprostadil and kaempferol or (b) alprostadil and inhibits p03 expression or activity is a p63 antibody or an pramoxine. antigen-binding fragment thereof an RNAi molecule that 0013. In another aspect, the invention features a method of inhibits p03 expression or a nucleic acid encoding the RNAi increasing SKP proliferation in a subject. The method molecule; or a polypeptide Substantially identical to a domi includes administering to the Subject a Sufficient amount of nant negative form of p63, a fragment thereof, or a nucleic (a) one or more compounds that inhibits p63 expression or acid encoding Such a polypeptide, where the polypeptide or activity; or (b) one or more compounds selected from the the fragment has dominant negative p63 activity. In particular group consisting of acyclovir, alprostadil, aristolochic acid, embodiments, the compounds are (a) alprostadil and carbadox, chlorpyrifos, cyclocreatine, 7,4'-dimethoxyisofla kaempferol or (b) alprostadil and pramoxine. vone, dorzolamide, S(-) eticlopride, evoxine, guaifenesin, 0016. In another aspect, the invention features a method of hydroxyprogesterone, kaempferol, linamarin, mexicanolide, improving skin health in a Subject for example, by reducing MG 624, pramoxine, tannic acid, targinine, alpha-methylly skin aging (or the appearance of aging) or reducing wrinkles. cacontine, mecamylamine hexamethonium, alsterpaullone, The method includes administering to the subject a sufficient and yohimbic acid, or an analog thereof. The compound can amount of (a) one or more compounds that inhibits p63 be administered topically, systemically, or by any route expression or activity or (b) one or more compounds selected described herein. In certain embodiments, the compound that from the group consisting of acyclovir, alprostadil, aris inhibits p63 expression or activity is a pé3 antibody or an tolochic acid, carbadox, chlorpyrifos, cyclocreatine, 7,4'- antigen-binding fragment thereof an RNAi molecule that dimethoxyisoflavone, dorzolamide, S(-)eticlopride, evoxine, inhibits p03 expression or a nucleic acid encoding the RNAi guaifenesin, hydroxyprogesterone, kaempferol, linamarin, molecule; or a polypeptide Substantially identical to a domi mexicanolide, MG 624, pramoxine, tannic acid, targinine, nant negative form of p63, a fragment thereof, or a nucleic alpha-methylycacontine, mecamylamine hexamethonium, acid encoding Such a polypeptide, where the polypeptide or alsterpaullone, and yohimbic acid, or an analog thereof. The the fragment has dominant negative p63 activity. In particular compound may be administered topically, systemically, or by embodiments, the compounds are (a) alprostadil and any route described herein. In certain embodiments, the com kaempferol or (b) alprostadil and pramoxine. pound that inhibits p03 expression or activity is a po3 anti 0014. In another aspect, the invention features a method of body or an antigen-binding fragment thereof, an RNAi mol promoting hair growth in a subject. The method includes ecule that inhibits p03 expression or a nucleic acid encoding administering to the Subject a Sufficient amount of (a) one or the RNAi molecule; or a polypeptide substantially identical to more compounds that inhibits p63 expression or activity or a dominant negative form of p63, a fragment thereof, or a (b) one or more compounds selected from the group consist nucleic acid encoding the polypeptide, where the polypeptide ing of acyclovir, alprostadil, aristolochic acid, carbadox, or the fragment has dominant negative p63 activity. In par chlorpyrifos, cyclocreatine, 7,4'-dimethoxyisoflavone, dor ticular embodiments, the compounds are (a) alprostadil and Zolamide, S(-)eticlopride, evoxine, guaifenesin, hydrox kaempferol or (b) alprostadil and pramoxine. yprogesterone, kaempferol, linamarin, mexicanolide, MG 0017. In any of the above methods, the subject may be a 624, pramoxine, tannic acid, targinine, alpha-methyllycacon human. tine, mecamylamine hexamethonium, alsterpaullone, and 0018. In another aspect, the invention features a method of yohimbic acid, or an analog thereof. The compound may be identifying a compound capable of altering cell proliferation, administered topically, systemically, or by any route the method including (a) contacting (e.g., in vitro) a SKP described herein. In certain embodiments, the compound that (e.g., human or non-human SKP. Such as a mouse or rat SKP) inhibits p63 expression or activity may be a pé3 antibody or with a candidate compound, and (b) determining the prolif an antigen-binding fragment thereof an RNAi molecule that eration rate of the SKP, where an alteration in the proliferation inhibits p03 expression or a nucleic acid encoding the RNAi rate of the SKP in the presence of the compound as compared molecule; or a polypeptide Substantially identical to a domi to in the absence of the compound, indicates that the com nant negative form of p63, a fragment thereof, or a nucleic pound alters the rate of cell proliferation. In certain embodi acid encoding the polypeptide, where the polypeptide or the ments, the compound is selected from a chemical library. fragment has dominant negative p63 activity. In particular 0019. In embodiments of the above aspects of the inven embodiments, the compounds are (a) alprostadil and tion where the Subject is administered acyclovir or analog kaempferol or (b) alprostadil and pramoxine. thereof, the subject may not suffer from, or be diagnosed with, 0.015. In another embodiment, the invention features a herpes (e.g., genital herpes) or chickenpox. In embodiments method of repairing skin in a Subject, the method including where the Subject is administered alprostadil or an analog administering a Sufficient amount of (a) one or more com thereof, the subject may not suffer from, or be diagnosed with pounds that inhibits p63 expression or activity or (b) one or erectile dysfunction or be in need of weight loss (e.g., be more compounds selected from the group consisting of acy overweight or obese). In embodiments where the subject is clovir, alprostadil, aristolochic acid, carbadox, chlorpyrifos, administered aristolochic acid or an analog thereof, the Sub cyclocreatine, 7,4'-dimethoxyisoflavone, dorzolamide, S(-) ject may not be in need of weight loss (e.g., be overweight or eticlopride, evoxine, guaifenesin, hydroxyprogesterone, obese). In embodiments where the subject is administered kaempferol, linamarin, mexicanolide, MG 624, pramoxine, dorzolamide or an analog thereof, the Subject may not have, tannic acid, targinine, alpha-methyllycacontine, mecamy or may not be diagnosed with, increased intraocular pressure lamine hexamethonium, alsterpaullone, and yohimbic acid, (e.g., ocular hypertension or open-angle glaucoma). In or an analog thereof. In certain embodiments, the Subject has embodiments where the Subject is administered guaifenesin a wound, and the compound is administered in an amount or an analog thereof, the Subject may not be in need of an Sufficient to improve healing of the wound. The compound expectorant (e.g., Suffering from a cold, an allergy, or airway US 2014/0234271 A1 Aug. 21, 2014

infection). In embodiments where the subject is administered 16 amino acids, preferably at least 20 amino acids, more hydroxyprogesterone, the Subject may not be suffering from, preferably at least 25 amino acids, and most preferably at or may not be diagnosed with, congenital adrenal hyperpla least 35 amino acids. For nucleic acid molecules, the length of sia, 21-hydroxylase deficiency, or breast neoplasms, or is not comparison sequences is at least 50 nucleotides, preferably at at risk of having a preterm birth. In embodiments where the least 60 nucleotides, more preferably at least 75 nucleotides, Subject is administered pramoxine, the Subject may not be in and most preferably at least 110 nucleotides. need of a topical anesthetic (e.g., due to itching, burning, or 0028. The term “antibody' is used in the broadest sense other pain). In embodiments where the Subject is adminis and specifically covers, for example, single monoclonal anti tered yohimbic acid, the subject may not be suffering from bodies against p63, antibody compositions with polyepitopic erectile dysfunction, panic disorder, alcoholism, or depres specificity, single chain antibodies, nanobodies, and frag S1O. ments of antibodies. Antibody' includes intact immunoglo 0020. In any of the above aspects, the SKP may express bulin orantibody molecules, polyclonal antibodies, multispe any one, two, three, four, five, or more of the markers for cific antibodies (e.g., bispecific antibodies formed from at SKPs described herein, or may not express any one, two three, least two intact antibodies), and immunoglobulin fragments four, five, or more of the markers not expressed by SKPs. In (such as Fab, F(ab'), or FV), so long as they exhibit any of the any of the above aspects, the analogs of the compounds may desired properties (e.g., antigen binding) described herein. be any analog described herein. 0029) “Antibody fragments’ comprise a portion of an 0021. By “decreasing expression of a gene or protein is intact antibody, generally the antigen binding or variable meant reducing (e.g., by 5%, 10%, 25%, 50%, 75%, 90%, region of the intactantibody. Examples of antibody fragments 95%, 99%, or 99.9%) in the amount the gene or protein include Fab, Fab'. F(ab')2, and Fv fragments, diabodies, produced. Decreased expression may occur, for example, by single chain antibody molecules, and multispecificantibodies a reduction in transcription, translation, or mRNA process formed from antibody fragments. 1ng. 0030 "Humanized forms of non-human (e.g., murine) 0022. By “decreased activity” is meant a reduction (e.g., antibodies are specific chimeric immunoglobulins, immuno by 5%, 10%, 25%, 50%, 75%, 90%, 95%, 99%, or 99.9%) by globulin chains, or fragments thereof (such as Fv, Fab, Fab', in the total activity of a protein in a cell. Reduction of activity F(ab') or other antigen-binding subsequences of antibodies) may result, for example, from direct inhibition of the protein which contain minimal sequence derived from non-human (e.g., by a compound that specifically binds the protein), immunoglobulin. increased degradation or processing, or decreased expression 0031. A “human antibody is one which possesses an of the protein. amino acid sequence which corresponds to that of an anti 0023. By a compound or composition that “selectively body produced by a human and/or has been made using any of inhibits’ a target protein (e.g., p63) is meant a compound or the techniques for making human antibodies known in the art. composition that decreases expression or activity of the target A "human antibody includes antibodies comprising at least protein and (a) binds specifically to the target protein (e.g., a one human heavy chain polypeptide or at least one human Small molecule or an antibody) and decreases its activity, (b) light chain polypeptide. binds specifically to an mRNA encoding the target protein, 0032. By “small molecule' is meant a molecule having a thereby decreasing expression of the protein, or (c) prevents molecular weight of less than about 1000 Da (e.g., less than the target protein from performing its normal function (e.g., 900, 800, 700, 600, 500, or 400 Da). by binding to a binding partner of the target protein). 0033 Compounds useful in the invention include those 0024. A compound which “specifically binds a target described herein in any of their pharmaceutically acceptable molecule is a compound which recognizes and binds the forms, including isomers such as diastereomers and enanti target, but which does not Substantially recognize and bind omers, salts, esters, amides, thioesters, Solvates, and poly other molecules. morphs thereof, as well as racemic mixtures and pure isomers 0025 By “subject' is meant a human or non-human ani of the compounds described herein. As an example, by mal (e.g., a mammal). “alprostadil is meant the free base as well as any pharma 0026. By a cell which does “not express' a protein or gene ceutically acceptable salt thereof. is meant that expression of the protein or gene cannot be 0034. The term “pharmaceutically acceptable salt' repre detected by standard methods. In the case of cell surface sents those salts which are, within the scope of sound medical markers, expression can be measured by flow cytometry, judgment, Suitable for use in contact with the tissues of using a cut-off value as obtained from negative controls (i.e., humans and lower animals without undue toxicity, irritation, cells known to lack the antigen of interest) or by isotype allergic response and the like, and are commensurate with a controls (i.e., measuring non-specific binding of the antibody reasonable benefit/risk ratio. Pharmaceutically acceptable to the cell). Thus, a cell that “does not express a marker salts are well known in the art. The salts can be prepared in appears similar to the negative control for that marker. For situ during the final isolation and purification of the com gene expression, a gene "does not express' if the presence of pounds of the invention, or separately by reacting the free its mRNA cannot be visually detected on a standard agarose base function with a suitable organic acid. Representative gel following standard PCR protocols. acid addition salts include acetate, adipate, alginate, ascor 0027. A nucleic acid molecule or polypeptide is said to be bate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, “substantially identical to a reference molecule if it exhibits, butyrate, camphorate, campherSulfonate, citrate, cyclopen over its entire length, at least 50% or 55% identity, preferably tanepropionate, digluconate, dodecylsulfate, ethane at least 60%. 65%, or 70% identity, more preferably at least Sulfonate, fumarate, glucoheptonate, glycerophosphate, 75% or 85% identity, and most preferably at least 90%, 95%, hemisulfate, heptonate, hexanoate, hydrobromide, hydro or 99% identity to the sequence of the reference molecule. For chloride, hydroiodide, 2-hydroxy-ethanesulfonate, isethion polypeptides, the length of comparison sequences is at least ate, lactobionate, lactate, laurate, lauryl Sulfate, malate, male US 2014/0234271 A1 Aug. 21, 2014

