Minoxidil adjuvant therapies

Abstract

Compositions and methods are disclosed herein for inducing (up-regulating) the expression and / or activity of sulfotransferases in the hair follicles, e.g., the scalp. The compositions include (i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person; (ii) an alkalinizing agent; (iii) liposomes encapsulating at least some of the alkalinizing agent; (iv) an antioxidant configured to prevent oxidative degradation of the liposomes; and (v) a buffer system configured to increase dermal penetration of the liposomes, and / or stabilize the liposome solution; and (vi) a solvent; and (vii) minoxidil or a salt thereof. Additional methods and compositions include the use of the topical solution containing an alkalinizing agent or an alkalinizing agent used with a penetration enhancer for up-regulating the sulfonating capacity of hair bearing skin, hair follicles, and / or keratinocyte cells. In addition, compositions and methods for increasing or decreasing the growth rate of hair follicles is disclosed by altering the intracellular pH. Additionally, methods for preparing a stable liposomal containing solution are described.

Description

MINOXIDIL ADJUVANT THERAPIESCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and the benefit of U.S. Provisional Application Serial No. 63 / 594,884 filed October 31, 2023, the entire contents of which is incorporated herein by reference in its entirety.FIELD

[0002] The present disclosure relates to compositions that increase the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, induce sulfotransferase in a hair follicle, induce expression of sulfotransferase, increase sulfotransferase activity, upregulate sulfotransferase, convert minoxidil non-responders to minoxidil responders, and / or improve minoxidil response. The compositions include minoxidil and (i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person; (ii) an alkalinizing agent; (iii) liposomes encapsulating at least some of the alkalinizing agent; (iv) an antioxidant configured to prevent oxidative degradation of the liposomes; (v) a buffer system configured to increase dermal penetration of the liposomes, and / or stabilize the liposome solution; and (vi) a solvent. The present disclosure further provides methods of making and using the compositions.BACKGROUND

[0003] In 1988 the US FDA approved 2% topical minoxidil solution as an OTC drug for the treatment of androgenetic alopecia (AGA). Since the FDA approval, minoxidil has become the mainstay therapy for AGA. However, the effectiveness of minoxidil in the general population is low, only 39% of patients respond to the drug (See Olsen E.A., et al., A multicenter, randomized, placebo-controlled, double-blind clinical trial of a novel formulation of 5% minoxidil topical foam versus placebo in the treatment of androgenetic alopecia in men. J Am Acad Dermatol. 2007 57(5):767-74). In the pivotal study submitted to the USFDA in support of the efficacy ofthe 5% topical minoxidil foam, no subjects had great improvement, 8% of the subjects had a moderate improvement, and 31% of the subjects had a slight improvement (Sec U.S. FDA Application 21-812 Medical Review).

[0004] Minoxidil is a pro-drug converted to its active form, minoxidil sulfate, by sulfotransferase enzymes present in the outer root sheath (ORS) of hair follicles (See Buhl A.E., et al., J. Invest. Dermatol. 1990 Nov; 95(5):553-7). Sulfotransferase enzymes catalyze the sulfate conjugation of many hormones, neurotransmitters, drugs, and xenobiotic compounds. These cytosolic enzymes are different in their tissue distributions and substrate specificities. The gene structure (number and length of exons) is similar among family members. The SULT1A1 enzyme is expressed in outer roots sheath of hair follicles. Minoxidil is converted to its active form (minoxidil sulfate) by SULT1 Al. See McCoy, J. et al., (2019) Journal of Biological Regulators and Homeostatic Agents 33 (3): 817-819. A large variability in sulfotransferase enzyme expression in hair is observed among people. Low sulfotransferase activity was found to be predictive to lack of response to topical minoxidil for hair re-growth. See Goren, A. et al., Dermatologic Therapy. 28 (1): 13-6. In a clinical study, a novel formula using a hypoxia mimetic pathway demonstrated to increase SULT1A1 activity in human subjects in-vivo. See Ramos, P.M., et al., (May 2020 Journal of the European Academy of Dermatology and Venereology. 34 (12): e799-e800.

[0005] It has been demonstrated that the activity of sulfotransferase in the ORS determines the clinical response to minoxidil (See Goren A. et al., Dermatol. Ther. 2014;27(3): 171-3). Sulfotransferase enzymes are expressed in abundance in the human liver. In the human liver, sulfotransferase is part of the Phase II enzymatic system that reduces xenobiotic toxicity (See Jancova P. et al., Biomed. Pap. Med. Fae Univ. Palacky Olomouc Czech Repub. 2010;154(2): 103-16).

[0006] Minoxidil sulfate is required for both the promotion of hair regrowth and the vasodilatory effects of minoxidil. Sulfotransferase enzymes are located in both the skin and the liver and are important Phase II xenobiotic metabolizing enzymes for a number of phenolic molecules including minoxidil (See Nimmagadda D. et al, Indian J. Exp. Biol. 2006; 44(3): 171-

[0007] Additionally, altering intracellular or extracellular pH is an important regulatory mechanism, which can influence cellular function and lead to cell differentiation in a range of stem cells (See Charruyer A., et al., Curr. Probl. Dermatol. 2018; 54:71-78). Cell differentiation is associated with the altered expression of many proteins including xenobiotic-metabolizing enzymes. Specifically, increased sulfotransferase is a marker for keratinocyte differentiation (See, Johnson G.A., et al., J. Invest. Dermatol. 1992 May; 98(5):730-3).

[0008] A colorimetric assay for detecting sulfotransferase in plucked hair samples described in U.S. Patent No. 8,691,518, which is incorporated herein in its entirety by reference.

[0009] Many methodologies exist for increasing the dermal penetration of active ingredients through the stratum comeum (SC). For example, the use of penetration enhancing chemical agents have been described (See Trammer H. et al., Skin Pharmacol. Physiol. 2006; 19(2): 106- 21). However, many of these agents alter the structure of the SC aggressively and can lead to irreversible damage to the SC. An alternative approach to the use of SC modifying agents is the use of encapsulation techniques (See Tiwari N. et al., Angew Chem. Int. Ed. Engl. 2022 Jan 17; 61(3):e202107960). Such techniques surround the active ingredients with carrier molecules sharing similar morphology to cellular membranes that interact more gently with the SC and can be used to deliver large payloads. Such systems are more biocompatible and offer advantages such as biodegradability and low toxicity. Many examples of such encapsulation techniques have been described - for example, liposomes, micelles, nanogels, lipid nanoparticles, selenium nanoparticles, bilosomes, dendrimers and carbon nanotubes. Unfortunately, the use of encapsulation technologies introduces additional challenges to the production of a stable topical formulas. Many of the encapsulating systems used are susceptible to hydrolysis, oxidation, and aggregation or breakdown of the macro encapsulation structure.

[0010] U.S. Patent No. 11,628,132 discloses topical solution compositions for inducing (up- regulating) the expression of sulfotransferases in the hair follicles, e.g., the scalp, that include an alkalinizing agent, a penetration enhancer and a minoxidil salt.

[0011] U.S. Patent Application Publication No. U.S. 2021 / 0059920 discloses topical solution compositions for inducing (up-regulating) the expression of sulfotransferases in the hair follicles, e.g., the scalp, that include an alkalinizing agent and a penetration enhancer.

[0012] Novel formulation technologies are required to successfully implement a stable and operational topical formula containing encapsulated actives.SUMMARY

[0013] In a first embodiment, the present disclosure provides a liposomal solution or emulsion Composition (Composition 1) for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfo transferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising:(i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person;(ii) an alkalinizing agent;(iii) liposomes encapsulating at least some of the alkalinizing agent;(iv) an antioxidant configured to prevent oxidative degradation of the liposomes; and(v) a buffer system configured to increase dermal penetration of the liposomes, and / or stabilize the liposome solution or emulsion; and(vi) a solvent; and(vii) minoxidil or a salt thereof; wherein the alkalinizing agent and the buffer system are the same, and / or comprise the same components.