ate, malonate, mesylate, methanesulfonate, C. alkynyls include, without limitation, ethynyl, 1-propy 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, nyl, 2-propynyl, 1-butynyl, 2-butynyl, and 3-butynyl. palmitate, pamoate, pectinate, persulfate, 3-phenylpropi onate, phosphate, picrate, pivalate, propionate, Stearate, suc 0040. By “C. heterocyclyl is meant a stable 5- to cinate, Sulfate, tartrate, thiocyanate, toluenesulfonate, unde 7-membered monocyclic or 7- to 14-membered bicyclic het canoate, Valerate salts, and the like. Representative alkali or erocyclic ring which is saturated, partially unsaturated, or alkaline earth metal salts include Sodium, lithium, potassium, unsaturated (aromatic), and which consists of 2 to 6 carbon calcium, magnesium, and the like, as well as nontoxic ammo atoms and 1, 2, 3, or 4 heteroatoms independently selected nium, quaternary ammonium, and amine cations, including, from N, O, and S and including any bicyclic group in which but not limited to ammonium, tetramethylammonium, tetra any of the above-defined heterocyclic rings is fused to a ethylammonium, methylamine, dimethylamine, trimethy benzene ring. The heterocyclyl group may be substituted or lamine, triethylamine, ethylamine, and the like. unsubstituted. Exemplary Substituents include alkoxy, ary 0035. In the generic descriptions of compounds of this loxy, sulfhydryl, alkylthio, arylthio, halide, hydroxy, fluoro invention, the number of atoms of a particular type in a alkyl, perfluoralkyl, amino, aminoalkyl, disubstituted amino, Substituent group is generally given as a range, e.g., an alkyl quaternary amino, hydroxyalkyl, carboxyalkyl, and carboxyl group containing from 1 to 4 carbon atoms or C alkyl. groups. The nitrogen and Sulfur heteroatoms may optionally Reference to such a range is intended to include specific be oxidized. The heterocyclic ring may be covalently attached references to groups having each of the integer number of via any heteroatom or carbon atom which results in a stable atoms within the specified range. For example, an alkyl group structure, e.g., an imidazolinyl ring may be linked at either of from 1 to 4 carbon atoms includes each of C, C, C, and C. the ring-carbon atom positions or at the nitrogen atom. A AC heteroalkyl, for example, includes from 1 to 12 car nitrogen atom in the heterocycle may optionally be quater bon atoms in addition to one or more heteroatoms. Other nized. Preferably when the total number of S and O atoms in numbers of atoms and other types of atoms may be indicated the heterocycle exceeds 1, then these heteroatoms are not in a similar manner. adjacent to one another. Heterocycles include, without limi 0036. As used herein, the terms “alkyl and the prefix tation, 1H-indazole, 2-pyrrolidonyl, 2H,6H-1.5.2-dithiazi “alk- are inclusive of both straight chain and branched chain nyl, 2H-pyrrolyl, 3H-indolyl, 4-piperidonyl, 4aH-carbazole, groups and of cyclic groups, i.e., cycloalkyl. Cyclic groups 4H-quinolizinyl, 6H-1,2,5-thiadiazinyl, acridinyl, azocinyl, can be monocyclic or polycyclic and preferably have from 3 benzimidazolyl, benzofuranyl, benzothiofuranyl, ben to 12 ring carbon atoms, inclusive. Exemplary cyclic groups Zothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimi groups. daZalonyl, carbazolyl, 4aH-carbazolyl b-carbolinyl, chroma 0037. By "C alkyl is meant a branched or unbranched nyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1.5. hydrocarbon group having from 1 to 4 carbon atoms. A Ca 2-dithiazinyl, dihydrofuroI2,3-btetrahydrofuran, furanyl, alkyl group may be substituted or unsubstituted. Exemplary furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-in Substituents include alkoxy, aryloxy, Sulfhydryl, alkylthio. dazolyl, indolenyl, indolinyl, indolizinyl, indolyl, isobenzo arylthio, halide, hydroxyl, fluoroalkyl, perfluoralkyl, amino, furanyl, isochromanyl, isolindazolyl, isoindolinyl, isoindolyl, aminoalkyl, disubstituted amino, quaternary amino, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl, naph hydroxyalkyl, carboxyalkyl, and carboxyl groups. Calkyls thyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1.2.3-Oxa include, without limitation, methyl, ethyl, n-propyl, isopro diazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadia pyl, cyclopropyl, cyclopropylmethyl, n-butyl, iso-butyl, sec Zolyl, oxazolidinyl, oxazolyl, oxazolidinylperimidinyl, butyl, tert-butyl, and cyclobutyl. phenanthridinyl, phenanthrolinyl, phenarsazinyl, phenazinyl, 0038. By “C. alkenyl is meant a branched or phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, unbranched hydrocarbon group containing one or more piperazinyl, piperidinyl, pteridinyl, piperidonyl, 4-piperido double bonds and having from 2 to 4 carbon atoms. A Ca nyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, alkenyl may optionally include monocyclic or polycyclic pyrazolinyl, pyrazolyl pyridazinyl, pyridooxazole, pyri rings, in which each ring desirably has from three to six doimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, members. The C- alkenyl group may be substituted or pyrrolidinyl, pyrrolinyl, pyrrolyl, quinazolinyl, quinolinyl, unsubstituted. Exemplary Substituents include alkoxy, ary 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, carbolinyl, tet loxy, sulfhydryl, alkylthio, arylthio, halide, hydroxyl, fluoro rahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinoli alkyl, perfluoralkyl, amino, aminoalkyl, disubstituted amino, nyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadia quaternary amino, hydroxyalkyl, carboxyalkyl, and carboxyl Zolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, groups. Calkenyls include, without limitation, vinyl, allyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl, thienoimi 2-cyclopropyl-1-ethenyl, 1-propenyl, 1-butenyl, 2-butenyl, dazolyl, thiophenyl, triazinyl, 1.2.3-triazolyl, 1,2,4-triazolyl, 3-butenyl, 2-methyl-1-propenyl, and 2-methyl-2-propenyl. 1,2,5-triazolyl, 1,3,4-triazolyl, and xanthenyl. Preferred 5 to 0039. By “C. alkynyl is meant a branched or 10 membered heterocycles include, but are not limited to, unbranched hydrocarbon group containing one or more triple pyridinyl, pyrimidinyl, triazinyl, furanyl, thienyl, thiazolyl, bonds and having from 2 to 4 carbon atoms. A C- alkynyl pyrrolyl pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, tetra may optionally include monocyclic, bicyclic, or tricyclic Zolyl, benzofuranyl, benzothiofuranyl, indolyl, benzimida rings, in which each ring desirably has five or six members. Zolyl, 1H-indazolyl, oxazolidinyl, isoxazolidinyl, benzotria The C alkynyl group may be substituted or unsubstituted. Zolyl, benzisoxazolyl, OXindolylbenzoxazolinyl, quinolinyl, Exemplary Substituents include alkoxy, aryloxy, Sulfhydryl, and isoquinolinyl. Preferred 5 to 6 membered heterocycles alkylthio, arylthio, halide, hydroxy, fluoroalkyl, perfluo include, without limitation, pyridinyl, pyrimidinyl, triazinyl, ralkyl, amino, aminoalkyl, disubstituted amino, quaternary furanyl, thienyl, thiazolyl pyrrolyl, piperazinyl, piperidinyl, amino, hydroxyalkyl, carboxyalkyl, and carboxyl groups. pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, and tetrazolyl. US 2014/0234271 A1 Aug. 21, 2014

0041. By "Caryl' is meant an aromatic group having a R", R", and R" are each independently an alkyl, alkenyl, ring system comprised of carbon atoms with conjugated at alkynyl, or aryl group. R may be an alkyl group linking the electrons (e.g., phenyl). The aryl group has from 6 to 12 quaternary amino nitrogen atom, as a Substituent, to another carbon atoms. Aryl groups may optionally include monocy moiety. The nitrogen atom, N, is covalently attached to four clic, bicyclic, or tricyclic rings, in which each ring desirably carbon atoms of alkyl, heteroalkyl, heteroaryl, and/or aryl has five or six members. The aryl group may be substituted or groups, resulting in a positive charge at the nitrogen atom. unsubstituted. Exemplary substituents include alkyl, 0055. Other features and advantages of the invention will hydroxy, alkoxy, aryloxy, Sulfhydryl, alkylthio, arylthio. be apparent from the following Detailed Description, the halide, fluoroalkyl, carboxyl, hydroxyalkyl, carboxyalkyl, drawings, and the claims. amino, aminoalkyl, monosubstituted amino, disubstituted amino, and quaternary amino groups. BRIEF DESCRIPTION OF THE DRAWINGS 0042. By “C, alkaryl is meant an alkyl substituted by an aryl group (e.g., benzyl, phenethyl, or 3,4-dichlorophen 0056 FIG. 1A is a photograph of a gel showing that wild ethyl) having from 7 to 14 carbon atoms. type SKPs express TAp63 mRNA, whereas SKPs from p63 0043. By “Coalkheterocyclyl is meant an alkyl substi null mice do not. tuted heterocyclic group having from 3 to 10 carbon atoms in 0057 FIG. 1B is photograph of a gel showing that SKPs addition to one or more heteroatoms (e.g., 3-furanylmethyl, from both wild-type and p63 null mice express p53, but do not 2-furanylmethyl, 3-tetrahydrofuranylmethyl, or 2-tetrahy express p73. GAPDH is shown as a control. drofuranylmethyl). 0.058 FIG. 1C is a set of photomicrographs showing that 0044. By “C, heteroalkyl is meant a branched or p63 is expressed in wild-type SKPs, but not in SKPs from p63 unbranched alkyl, alkenyl, or alkynyl group having from 1 to null mice (top panels). Hoescht stained cells are shown in the 7 carbon atoms in addition to 1, 2, 3, or 4 heteroatoms inde corresponding lower panels. pendently selected from the group consisting of N, O, S, and 0059 FIG. 1D is a set of photomicrographs showing that P. Heteroalkyls include, without limitation, tertiary amines, both SKPs from both wild-type and p63 null mice express the secondary amines, ethers, thioethers, amides, thioamides, SKP markers fibronectin, nestin, vimentin, and SMA. carbamates, thiocarbamates, hydrazones, imines, phosphodi 0060 FIG. 1E is a set of photomicrographs showing that esters, phosphoramidates, Sulfonamides, and disulfides. A the proliferation marker Ki67 is expressed at higher levels in heteroalkyl may optionally include monocyclic, bicyclic, or SKPs from p63 null mice than in wild-type SKPs. Results tricyclic rings, in which each ring desirably has three to six using whisker and back cells from neonatal mice, as well as members. The heteroalkyl group may be substituted or using whisker cells from one-month-old mice, are shown. unsubstituted. Exemplary Substituents include alkoxy, ary 0061 FIG. 1F is graph showing increasing Ki67 expres loxy, sulfhydryl, alkylthio, arylthio, halide, hydroxyl, fluoro sion in p63 null mice as compared to wild-type mice in SKPs alkyl, perfluoralkyl, amino, aminoalkyl, disubstituted amino, taken from E18, P1, and P30 mice. quaternary amino, hydroxyalkyl, hydroxyalkyl, carboxy 0062 FIG.1G is a graph showing that newborn TAp63-/- alkyl, and carboxyl groups. Examples of C, heteroalkyls and E18 pé3-/- SKPs self-renewed approximately 4 times include, without limitation, methoxymethyl and ethoxyethyl. more robustly than did wild-type SKPs and SKPs from one 0045. By “halide' or “halogen' is meant bromine, chlo month-old mice proliferated and self-renewed about 1.5 fold rine, iodine, or fluorine. more than wild-type SKPs. 0046 By “fluoroalkyl is meant an alkyl group that is 0063 FIG. 1H is a photograph of a gel showing that the substituted with a fluorine atom. p57 mRNA expression is reduced in SKPs from p63 null mice 0047. By “perfluoroalkyl is meant an alkyl group consist as compare to those from wild-type mice. ing of only carbon and fluorine atoms. 0064 FIG. 1I is a set of photomicrographs showing p57 0048. By “carboxyalkyl is meanta chemical moiety with expression in wild-type, p63-/-, and TAp63-/- cells. the formula—(R)—COOH, wherein R is selected from C, 0065 FIG. 1J is a photograph of a gel showing that, using alkyl, C-7 alkenyl, C-7 alkynyl, Cheterocyclyl, Caryl, RT-PCR, p63 is was bound to a previously defined binding C- alkaryl, Coalkheterocyclyl, or C, heteroalkyl. site in the p57 promoter. 0049. By “hydroxyalkyl is meant a chemical moiety with 0.066 FIG. 1K is a graph showing the percentage of p57 the formula —(R)—OH, wherein R is selected from C, positive cells in wild-type and in Tap63-/- cells. alkyl, C2-7 alkenyl, C2-7 alkynyl, C-heterocyclyl, C-2 aryl, 0067 FIG. 1L is a set of photomicrographs showing Cza alkaryl, Coalkheterocyclyl, or C, heteroalkyl. immunostaining for p57Kip2 and Ki67 in TAp63-/- SKPs. 0050. By “alkoxy” is meant a chemical substituent of the plated on an adherent 4-well chamber slides, two days after formula —OR, wherein R is selected from C, alkyl, C-7 transfection with or without a pCMV-p57Kip2 expression alkenyl, C2-7 alkynyl, C2-a heterocyclyl, C-2 aryl, C7-14 vector. Cells were counterstained with Hoechst33258. Trans alkaryl, Coalkheterocyclyl, or C-7 heteroalkyl. fected p57-positive cells were never double-labelled for 0051. By “aryloxy' is meant a chemical substituent of the Ki67. The images are taken at 200x magnification with n=3 formula —OR, wherein R is a C-2 aryl group. independent experiments. 0052 By “alkylthio’ is meant a chemical substituent of 0068 FIG. 2A is a set of photomicrographs showing that the formula—SR, wherein R is selected from C, alkyl, C-7 wild-type and p63-/- SKPs are capable of differentiating to alkenyl, C2-7 alkynyl, C2-a heterocyclyl, C-2 aryl, C7-14 neural and mesoderm Subtypes. Left, neurofilament medium alkaryl, Coalkheterocyclyl, or C, heteroalkyl. (NFM) expression; center, smooth muscle actin (SMA) 0053. By “arylthio’ is meant a chemical substituent of the expression: right, glial fibrillary acidic protein (GFAP) and formula—SR, wherein R is a Caryl group. S100B expression. 0054 By “quaternary amino” is meant a chemical sub 0069 FIG. 2B is a set of photomicrographs showing that stituent of the formula—(R) N(R')(R") (R")", wherein R, both wild-type and p63-/- SKPs, when transplanted into the US 2014/0234271 A1 Aug. 21, 2014

neural crest migratory stream of HH stage 18 chicks in ovo, I0084 FIG. 15 is a series of graphs summarizing in vitro migrated into neural crest targets. sphere assay data demonstrating that certain compounds pro 0070 FIG. 3 is a schematic diagram of an exemplary mote self-renewal of both human and mouse SKPs. screening procedure described herein. I0085 FIG.16 is a table showing that topical application of 0071 FIG. 4A shows a schematic diagram of the sphere certain compounds promotes hair growth. Mice were treated formation assay employed to confirm the hits identified in the topically with the indicated compounds. At specific time SCC. points, approximately 30-40 hairs were plucked from each 0072 FIG. 4B is a graph showing the effects of increasing mouse and their length was measured. Three mice were used concentrations of rapamycin on sphere formation in various to test each compound. cell lines. I0086 FIGS. 17 and 18 are each a series of graphs showing quantification of hair length on day 23. Hair length promoted 0073 FIG. 4C is a graph showing inhibition of tumor by compounds is shown in distribution histograms relative to formation by rapamycin and vinblastine. the control group. Hair length was binned in classes of 200um 0074 FIG. 5 is a graph showing the effects of MG 624 on and each class was expressed as a percentage of total hair SKPs (left) and neuroblastoma (right) proliferation at various population. The distribution histograms show a shift versus concentrations. At 200 nM, SKP proliferation was increased. longer hair length, particularly in the groups treated with 0075 FIGS. 6A-6E are graphs showing dose-response alprostadil or kaempferol. curves for proliferation of human SKPs in response to I0087 FIG. 19 is a series of photomicrographs and graphs MG624 (FIG. 6A), GSK3B (FIG. 6B), pramoxine (FIG. 6C), showing that topical application of certain compounds kaempferol (FIG. 6D), and alprostadil (FIG. 6E) in presence induces dermal thickness. Dorsal skin of animals topically of EGF and FGF2. treated with various compounds was isolated, cryosectioned, 0076 FIG. 7 is a schematic illustration of the in vitro and stained with hematoxylin and eosin. Dermal thickness sphere assay. SKPs were isolated from mouse back, whisker was manually measured randomly through the slices. A mean and human foreskin and allowed to grow in spheres. After 2-3 of three measurements through the slice were performed and passages, spheres were dissociated and 3000 cells were plated about 20-30 slices were analyzed for each mouse within each in each well of a 96-well plate. Cells were then treated in compound group. triplicate with different concentrations of drugs or only with I0088 FIG. 20 is a series of photomicrographs and graphs vehicle DMSO. Drugs were applied again on day 3 and the showing topical application of certain compounds induces number of spheres was analyzed on day 7. anagenhair cycle and follicle density. Skin Samples were also 0077 FIG. 8 is a series of graphs showing that various analysed for density and morphology of hair follicles. The compounds promote human SKPs self-renewal. Graphs show density is expressed as number of follicles per mm whereas pooled data from two set of experiments. Four different cell the anagen hair follicles are determined by morphology. lines were used. I0089 FIG. 21 is a series of graphs showing the effect of 0078 FIG.9 is a graph showing that various compounds at certain compounds and compound combinations on human 100 nM promote human SKPs self-renewal. Pooled data from SKP sphere size in vitro. The first graph (left, top) shows two different experiments show robustincrease of the number mean sphere size following treatment with the indicated com of spheres formed after drug treatments at 100 nM. Results pounds, compound combinations, or a DMSO control. The indicate that these specific drugs promote the ability of stem other two graphs are histograms showing the shift in human cells to produce another stem cell. SKP sphere size when treated by alprostadil and pramoxine 007.9 FIG. 10 is a series of photomicrographs showing (right, top) or alprostadil and kaempferol (left, bottom). Alp. that alprostadil and kaempferol increase the size of human =alprostadil; Kae.-Kaempferol: MG=MG 624; SKP spheres. Human SKPs were treated with the indicated Pram pramoxine: Alsterp. alsterpaullone; Kenp.-ken compound or vehicle for seven days, as indicated in FIG.7. At paullone. the end of the treatment period, cells were stained with (0090 FIGS. 22A-22C are graphs showing the effect of in Hoechst. An increase in the size of the spheres was observed vivo hair growth on topical treatment with either kaempferol in the alprostadil- and kaempferol-treated cells compared to or alprostadil. FIG. 22A shows the length of hair in mice vehicle control, indicating that these drugs increase cell pro treated with either kaempferol, alprostadil, or a vehicle con liferation within a sphere. trol at days 16, 19, and 23. FIGS. 22B and 22C show the hair 0080 FIG. 11 is a series of photomicrographs of human length distribution on day 23 from mice treated with either SKP spheresformed after 7 days of treatment with alprostadil kaempferol or alprostadil as compared to control mice. or vehicle respectively. 0081 FIG. 12 is a series of graphs showing that various DETAILED DESCRIPTION compounds that promote self-renewal of human SKPs also (0091. We have discovered that the p63 pathway is function on mouse SKPs. SKPs were isolated from mouse involved in inhibition of proliferation of skin-derived precur back and whisker and treated with the indicated concentration sors (SKPs). The invention thus features methods of increas of drugs. Results are pooled data from two experiments. A ing proliferation of a cell (e.g., a stem cell Such as a SKP) by total of four whisker and four back samples were used. inhibiting p63 expression or activity. 0082 FIG. 13 is a series of graphs showing that various 0092. In addition, we have performed a screen to identify compounds promote self-renewal of mouse SKPs. agents useful for enhancing cellular proliferation. This screen 0083 FIG. 14 is a graph showing that various compounds resulted in identification compounds including acyclovir, at 100 nM promote mouse SKPs self-renewal. Pooled data alprostadil, aristolochic acid, carbadox, chlorpyrifos, cyclo from two experiments show robust increase of the number of creatine, 7,4'-dimethoxyisoflavone, dorzolamide, S(-)eticlo spheres formed for both back and whisker cells after drug pride, evoxine, guaifenesin, hydroxyprogesterone, treatment. kaempferol, linamarin, mexicanolide, MG 624, pramoxine, US 2014/0234271 A1 Aug. 21, 2014 tannic acid, targinine, and yohimbic acid. On this basis, these Lane), 10 ng/ml BDNF (Peprotech), and 10 ng/ml NT3 (Pep compounds, or analogs of these compounds, can be used to rotech). For Schwann cell differentiation, dissociated spheres increase proliferation of cells, including SKPs or neuroblas were cultured in DMEM-F12 3:1 supplemented with 10% toma cells. As SKPs are known to be involved in skin regen FBS for 7 days, then switched to the same medium supple eration and hair growth, stimulation of SKPS using these mented with 4 uM forskolin (Sigma). compounds can enhance skin repair (e.g., wounded skin), Compounds that Decrease p63 Expression or Activity improve dermal maintenance or skin health, or promote hair 0098. The compositions, methods, and kits of the inven growth in a subject. tion may employ a compound that decreases p63 expression or activity. These compounds can be used either in place of or Cells in addition to other growth factors (e.g., FGF2 or EGF). In 0093. The compounds identified herein may be used to certain embodiments, the addition of a compound that increase proliferation of any cell that is capable of prolifera decreases p63 expression or activity allows for the use of a tion, such as tumor cell lines (e.g., neuroblastoma) or stem reduced concentration of another growth factor or factors cells. In certain embodiments, the cell is a stem cell Such as a (e.g., FGF2 or EGF). Exemplary compounds include RNAi SKP cell. Other stem cells include embryonic stem cells and molecules that target p63 mRNA, antibodies that specifically adult stem cells such as mesenchymal cells and hematopoi bind p63, and dominant negative forms of p83. etic stem cells. 0099 p63 RNAi Molecules 0.100 RNAi molecules such as siRNA molecules that tar 0094 SKPs are described in U.S. Patent Application Pub get p63 are known in the art or can be designed and tested for lication NOS. 2004/0033597 and 2007/0248574. SKPS can RNA interference activity. p63 RNAi molecules are commer express at least one, two, three, or more of the following cially available (e.g., from Santa Cruz, Biotechnology, Inc., molecular markers: nestin, WNT-1, vimentin, versican, Santa Cruz, Calif., Catalog No. sc-36161). Additional RNAi fibronectin, 51 0013, slug, snail, twist, Pax3, Sox9. Dermo-1, molecules (e.g., siRNA molecules) can be designed based on and Sox2. SKPs may also express increased levels of slug, the human pé3 mRNA sequence (NCBI accession Nos. snail, twist, and Pax3 relative to central nervous system neural NM 003722, NM 001114978, NM 001114979, stem cells. Desirably, the multipotent stem cells of the inven NM 001114980, NM 001114981, and NM 001114982) tion do not express measurable levels of at least one, two, or the corresponding mouse (NCBI accession Nos. three, or more of the following molecular markers: tyrosi NM 011641, NM 001 127259, NM 001 127260, nase, c-kit, tryp-1, and DCT, which are markers of melano NM 001 127261, NM 001 127262, NM 001 127263, blasts and melanocytes. The multipotent stem cells also may NM 001 127264, NM 001 127265) or rat (NCBI accession not express of one or more of the following markers of Nos. NM 001 127339, NM 001127341, NM 001127342, Schwann cells: MBP P0, p75NTR, and Sox10. NM 001 127343, NM 001 127344, and NM 019221) 0095 SKPs are capable of differentiating into various Sequences. non-neural cells (e.g., hair follicle cell, bone cell, Smooth 0101 pé3 Antibodies muscle cell, or adipocyte) and neural cells (e.g., a neuron, 0102 Anti-p63 antibodies may be employed in the com astrocyte, Schwann cell, or oligodendrocyte). positions, methods, and kits of the invention. Any antibody or 0.096 SKPs can be isolated as described in the art. In one antibody variant (e.g., monoclonal, polyclonal, human, example, dorsal or facial skin from mouse embryos (E15-19), humanized, single chain), a fragment thereof, or a nanobody mouse or rat neonates (P2-P6), or adults (3 weeks and older) can be employed. was dissected from the animal and cut into 2-3 mm pieces. 0103 Such antibodies are commercially available (e.g., Tissue was digested with 0.1% trypsin for 10-45 min at 37° from Abcam, Inc., Cambridge, Mass., Catalog No. ab735, or C., mechanically dissociated and filtered through a 40 um cell from Santa Cruz, Biotechnology, Inc., Catalog No. sc-8431). strainer (Falcon). Additional antibodies (polyclonal or monoclonal) against p63 or regions of p63 can also be generated using methods Cell Culture known in the art. 0097. The cells (e.g., SKPs) may be cultured under stan 0104 Humanized antibodies can be generated following dard cell culture conditions, such as those described herein or the method of Winter and co-workers (Jones et al., Nature, known in the art. In one example, SKPs are cultured as 321:522-525, 1986; Riechmann et al., Nature, 332:323-327 described in Toma et al. (Nat. Cell Biol. 3:778-784, 2001). (1988); Verhoeyenet al., Science, 239:1534-1536 (1988)), by Dissociated cells (e.g., as described above) were pelleted and substituting rodent CDRs or CDR sequences for the corre plated in DMEM-F12, 3:1 (Invitrogen), containing 20 ng/ml sponding sequences of a human antibody. EGF and 40 ng/ml FGF2 (both from Collaborative Research), 0105. Accordingly, such “humanized' antibodies are chi hereafter referred to as proliferation medium. Cells were cul meric antibodies wherein Substantially less than an intact tured in 25 cm tissue culture flasks (Falcon) in a 37° C., 5% human variable domain has been substituted by the corre CO tissue culture incubator. SKPs were passaged by sponding sequence from a non-human species. In practice, mechanically dissociating spheres and splitting 1:3 with 75% humanized antibodies are typically human antibodies in new medium and 25% conditioned medium from the initial which some CDR residues and possibly some FR residues are flask. For neuronal differentiation, SKP spheres or primary Substituted by residues from analogous sites in rodent anti dissociated skin cells were mechanically dissociated and bodies. plated on chamber slides (Nunc) coated with poly-D-lysine/ 0106 Human monoclonal antibodies can be made by the laminin in DMEM-F 12 3:1 supplemented with 40 ng/ml hybridoma method, as is known in the art (see, e.g., Kozbor, FGF2 and 10% FBS (BioWhittaker) for 5-7 days. Cells were J. Immunol. 133,3001, 1984, and Brodeur et al., Monoclonal then cultured an additional 5-7 days in the same medium Antibody Production Techniques and Applications, pp. 51-63 without FGF2 but with the addition of 10 ng/ml NGF (Cedar (Marcel Dekker, Inc., New York, 1987)). US 2014/0234271 A1 Aug. 21, 2014

0107 Nanobodies can be generated by immunization of DN p63 beta (NCBI accession Nos. AAG45611 and an animal (e.g., a camel or llama) which produces nanobod AAC62638 (human): AAP87986 and AAC62643 (mouse)); ies, which can then be purified using standard techniques. and DN p63 gamma (NCBI accession Nos. AAG45612 and 0108 Single chain Fv fragments may be produced such as AAC62634 (human); AAP87987 and AAC62642 (mouse)). described in Iliades et al., FEBS Letters, 409:437-441 (1997). Other dominant negative p63 polypeptides can sequences Coupling of such single chain fragments using various linkers Substantially identical to those sequences, or may contain is described in Kortt et al., Protein Engineering, 10:423-433 chemical modifications, as is known in the art. (1997). 0114 Gene Therapy 0109 The compositions, methods, and kits of the inven 0115 Decreases in p63 expression or activity may also be tion can also include a p63 antibody fragment (e.g., a frag achieved through introduction of a gene vector into a Subject. ment of a murine, human or humanized antibody, and anti To increase cellular proliferation, enhance skin repair or skin body variants). Such fragments can be derived by proteolytic health, or promote hair growth, p63 expression or activity digestion of intact antibodies (see, e.g., Morimoto et al., J. may be decreased, for example, by administering to a subject Biochem. Biophys. Methods 24:107-117 (1992)and Brennan a vector containing a polynucleotide sequence encoding a et al., Science 229:81, 1985) or can be produced directly by dominant negative form of p83 or an RNAi molecule that recombinant host cells. For example, Fab'-SH fragments can targets p03 mRNA, operably linked to a promoter capable of be directly recovered from E. coli and chemically coupled to driving expression in targeted cells. Any standard gene form F(ab') fragments (Carter et al., Bio/Technology therapy vector and methodology may be employed for Such 10:163-167, 1992). Single chain Fv fragments may be pro administration. duced such as described in Iliades et al., FEBS Letters, 409: 437-441 (1997). Coupling of such single chain fragments Compounds using various linkers is described in Kortt et al., Protein Engineering, 10:423-433, 1997. 0116 Based on the results of the screen described herein, 0110 Triabodies can also be used in the compositions, we have identified compounds capable of increasing prolif methods, and kits of the invention. Such antibodies are eration. Accordingly, these compounds, or analogs of these described for instance in Iliades et al., Supra and Kortt et al., compounds, may be used in methods for increasing prolifera Supra. tion (e.g., without affecting differentiation) in vivo or in vitro 0111. The antibodies of the present invention may be of any type of cell capable of proliferation (e.g., stem cells, modified by conjugating the antibody to an agent (e.g., any of Such as SKPs). The compounds may be used in conjunction the compounds described herein). Further antibody modifi with any cell culture techniques known in the art. Use of these cations are also contemplated. For example, the antibody may compounds may allow for reduced concentrations of other be linked to one of a variety of nonproteinaceous polymers, growth factors. In addition, the compounds can also be used e.g., polyethylene glycol, polypropylene glycol, polyoxy to treat a disease or condition where increases in SKP prolif alkylenes, or copolymers of polyethylene glycol and polypro eration are desirable (e.g., those described herein). pylene glycol. The antibody also may be entrapped in micro 0117. Acyclovir capsules prepared, for example, by coacervation techniques 0118. The compositions, methods, and kits of the inven or by interfacial polymerization (for example, hydroxymeth tion may include acyclovir or an analog thereof. Acyclovir ylcellulose or gelatin-microcapsules and poly-(methyl has the structure: methacylate) microcapsules, respectively), in colloidal drug delivery systems (for example, liposomes, albumin micro spheres, microemulsions, nano-particles and nanocapsules), or in macroemulsions. Such techniques are disclosed in Rem ington. The Science and Practice of Pharmacy, 20th edition, 2000, ed. A. R. Gennaro, Lippincott Williams & Wilkins, Philadelphia. To increase the serum half-life of the antibody, one may incorporate a salvage receptor binding epitope into ar) the antibody (especially an antibody fragment), as described 2 ls OH in U.S. Pat. No. 5,739,277, for example. A “salvage receptor 1N1 binding epitope' is an epitope of the Fc region of an IgG molecule (e.g., IgG1, IgG2, IgG3, or IgG4) that is responsible 0119 Analogs of acyclovir are described in U.S. Pat. No. for increasing the in vivo serum half-life of the IgG molecule. 4,199,574 and have the structure: 0112 Dominant Negative p63 0113 Dominant negative forms of p83, polypeptides sub stantially identical to Such dominant negative forms and hav ing dominant negative activity, or fragments thereof having dominant negative activity can be used in the compositions, methods, and kits of the invention. Forms of p83 with the N-terminal transactivation domain are termed TAp63, whereas those lacking this domain (AN-p63 isoforms) have dominant negative activity. See, e.g., Ghioni et al., Mol Cell Biol 22:8659-8668, 2002. Exemplary sequences encoding dominant negative proteins are known in the art and include DN p63 alpha (NCBI accession Nos. AAG45610 and AAC62636 (human): AAP87985 and AAC62644 (mouse)); US 2014/0234271 A1 Aug. 21, 2014