[0014] The present disclosure further provides the following compositions:1.1 Composition 1 , wherein the agent configured to increase in the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is an inorganic sulfur source;1.2 Composition 1 or 1.1, wherein the agent configured to increase in the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is cysteine, L-cysteine, hydrogen sulfide, elemental sulfur, sulfite, thiosulfate, polythionates, magnesium sulfate, sodium sulfate or sodium metabisulfite; for example sodium metabisulfite;1.3 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the agent configured to increase in the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells issodium metabisulfite;1.4 Any of the foregoing Compositions 1 or 1.1 ct scq., wherein the alkalinizing agent comprises a pH regulating agent;1.5 Composition 1.4, wherein the pH regulating agent comprises or consists of a buffer system selected from bicarbonate, acetate, citrate, TRIS, or HEPES;1.6 Composition 1.4, wherein the pH regulating agent comprises or consists of TRIS / TRIS HC1 buffer at a pH above 7; for example from 7-10; for example from 7-9; for example from 8 to 9;1.7 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the buffer system is selected from one or more of carbonate, acetate, citrate, TRIS or HEPES;1.8 Composition 1.7, wherein the buffer system is selected from one or more of sodium bicarbonate / sodium carbonate, acetic acid / sodium acetate, mono-, di- or trisodium citrate, and TRIS / TRIS HC1;1.9 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the buffer system comprises TRIS / TRIS HC1 at a pH of from 8 to 9;1.10 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the alkalinizing agent and the buffer system both comprise TRIS, e.g. TRIS / TRIS HC1;1.11 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the antioxidant is selected from ascorbic acid, monothioglycerol, potassium metabisulfite, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium sulfite, sodium thiosulfate, tocopherols or ester derivatives thereof such as tocopherol acetate, sodium metabisulfite, butylated hydroxy toluene (BHT), butylated hydroxyanisole (BHA), ascorbyl palmitate, propyl gallate, acai oil, alpha lipoic acid, green tea extract, retinal, vitamin C, coenzyme Q10 (Co Q-10), isoflavones, polyphenols, curcumin, turmeric, pomegranate, rosemary extract, glutathione, selenium, zinc, a chelating molecule, ethylenediaminetetraacetic acid (EDTA), disodium EDTA, tetrasodium EDTA, pentasodium penetrate, sodium metasilicate and phosphate derivatives, etidronic acid and / or its derivatives, and / or galactaric acid; tocopherols, e.g. tocopherol acetate.1.12 Composition 1.11, wherein the antioxidant comprises tocopherol acetate.1.13 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the Composition further comprises a penetration enhancer1.14 Composition 1.13, wherein the penetration enhancer is selected from diethylene glycol monocthyl ether, and a polyethylene glycol derivative of a mixture of mono-, di-, and triglycerides of caprylic and capric acids with an average of 6 moles of ethylene oxide; wherein the penetration enhancer is selected from longer chain 1,2-glycols, e.g., Arachidyl Glycol (1,2- Eicosanediol, CAS 39825-93-9), Cetyl Glycol (1,2-Dihydroxyhexadecane, 1 ,2-Hexadecanediol, 1,2-Hexadecylene Glycol, 2-Hydroxycetyl Alcohol, CAS 6920-24-7), Hexacosyl Glycol (Hexacosil glycol), Lauryl Glycol (1,2-Dihydroxy dodecane, 1,2-Dodecanediol, 1,2-Dodecylene Glycol, CAS 1119-87-5), Myristyl Glycol ( 1 ,2-Tetradecanediol, CAS 21129-09-9), Octacosanyl Glycol (1,2-Octacosanediol, CAS 97338-11-9), Stearyl Glycol (1,2-Dihydroxyoctadecane, 1,2- Octadecanediol, CAS 20294-76-2), Caprylyl Glycol (Capryl Glycol, 1 ,2-Dihydroxyoctane, 1,2- Octanediol, 1,2-Octylene Glycol, CAS 1117-86-8), Decylene Glycol (1,2-Decanediol, CAS 1119-86-4), Pentylene Glycol (1,2-Dihydroxypentane, 1,2-Pentanediol, CAS 5343-92-0), 1,2- Butanediol (1,2-Butylene Glycol, 1 ,2-Dihydroxybutane, CAS 584-03-2), 1 ,2-Hexanediol (1,2- Dihydroxyhexane, CAS 6920-22-5), C14-18 Glycol (Ethylene Glycol Fatty Acid Ester), C15-18 Glycol (Alkylene (15-18) Glycol, Cetyl Stearyl Vicinal Glycol, Glycols C15-18, CAS 70750-40- 2, CAS 92128-52-4), C18-30 Glycol (Ethylene Glycol Fatty Acid Ester), C20-30 Glycol (Alkylene (20-30) Glycol).1.15 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the Composition further comprises a chelating agent;1.16 Composition 1.15, wherein the chelating agent is tetrasodium EDTA;1.17 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the Composition further comprises a preservative;1.18 Composition 1.17, wherein the preservative is an antibacterial and / or antifungal agent;1.19 Composition 1.18, wherein the preservative includes or consists of phenoxyethanol and ethylhexylglycerin, e.g. EUXYL PE 9010, or the preservative includes or consists of caprylhydroxamic acid and 1 ,2-hexanediol and propanediol, e.g., Spectrastat PHL;1.20 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the Composition further comprises a surfactant;1.21 Composition 1.20, wherein the surfactant comprises polysorbate 80;1 .22 Any of the foregoing Compositions 1 or 1 .1 et seq., wherein the Composition further comprises glycerin;1.23 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the Composition further comprises a fragrance compound;1.24 Composition 1, wherein the Composition further comprises one or more of: a preservative, for example an antibacterial / antifungal agt., e.g. EUXYLPE 9010 and / or Spectrastat PHL; a fragrance compound; a surfactant, e.g., polysorbate 80; and glycerol.1.25 Composition 1, wherein: the agent configured to increase in the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is an inorganic sulfur source, e.g. sodium metabisulfite; the alkalinizing agent and the buffer system each comprise TRIS / TRIS HC1; the antioxidant comprises tocopherol; e.g. tocopherol acetate; the solvent is water; and the liposomes comprise phosphatidylcholine.1.26 Composition 1.25, wherein the Composition further comprises: a chelating agent that is tetrasodium EDTA; an antibacterial / antifungal agt., e.g. EUXYL PE 9010; a surfactant, e.g., polysorbate 80; glycerol; and optionally, a fragrance compound;1.27 Composition 1, wherein the Composition comprises the following components:1.28 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the minoxidil is present as a salt, e.g., minoxidil tromethamine, minoxidil carbonate, minoxidil acetate, or minoxidil citrate;1.29 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the minoxidil is present in the liposomes;1.30 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the minoxidil is present outside of the liposomes;1.31 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the minoxidil is present both inside and outside of the liposomes;1.32 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the composition is a mixture, an emulsion, or a solution;1.33 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the minoxidil is added to the other ingredients of the composition at the point of use to form the composition;1.34 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the composition is configured to be administered by applying the composition at a predetermined frequency;1.35 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the composition is formulated as a cosmetic product;1 .36 Any of the foregoing Compositions 1 or 1 .1 et seq., wherein the composition is formulated as any one or combination of a time release vehicle, a cream, a solution, an emulsion, a lotion, a serum, an ointment, a spray, an aerosol medium, a capsule, a shampoo, a gel, a foam, a cosmetic, a hair conditioner, a hair care product, a hair styling product (e.g., a styling gel, a styling foam or a hair conditioner), a hair mask, a deodorant, an antiperspirant, a moisturizer, or a shaving cream or gel;1.37 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the minoxidil is presentin a concentration of 2-15%.1.38 Any of the foregoing Compositions 1 or 1.1 ct scq., wherein the composition is a therapeutic product.1.39 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the composition is a cometic product.1.40 Any of the foregoing Compositions 1 or 1.1 et seq., further comprising SULT1A1 protein.1.41 Composition 1.40, wherein the SULT1A1 protein is human SULT1A1 protein1.42 Composition 1.40 or 1.41, wherein the SULT1A1 protein is added as a solution in a buffer, or as a solid protein.