where X is Sulfur or oxygen, R is H. halogen, OH. C. alkoxy, azide, thio. C alkylthio, amino, Calkylamino or dialkylamino; R is H. halogen, C alkylthio, acylamino, amino or azide; R is H. Straight or branch chain or cyclic C alkyl, C. hydroxyalkyl, benzyloxyalkyl, or phenyl; R is H. OH, or alky; Rs is H, OH, NH, C alkyl, Ca hydroxyalkyl, benzyloxy, benzoyloxy, benzoyloxymethyl, Sulfamoyloxy, phosphate carboxypropiamyloxy, straight chain or cyclic acyloxy having from 1 to 8 carbon atoms e.g., acetoxy or substituted carbamoyl group of formula NH.CO—Z wherein Z is alkyl, aryl or aralkyl optionally substituted by one or where R is hydrogen or a Cls alkyl group, R is a member selected from the group consisting of H. Cls alkyl, and the more of Sulfonyl, amino, carbamoyl or halogen; R is H or acyl group of a hydrocarbon carboxylic acid of 2 to 18 carbon alkyl (e.g., when X is O and R. R. R. and Rare H. R. is not atoms; R is OH, n is 3, 4, or 5, and OR and R may have an amino or methylamino when Rs is hydrogen or hydroxy), or C. or B configuration. Exemplary of lower alkyl containing a salt thereof. from 1 to 8 carbon atoms suitable for include branched or 0120. Other analogs of acyclovir include 1-O-hexadecyl straight chain hydrocarbon groups such as methyl, ethyl, propanediol-3-p-acyclovir, 1-O-hexadecylpropanediol-3- propyl, isopropyl, butyl, tert-butyl, pentyl, and hexyl. Exem phosphoacyclovir, 1-O-octadecyl-sn-glycero-3-phospho plary acyl groups include acetyl, propionyl, butyryl, hex acyclovir. 2'-O-glycyl acyclovir, 2,6-diamino-9-(2- anoyl, octanoyl, lauroyl, palmitoyl, Stearoyl, and oleoyl. hydroxyethoxymethyl)purine, 3,9-dihydro-3-((2- Otheralprostadil analogs are described in U.S. Pat. No. 5.219, hydroxyethoxy)methyl)-6-ethyl-9-oxo-5H-imidazo 1,2-a 885. Other prostaglandins that can be used in the invention are purine, 3-methyl-cyclosal-PCVMP 6-deoxypenciclovir, described below. 8-fluoropenciclovir, 9-(1,3-dihydroxy-2-propylthio)me 0.124. Other prostaglandins include 2-chloro-4-hydroxy thyl)guanine, 9-((2-aminoethoxy)methyl)guanine, 9-(2-(9"- 5-(6-methoxycarbonyl-2-hexyenylidene)-4-n-octyl-2-cyclo octadecenoyloxy)ethoxymethyl)guanine, 9-(3,4-dihydroxy pentanone, 4-fluoroenisoprost, 7-hydroxy-5,11-dioxotetran butyl)guanine, 9-(4-hydroxy-2-(hydroxymethyl)butyl)- orprostane-1,16-dioic acid, AY 22093, bromovulone III, guanine triphosphate, 9-(4-hydroxybutyl)guanine, acyclovir carijenone, claviridenone E. claviridenone F, claviridenone G. clavubicyclone, MR 256, NEPP11 compound, NEPP6 B-glucoside, acyclovir diphosphate dimyristoylglycerol, acy compound, NEPP6-biotin, prostaglandin endoperoxides, clovir fluorophosphate, acyclovir monophosphate, acyclovir prostaglandins A, prostaglandins B, prostaglandins D, pros monophosphate-lactosaminated serum albumin conjugate, taglandins E. prostaglandins F. prostaglandins I (e.g., epo acyclovir phosphite, acyclovir triphosphate, acyclovir-5'- prostenol), RS 61756-007, S 1033, tricycloclavulone. (phenyl methoxy alaninyl)phosphate, BIOLF 143, bis(2- (guanin-9-ylmethoxy)ethoxy)-4-(methylsulfonyl)phenyl 0.125 Prostaglandins A include 13, 14-dihydro-15-deoxy A-prostaglandin-A1-methyl ester, 13, 14-dihydro-A7-pros phosphate, BRL 42359, bucyclovir triphosphate, desciclovir, taglandin A1 methyl ester, 15-keto-13, 14-dihydroprostaglan diamminechloro(9-(2-hydroxyethoxymethyl)guan-7-yl) din A2, 16, 16-dimethylprostaglandin A2 methyl ester, 5-(4- platinum(II), famciclovir, Y-glutamylacyclovir, ganciclovir, N,N-dimethylaminophenylmethylene)-4-hydroxy-2-(1- guanin-9-yl methyloxy-2-ethyl bis(S-acetyl-2-thioethyl) methylimidazol-2-ylthio)-4-(4-phenylbutyl)-2- phosphate, guanin-9-yl methyloxy-2-ethyl bis(S-pivaloyl-2- cyclopentenone, 8-isoprostaglandin A2, chloroVulone I. thioethyl) phosphate, penciclovir, penciclovir triphosphate, clavulone II, clavulones, prostaglandin A1, prostaglandin A2, tyrosylacyclovir, Val-Valacyclovir, and Valacyclovir. prostaglandin A2 isopropyl ester, and TEI 3313. 0121 Additional analogs are described in U.S. Pat. Nos. 0.126 Prostaglandins B include 15-ketoprostaglandin B1, 4,806,642, 4,957,924, 5,580,571, 6,214,811, and 6,031,096. 16, 16-dimethyl-15-dehydroprostaglandin B1 trimer, 19-hy droxyprostaglandin B2, di-Calciphor, OC 5181. OC 5186, 0122) Prostaglandins prostaglandin B1, prostaglandin B2, and prostaglandin BX. 0123. The compositions, methods, and kits of the inven I0127 Prostaglandins D include 9-chloro-15-cyclohexyl tion may include a prostaglandin Such as alprostadil (prostag 11, 15-dihydroxypentanor-5,13-prostadienoic acid, 9-fluoro landin E1), or an analog thereof. Alprostadil has the formula: 15-cyclohexyl-11, 15-dihydroxypentanor-5,13-prostadienoic acid, 9-fluoro-15-hydroxy-16-phenoxy-17, 18,19,20-tetra nor-5,13-prostadienoic acid, 9-hydroxy-11, 15-dioxo-2.3.18. 19-tetranorprost-5-ene-1,20-dioic acid, prostaglandin D2, and prostaglandin D3. I0128 Prostaglandin D2 analogs include 11, 15-dioxo-9- hydroxy-2,3,4,5-tetranorprostan-120-dioic acid, 11-meth Oxime prostaglandin D2.11-methyleneprostaglandin D2, 13, 14-dihydro-15-ketoprostaglandin D2, 15-deoxy-A(12. 14)-prostaglandin J2, 15-deoxyprostaglandin J2, 15-methyl prostaglandin D2.9-deoxy-9,10-didehydro-12,13-didehy dro-13, 14-dihydroprostaglandin D2,9-deoxy-A-9- prostaglandin D2, anhydrolevulgandin D2, A(12)- Analogs of alprostadil are described in U.S. Pat. No. 3,735, prostaglandin J(2), prostaglandin D2 4-azidophenacyl ester, 005 and have the formula: prostaglandin D2 methyl ester, and TS-002. US 2014/0234271 A1 Aug. 21, 2014

0129. Prostaglandins E include 17-isolevuglandin E4. hydroprostaglandin E2, 8, 12-epi-prostaglandin E2, 8-iso 6(9)-oxy-11, 15-dihydroxy-prosta-7,13-dienoic acid, anhy prostaglandin E2, 9-deoxy-9-chloro-15-deoxy-16-hydroxy drolevuglandin E2, levuglandin E2, lymphocytic thyroid 17, 17-trimethylene-19.20-didehydroprostaglandin E2, stimulator (non-immunoglobulin), alprostadil, dinoprostone, 9-enol-prostaglandin E2 methyl ester trimethylsilyl ether, prostaglandin-inositol cyclic phosphate, Ro 22-1327, and FCE 20700, HOE 260, N-acetylprostaglandin E2 carboxam Suppressor active peptide (thermal injury.) ide, ONO AE 248, prostaglandin D2 ethanolamide, prostag 0130 Prostaglandin E1 analogs include 11, 15-bisdeox landin E2 azidophenacyl ester, prostaglandin E2 ethanola yprostaglandin E1, 11-deoxy-10-hydroxyprostaglandin E1 mide, prostaglandin E2 glyceryl ester, prostaglandin E2 methyl ester, 11-deoxy-13, 14-dihydro-8-azaprostaglandin methyl ester, prostaglandin E2 methyl oxime, prostaglandin E(1), 11-deoxy-16, 16-trimethyleneprostaglandin E1. F2C, ethanolamide, prostamide H2, Sulprostone, trimoprostil, 11-deoxy-16-phenoxy-17, 18,19,20-tetranorprostaglandin and Viprostol. E1, 11-deoxy-4,4-dimethyl-4-Silaprostaglandin E1, I0132) Prostaglandins F analogs include (5Z)-7-((1R,2R, 11-deoxy-8-azaprostaglandin E(1), 11-deoxyprostaglandin 3R,5S)-2-((1E)-3,3-difluoro-4-phenoxy-1-butenyl)-3,5-di E1, 11-deoxyprostaglandin E1 4-hydroxyphenyl ester, 13.14 hydroxycyclopentyl)-5-heptenoic acid, 15-cis-(4-n-propyl dihydro-15-ketoprostaglandin E1, 13.14-dihydroprostaglan cyclohexyl)-16, 17, 17.19.20-pentanor-9-deoxy-6.9-O- din E1, 15(S)-15-methylprostaglandin E1, 15-cyclohexyl nitriloprostaglandin F1, 2,3-dinor-6-oxoprostaglandin F1B, omega-pentanor-7-thiaprostaglandin E1, 15-fluoro-11, 15 5(6)-epoxyprostaglandin F1C., 5,6-dihydroxyprostaglandin dideoxyprostaglandin E1, 15-keto-13, 14-dihydro-6- F1, 5,7-dihydroxy-11-ketotetranorprostanoic acid, alfapros ketoprostaglandin E1. 15-keto-prostaglandin E0. tol, butyryl prostaglandin F1 butyl ester, C22-prostaglandin 15-ketoprostaglandin E, 16, 16-dimethyl-A2-prostaglandin F4C. 6-Ketoprostaglandin F1C., lymphocytic thyroid stimu E1, 16, 18-ethano 20-ethyl-6-oxoprostaglandin E1, 16, 18 lator (non-immunoglobulin), Dinoprost, prostaglandin ethano-20-ethyl-6-oxoprostaglandin E1 leucinamide, 17, 18 F-main urinary metabolite, prostaglandin F1, prostaglandin dehydroprostaglandin E1, 18-hydroxyprostaglandin E1. F1B, prostaglandin F2B, prostaglandin F3C, and tafluprost. 19-hydroxyprostaglandin E1.20-dimethyl-7-thiaprostaglan 0.133 6-Ketoprostaglandin F1C. analogs include 2,3-di din E1 methyl ester, 20-hydroxyprostaglandin E1.3,7-dithia nor-6-ketoprostaglandin F1C, 5-hydroxy-6-ketoprostaglan (11C.,13E,15S)-11, 15-Dihydroxy-9-oxoprost-13-en-1-oic din F1C., 6.15-diketo-13, 14-dihydroprostaglandin F1C., 6.15 acid, 3-oxa-4.5,6-nor-(3.7-inter)-3-phenyleneprostaglandin diketoprostaglandin F1C, 6-ketoprostaglandin F1C.- E1 methyl ester, 3-oxa-4,5,6-nor-3.7-inter-3-phenylenepros thyroglobulin conjugate, 6-ketoprostaglandin F1 C-tyramide, taglandin E1, 3-oxa-4,5,6-trinor-3.7-inter-3-phenylenepros A(17)-6-ketoprostaglandin F1C, and endothelin-1,2-6-keto taglandin E1 amide, 4-thiaprostaglandin E1, 5(6)-epoxypros prostaglandin F1C. taglandin E1C, 5,6-dihydroxyprostaglandin E1, 0.134 Dinaprost (prostaglandin F2C.) analogs include 6-ketoprostaglandin E1, 7-keto-9.9-ethylenedioxipros 1-ethyl (5Z)-7-((1R,2R,3R,5S)-2-(1E)-3,3-difluoro-4-phe tanoid, 7-oxo-15-methylprostaglandin E1 methyl ester, noxy-1-butenyl)-3,5-dihydroxycyclopentyl)-5-heptenoate, 8-azaprostaglandin E1, alibra, AY 23626, butaprost, 8-17 1-methyl (5Z)-7-((1R,2R,3R,5S)-2-(1E)-3,3-difluoro-4- tetranorprostaglandin E1, enisoprost, gemeprost, isoprostag phenoxy-1-butenyl)-3,5-dihydroxycyclopentyl)-5-hep landin E1, limaprost, limaprost-alfadex, lubiprostone, MDL tenoate, 11-fluoro-11-dehydroxyprostaglandin F2C, 646, MR 356, NP 01A, NP 07A, NP 13 A, ONO 1082, ONO 11-fluoro-1'-deoxyprostaglandin F2C, 11-ketotetranorpros 1206, ONO-DI-004, ornoprostil, prostaglandin E0, prostag taglandin F2C, 13, 14-dihydroprostaglandin F2C, 13, 14-di landin E1 ethyl ester, prostaglandin E1 methyl ester, prostag hydroxy-15-ketoprostaglandin F2C, 15-fluoro-15-deox landin El C-cyclodextrin, prostaglandin E1-hexyl-Sepharose, yprostaglandin F2C, 15-keto-13, 14-dihydroprostaglandin prostaglandin E3, SC 29169, SC 31391, SC-46275, SPM F2C, 15-keto-17-phenyl-18,19,20-trimorprostaglandin F2C.- 206, tetranorprostaglandin E1. TFC 612, and TR4161. 1-isopropyl ester, 15-ketoprostaglandin F2C, 15-propionat 0131 Dinoprostone (prostaglandin E2) analogs include prostaglandin F2C-isopropyl ester, 16, 16-dimethylprostag 11-deoxy, 16, 16-dimethyl prostaglandin E2, 11-deoxy-11, landin F2C, 16-(3-chlorophenoxy)-17, 18, 19.20 12-methanoprostaglandin E2, 11-deoxy-11 C-(2-hydroxyeth tetranorprostaglandin F2C, 16-aminoprostaglandin F2C. ylthio)-prostaglandin E2 methyl ester, 11-deoxy-15-keto-13, methyl ester, 16-fluoromethyleneprostaglandin F2C, 17-aza 14-dihydro-11 beta, 16-cycloprostaglandin E2, prostaglandin F2C. 17-phenyl-18,19,20-trinor-prostaglandin 11-deoxyprostaglandin E2-1-alcohol, 12-isoprostaglandin F2C-1-isopropyl ester, 17-phenyl-18,19,20-trinorprostaglan F(2C.), 13, 14-didehydroprostaglandin E2, 13, 14-dihydro-16 din F2C, 17-phenylprostaglandin F2C., 18,19,20-trinor-17 phenyl-S2-tetranorprostaglandin E2, 13, 14-dihydroprostag cyclohexyl-13, 14-dehydroprostaglandin F2C. methyl ester, landin E2, 15-deoxy-16-hydroxy-16-vinylprostaglandin E2. 19-hydroxyprostaglandin F 1a, 1b-dihomoprostaglandin F2. 15-fluoro-15-deoxyprostaglandin E2, 15-hydroperoxypros 2,3-dinor-5,6-dihydro-15-F-isoprostane, 2,3-dinor-5,6-dihy taglandin E2, 15-keto-13, 14-dihydroprostaglandin E2. dro-8-iso-prostaglandin F2C, 2,3-dinor-5,6-dihydroisopros 15-keto-13, 14-dihydroprostaglandin E2-thyroglobulin con tane F2C-III, 2-decarboxy-2-(P-methylphosphinico)-16 jugate, 15-ketoprostaglandin E2, 16-methyl prostaglandin phenoxytetranorprostaglandin F2C, 20-methyl-13, 14 E2, 16-methyl-13, 14-didehydroprostaglandin E2, 16-me (didehydroprostaglandin) F2C, 3-(3,5-dihydroxy-2- thyl-16-methoxyprostaglandin E2, 16-methyl-20-methoxy (oxodecyl)cyclopentyl)propionic acid, 6-keto-prostaglandin prostglandin E2, 17,17-dimethylprostaglandin E2, 17-(4-azi F2C, 7-(3,5-dihydroxy-2-(3-oxodecyl)cyclopentyl)hept-5- dophenyl)-18,19,20-trinorprostaglandin E2, enoic acid, 8, 12-iso-isoprostane F2C-III, 8, 12-iso-isopros 17-phenyltrinorprostaglandin E2, 18,18,20-trimethylpros tane F2C-VI. 8-epi-prostaglandin F2C, 9-chloro-15-cyclo taglandin E2, 18-hydroxyprostaglandin E2, 19.20-dehydro hexyl-11, 15-dihydroxy-3-oxa-16, 17, 18, 19.20-pentanor-5- prostaglandin E2, 19-hydroxyprostaglandin E2, 1a, 1b-diho prostenoic acid, AFP-157, AL 6598, AL 8810, AL 8810 moprostaglandin E2, 20-hydroxyprostaglandin E2. ethylamide, AL-3138, AY 24366, delprostenate, dinoprost 20-isopropylidene prostaglandin E2, 20-methyl-13, 14-dide tromethamine, etiproston, etiproston tromethamine, flupros US 2014/0234271 A1 Aug. 21, 2014 tinol, ICI-79939, isopropyl unoprostone, N-dimethylamino CL 115574, CL 116069, CP 48630, 16, 16-dimethylprostag prostaglandin F2C, PhXa 85, prostaglandin F2 ethyl ester, landin E2 or an analog thereof (e.g., 11-methyl-16, 16-dim prostaglandin F2 isopropyl ester, prostaglandin F2 methyl ethylprostaglandin E2, 16, 16-dimethylprostaglandin E2 ester, prostaglandin F2C. 11-methyl ether, prostaglandin F2C. (4-acetamidobenzamido)phenyl ester, 16, 16-dimethypros 15-methyl ether, prostaglandin F2C.9-methyl ether, prostag taglandin E2 4-benzaldehyde semicarbazone ester, 19-hy landin F2C. N-dimethylamide, ZK 110841, ZK 118182, and droxy-16, 16-dimethylprostaglandin E2,9-deoxy-16, 16-dim ZK 71677. ethyl-tetranor-9-methyleneprostaglandin E2, and 0135 Epoprostenol analogs include 10,10-difluoro-13 meteneprost), enprostil, GR 63799x, arbaprostill or a analog dehydroprostacyclin, 11-desoxyprostacyclin, 13.14-dehy thereof (e.g., 15-deoxy-16-methyl-16-hydroxy-3,4-didehy droprostaglandin I2, 13.14-dehydroprostaglandin I2 methyl droprostaglandin E2 methyl ester and 15-methylprostaglan ester, 13, 14-didehydro-20-methylcarboprostacyclin, 13.14 din E2 methyl ester), misoprostol or an analog thereof (e.g., dinor-inter-p-phenylene carbacyclin, 15-cyclopentyl-7-OXo 2-demethoxycarbonyl-2-ethoxycarbonyl-11-deoxymiso prostaglandin I2-ephedrine, 15-deoxy-(16-m-tolyl)-17, 18. prostol, Arthrotec, diclofenac-misoprostol, misoprostol acid, 19,20-tetranorisocarbacyclin methylester, 15-deoxy-16-m- and SC 53450), ONO-747, rioprostil, Ro 22-6923, RS 20216, tolyl-17, 18,19,20-tetranorisocarbacyclin, 15-fluoro-13, 14 RS 61565, and Wy 17186. dehydrocarbacyclin, 15-ketoprostaglandin 12, 16-tolyl-17, 0.137 Aristolochic Acid 18,19,20-tetranorisocarbacyclin, 1720 0.138. The compositions, methods, and kits of the inven dimethylisocarbacyclin, 19-(3-azidophenyl)-20 tion may include the use of aristolochic acid, or an analog norisocarbacyclin, 2.2,10,10-tetrafluoro-13 thereof. Aristolochic is a phospholipase A inhibitor. The dehydroprostacyclin, 20-methyl-13, 14-didehydro-2,4-inter structure of aristolochic acid is: 3-phenylene prostaglandin I2, 3-oxa-9(O)-methano-delta(6. 9)prostaglandin I(1), 3-oxacarbacyclin, 3-oxahomoisocarbacyclin, 4,5-didehydroisocarbacyclin, O 5,6-dihydroprostacyclin, 5-hydroxyprostaglandin I, 5-meth O yleneisocarbacyclin, 5-nitroprostaglandin I1, 5-nitroprostag OH landin I2, 6.9-thiaprostacyclin, 6a-carbaprostaglandin I3. { O 7-fluoroprostacyclin, 7-oxo-cyclopentyl-prostaglandin I2, O 4. 7-OXo-prostaglandin I2-ephedrine, 7-oxoprostaglandin I2, \ 7a-homo-2-norprostacyclin, 9-O-methanoprostaglandin I, O AFP 03, AFP 06, AFP 07, APS 306, benzodioxane prostacy clin, beraprost, bicyclo(4.3.0)non-2-ene homoisocarbacy clin, carbaprostacyclin, carboprostacyclin, CG 4303, Chinoin o1 7284, Chinoin 7384, cicaprost, ciprostene, CL 115999, dehy dro-15-cyclohexylcarbaprostacyclin, dihomo-prostaglandin I(2), FCE 21258, HOE 892, homoisocarbacyclin, KP 10614, 0.139 Analogs of aristolochic acid include 7-(deoxyad MM 706, naxaprostene, nileprost, nitriloprostaglandin 12, enosin-N(6)-yl)aristolactam II, 7-(deoxyadenosin-N(6)-yl) ONO 41483, OP 2507, OP 41483-O-cyclodextrin, piriprost, aristolactam 1.7-(deoxyguanosin-N(2)-yl)aristolactam II, prostaglandin I2 11-methyl ether, prostaglandin I2 15-methyl 7-(deoxyguanosin-N(2)-yl)aristolactam 1.7-methoxyaris tolochic acid A, 7-methoxyaristololactam IV. 9-ethoxyaristo ether, prostaglandin I2 methyl ester, prostaglandin I3, R lactone, 9-ethoxyaristololactam, 9-hydroxy aristolochic acid 59274, SC 39902, SM 10902, SM 10906, taprostene, TEI I, aristolactam I, aristolic acid, aristolochic acid C, aris 1324, TEI 3356, TEI 4343, TEI 9090, TFC 132, tilsuprost, tolochic acid E, aristolochic acid II, aristololactam BII, aris treprostinil, TRY 200, TTC 909, TY 10957, TY 11223, U. tololactam IVa, aristololactam-glucoside, aristoloside, 56467, U 68215, and U 72382. methyl aristolate, and N-((6'-p-coumaroyl)glucopyranosyl) 0.136 Synthetic prostaglandin analogs include (+)(-)-8, aristolactam. 12-trans-9-oxo-prosta-5,14-dienoic acid, 11-methoxime prostaglandin D2, 9C,11C, 15C-trihydroxy-16-phenoxy-17, 0140 Carbadox 18,19,20-tetranorprosta-4,5,13-trienoic acid, 9-chloro-15 0.141. The compositions, methods, and kits of the inven cyclohexyl-11, 15-dihydroxypentanor-5,13-prostadienoic tion may include the use of carbadox, or an analog thereof. acid, 9-fluoro-15-cyclohexyl-11, 15-dihydroxypentanor-5, The structure of carbadox is: 13-prostadienoic acid, AL-12182, EMD 33290, EP 045, EP 092, GIF 0010, GIF 0037, Iloprost, synthetic prostaglandin endoperoxides, synthetic prostaglandins A, synthetic pros taglandins E, synthetic prostaglandins F, and U 62840. Ilo prost analogs include 2,6-dichloro-4-aminophenol iloprost, eptaloprost, and iloprost phenacyl ester. Synthetic prostag sNH landin endoperoxides include 9,11-azo-13-oxa-15-hydrox O N yprostanoic acid, 9,11-iminoepoxyprosta-5,13-dienoic acid, prostaglandin H29-cyclic ether, prostaglandin H2 methyl N ester, and U 44069. Synthetic prostaglandins A include 16, 16-dimethylprostaglandin A1, 9-oxo-15-hydroxy-A7,10, 13-prostatrienoic acid methyl ester, GSH-prostaglandin A1, HR 546, punaglandin III, and TEI9826. Synthetic prostaglan dins E include 16, 16-dimethylprostaglandin E, 16-hydroxy 16-methyl-9-oxo-prosta-10,13-dien-1-oic acid methyl ester, US 2014/0234271 A1 Aug. 21, 2014 12

0142 Analogs of carbadox are described in U.S. Pat. No. 0148 Creatine analogs are described in U.S. Pat. No. 3,371,090 and have the formula: 5,998,457 and have the structure: . Z N R )-x-A-Y z: ---2 R. wherein O 0149 Y is COOHNHOH, NO, SOH, C(=O) NHSOJ, or - P(=O)(OH)(OJ), where J is H, C alkyl, C2-alkenyl, or aryl; where each of R and R is individually selected from the group consisting of H and Cls alkyl; R is selected from the 0150 A is C, CH, Cs alkyl, Cs alkenyl, Cs alkynyl, or group consisting of NHCONH2, NHC(S)NH2, NHCNHNH2, Cs alkoyl, each having 0-2 substituents selected indepen NHR, NHCOOR, NHCOR, OR, where R is selected dently from K, where K is C alkyl, Ce alkenyl, C. from the group consisting of Cs alkyl, phenyl, benzyl, and straight alkoyl, and Cabranched alkoyl, having 0-2 substitu hydroxyalkyl containing from 2 to 4 carbon atoms; Rs is ents independently selected from Br, Cl, epoxy, and acetoxy; selected from the group consisting of lower alkyl, hydroxy a 1-2 ring aryl group optionally heterocyclic having 0-2 Sub alkyl containing from 2 to 4 carbon atoms, and haloalkyl stituents independently selected from —CHL and containing from 2 to 4 carbon atoms; R is selected from the —COCHL, where L is Br, Cl, epoxy, or acetoxy; and 3) group consisting of Cs alkyl and phenyl; and R, is selected —NH-M, where M is H. C. alkyl, C. alkenyl, Ca from the group consisting of hydrogen and Cls alkyl. alkoyl, and C branched alkoyl; 0151 X is NR, CHR, CR, O, or S, where R is H; K, 0143 Chlorpyrifos where K is C alkyl, Calkenyl, C. Straight alkoyl, and 0144. The compositions, methods, and kits of the inven Ce branched alkoyl, having 0-2 substituents independently tion may include the use of chlorpyrifos, oran analog thereof. selected from Br, Cl, epoxy, and acetoxy; a 1-2 ring aryl group The structure of chlorpyrifos is: optionally heterocyclic having 0-2 substitutions of —CHL and COCHL where L is Br, Cl, epoxy, or acetoxy; a C C.-amino-()-methyl-(1)-adenosylcarboxylic acid attached by C the ()-methyl carbon; a Cso C.-amino-(D-aza-J-methyl-f-ad enosylcarboxylic acid attached by the ()-methyl carbon; and C a Cso C.-amino-(o-thia-()-methyl-(1)-adenosylcarboxylic acid attached by the co-methyl carbon; 0152 Z and Z are chosen independently from the group C consisting of: —O. —NHR, —CHR, —NROH; where Z. and Z are not both=O and where R is H. K, where K is C. alkyl, C- alkenyl, C. Straight alkoyl, and Ce branched 0145 Analogs of chlorpyrifos are described in U.S. Pat. alkoyl and contains 0-2 Substituents independently selected No. 3,244,586 and have the structure: from Br, Cl, epoxy, and acetoxy; a 1-2 ring aryl group option ally heterocyclic containing 0-2 Substituents independently selected from —CHL and —COCHL where L is Br, Cl, epoxy, or acetoxy; a Cas C-amino-carboxylic acid attached f R R-O / via the co-carbon; B, where B represents —COH NHOH, N - SOH, NO OP(=O)(OH)(OJ), or P(=O)(OH) R (OJ), where J is H. C. alkyl, Calkenyl, or aryl, and B is optionally connected to the nitrogen by C. alkyl, Calkenyl, where R is pyridyl optionally substituted with one or more or Calkoyl; -D-E, where D is C alkyl, C- alkenyl, C (e.g., 2, 3, 4) halogen groups, Z is O or S, and each R' straight alkoyl, aryl, or aroyl; and E is —(PO)NMP, where independently represents Cls alkoxy, amino, or Cs alky n is 0-2 and NMP is ribonucleotide monophosphate con lamino. nected by the 5'-phosphate, 3'-phosphate, or the aromatic ring Cyclocreatine of the base;—P(=O)(OCH)(O)-Q, where m is 0-3 and Q 014.6 is a ribonucleoside connected via the ribose or the aromatic 0147 Creatines, such as cyclocreatine, may be used in the ring of the base; —P(=O)(OH)(CH)-Q, where m is 0-3 compositions, methods, and kits of the invention. Cyclocre and Q is a ribonucleoside connected via the ribose or the atine has the structure: aromatic ring of the base; and an aryl group containing 0-3 Substituents chosen independently from Cl, Br, epoxy, acetoxy, —OG, —C(=O)C, and —COG, where G repre sents C- alkyl, C2- alkenyl, Co straight alkoyl, Cao branched alkoyl, and where E may be attached to any point to D, and if D is alkyl or alkenyl, D may be connected at either "Sr.O S. or both ends by an amide linkage; and —E, where E repre sents —(PO)NMP, where n is 0-2 and NMP is a ribonucle otide monophosphate connected by the 5'-phosphate,