[0015] The present disclosure further provides, in a second embodiment, a solution or emulsion Composition (Composition 2) for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising:(i) minoxidil;(ii) an alkalinizing agent; and(iii) a penetration enhancer.

[0016] The present disclosure further provides the following compositions:2.1 Composition 2, wherein the alkalinizing agent comprises a pH regulating agent that maintains the pH of the Composition at a pH of 7 or greater; for example at a pH of from 7-10; for example at a pH of from 7-9; for example at a pH of from 8-9;2.2 Composition 2.1, wherein the pH regulating agent comprises or consists of a buffer system selected from bicarbonate, acetate, citrate, TRIS, or HEPES;2.3 Composition 2.1, wherein the pH regulating agent comprises or consists of TRIS / TRIS HC1 buffer at a pH of from 7 to 9; preferably from 8 to 9;2.4 Any of the foregoing Compositions 2-2.3, wherein the penetration enhancer is selected from diethylene glycol monoethyl ether, and a polyethylene glycol derivative of a mixture of mono-, di-, and triglycerides of caprylic and capric acids with an average of 6 molesof ethylene oxide;2.5 Any of the foregoing Compositions 2-2.3, wherein the penetration enhancer is selected from longer chain 1,2-glycols, e.g., Arachidyl Glycol (1,2-Eicosanediol, CAS 39825-93- 9), Cetyl Glycol (1,2-Dihydroxyhexadecane, 1 ,2-Hexadecanediol, 1 ,2-Hexadecylene Glycol, 2- Hydroxycetyl Alcohol, CAS 6920-24-7), Hexacosyl Glycol (Hexacosil glycol), Lauryl Glycol (1,2-Dihydroxy dodecane, 1,2-Dodecanediol, 1,2-Dodecylene Glycol, CAS 1119-87-5), Myristyl Glycol ( 1 ,2-Tetradecanediol, CAS 21129-09-9), Octacosanyl Glycol (1,2-Octacosanediol, CAS 97338-11-9), Stearyl Glycol (1,2-Dihydroxyoctadecane, 1,2-Octadecanediol, CAS 20294-76-2), Caprylyl Glycol (Capryl Glycol, 1,2-Dihydroxyoctane, 1,2-Octanediol, 1,2-Octylene Glycol, CAS 1117-86-8), Decylene Glycol ( 1 ,2-Decanediol, CAS 1119-86-4), Pentylene Glycol (1,2- Dihydroxypentane, 1,2-Pentanediol, CAS 5343-92-0), 1,2-Butanediol (1,2-Butylene Glycol, 1,2- Dihydroxybutane, CAS 584-03-2), 1 ,2-Hexanediol ( 1 ,2-Dihydroxyhexane, CAS 6920-22-5), C14-18 Glycol (Ethylene Glycol Fatty Acid Ester), C15-18 Glycol (Alkylene (15-18) Glycol, Cetyl Stearyl Vicinal Glycol, Glycols C15-18, CAS 70750-40-2, CAS 92128-52-4), C18-30 Glycol (Ethylene Glycol Fatty Acid Ester), C20-30 Glycol (Alkylene (20-30) Glycol).2.6 Any of the foregoing Compositions 2-2.5, wherein the Composition further comprises an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells;2.7 Composition 2.6, wherein the agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is an inorganic sulfur source;2.8 Composition 2.6, wherein the agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is an inorganic sulfur source selected from cysteine, L-cysteine, hydrogen sulfide, elemental sulfur, sulfite, thiosulfate, poly thionates, magnesium sulfate, sodium sulfate or sodium metabisulfite; for example sodium metabisulfite;2.9 Any of the foregoing Compositions 2.6-2.8, wherein the agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is sodium metabisulfite;2.10 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the Composition further comprises an antioxidant;2.11 Composition 2.10, wherein the antioxidant is selected from ascorbic acid,monothioglycerol, potassium metabisulfite, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium sulfite, sodium thiosulfate, tocopherols or ester derivatives thereof such as tocopherol acetate, sodium metabisulfite, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), ascorbyl palmitate, propyl gallate, acai oil, alpha lipoic acid, green tea extract, retinal, vitamin C, coenzyme Q10 (Co Q-10), isoflavones, polyphenols, curcumin, turmeric, pomegranate, rosemary extract, glutathione, selenium, zinc, a chelating molecule, ethylenediaminetetraacetic acid (EDTA), disodium EDTA, tetrasodium EDTA, pentasodium penetrate, sodium metasilicate and phosphate derivatives, etidronic acid and / or its derivatives, and / or galactaric acid; tocopherols, e.g. tocopherol acetate.2.12 Composition 2.11, wherein the antioxidant comprises tocopherol acetate.2.13 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the Composition further comprises a chelating agent;2.14 Composition 2.13, wherein the chelating agent is tetrasodium EDTA;2.15 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the Composition further comprises a preservative;2.16 Composition 2.15, wherein the preservative is an antibacterial and / or antifungal agent;2.17 Composition 2.16, wherein the preservative includes or consists of phenoxyethanol and ethylhexylglycerin, e.g. EUXYL PE 9010, or the preservative includes or consists of caprylhydroxamic acid and 1 ,2-hexanediol and propanediol, e.g., Spectrastat PHL;2.18 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the Composition further comprises a surfactant;2.19 Composition 2.18, wherein the surfactant comprises polysorbate 80;2.20 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the Composition further comprises a solvent;2.21 Composition 2.20, wherein the solvent comprises or consists of water;2.22 Any of the foregoing Compositions 1 or 1.1 et seq., wherein the Composition further comprises glycerin;2.23 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the Composition further comprises a fragrance compound;2.24 Any of the foregoing Compositions 2 or 2.1 -2.14, wherein the Composition further comprises one or more of: a preservative, for example an antibacterial / antifungal agt., e.g. EUXYL PE 9010 and / or Spectrastat PHL; a fragrance compound; a surfactant, e.g., polysorbate 80; and glycerol.2.25 Composition 2, wherein: the alkalinizing agent comprises TRIS / TRIS HC1; and the penetration enhancer is selected from longer chain 1,2-glycols.2.26 Composition 2.25, wherein the Composition further comprises: an antioxidant that comprises tocopherol; e.g. tocopherol acetate; and optionally, an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells; for example an inorganic sulfur source; for example sodium metabisulfite;2.27 Composition 2.26, wherein the Composition further comprises one or more of: a chelating agent that is tetrasodium EDTA; an antibacterial / antifungal agt., e.g. EUXYL PE 9010; a surfactant, e.g., polysorbate 80; glycerol; and a fragrance compound;2.28 Composition 2, wherein the Composition comprises the following components:Composition 2.28, further comprising one or more of the following components:Composition 1.29, further comprising one or both of the following components:2.31 Any of the foregoing Compositions 2 or 2.1 ct scq., wherein the minoxidil is present as a salt, e.g., minoxidil tromethamine, minoxidil carbonate, minoxidil acetate, or minoxidil citrate:2.32 Any of the foregoing Compositions 2 or 2.1-2.30, wherein the minoxidil is present in non-salt form;2.33 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the composition is a mixture, an emulsion, or a solution;2.34 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the Composition comprises liposomes encapsulating at least some of the alkalinizing agent;2.35 Composition 2.34, wherein the liposomes comprise or consist of phosphatidylcholine or lecithin;2.36 Composition 2.34, wherein the liposomes comprise or consist of phosphatidylcholine;2.37 Any of the foregoing Compositions 2.28-2.30, further comprising the following component:2.38 Any of the foregoing Compositions 2.34 et seq., wherein the minoxidil is present in the liposomes;2.39 Any of the foregoing Compositions 2.34 et seq., wherein the minoxidil is present outside of the liposomes;2.40 Any of the foregoing Compositions 2.34 et seq., wherein the minoxidil is present both inside and outside of the liposomes;2.41 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the Composition further comprises a buffer system configured to increase dermal penetration of the alkalinizing agent and / or the liposomes, and / or stabilize the liposome in the solution or emulsion;2.42 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the minoxidil is added to the other ingredients of the composition at the point of use to form the composition;2.43 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the composition is configured to be administered by applying the composition at a predetermined frequency;2.44 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the composition is formulated as a cosmetic product;2.45 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the composition is formulated as any one or combination of a time release vehicle, a cream, a solution, an emulsion, a lotion, a serum, an ointment, a spray, an aerosol medium, a capsule, a shampoo, a gel, a foam, a cosmetic, a hair conditioner, a hair care product, a hair styling product (e.g., a styling gel, a styling foam or a hair conditioner), a hair mask, a deodorant, an antiperspirant, a moisturizer, or a shaving cream or gel;2.46 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the minoxidil is present in a concentration of 2-15%.2.47 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the composition is a therapeutic product.2.48 Any of the foregoing Compositions 2 or 2.1 et seq., wherein the composition is a cometic product.2.49 Any of the foregoing Compositions 2 or 2.1 et seq., further comprising SULT1A1 protein.2.50 Composition 1.40, wherein the SULT1A1 protein is human SULT1A1 protein2.51 Composition 1.40 or 1.41, wherein the SULT1A1 protein is added as a solution in a buffer, or as a solid protein.