US 2014/0234271 A1 Aug. 21, 2014 14 drolicoisoflavone, duartin, echinoisoflavanone, EMD 16795, equol. eriotriochin, ery cristagallin, euchrenone b10, eury carpin A. eurycarpin B, formononetin, fremontin, fremon A tone, fujiflavone P40, furowanin B, gancaonin A, genistein, genistein 4'.7'-bis(6-deoxytalopyranoside), genistein 7-(6- ( ) deoxytalopyranoside), genistein 7-O-alpha-L-rhamnopyra B N1 O noside-4'-O-(6"-O-alpha-L-rhamnopyranosyl)-beta Sophoroside, genistein 7-O-beta-D-glucopyranoside-4'-O- wherein A together with the two carbon atoms denoted as C. (6"-O-alpha-L-rhamnopyranosyl)-beta-sophoroside, and B is the group: genistin, glabrene, glabrizoflavone, glaziovianin A, gli coisoflavanone, glisoflavanone, glycitein, glycitin, hagininA, hydroxyethylpuerarin, indicanine C, intricatinol, ipriflavone, irigenin, irilin A, irilin B, irilin D, irilone, irisone A, irisone B. isocytisoside, isocytisoside-7-O-glucopyranoside, isofla Vanone, isoVitexin 2'-O-glucoside, isovitexin-4'-O-gluco side, judaicin (isoflavone), kakkalide, kievitone, kraussian one 1, kraussianone-1, kraussianone-2, kwakhurin, lanceolarin, licoisoflavone A, licoisoflavone B, lupinalbisone wherein A. lupinalbisone B. mahuangchiside, malonylgenistin, manuifolin D, manuifolin E, manuifolin F, manuifolin Q. X is S, SO, SO, or CH: menoflavon, methylaminium 4,7-dihydroxyisoflavone-3- 0162 Y is S. O, or NR where R is H. C. alkyl, or Sulfonate, methylophiopogonanone B. millewanin G. mille benzyl: wanin H, munetone, muscomin, N99-596 A, N99-596 B, n is 1 or 2; nigrolineaisoflavone A, O-desmethylangolensin, O-methyl R. R. R. R. are independently: claussequinone, ormosidin, osajaxanthone, osajin, pallidiflo (0163 H: ric acid methyl ester, pallidiflorin, pentandrin, pentandrin 0164 OR where Rs is H. Cls alkyl optionally substituted glucoside, phaseollin (isoflavan), phenoXodiol, phytosoya, with OH or NRR, where R and R, are independently Hor promensil, prunetin, prunetin-4'-O-beta-D-gentiobioside, Cs alkyl or are joined together form a heterocycle with the psi-baptigenin, psi-tectorigenin, Pterocarpans, puerarin, nitrogen to which they are attached (e.g., piperidino, mor pholino, or piperazino), Cs alkanoyl optionally substituted robustic acid, rotenone, sativan, Scandenone, Senegalensin, with OH, NRR, NHCORs, or CORs where R is OH, sigmoidin I, sigmoidin J. Sigmoidin K. Smiranicin, Sophorai NRR, or Cs alkoxy; or COR, where R is NRR, or a 5 Soflavanone D, Sophoraisoflavone A, Sophorol, soy isoflavone or 6-membered aromatic heterocycle Such as pyridyl, imida aglycone, tectoridin, tectorigenin, tectorigenin 7-O-(api Zolyl, pyrazinyl, thiazolyl, thienyl, or oxazolyl, ofuranosyl-(1-6)-glucopyranoside), tetrahydrodaidzein, tha (0165 NRR: tancuayin, torvanol A, triquetrumone A, triquetrumone B. (0166 NHR wherein Rio is SONRR, SOR where triquetrumone C, triquetrumone D, ulexin C, ulexin D, R is Cls alkyl, or CONRR-7, Cls alkyl, optionally Substi Vavain, Vavain 3'-O-glucoside, Vogelin A, Vogelin B, Vogelin tuted with ORs, CN, NRR, or CORs, C, Vogelin H. Vogelin I, Vulgarin (isoflavones), warangalone, (0167. SOR: warangalone-4'-methyl ether, and yufengningXin. Analogs (0168 SONRR, or are also described in U.S. Pat. Nos. 2,764,596, 2,892,845, (0169 halo, such as Cl, Br, or F: 2,892,846, 3,755,372, 3,907,830, 3,864,362, 4,841,077, and R and R, or R and R taken together represent a double 4,264,509. bond; R and R, or R and R taken together represent=O. or —NOR, where R is Hor C-alkyl; and one of the CH 0159. Dorzolamide groups of (CH), is optionally Substituted with —CORs. 0160 The compositions, methods, and kits of the inven —CHRs, or —CHCORs. tion may use dorzolamide oran analog thereof. Dorzolamide (0170 S(-)eticlopride has the structure: 0171 S(-)eticlopride, or an analog thereof can be used in the compositions, methods, and kits of the invention. S(-) eticlopride has the structure:

C OS C y New N OH O N

0161 Analogs of dorzolamide are described in U.S. Pat. 0172 Analogs of eticlopride are described in U.S. Pat. No. No. 4,677,115 and have the structure: 4,789,683 and have the formula: US 2014/0234271 A1 Aug. 21, 2014 15

R OA

H N R OA2 O R3 where R and R are independently H, a halogen, CN, C alkyl, or acyl: R is C alkyl, a C- alkenyl, or C-2 aryl 0181 Other hydroxyprogesterone compounds include optionally substituted with F, Cl, Br, CF, C alkyl, or C. 11C.-bromoacetoxyprogesterone, 11,14-dihydroxypregn-4- alkoxy, A and Aare independently H.C. alkyl group, acyl, ene-320-dione, 11-hydroxyprogesterone, 11-hydrox Calkoxycarbonyl, or a dialkylcarbamyl group, (e.g., pro yprogesterone 11-glucuronide, 12C-hydroxyprogesterone, vided that when A and A are the same lower alkyl group and 14-hydroxypregna-1,4-diene-320-dione, 14-hydrox R is ethyl, R, R2, or both are selected among CN, C-alkyl, yprogesterone, 15,17-dihydroxyprogesterone, 15-hydrox and acyl); or a physiologically acceptable salt or optical iso yprogesterone, 16C, 17O-isopropylidenedioxyprogesterone, mers thereof. 16 O-bromoacetoxyprogesterone, 16,17-epoxy-3-hydrox 0173 Evoxine ypregn-5-en-20-one, 16-hydroxyprogesterone, 16-methyl 0.174 Evoxine, or an analog thereof, may be used in the ene-17 C-acetoxyprogesterone, 16-methylene-17 C-hydrox compositions, methods, and kits of the invention. Evoxine has yprogesterone, 17CL.20B-dihydroxypregn-4-en-3-one, 17 the structure: O-acetoxy-2,2'.6.f3-trifluoro-6(3.7 B-dihydro-16-methyl enecyclopropa (6.7)progesterone, 17.20-dihydroxy-4-preg nen-3-one, 17-(bromoacetoxy)progesterone, 17-acetoxy-11 No oxaprogesterone, 17-acetoxy-7-Oxaprogesterone, 17-O- hydroxy-progesterone caproate, 17-hydroxypregn-4-ene-3- One, 17-hydroxyprogesterone 17-(9-oxo-10 chlorodecanoate), 17-hydroxyprogesterone heptanoate, 17C.- hydroxy-6-methylene-progesterone, ck- 4No 18-hydroxyprogesterone, 19-hydroxyprogesterone, 2-bro OH CO)19 moacetoxyprogesterone, 21-amino-17-hydroxyprogester one, 21-bromoacetoxyprogesterone, 3-O-glucopyranosyl-3, 15-dihydroxypregn-5-en-20-one, 4-pregnen-20,21-diol-3- one, 63-acetoxyprogesterone, 6,17.20-trihydroxypregn-4- 0175 Evoxine acts as a sedative and is an antagonist of ene-3-one, 6-bromoacetoxyprogesterone, 6-hydroxy-6- Strychnine and pentylenetrazole. methyl-17-acetoxyprogesterone, 6-hydroxyprogesterone, (0176 Guaifenesin 6-trifluoromethyl-16-methylene-17 C.-hydroxy-4,6-pregna 0177 Guaifenesin, or an analog thereof, may be used in diene-320-dione 17-acetate, 7 C-carboxymethyl-17-hydrox the compositions, methods, and kits of the invention. yprogesterone, 73-carboxymethyl-17-hydroxyprogesterone, Guaifenesin has the structure: 7.14-dihydroxypregn-4-ene-320-dione, 7-hydroxyprogest erone, caprovestrol, deluteval, flumedroxone, 17O-acetoxy 3B-butanoyloxy-6-methyl-pregna-4,6-dien-20-one, 17C.-ac etoxy-3f-isopropyloxy-6-methylpregna-4,6-dien-20-one, 17C.-acetoxy-3B-phenylpropionyloxy-6-methylpregna-4,6- dien-20-one, 17-O-hydroxyprogesterone. Injectable No. 1, arro methylene-dehydroacetoxy-progesterone, pregna-1,4-diene OH O 11-ol-320-dione, primosiston, progesterone 11-glucu N ronide-alkaline phosphatase conjugate, progesterone 11-hemisuccinate, progesterone 11-hemisuccinate-(2-io 0.178 Gauifenesin is used as an expectorant. dohistamine), thymine-hydroxyprogesterone, and trophobo 0179 Hydroxyprogesterones len. Kaempferol 0180 17o-Hydroxyprogesterone (CAS No. 68-96-2), or 0182 an analog thereof, may be used in the compositions, methods, 0183 Kaempferol, or an analog thereof, may be used in and kits of the invention. 17C.-Hydroxyprogesterone has the the compositions, methods, and kits of the invention. The Structure: structure of kaempferol is:

US 2014/0234271 A1 Aug. 21, 2014 17

0193 Analogs of MG 624 are described in Mantegazza et al., Arch. Int. Pharmacodyn., 4:371, 1955 and Gotti et al., Br. J. Pharmacol. 124:1197, 1998. 0.194 Pramoxine 0.195 Pramoxine, oran analog thereof, may be used in the compositions, methods, and kits of the invention. The struc ture of pramoxine is:

0.190 Mexicanolide is found in the Spanish cedar (Ce drella odorata L). / / (0191) MG 624 0.192 MG 624, or an analog thereof, may be used in the compositions, methods, and kits of the invention. The struc ture of MG 624 is: 0196. Pramoxine is used as a topical anesthetic, often in treatment of insect bites. 0.197 Tannic Acid 0198 Tannic acid or tannic complexes may be used in the compositions, methods, and kits of the invention. Tannic acid is a polymer made up of a glucose molecule attached by ester linkages and to one or more gallic acid units, which, in turn, may be further attached to additional gallic acid units. Tannic acid can have the structure:

O OH OH O

HO OH O O O O O OH OH HO O HO O OH OH HO US 2014/0234271 A1 Aug. 21, 2014

0199 Tannic acid is used topically to treat injuries such as 0203 Yohimbic Acid burns, and has been shown to have some antineoplastic 0204 Yohimbic acid, oran analog thereof, may be used in effects. the compositions, methods, and kits of the invention. The (0200 Targinine structure of yohimbic acid is: 0201 Targinine, or an analog thereof, may be used in the

compositions, methods, and kits of the invention. The struc ture of targinine is:

HN OH

O s OH OH HN Analogs of yohimbic acid are described in U.S. Pat. Nos. HN 1. 2,796,420, 2,841,586, 2,977,367, 2,998,556, 3,120,532, 3,126,390, 3,139.434, 3,337,559, and 3,940,387. Analogs of yohimbic acid include yohimbine, 1-methylyohimbine, 10-hydroxyyohimbine, 11-hydroxy-yohimbine, 11-hydrox 0202 Analogs of targinine are described in U.S. Pat. No. yyohimbine, 14-(4-nitrobenzoyloxy)yohimbine, 14-benzoy 4,698,442 and have the structure: loxyyohimbine, 17-hydroxy-20-yohimban-1 6-(N-(4-azido 3-iodo)phenyl)carboxamide, 5-carboxytetrahydroalstonine, 9-methoxy-3-epi-alpha-yohimbine, raubasine, apoyohim bine, iodinated rauwolscine, NMI 187, rauwolscine 4-ami nophenylcarboxamide, rauwolscine, Reserpine or an analog thereof (e.g., 16, 18-reserpinediol, adelphan-esidrex, adel phane, aldatense, Bendigon, bietaserpine, Briserin, bromore serpine, butiserpazide, caprinol, CD 3400, Crystepin CH, deserpidine, dibromoreserpine, enderpins, FH 109 C. Hyparez, meprocalm, methyl reserpate, Nortensin, Regroton, rescinamine, rescinnamine, reserpic acid, reserpine methoni trate, Sinepres, and Syrosingopine), and trimethoxybenzoy where n is 1-5; R is C. alkyl, Chalogen Substituted lyohimbine. alkyl, or NHRs where R is C. alkyl, C. cycloalkyl, phenyl, benzyl, Chalogen Substituted alkyl, morpholino, Treatment of a Subject or (CH)N(R) where n is 1-5 and R is C alkyl; R is H 0205 The invention also provides methods for increasing or R, or R and R comprise a ring represented by the fol cellular proliferation, enhancing skin repair or skin health, lowing structural formulas: and promoting hair growth by administration of a compound that decreases expression or activity of p63 (e.g., dominant negative forms of p63, a p63 antibody, or a compound selected from the group consisting of acyclovir, alprostadil, aristolochic acid, carbadox, chlorpyrifos, cyclocreatine, 7,4'- 1. dimethoxyisoflavone, dorzolamide, S(-)eticlopride, evoxine, guaifenesin, hydroxyprogesterone, kaempferol, linamarin, and . HN ul NH mexicanolide, MG 624, pramoxine, tannic acid, targinine, \ / \ / and yohimbic acid, or an analog thereof, to a Subject in need thereof. The compounds can be further modified using stan X dard chemical, physical, or biochemical methods. Formulation of Pharmaceutical Compositions HN 1.SN 0206. The administration of any compound or composi tion described herein may be by any suitable means that A (CH2), results in a concentration of the compound that increases A cellular proliferation, enhances skin repair or skin health, or B B promotes hair growth. The compound may be contained in any appropriate amount in any suitable carrier Substance, and where m is 0-6: A and B are independently H, C alkyl, or is generally present in an amount of 1-95% by weight of the C-2 cycloalkyl, and X is halo or A (e.g., provided that total weight of the composition. The composition may be whereinn is 3 and R is NHR, wherein R is methyl and R provided in a dosage form that is suitable for the oral, skin is H or methyl are excluded). (e.g., topical or by patch), cutaneous, parenteral (e.g., intra US 2014/0234271 A1 Aug. 21, 2014

venous or intramuscular), rectal, nasal, vaginal, inhalant, ocu gel, or other solid or semisolid composition, once, twice, lar, or intracranial administration route. Thus, the composi three times, or more than three times per day. These formu tion may be in the form of, e.g., tablets, capsules, pills, lations can be made according to known and conventional powders, granulates, Suspensions, emulsions, solutions, gels methods for preparing Such formulations. including hydrogels, pastes, ointments, creams, plasters, 0210 For compounds of the invention that are not highly drenches, osmotic delivery devices, Suppositories, enemas, soluble in water at physiological conditions, a solubilizing injectables, implants, sprays, or aerosols. The pharmaceutical excipient may be used to increase solubility. Solubilization is compositions may be formulated according to conventional taken to mean an improvement in the solubility by virtue of pharmaceutical practice (see, e.g., Remington. The Science Surface-active compounds that can convert Substances that and Practice of Pharmacy, 20th edition, 2000, ed. A. R. are insoluble or virtually insoluble in water into clear, or Gennaro, Lippincott Williams & Wilkins, Philadelphia, and opalescent, aqueous solutions without changing the chemical Encyclopedia of Pharmaceutical Technology, eds. J. Swar structure of these Substances in the process. Excipients used brick and J. C. Boylan, 1988-1999, Marcel Dekker, New for this purpose are restricted to those that are safe for admin York). istration to humans. Typically Such co-solvents are employed 0207 Pharmaceutical compositions may be formulated to at a level of about 0.01% to 2% by weight. release the active compound immediately upon administra 0211 A variety of solubilizing excipients may be used for tion or at any predetermined time or time period after admin the formulation of a compound of the invention, including istration. The latter types of compositions are generally compounds belonging to the following classes: polyethoxy known as controlled release formulations, which include (i) lated fatty acids, PEG-fatty acid diesters, PEG-fatty acid formulations that create Substantially constant concentrations mono-ester and di-ester mixtures, polyethylene glycol glyc of the agent(s) of the invention within the body over an erol fatty acid esters, alcohol-oil transesterification products, extended period of time; (ii) formulations that after a prede polyglycerized fatty acids, propylene glycol fatty acid esters, termined lag time create Substantially constant concentra mixtures of propylene glycol esters and glycerol esters, tions of the agents of the invention within the body over an mono- and diglycerides, sterol and sterol derivatives, poly extended period of time; (iii) formulations that sustain the ethylene glycol sorbitan fatty acid esters, polyethylene glycol agent(s) action during a predetermined time period by main alkyl ethers, Sugar esters, polyethylene glycol alkyl phenols, taining a relatively constant, effective level of the agent(s) in polyoxyethylene-polyoxypropylene block copolymers, Sor the body with concomitant minimization of undesirable side bitan fatty acid esters, lower alcohol fatty acid esters, or ionic effects associated with fluctuations in the plasma level of the Surfactants. agent(s) (sawtooth kinetic pattern); (iv) formulations that 0212. The compounds described herein can also beformu localize action of agent(s), e.g., spatial placement of a con lated as part a lotion Such as a moisturizing lotion. Exemplary trolled release composition adjacent to or in the diseased lotion formulations are described in U.S. Pat. Nos. 4,595,586: tissue or organ; (V) formulations that achieve convenience of 4,459,285; 3,867,528; 3,265,571; 4,512,978; 4,564,462: dosing, e.g., administering the composition once per week or 4,165,385; 3,062,721: 3,949,071; 4,482,537; 4,295,985; once every two weeks; and (vi) formulations that target the 2,507,236; and 3,987,775. action of the agent(s) by using carriers or chemical derivatives to deliver the compound to a particular target cell type. Solid Dosage Forms for Oral Use Administration of the compound in the form of a controlled 0213 Formulations for oral use include tablets containing release formulation is especially preferred for compounds the active ingredient(s) in a mixture with non-toxic pharma having a narrow absorption window in the gastro-intestinal ceutically acceptable excipients, and Such formulations are tract or a relatively short biological half-life. known to the skilled artisan (e.g., U.S. Pat. Nos. 5,817.307. 0208 Any of a number of strategies can be pursued in 5,824,300, 5,830,456, 5,846,526, 5,882,640, 5,910,304, order to obtain controlled release in which the rate of release 6,036,949, 6,036,949, 6,372,218, hereby incorporated by ref. outweighs the rate of metabolism of the compound in ques erence). These excipients may be, for example, inert diluents tion. In one example, controlled release is obtained by appro or fillers (e.g., Sucrose, Sorbitol, Sugar, mannitol, microcrys priate selection of various formulation parameters and ingre talline cellulose, starches including potato starch, calcium dients, including, e.g., various types of controlled release carbonate, sodium chloride, lactose, calcium phosphate, cal compositions and coatings. Thus, the compound is formu cium Sulfate, or sodium phosphate); granulating and disinte lated with appropriate excipients into a pharmaceutical com grating agents (e.g., cellulose derivatives including microc position that, upon administration, releases the compound in rystalline cellulose, starches including potato starch, a controlled manner. Examples include single or multiple unit croScarmellose Sodium, alginates, or alginic acid); binding tablet or capsule compositions, oil solutions, Suspensions, agents (e.g., Sucrose, glucose, Sorbitol, acacia, alginic acid, emulsions, microcapsules, molecular complexes, micro Sodium alginate, gelatin, starch, pregelatinized starch, micro spheres, nanoparticles, patches, and liposomes. crystalline cellulose, magnesium aluminum silicate, car boxymethylcellulose sodium, methylcellulose, hydroxypro Topical Formulations pyl methylcellulose, ethylcellulose, polyvinylpyrrolidone, or 0209 Pharmaceutical compositions according to the polyethylene glycol); and lubricating agents, glidants, and present invention can be formulated for topical administra anti-adhesives (e.g., magnesium Stearate, Zinc Stearate, tion. Subjects can be administered effective amounts of a Stearic acid, silicas, hydrogenated vegetable oils, or talc). compound described herein by means of a solution (e.g., Other pharmaceutically acceptable excipients can be colo drops), ointment, gel, or aerosol (e.g., nebulizer). The com rants, flavoring agents, plasticizers, humectants, buffering position is typically administered to the affected area by agents, and the like. topically applying, for example, one to four drops of a solu 0214. The tablets may be uncoated or may be coated by tion or Suspension, or a comparable amount of an ointment, known techniques, optionally to delay disintegration and US 2014/0234271 A1 Aug. 21, 2014 20 absorption in the gastrointestinal tract and thereby providing composition may include Suspending, Solubilizing, stabiliz a Sustained action over a longer period. The coating may be ing, pH-adjusting agents, tonicity adjusting agents, and/or adapted to release the compound in a predetermined pattern dispersing agents. (e.g., in order to achieve a controlled release formulation) or 0220. As indicated above, the pharmaceutical composi it may be adapted not to release the agent(s) until after pas tions according to the invention may be in a form Suitable for sage of the stomach (enteric coating). The coating may be a sterile injection. To prepare such a composition, the Suitable Sugar coating, a film coating (e.g., based on hydroxypropyl active agent(s) are dissolved or Suspended in a parenterally methylcellulose, methylcellulose, methylhydroxyethylcellu acceptable liquid vehicle. Among acceptable vehicles and lose, hydroxypropylcellulose, carboxymethylcellulose, acry Solvents that may be employed are water, water adjusted to a late copolymers, polyethylene glycols, and/or polyvinylpyr suitable pH by addition of an appropriate amount of hydro rolidone), or an enteric coating (e.g., based on methacrylic chloric acid, sodium hydroxide or a suitable buffer, 1,3-bu acid copolymer, cellulose acetate phthalate, hydroxypropyl tanediol, Ringer's Solution, dextrose solution, and isotonic methylcellulose phthalate, hydroxypropyl methylcellulose Sodium chloride solution. The aqueous formulation may also acetate Succinate, polyvinyl acetate phthalate, shellac, and/or contain one or more preservatives (e.g., methyl, ethyl, or ethylcellulose). Furthermore, a time delay material such as, n-propyl p-hydroxybenzoate). In cases where one of the com e.g., glyceryl monostearate or glyceryl distearate, may be pounds is only sparingly or slightly soluble in water, a disso employed. lution enhancing or solubilizing agent can be added, or the 0215. The solid tablet compositions may include a coating solvent may include 10-60% w/w of propylene glycolor the adapted to protect the composition from unwanted chemical like. changes, (e.g., chemical degradation prior to the release of the active Substances). The coating may be applied on the Solid Dosages dosage form in a similar manner as that described in Ency 0221) The dosage of any compound described herein clopedia of Pharmaceutical Technology, Supra. depends on several factors, including: the administration 0216. The compositions of the invention may be mixed method, the amount of increase in cellular proliferation, skin together in the tablet, or may be partitioned. In one example, repair, hair growth, or skin health enhancement desired, and a first agent is contained on the inside of the tablet, and a the age, weight, and health of the subject to be treated. second agent is on the outside. Such that a substantial portion 0222. With respect to the treatment methods of the inven of the second agent is released prior to the release of the first tion, it is not intended that the administration of a compound agent. to a subject be limited to a particular mode of administration, 0217 Formulations for oral use may also be presented as dosage, or frequency of dosing; the present invention contem chewable tablets, or as hard gelatin capsules wherein the plates all modes of administration, including oral, cutaneous, active ingredient is mixed with an inert Solid diluent (e.g., Subcutaneous, intramuscular, intravenous, intraperitoneal, potato starch, lactose, microcrystalline cellulose, calcium intravesicular, intraarticular, intralesional, or any other route carbonate, calcium phosphate, or kaolin), or as soft gelatin Sufficient to provide a dose adequate to increase cellular pro capsules wherein the active ingredient is mixed with water or liferation, enhance skin repair or skin health, or promote hair an oil medium, for example, peanut oil, liquid paraffin, or growth treat. The compound may be administered to the olive oil. Powders and granulates may be prepared using the Subject in a single dose or in multiple doses. For example, a ingredients mentioned above under tablets and capsules in a compound described herein may be administered once a conventional manner using, e.g., a mixer, a fluid bed appara week for, e.g., 2, 3, 4, 5, 6, 7, 8, 10, 15, 20, or more weeks. It tus, or spray drying equipment. is to be understood that, for any particular Subject, specific dosage regimes should be adjusted overtime according to the Parenteral Compositions individual need and the professional judgment of the person 0218. A composition containing a compound described administering or Supervising the administration of the com herein oridentified using the methods of the invention may be pound. For example, the dosage of a compound can be administered parenterally by injection, infusion, or implan increased if the lower dose does not provide sufficientactivity tation (Subcutaneous, intravenous, intramuscular, intraperito in the treatment of a metabolic disorder (e.g., diabetes or neal, or the like) in dosage forms, formulations, or via Suitable obesity). Conversely, the dosage of the compound can be delivery devices or implants containing conventional, non decreased if the metabolic disorder is reduced or eliminated. toxic pharmaceutically acceptable carriers and adjuvants. 0223) While the attending physicianultimately will decide The formulation and preparation of Such compositions are the appropriate amount and dosage regimen, a therapeutically well known to those skilled in the art of pharmaceutical effective amount of a compound described herein may be, for formulation. example, in the range of 0.0035 ug to 20 g/kg body weight/ 0219 Compositions for parenteral use may be provided in day or 0.010 ug to 140 ug/kg body weight/week. Desirably a unit dosage forms (e.g., in single-dose ampoules), or in vials therapeutically effective amount is in the range of 0.025ug to containing several doses and in which a suitable preservative 10 ug/kg, for example, at least 0.025, 0.035, 0.05, 0.075, 0.1, may be added (see below). The composition may be in form 0.25, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 7.0, 8.0, or of a solution, a Suspension, an emulsion, an infusion device, 9.0 ug/kg body weight administered daily, every other day, or ora delivery device for implantation, or it may be presented as twice a week. In addition, a therapeutically effective amount a dry powder to be reconstituted with water or another suit may be in the range of 0.05 ug to 20 ug/kg, for example, at able vehicle before use. Apart from the active agent(s), the least 0.05, 0.7, 0.15, 0.2, 1.0, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, composition may include Suitable parenterally acceptable 10.0, 12.0, 14.0, 16.0, or 18.0 g/kg body weight adminis carriers and/or excipients. The active agent(s) may be incor tered weekly, every other week, or once a month. Further porated into microspheres, microcapsules, nanoparticles, more, a therapeutically effective amount of a compound may liposomes, or the like for controlled release. Furthermore, the be, for example, in the range of 100 ug/m to 100,000 g/m US 2014/0234271 A1 Aug. 21, 2014

administered every other day, once weekly, or every other cell line (Beretta et al., Cell Cycle. 4:1625-1631, 2005), we week. In a desirable embodiment, the therapeutically effec performed chromatin immunoprecipitations using an anti tive amount is in the range of 1000ug/m to 20,000 g/m, for body for p63. RT-PCR of the associated DNA demonstrated example, at least 1000, 1500, 4000, or 14,000 ug/m of the that pó3 was bound to a previously defined binding site in the compound administered daily, every other day, twice weekly, p57 promoter (FIG. 1J). Thus, TAp63 regulation of SKPs weekly, or every other week. self-renewal likely involves regulation of p57. 0228 To quantify the number of p57 positive cells in Example 1 TAp63-/- cells and in wild-type cells, both types of cells 0224 pé3 Inhibits Cellular Proliferation were stained as shown in FIG. 1I, and the percentage of p57 0225. To determine whether there is a cell-autonomous positive cells was calculated. From these results, approxi requirement for tumor antigen p63 (TAp63) in the mainte mately 50% of the wild-type cells were identified as being nance of dermal stem cells, we isolated and analyzed SKPs p57 positive, whereas only 18% of the TAp63-/- cells were from wild-type and p63 null mice. We used RT-PCR to char p57 positive (FIG. 1K). acterize the p53 family members expressed in cultured wild 0229 We also determined whether the hyperproliferation type SKPs. Consistent with the expression of p83 in dermal phenotype in TAp63-/- SKPs could be rescued by p57Kip2 sheath cells in vivo, SKPs expressed TAp63 mRNA, whereas (p57). To do this, TAp63-/- SKPs were plated on an adherent p63 expression was not observed in cells taken from p63-/- substrate, transfected with either the empty vector or a mice (FIG. 1A). These cells did not express dominant nega p57Kip2 expression vector, and immunostained two days tive pé3 (dNp63) or p73, and p53 levels were similar in later for p57Kip2 and Ki67. Under these conditions, approxi p63-/- and p63+/+SKPs (FIG. 1B). Immunostaining with mately 45% of TAp63-/- cells transfected with the empty anti-p63 confirmed that wild-type, but not p63-/-, SKPs vector were dividing, as monitored by Ki67 expression, and expressed detectable levels of p63 protein (FIG.1C). Thus, of none of them expressed p57Kip2 (FIG. 1L). By contrast, the p53 family members, SKPs express only TAp63 and p53. approximately 20% of the cells transfected with p57Kip2 0226 To determine whether TAp63 is necessary for main expressed detectable levels of this protein, and none of these tenance of SKPs, the cells were isolated from the skin of transfected cells coexpressed Ki67 (FIG.1L). Thus, p57Kip2 TAp63-/- and TAp63+/+ mice at various times postnatally. completely rescued the hyperproliferation, indicating that Immunocytochemical analysis of SKP spheres for fibronec TAp63 regulates SKPs self-renewal at least in part by regu tin, nestin, vimentin, and versican demonstrated that there lating p57Kip2. were no overt differences in expression of these markers for SKPs with the loss of TAp63 (FIG. 1D). By contrast, immu Example 2 nostaining for the proliferation marker Ki67 demonstrated that TAp63-/- neonatal SKPs proliferated approximately Screen for Compounds that Enhance Cellular 2-3-fold more than their wild-type counterparts. Similar Proliferation results were obtained when SKPs were isolated from the rudimentary dermis that is present in E18 pé3-/- mice (FIGS. 0230. To identify molecules (e.g., small molecules) that 1E and 1F). To determine whether this increased proliferation promote proliferation of SKPs, a simple, reproducible and reflected increased self-renewal, SKPs were dissociated to robust assay that measures cell proliferation using Alamar single cells, plated at low density in medium containing meth Blue(R) dye, which yields a fluorescent signal in a response to ylcellulose, and the percentage of cells that initiated a new metabolic activity, was developed. Compounds (5uM) were sphere was determined. As seen in the proliferation assay, added in singlet to 96-well uncoated plates, 3000 early pas newborn TAp63-/- and E18 pé3-/- SKPs self-renewed sage dissociated sphere cells were robotically seeded, and approximately 4 times more robustly than did wildtype SKPs plates were incubated in basal growth medium. After 30 (FIG. 1G). When TAp63-/- SKPs were isolated from 1 hours, Alamar Blue was added and its reduction assessed after month old mice, they still proliferated and self-renewed more another 24 hours. Typically, there is an 8-10 fold difference in than did wild-type SKPs, although the magnitude of the Alamar increase was reduced to approximately 1.5-fold (FIG.1G). In 0231 Blue reduction between positive and negative con spite of this hyperproliferation, TAp63-/- and TAp63+/+ trols. A compound was identified as a hit if Alamar Blue SKPs were both able to differentiate into both neural and reduction is increased by three standard deviations from the mesodermal cell types under previously defined conditions mean of all the compounds in aparticular screen. In our assay, (FIG.2A), and, when transplanted into the neural crest migra the variability of signals are low, with CV values ranging from tory stream of cHH stage 18 chicks in ovo, both populations 3.5-4.5% and the dimensionless statistical parameters Zand of SKPs migrated into neural crest targets (FIG. 2B). Thus, Z factors >0.5, indicative of an excellent assay. The chemical TAp63 regulates SKP proliferation and self-renewal, but does libraries we used include the LOPAC, Prestwick, Biomol, and not overtly affect their phenotype or differentiation capacity. Spectrum collections, which comprise 3,500 unique low-mo 0227 Thus, TAp63 may normally function to dampen the lecular weight compounds, including both natural products self-renewal rate of SKPs, potentially as a mechanism to and synthetic chemicals, known drugs and drug-like com ensure that the SKPs last for the lifetime of the animal. One pounds, and phosphatase and kinase inhibitors. These screens p63 target that decreases cellular proliferation is p57. RT were done at the same time as high-throughput screens using PCR analysis demonstrated that p57 mRNA levels were human neuroblastoma tumor cells with the goal of identifying reduced in TAp63-/- and p63-/- SKPs (FIG. 1H). Immuno drugs that are cytotoxic for cancer but not for normal cells cytochemistry for p57 confirmed this result, and demon (SKPs) (FIG. 3). strated that p57 protein levels were decreased in SKPs in the 0232. Using this screen, we identified several compounds absence of TAp63 (FIG. 1I). To confirm that p57 is a direct that enhance the proliferation of human SKPs, shown in the target of p83 in SKPs, as it is in human keratinocyte HaCat Table below: US 2014/0234271 A1 Aug. 21, 2014 22