[0017] The present disclosure further provides, in a third embodiment, a solution or emulsion Composition (Composition 3) for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising:(i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person;(ii) an alkalinizing agent;(iii) inorganic source of sulfur;(iv) a penetration enhancer meant to aid the delivery of an alkalinizing agent and inorganic source of sulfur;(v) a solvent; and(vi) minoxidil or a salt thereof.

[0018] The present disclosure further provides the following compositions:3.1 Composition 3, wherein the alkalinizing agent comprises a pH regulating agent that maintains the pH of the Composition at a pH of 7 or greater; for example at a pH of from 7-10; for example at a pH of from 7-9; for example at a pH of from 8-9;3.2 Composition 3.1, wherein the pH regulating agent comprises or consists of a buffer system selected from bicarbonate, acetate, citrate, TRIS, or HEPES;3.3 Composition 3.1, wherein the pH regulating agent comprises or consists of TRIS / TRIS HC1 buffer at a pH of from 7 to 9; preferably from 8 to 9;3.4 Any of the foregoing Compositions 3-3.3, wherein the penetration enhancer is selected from diethylene glycol monoethyl ether, and a polyethylene glycol derivative of a mixture of mono-, di-, and triglycerides of caprylic and capric acids with an average of 6 moles of ethylene oxide;3.5 Any of the foregoing Compositions 3-3.3, wherein the penetration enhancer is selected from longer chain 1,2-glycols, e.g., Arachidyl Glycol (1,2-Eicosanediol, CAS 39825-93- 9), Cetyl Glycol (1,2-Dihydroxy hexadecane, 1 ,2-Hexadecanediol, 1,2-Hexadecylene Glycol, 2- Hydroxycetyl Alcohol, CAS 6920-24-7), Hexacosyl Glycol (Hexacosil glycol), Lauryl Glycol( 1 ,2-Dihydroxy dodecane, 1,2-Dodecanediol, 1 ,2-Dodecylene Glycol, CAS 1119-87-5), Myristyl Glycol (1,2-Tetradecanediol, CAS 21129-09-9), Octacosanyl Glycol (1,2-Octacosanediol, CAS 97338-11-9), Stearyl Glycol (1,2-Dihydroxyoctadecane, 1,2-Octadecanediol, CAS 20294-76-2), Caprylyl Glycol (Capryl Glycol, 1,2-Dihydroxyoctane, 1,2-Octanediol, 1,2-Octylene Glycol, CAS 1117-86-8), Decylene Glycol (1,2-Decanediol, CAS 1119-86-4), Pentylene Glycol (1,2- Dihydroxypentane, 1,2-Pentanediol, CAS 5343-92-0), 1,2-Butanediol (1,2-Butylene Glycol, 1,2- Dihydroxybutane, CAS 584-03-2), 1 ,2-Hexanediol ( 1 ,2-Dihydroxy hexane, CAS 6920-22-5), C14-18 Glycol (Ethylene Glycol Fatty Acid Ester), C 15- 18 Glycol (Alkylene (15-18) Glycol, Cetyl Stearyl Vicinal Glycol, Glycols C15-18, CAS 70750-40-2, CAS 92128-52-4), C18-30 Glycol (Ethylene Glycol Fatty Acid Ester), C20-30 Glycol (Alkylene (20-30) Glycol).3.6 Any of the foregoing Compositions 3-3.5, wherein the inorganic sulfur source is selected from cysteine, L-cystcinc, hydrogen sulfide, elemental sulfur, sulfite, thiosulfate, polythionates, magnesium sulfate, sodium sulfate or sodium metabisulfite; for example sodium metabisulfite;3.7 Any of the foregoing Compositions 3-3.5, wherein the inorganic sulfur source comprises sodium metabisulfite;3.8 Any of the foregoing Compositions 3-3.5, wherein the agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is SULT1A1 protein;3.9 Composition 3.8, wherein the SULT1A1 protein is human SULT1A1 protein3.10 Composition 3.8 or 3.9, wherein the SULT1A1 protein is added as a solution in a buffer, or as a solid protein.3.11 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the Composition further comprises an antioxidant;3.12 Composition 3.11, wherein the antioxidant is selected from ascorbic acid, monothioglycerol, potassium metabisulfite, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium sulfite, sodium thiosulfate, tocopherols or ester derivatives thereof such as tocopherol acetate, sodium metabisulfite, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), ascorbyl palmitate, propyl gallate, acai oil, alpha lipoic acid, green tea extract, retinal, vitamin C, coenzyme Q10 (Co Q-10), isoflavones, polyphenols, curcumin, turmeric, pomegranate, rosemary extract, glutathione, selenium, zinc, a chelating molecule, ethylenediaminetetraacetic acid (EDTA), disodium EDTA, tetrasodium EDTA, pentasodium penetrate, sodium metasilicate and phosphate derivatives, etidronic acid and / or its derivatives, and / or galactaric acid; tocopherols, e.g. tocopherol acetate.3.13 Composition 3.12, wherein the antioxidant comprises tocopherol acetate.3.14 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the Composition further comprises a chelating agent;3.15 Composition 3.14, wherein the chelating agent is tetrasodium EDTA;3.16 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the Composition further comprises a preservative;3.17 Composition 3.16, wherein the preservative is an antibacterial and / or antifungal agent;3.18 Composition 3.17, wherein the preservative includes or consists of phenoxyethanol and ethylhexylglycerin, e.g. EUXYL PE 9010, or the preservative includes or consists of caprylhydroxamic acid and 1 ,2-hexanediol and propanediol, e.g., Spectrastat PHL;3.19 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the Composition further comprises a surfactant;3.20 Composition 3.18, wherein the surfactant comprises polysorbate 80;3.21 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the Composition further comprises a solvent;3.22 Composition 3.21, wherein the solvent comprises or consists of water;3.23 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the Composition further comprises glycerin;3.24 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the Composition further comprises a fragrance compound;3.25 Any of the foregoing Compositions 3 or 3.1 et seq, wherein the Composition further comprises one or more of: a preservative, for example an antibacterial / antifungal agt., e.g. EUXYL PE 9010 and / or Spectrastat PHL; a fragrance compound; a surfactant, e.g., polysorbate 80; and glycerol.3.26 Composition 3, wherein: the alkalinizing agent comprises TRIS / TRIS HC1; and the penetration enhancer is selected from longer chain 1,2-glycols.3.27 Composition 3.26, wherein the Composition further comprises: an antioxidant that comprises tocopherol; e.g. tocopherol acetate; and optionally, an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells; for example an inorganic sulfur source; for example sodium metabisulfite;Composition 3.27, wherein the Composition further comprises one or more of: a chelating agent that is tetrasodium EDTA; an antibacterial / antifungal agt., e.g. EUXYL PE 9010; a surfactant, e.g., polysorbate 80; glycerol; and a fragrance compound; Composition 3, wherein the Composition comprises the following components:Composition 3.29, further comprising one or more of the following components:3.31 Composition 3.30, further comprising one or both of the following components:3.32 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the minoxidil is present as a salt, e.g., minoxidil tromethamine, minoxidil carbonate, minoxidil acetate, or minoxidil citrate;3.33 Any of the foregoing Compositions 3 or 3.1-3.31, wherein the minoxidil is present in non-salt form;3.34 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the composition is a mixture, an emulsion, or a solution;3.35 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the Composition comprises liposomes encapsulating at least some of the alkalinizing agent;3.36 Composition 3.33, wherein the liposomes comprise or consist of phosphatidylcholineor lecithin;3.37 Composition 3.35, wherein the liposomes comprise or consist of phosphatidylcholine;3.38 Any of the foregoing Compositions 3.29-3.37, further comprising the following component:3.39 Any of the foregoing Compositions 3.35 et seq., wherein the minoxidil is present in the liposomes;3.40 Any of the foregoing Compositions 3.35 et seq., wherein the minoxidil is present outside of the liposomes;3.41 Any of the foregoing Compositions 3.35 et seq., wherein the minoxidil is present both inside and outside of the liposomes;3.42 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the Composition further comprises a buffer system configured to increase dermal penetration of the alkalinizing agent and / or the liposomes, and / or stabilize the liposome in the solution or emulsion;3.43 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the minoxidil is added to the other ingredients of the composition at the point of use to form the composition;3.44 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the composition is configured to be administered by applying the composition at a predetermined frequency;3.45 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the composition is formulated as a cosmetic product;3.46 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the composition is formulated as any one or combination of a time release vehicle, a cream, a solution, an emulsion, a lotion, a serum, an ointment, a spray, an aerosol medium, a capsule, a shampoo, a gel, a foam, a cosmetic, a hair conditioner, a hair care product, a hair styling product (e.g., a styling gel, a styling foam or a hair conditioner), a hair mask, a deodorant, an antiperspirant, a moisturizer, or a shaving cream or gel;3.47 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the minoxidil is presentin a concentration of 2-15%.3.48 Any of the foregoing Compositions 3 or 3.1 ct scq., wherein the composition is a therapeutic product.3.49 Any of the foregoing Compositions 3 or 3.1 et seq., wherein the composition is a cometic product.

[0019] The present disclosure also provides a kit (Kit 1) for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, the kit comprising a composition and a dispenser, implant, or pill; wherein the composition comprises a composition according to any of the foregoing Compositions 1 or 1.1, et seq., or 2 or 2.1 et seq., or 3 and 3.1 et seq.

[0020] The present disclosure further provides the following kits:1.1 Kit 1, wherein the composition is formulated as any one or combination of a time release vehicle, a cream, a solution, an emulsion, a lotion, a serum, an ointment, a spray, an aerosol medium, a capsule, a shampoo, a gel, a foam, a cosmetic, a hair conditioner, a hair care product, a hair styling product (e.g., a styling gel, a styling foam or a hair conditioner), a hair mask, a deodorant, an antiperspirant, a moisturizer, or a shaving cream or gel;1.2 Kit 1.1, wherein the composition is a composition according to any of Compositions 1.27 or 2.28-2.37 or 3.29-3.38.1.3 Kit 1.1 or 1.2, wherein the kit further comprises a hair brush or comb, particularly a brush or comb that provides exfoliating effect on the scalp such that there is light abrasion after its use that enhances penetration of the therapeutic agent to the AP muscle and / or hair follicle.1.4 Any of the foregoing Kits 1 or 1.1 et seq., wherein at least the alkalinizing agent and the minoxidil is provided in a metered dose applicator that provides for a fixed volume of the composition to be administered with each administration, such as 1 ml of the topical composition per administration.

[0021] The present disclosure further provides a method [Method 1] for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity,upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising administering a Composition according to any of the foregoing Compositions 1 or 1.1, et seq., or 2 or 2.1 et seq., or 3 and 3.1 et seq. to a patient or subject in need thereof.

[0022] The present disclosure further provides the following Methods:1.1 Method 1, wherein the method induces or up-regulates the expression of a sulfotransferase;1.2 Method 1.1, wherein the sulfotransferase is sulfotransferase (SULT) 1A1 and / or 2A1;1.3 Method 1.1, wherein the sulfotransferase is sulfotransferase (SULT) 1A1;1.4 Any of the foregoing Methods 1-1.3, wherein the composition is applied manually;1.5 Any of the foregoing Methods 1-1.3, wherein the composition is applied with a sprayer;1.6 Any of the foregoing Methods 1-1.3, wherein the composition is applied as a mist;1.7 Any of the foregoing Methods 1-1.3, wherein the composition is a time release vehicle, a cream, a solution, an emulsion, a lotion, a serum, an ointment, a spray, an aerosol medium, a capsule, a shampoo, a gel, a foam, a cosmetic, a hair conditioner, a hair care product, a hair styling product (e.g., a styling gel, a styling foam or a hair conditioner), a hair mask, a deodorant, an antiperspirant, a moisturizer, or a shaving cream or gel.

[0023] The present disclosure further provides a method [Method 2] for preparing a liposomal solution or emulsion composition for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, the composition comprising:(i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person;(ii) an alkalinizing agent;(iii) liposomes encapsulating at least some of the alkalinizing agent;(iv) an antioxidant configured to prevent oxidative degradation of the liposomes; and(v) a buffer system configured to increase dermal penetration of the liposomes, and / or stabilize the liposome solution or emulsion; and(vi) a solvent; and(vii) minoxidil; optionally, a penetration enhancer; optionally, a chelating agent; wherein the liposomes comprise phosphatidylcholine and / or lecithin; and optionally wherein the alkalinizing agent and the buffer system are the same, or comprise the same components; wherein the method comprises: a) encapsulating at least some of each of one or more of the agent configured to increase sulfonating ability; the alkalinizing agent; the antioxidant; the penetration enhancer (if present) and the chelating agent (if present) in liposomes to form a liposome solution or emulsion; b) adding a preservative, and optionally a fragrance and optionally a surfactant to the liposome solution or emulsion; and c) adding minoxidil; wherein the liposomes comprise phosphatidylcholine and / or lecithin.