human SKPs self-renewal in a dose-response manner. The Acyclovir Dorzolamide hydrochloride Mexicanolide graphs show pooled data from two set of experiments, in Alprostadil S(-)Eticlopride hydrochloride MG 624 which four different cell lines were used. Aristolochic acid Evoxine Pramoxine hydrochloride 0238 FIG. 9 is a graph that shows that compounds at 100 Carbadox Guaifenesin Tannic acid Chlorpyrifos Hydroxyprogesterone Targinine nM promote human SKPs self-renewal. Pooled data from two hydrochloride different experiments show robust increase of the number of Cyclocreatine Kaempferol Yohimbic acid spheres formed after drug treatments at 100 nM. The results 7,4'- Linamarin indicate that these compounds promote the ability of stem Dimethoxyisoflavone cells to produce another stem cell. 0239 FIGS. 10 and 11 are each a series of photographs 0233. To confirm these hits dose-response curves (IC50s) showing that alprostadil and kaempferol increase the size of were determined using human neuroblastoma tumor cells in spheres. Human SKPs were treated with the indicated com our standard Alamar Blue proliferation assay. In addition, we pound or vehicle for seven days, as described above. At the monitored proliferation over one week by quantifying cell end of the treatment period, cells were stained with Hoechst. numbers with trypan blue, and further, examined sphere for An increase in the size of the spheres was observed in the mation over one week (as described in FIG. 4) with at least 3 alprostadil- and kaempferol-treated cells compared to vehicle independent human SKP cell isolates. The high-throughput control, indicating that these drugs increase cell proliferation screens with human neuroblastoma tumor cells identified within a sphere. several small molecules that are effective in inhibiting prolif 0240 FIGS. 12 and 13 are each a series of graphs showing eration of multiple neuroblastoma patient samples in vitro that certain compounds that promote self-renewal of human and Xenograft neuroblastoma tumors in vivo (FIG. 4). SKPs also function on mouse SKPs. SKPs were isolated from mouse back and whisker and treated with different concen Example 3 tration of the indicated compounds. The results are pooled data from two experiments. A total of four whisker and four Characterization of MG 624 back samples were used. 0234. The proliferative effects of MG 624 on SKPs and 0241 FIG. 14 is a graph showing that certain compounds neuroblastomatumor initiating cells have also been studied in at 100 nM promote mouse SKPs self-renewal. Pooled data detail. Dose response curves for proliferation of human SKPs from two experiments show robust increase of the number of and neuroblastoma tumor initiating cells in response to MG spheres formed for both back and whisker cells after drug 624 were generated in the presence of EGF and FGF2. At 200 treatments at 100 nM. nM. MG 624, SKP cell proliferation is enhanced (FIG. 5). 0242 FIG. 15 is a series of graphs that summarize the in Example 4 vitro sphere assay data. Dose-Response for Proliferation of Human SKPs Example 6 0235. To characterize enhancement of proliferation Assays Demonstrating that Topical Application of brought about by the agents identified above, dose-response Certain Compounds Induces Anagen Hair Cycle and curves for MG624, GSK313, pramoxine, kaempferol, and Greater Follicle Density alprostadil in presence of EGF and FGF2 were generated (FIGS. 6A-6E). In all cases, enhanced proliferation at 100 nM 0243 We tested compounds to determine whether their concentration of each agent was observed as compared to topical application could induce anagen hair cycle and fol cells grown in the absence of the agent. licle density. Briefly, seven-week old C57BL/6 mice were used. Their dorsal skin was pinkish in color, indicating that Example 5 the hair was intelogen (resting) phase. Mice were induced to enter anagen phase by being depilated in the dorsal region. Assays Demonstrating that Certain Compounds can Compounds (100 mL) were topically applied onto the dorsal Induce Proliferation of Human and Mouse SKPs skin once daily for 3 weeks. Three mice were used for each 0236 FIG. 7 is a schematic presentation of an in vitro compound group. Approximately 30 hairs from each mouse sphere assay employed to demonstrate the ability of com were plucked at specific days (day 16, day 19, and day 23) and pounds to promote proliferation of human and mouse SKPs the hair length was measured. Additionally, skin samples for we performed a secondary drug screen. SKPs were isolated cryosectioning were obtained from the treated dorsal region from mouse back, whisker, and human foreskin and allowed of each mouse. to grow in spheres. After 2-3 passages, spheres were dissoci 0244 All compounds were resuspended in vehicle solu ated and 3000 cells were plated in each well of a 96 well plate. tion (1.2 propanediol:EtOH:H20 (40:20:40)). Compound Cells were then treated in triplicate with different concentra concentrations were as follows: alprostadil 40 ug/mL, tions of drugs or with vehicle (DMSO) only. Drugs were kaempferol 28.6 ug/mL, pramoxine 329.8 ug/mL: MG 624 applied again on day 3 and number of spheres was analyzed 45.13 g/mL. Also tested was Gold Bond topical anesthetic on day 7. cream. Latanoprost (150 g/mL) was used as a positive con 0237 FIG. 8 depicts quantification of the percentage of trol and AKT inhibitor A443654 (100 uM) was used as a spheres formed after treatments of human SKPs with differ negative control. ent concentration of drugs. The sphere assay was performed 0245 FIG.16 is a table showing that topical application of on human SKPs treated with specific drugs or vehicle for 7 certain compounds promote hair growth. Mice were treated days, as described above. Certain compounds promoted topically with compounds described above. At specific time US 2014/0234271 A1 Aug. 21, 2014 points, approximately 30-40 hairs were plucked from each Example 8 mouse and their length was measured. Three mice were used to test each compound. Single Agent In Vivo Data 0246 FIGS. 17 and 18 are each a series of graphs showing 0252. The efficacy of alprostadil and kaempferol in quantification of hair length on day 23. Hair length promoted increasing length of hair growth in mice has been demon by compounds is shown in distribution histograms relative to strated. Sox2-EGFP mice, 7-8 weeks old, were shaved on the the control group. Hair length was binned in classes of 200um dorsal back to synchronize the hair cycle, and kaempferol and each class was expressed as a percentage of total hair (Kae) oralprostadil (Alp) was topically applied daily. Control population. The distribution histograms show a shift versus mice were treated with vehicle only. At days 16, 19, and 23, longer hair length, particularly in the groups treated with about 30-40 hairs were plucked from each mouse, and their alprostadil or kaempferol. length was measured. Three or four mice were used in each 0247 FIG. 19 is a series of photomicrographs and graphs group. An increase in hair length is observed in the alpros showing that topical application of the indicated compounds tadil- and kaempferol-treated mice as compared to control induces dermal thickness. Dorsal skin of animals topically mice (FIG.22A). Hair length promoted by the compounds on treated with compounds described above, then isolated, cryo day 23 is shown in distribution histograms relative to the sectioned and stained with hematoxylin and eosin. Dermal control group. Hair length was binned in classes of 200 um, thickness was manually measured randomly through the and each class was expressed as a percentage of total hair slices. A mean of three measurements through the slice were population. The distribution histograms show a shift to longer performed and about 20-30 slices were analyzed for each hair lengths in the groups treated with kaempferol (FIG.22B) mouse within each compound group. and alprostadil (FIG.22C), as compared to the control mice. 0248 FIG. 20 is a series of photomicrographs and graphs Other Embodiments showing that alprostadil, kaempferol, pramoxine, or MG 624 induces anagen hair cycle and follicle density. The density is 0253 All patents, patent applications, and publications expressed as number of follicles per mm whereas the anagen mentioned in this specification are herein incorporated by hair follicles are determined by morphology. reference, including U.S. Provisional Application No. 61/101443, filed Sep. 30, 2008, and International Applica tion No. PCT/US2009/058723, filed Sep. 29, 2009, to the Example 7 same extent as if each independent patent, patent application, or publication was specifically and individually indicated to Combinations of Compounds for Inducing Anagen be incorporated by reference. Hair Cycle and Greater Follicle Density What is claimed is: 1. A method of increasing proliferation of a cell, said 0249. The use of combinations of compounds for inducing method comprising contacting said cell with a sufficient anagen hair cycle and greater follicle density is also an aspect amount of one or more of: of the invention. Any combination of two or more compounds (a) a compound that decreases p63 expression or activity, described herein can be used for this purpose, including O alprostadil and kaempferol; alprostadil and pramoxine; (b) a compound selected from the group consisting of kaempferol and pramoxine; alprostadil and MG 624; acyclovir, alprostadil, aristolochic acid, carbadox, chlo kaempferol and MG 624; and pramoxine and MG 624. rpyrifos, cyclocreatine, 7,4'-dimethoxyisoflavone, dor 0250 Combinations of (a) alprostadil and kaempferol and Zolamide, S(-)eticlopride, evoxine, guaifenesin, (b) alprostadil and pramoxine were demonstrated to increase hydroxyprogesterone, kaempferol, linamarin, mexi SKP sphere size invitro more effectively than single agents or canolide, MG 624, pramoxine, tannic acid, targinine, a DMSO control (FIG. 21). Sphere size is corrected with, and alpha-methylycacontine, mecamylamine hexametho is thus a measure of cell proliferation. SKPs were isolated nium, alsterpaullone, and yohimbic acid, or an analog from human foreskin and allowed to grow in spheres. After thereof, under conditions that support cell proliferation. 2-3 passages, spheres were dissociated and 3000 cells were 2. The method of claim 1, wherein said cell is from a tumor plated in each well of a 96 well plate. Cells were then treated cell line, is a stem cell, or is a skin-derived precursor (SKP). in triplicate with (a) 100 nM of a single drug, (b) a combina 3. The method of claim 1, wherein said cell is contacted tion of drugs in which each drug is present at a concentration with (a) alprostadil and kaempferol or (b) alprostadil and of 100 nM, or (c) vehicle (DMSO). Drugs were applied again pramoxine. on day 3. To assess how well cells proliferate within the 4. The method of claim 1, wherein said cell is being cul sphere, the size of spheres was analyzed on day 7. Positive tured in vitro. controls are kenpaullone, an inhibitor of GSK313, and alster 5. The method of claim 4, wherein said cell culture further paullone, an inhibitor of GSK3 B, CDK5/p25, and CDK1/ comprises an additional growth factor or said cell culture cyclin B. further comprises FGF2 or EGF. 0251 Sphere size in the presence of either (a) alprostadil 6. A composition comprising: and kaempferol or (b) alprostadil and pramoxine is shown in (a) an isolated stem cell; and distribution histograms relative to the control group (FIG. (b) one or more compounds that decrease p63 expression or 21). Sphere size was binned in classes of 20 Lum, and each activity and/or one or more compounds selected from class was expressed as a percentage of total sphere popula the group consisting of acyclovir, alprostadil, aris tion. The distribution histograms of the compound combina tolochic acid, carbadox, chlorpyrifos, cyclocreatine, tions show a shift to bigger sphere sizes in the presence of the 7,4'-dimethoxyisoflavone, dorzolamide, S(-) eticlo drugs as compared to the control samples. pride, evoxine, guaifenesin, hydroxyprogesterone, US 2014/0234271 A1 Aug. 21, 2014 24

kaempferol, linamarin, mexicanolide, MG 624, (a) one or more compounds that inhibit p63 expression or pramoxine, tannic acid, targinine, alpha-methyllycacon activity; and/or tine, mecamylamine hexamethonium, alsterpaullone, (b) one or more compounds selected from the group con and yohimbic acid, or an analog thereof, wherein said sisting of acyclovir, alprostadil, aristolochic acid, carba compound is present in an amount Sufficient to promote dox, chlorpyrifos, cyclocreatine, 7,4'-dimethoxyisofla proliferation of said stem cell. vone, dorzolamide, S(-)eticlopride, evoxine, guaifenesin, hydroxyprogesterone, kaempferol, linama 7. The composition of claim 6, wherein said stem cell is a rin, mexicanolide, MG 624, pramoxine, tannic acid, SKP. targinine, alpha-methyllycacontine, mecamylamine 8. The composition of claim 6 further comprising a growth hexamethonium, alsterpaullone, and yohimbic acid, or factor, FGF2, or EGF. an analog thereof. 9. The composition of claim 6, wherein said compounds 14. The method of claim 13, wherein said subject has a are (a) alprostadil and kaempferol or (b) alprostadil and wound, and said compound is administered in an amount pramoxine. Sufficient to improve healing of said wound. 10. A composition comprising: 15. The method of claim 13, wherein said compounds are (a) one or more compounds that decrease p63 expression or (a) alprostadil and kaempferol or (b) alprostadil and pramox activity and/or one or more compounds selected from 1C. the group consisting of acyclovir, alprostadil, aris 16. The method of claim 13, wherein said compound is tolochic acid, carbadox, chlorpyrifos, cyclocreatine, administered topically or systemically. 7,4'-dimethoxyisoflavone, dorzolamide, S(-)eticlo 17. The method of claim 13, wherein said administration of pride, evoxine, guaifenesin, hydroxyprogesterone, said one or more compounds reduces wrinkles in the skin of kaempferol, linamarin, mexicanolide, MG 624, said Subject. pramoxine, tannic acid, targinine, alpha-methyllycacon 18. The method of claim 13, wherein said subject is a tine, mecamylamine hexamethonium, alsterpaullone, human. and yohimbic acid, or an analog thereof, and 19. A method of identifying a compound capable of alter (b) a carrier suitable for topical administration. ing cell proliferation, said method comprising: 11. The composition of claim 10, wherein said carrier is a (a) contacting a SKP with a candidate compound; and lotion. (b) determining the proliferation rate of said SKP, wherein 12. The composition of claim 10, wherein said compounds an alteration in the proliferation rate of said SKP in the are (a) alprostadil and kaempferol or (b) alprostadil and presence of Said compound as compared to in the pramoxine. absence of said compound, indicates that said com 13. A method of increasing SKP proliferation in a subject, pound alters the rate of cell proliferation. promoting hair growth, repairing skin, or improving skin 20. The method of claim 19, wherein said SKP cell is a health in a Subject, said method comprising administering a human, mouse, or rat cell. sufficient amount of: k k k k k