[0024] The present disclosure further provides the Methods:2.1 Method 2, wherein the product of the method is a Composition according to any of the foregoing Compositions 1 or 1.1, et seq., or 2 or 2.1 et seq., or 3 and 3.1 et seq.;2.2 Method 2, wherein the product of the method is a Composition according to Composition 1 or 1.1 et seq.

[0025] The present disclosure further provides a method [Method 3] for preparing a solution or emulsion composition for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, the composition comprising:(i) minoxidil;(ii) an alkalinizing agent; and(iii) a penetration enhancer; wherein the method comprises: a) combining the alkalinizing agent and the penetration enhancer in a solvent; and adding the minoxidil to form the solution or emulsion.

[0026] The present disclosure further provides the Methods:3.1 Method 3, wherein the alkalinizing agent is selected from those in any of Compositions 2.1-2.3;3.2 Method 3 or 3.1, wherein the penetration enhancer is selected from those in any of Compositions 2.4-2.5;3.3 Any of Methods 3 or 3.1 et seq., further comprising the step of adding an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells as described in any of Compositions 2.6-2.9;3.4 Any of Methods 3 or 3.1 et seq., further comprising the step of adding an antioxidant as described in any of Compositions 2.10-2.12;3.5 Any of Methods 3 or 3.1 et seq., further comprising the step of adding a chelating agent as described in any of Compositions 2.13-2.14;3.6 Any of Methods 3 or 3.1 et seq., further comprising the step of adding a preservative as described in any of Compositions 2.15-2.17;3.7 Any of Methods 3 or 3.1 et seq., further comprising the step of adding a surfactant as described in any of Compositions 2.18-2.19;3.8 Any of Methods 3 or 3.1 et seq., wherein the solvent comprises or consists of water;3.9 Any of Methods 3 or 3.1 et seq., further comprising the step of adding glycerin;3.10 Any of Methods 3 or 3.1 et seq., further comprising the step of adding a fragrance compound;3.11 Method 3, wherein the product of the method is a Composition according to any of the foregoing Compositions 2 or 2.1 et seq.;

[0027] The present disclosure further provides a method [Method 4] for preparing a solutionor emulsion composition for increasing the sulfonating capacity of a hair follicle cell and / or kcratinocytc cell, inducing sulfotransfcrasc in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, the composition comprising:(i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person;(ii) an alkalinizing agent;(iii) inorganic source of sulfur;(iv) a penetration enhancer meant to aid the delivery of an alkalinizing agent and inorganic source of sulfur;(v) a solvent; and(vi) minoxidil or a salt thereof; wherein the method comprises: a) combining the agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person, the alkalinizing agent, the inorganic source of sulfur and the penetration enhancer in a solvent; and adding the minoxidil or salt thereof to form the solution or emulsion.

[0028] The present disclosure further provides the Methods:4.1 Method 4, wherein the alkalinizing agent is selected from those in any of Compositions 3.1-3.3;4.2 Method 4 or 4.1, wherein the penetration enhancer is selected from those in any of Compositions 3.4-3.5;4.3 Any of Method 4 or 4.1 et seq, wherein the inorganic source of sulfur is selected from those in any of Compositions 3.6-3.7;4.4 Any of Methods 4 or 4.1 et seq., wherein the agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person is SULT1A1 protein as described in Compositions 3.8, 3.9 and 3.10;4.5 Any of Methods 4 or 4.1 et seq., further comprising the step of adding an antioxidant as described in any of Compositions 3.11-3.13;4.6 Any of Methods 4 or 4.1 et seq., further comprising the step of adding a chelating agent as described in any of Compositions 3.14-3.15;4.7 Any of Methods 4 or 4.1 et seq., further comprising the step of adding a preservative as described in any of Compositions 3.16-3.18;4.8 Any of Methods 4 or 4.1 et seq., further comprising the step of adding a surfactant as described in any of Compositions 3.19-3.20;4.9 Any of Methods 4 or 4.1 et seq., wherein the solvent comprises or consists of water;4.10 Any of Methods 4 or 4.1 et seq., further comprising the step of adding glycerin;4.11 Any of Methods 4 or 4.1 et seq., further comprising the step of adding a fragrance compound;4.12 Method 4, wherein the product of the method is a Composition according to any of the foregoing Compositions 3 or 3.1 et seq.DETAILED DESCRIPTION

[0029] The compositions and methods of the present disclosure induce (up-regulate) the expression or activity of sulfotransferases, e.g. SULT1A1, in hair bearing skin, hair follicles, and / or keratinocyte cells. Increasing sulfotransferase is beneficial for metabolizing pro-drugs that require sulfation to be activated, in particular the sulfation of minoxidil to its active metabolite, minoxidil sulfate. Thus, the present compositions are useful for treating conditions of hair bearing skin, hair follicles, and / or keratinocyte cells that are amendable to treatment with minoxidil, such as androgenetic alopecia (AGA) and other hair loss disorders.

[0030] Androgenetic alopecia (AGA) is a common dermatological condition affecting approximately 50% of the population by the age of 50. Currently, the only drug approved by the US Food and Drug Administration (FDA) for the treatment of AGA in both men and women is topical minoxidil. Clinical trials have demonstrated that following 16 weeks of 5% minoxidil therapy approximately 30-40% of patients regrow hair.

[0031] While the exact mechanism of action of minoxidil in the treatment of AGA is not completely understood, research has demonstrated that minoxidil sulfate is the active compound that stimulates hair follicles. Minoxidil is converted to its active form, minoxidil sulfate, in theouter root sheath of the hair follicle by endogenous sulfotransferase enzymes utilizing 3'- phosphoadcnosinc 5'-phosphosulfatc (PAPS). PAPS is produced in cells utilizing 3'- phosphoadenosine 5'-phosphosulfate synthase (PAPSS). Several studies have reported a correlation between sulfotransferase activity in plucked hair follicles and minoxidil response for AGA patients. In theory, increasing sulfotransferase or PAPS in the scalp would increase the likelihood that a subject will respond to topical minoxidil (thereby increasing the efficacy of a topical minoxidil); however, this has not been demonstrated in clinical studies. Further, in medicine, the induction of a deficient enzyme frequently does not result in a clinical benefit. For example, the pro-drug acyclovir used for the treatment of HSV is activated by the human thymidine kinase enzyme; however, while the induction of the thymidine kinase enzyme in-vitro activates acyclovir, in human studies thymidine kinase enzyme induction does not convert nonresponders to acyclovir into responders. As noted herein, minoxidil compositions can be used to treat forms of alopecia. Thus, increasing the efficacy of minoxidil (which can involve converting androgenetic alopecia patients who are non-responders into responders to minoxidil) can improve hair growth (including hair diameter) of subjects using topical minoxidil. In addition, methods and compositions disclosed herein for increasing the efficacy of minoxidil for treatment of forms of alopecia can accelerate hair growth of subjects using topical minoxidil.

[0032] Biological sulfation is the conversion of the very stable oxy-anion sulfate to the high- energy sulfate donor 3'-phospho-adenosine-5'-phosphosulphate (PAPS). Sulfation of a variety of biomolecules depend on availability of the precursor PAPS, which is rate-limiting. In mammals, PAPS is synthesized in two steps by a bi-functional enzyme called PAPS synthetase (PAPSS). The synthesis of PAPS from inorganic sulfate and ATP is catalyzed by PAPSS. The sources of inorganic sulfur in nature are broad but include cysteine, 1 -cysteine, hydrogen sulfide, elemental sulfur, sulfite, thiosulfate, and various polythionates (e.g., tetrathionate).

[0033] The compositions and methods of the present disclosure induce (up-regulate) the expression or activity of sulfotransferases in hair bearing skin, hair follicles, and / or keratinocyte cells. Increasing sulfotransferase is beneficial for metabolizing pro-drugs that require sulfation to be activated, e.g., the sulfation of minoxidil to its active metabolite, minoxidil sulfate. Thus, the present compositions, which contain minoxidil in a formulation that increases the expression oractivity of sulfotransferases, provides a significant increase in minoxidil efficacy over topical administration of minoxidil alone.

[0034] While not wishing to be bound by a particular theory, it is believed that the present compositions, by providing in a single composition an inorganic sulfate substrate for PAPSS, an alkalinizing agent (for example encapsulated in liposomes), and minoxidil, will result in increased synthesis of PAPS, and increased sulfation of minoxidil to provide the active metabolite minoxidil sulfate, resulting in significantly increased efficacy of minoxidil relative to administration of topical minoxidil alone, or administration of topical minoxidil prior to or after application of an alkalinizing agent and / or inorganic source of sulfur..

[0035] It also is believed that the present compositions, by modifying hair follicle stem cells intracellular pH, can control hair follicle stem cell differentiation. In some instances, increase of hair follicle stem cells intracellular pH induces and / or increases the sulfotransferase enzymatic activity in hair follicle cells. Further, induction of the sulfotransferase enzyme in hair follicles increases the sulfonation capacity of minoxidil; thus, increasing the response level to oral and topical minoxidil, for example in the treatment of alopecia.

[0036] Thus, in some embodiments the present disclosure provides more efficacious treatments for conditions treated by prodrugs that are sulfated in vivo, and / or alleviation of the symptoms of those conditions. For example, embodiments of the methods and compositions disclosed herein can be used to increase the metabolism of minoxidil (which can result in the increase of bioavailable minoxidil sulfate) in hair follicles of patients suffering from various forms of alopecia, for example female pattern hair loss or androgenetic alopecia.

[0037] Accordingly, in a first embodiment, the present disclosure provides compositions for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising:(i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person;(ii) an alkalinizing agent;(iii) liposomes encapsulating at least some of the alkalinizing agent;(iv) an antioxidant configured to prevent oxidative degradation of the liposomes; and(v) a buffer system configured to increase dermal penetration of the liposomes, and / or stabilize the liposome solution or emulsion; and(vi) a solvent; and(vii) minoxidil or a salt thereof; wherein the alkalinizing agent and the buffer system are the same, and / or comprise the same components.

[0038] The present disclosure further provides, in a second embodiment, a solution or emulsion Composition (Composition 2) for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising:(i) minoxidil;(ii) an alkalinizing agent; and(iii) a penetration enhancer.

[0039] In further aspects of the second embodiment, the Composition further comprises one or more of an inorganic sulfur source; a chelating agent; an antioxidant; a surfactant; and a preservative I antibacterial / antifungal agent. In further aspects, the Composition further comprises one or both of glycerol and a fragrance compound. In further aspects, the Composition further comprises liposomes that encapsulate at least a portion of the alkalinizing agent, preferably wherein the liposomes comprise or consist of phosphatidylcholine or lecithin.

[0040] Additionally, the present disclosure provides compositions for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising:(i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person;(ii) an alkalinizing agent;(iii) inorganic source of sulfur;(iv) a penetration enhancer meant to aid the delivery of an alkalinizing agent and inorganic source of sulfur;(v) a solvent; and(vi) minoxidil or a salt thereof.

[0041] Agents that are configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person include compounds that promote biological sulfation of minoxidil. Biological sulfation is the conversion of the very stable oxy-anion sulfate to the high-energy sulfate donor 3'-phospho-adenosine-5'-phosphosulphate (PAPS). While not wishing to be bound by a particular theory, it is believed that sulfation of a variety of biomolecules, including minoxidil, depend on availability of the precursor PAPS, which is ratelimiting. In mammals, PAPS is synthesized in two steps by a bi-functional enzyme called PAPS synthetase (PAPSS). The synthesis of PAPS from inorganic sulfate and ATP is catalyzed by PAPSS. The sources of inorganic sulfur in nature are broad but include cysteine, L-cysteine, hydrogen sulfide, elemental sulfur, sulfite, thiosulfate, and various polythionates (e.g., tetrathionate).

[0042] Accordingly, in some embodiments, the agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person comprises or consists of an inorganic sulfur source. Thus, in one embodiment, the present invention concerns up-regulating the sulfonating capacity of hair bearing skin by applying a topical solution containing, inter alia, a source of inorganic sulfur to increase the concentration of PAPS. Examples of suitable inorganic sulfur sources include but are not limited to cysteine, L-cysteine, hydrogen sulfide, elemental sulfur, sulfites, thiosulfates, poly thionates, magnesium sulfate, sodium sulfate and metabisulfites, for example sodium metabisulfite. In some embodiments, the present compositions include the inorganic sulfur source, for example sodium metabisulfite, at an amount of 0.01-0.5 wt%; e.g. 0.05-0.20 wt%; 0.075-0.15 wt%; e.g. 0.1 wt%.

[0043] In some embodiments, the agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person can include or consist of SULT1A1 protein. In some embodiments, the SULT1A1 protein is human SULT1A1 protein. In some embodiments, the SULT1A1 protein is added as a solution in a buffer, or as a solidprotein. In some embodiments, the SUMLTA1 protein is added in sufficient amount to provide at least 0.1 pmol / min / pg of SULT1A1 activity as measured by transfer sulfate from PAPS to 1- Napthol. SULT1A1 protein can be obtained from, for example, Bio-Techne R&D Systems, Minneapolis, MN.

[0044] Alkalinizing Agents

[0045] As used herein the term "alkalinizing agent" means a pH regulating agent; i.e., an agent that either: (i) directly increase the intracellular pH; (ii) indirectly increase the intracellular pH by activating or inhibiting the various ion carriers that regulate cellular pH; (iii) indirectly increase the intracellular pH by upregulating or downregulating the various ion carriers that regulate cellular pH; or (iv) increase the intracellular pH by changing the extracellular pH. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment. In some embodiments, the alkalinizing agent increases intracellular pH to an alkaline pH range of 7.4-11.0. In some embodiments, the alkalinizing agent is present in the compositions of the present disclosure in an amount of 2-12 wt.%; e.g. 4-10 wt.%; e.g. 6-8 wt.%; e.g. 7-7.2 wt.%; e.g. 7.1 wt.%; e.g. 7.09 wt.%.

[0046] In some embodiments, the compositions of the present disclosure provide the upregulation of the sulfonating capacity of hair bearing skin, hair follicles, and / or keratinocyte cells by applying a topical composition according to the present disclosure, which contains, inter alia, an alkalinizing agent that will raise the intracellular pH of cells in the outer root sheath of the hair follicle; and / or raise the extracellular pH of cells in the outer root sheath of the hair follicle.

[0047] In some embodiments of the compositions of the present disclosure, the pH range and alkalinizing agent are chosen such that it is optimal for preventing degradation of a liposomal solution and simultaneously within a range of pH optimal for inducing the upregulation of SULTIAI. In some embodiments, a pH range of 8.0-9.0 is used to both stabilize a liposomal solution and induce the expression of SULT I Al.

[0048] Liposomes

[0049] In some embodiments, at least a portion of the alkalinizing agent is encapsulated in liposomes. Such encapsulation can be used to increase or decrease water solubility, prevent oxidative degradation, and / or increase penetration through the stratum comeum of the alkalinizing agent and other encapsulated components of the composition. As used herein the term "liposome" means a small artificial vesicle, spherical in shape, having at least one lipid bilayer. The liposomes may be one or more of multilamellar vesicles (MLVs), which have several lamellar phase lipid bilayers; small unilamellar vesicles (SUVs), which contain a single lipid bilayer; large unilamellar vesicle (LUVs), which contain a single lipid bilayer; and cochleate vesicles. In some embodiments, the phospholipid of the liposomes of the present compositions include or consist of phosphatidyl choline. In some embodiments, the phospholipid of the liposomes of the present compositions include or consist of lecithin.Lecithin is a mixture of glycerophospholipids including phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine and phosphatidic acid.

[0050] In some embodiments, the phospholipid liposomes of the compositions of the present disclosure are present in the composition at a concentration of 0.10-5.0 wt.%; e.g. 0.5-4.0 wt.%; 1.00-3.00 wt.%; e.g. 1.50-2.50 wt.%; e.g. 2.00 wt.%. In one embodiment, liposomes comprising or consisting of phosphatidylcholine (e.g., available under the trade name: Phospholipon 90G) are used to encapsulate at least a portion of the alkalinizing agent, for example TRIS, for example at pH 8-9; for example at pH 8.

[0051] Formulas utilizing the abovementioned liposome encapsulation techniques to increase dermal penetration can be optimized by adjusting pH, temperature, and particle size (extrusion).

[0052] In some embodiments, one or more components of the present composition are in liposomes as described herein, to overcome poor water solubility, and / or to prevent oxidative degradation, and / or to facilitate dermal penetration.

[0053] In one embodiment of the present invention, a stable liposomal solution of encapsulated Tris buffer is used to induce the expression of SULTIAI in the outer root sheath of hair follicles. In another embodiment, a stable liposomal solution of encapsulated Tris buffer is prepared using Phosphatidylcholine (Phospholipon 90G). In another embodiment, a stable liposomal solution of encapsulated Tris buffer is prepared using Phosphatidylcholine (Phospholipon 90G) in an amount between I and 15% (w / w). In yet another embodiment, astable liposomal solution of encapsulated Tris buffer is prepared using 2% (w / w) Phosphatidylcholine (Phospholipon 90G). In one embodiment of the present invention, a liposomal solution containing Phosphatidylcholine (Plospholipon 90G) is protected from hydrolysis and oxidation by adding a Tris buffer at a pH range of 8.0-8.7. In yet another embodiment of the present invention, a liposomal solution containing Phosphatidylcholine (Plospholipon 90G) is stabilized using a Tris buffer at a concentration of 100-500 mM. In another embodiment, a Tris buffer at a concentration of 500 mM - 1 M is used. In another embodiment of the present invention, a liposomal solution containing Phosphatidylcholine (Plospholipon 90G) is stabilized using a Tris buffer at a concentration of 250 mM or 3% (w / w). In another embodiment, a liposomal solution containing Phosphatidylcholine (Phospholipon 90G) and 250 mM or 3% (w / w) Tris is used to induce the expression of SULT1A1 in the outer root sheath of hair follicles.[00541 Minoxidil

[0055] In some preferred embodiments, the present compositions include minoxidil in nonsalt form. Preferably, the minoxidil is present in a concentration of 2-15% w / w. In some embodiments, liposomes encapsulate at least some of the components of the present compositions, as discussed herein. In some such embodiments, at least some of the minoxidil is encapsulated in the liposomes. In some such embodiments, substantially all of the minoxidil is encapsulated in the liposomes. In some such embodiments, the minoxidil is present both inside the liposomes and outside the liposomes.

[0056] Solvents

[0057] In some embodiments, the solvent used in the compositions of the present disclosure is water, for example at 75-90 wt.%; e.g. 80-90 wt.%; e.g. 85-90 wt.%; e.g. 86-87 wt.%; e.g. 86.45 wt.%. In some embodiments, the composition can further include one or more additional solvent or lipid components to aid in obtaining the optimum viscosity of the solution or emulsion and / or to aid in providing the optimal lipid environment for liposome formation and stability. Examples of such suitable solvent or lipid components include poly -hydroxyl compounds such as glycerol, cetyl alcohol, stearic acid, carnauba wax, hydroxyethyl cellulose, guar gum, xanthangum, gelatin, magnesium aluminum silicate, silica, bentonite, carbomer (acrylic acid polymer), cetyl palmitate, and ammonium acryloyldimcthyltauratc, and combinations thereof. In some embodiments, the one or more additional solvent or lipid components arc present in the composition at a concentration of 0.10-4.00 wt.%; e.g. 0.50-3.00 wt.%; e.g. 1.00-2.50 wt.%; e.g. 1.50-2.00 wt.%; e.g. 1.75 wt.%. In some embodiments, the compositions include glycerol, for example at 0.10-4.00 wt.%; e.g. 0.50-3.00 wt.%; e.g. 1.00-2.50 wt.%; e.g. 1.50-2.00 wt.%; e.g. 1.75 wt.%.

[0058] Buffer Systems

[0059] In some embodiments, the compositions of the present disclosure contain a buffer system that functions to maintain the pH of the composition at a desired value, and aid in stabilizing the liposomes of the compositions. In one embodiment, the liposomal solution or emulsion is stabilized by adjusting the pH to a range of 7-11. In another embodiment the pH is adjusted to a range of 7.5 to 10. In yet another embodiment, the liposomal solution of emulsion is stabilized by adjusting the pH to a range of 8.0-9.0, e.g.,8.0-8.7. In one embodiment the buffering solution, for example, TRIS, acetate, citrate, or HEPES, is used to stabilize the pH of the liposomal solution or emulsion. In another embodiment, a TRIS-TRIS HC1 buffer is used. In another embodiment, a carbonate buffering or a phosphate buffering system is used. In one embodiment, the liposomal solution or emulsion is stabilized by adjusting the pH to a range of 8.0-9.0 using a TRIS buffer. In some embodiments, the buffer system is present in the compositions of the present disclosure in an amount of 2-12 wt.%; e.g. 4-10 wt.%; e.g. 6-8 wt.%; e.g. 7-7.2 wt.%; e.g. 7.1 wt.%; e.g. 7.09 wt.%.

[0060] In some embodiments, the buffer system and alkalinizing agent can comprise the same components - i.e., their components can serve both functions. For example, an alkalinizing agent comprising or consisting of TRIS buffer solution at pH 8-9 can be encapsulated in liposomes, e.g. liposomes composed of phosphatidylcholine, and the same TRIS buffer solution at pH 8-9 can be utilized as the buffer in the solution or emulsion.

[0061] Antioxidants

[0062] In some embodiments, oxidative degradation of the liposomal solution or emulsion is prevented by the addition of an antioxidant. In one embodiment, oxidative degradation of a liposomal solution or emulsion is prevented by the addition of ascorbic acid, monothioglycerol, potassium metabisulfite, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium sulfite, sodium thiosulfate, a tocopherol or an ester derivative thereof such as tocopherol acetate, sodium metabisulfite, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), ascorbyl palmitate, or propyl gallate. In some embodiments, oxidative degradation of a liposomal solution is prevented by the addition of acai oil, alpha lipoic acid, green tea extract, retinal, vitamin C, coenzyme Q10 (Co Q-10), isoflavones, polyphenols, curcumin, turmeric, pomegranate, rosemary extract, glutathione, selenium, or zinc, and chelating agents as described below. In some embodiments, the antioxidant(s) are present in the compositions of the present disclosure in an amount of 0.01-0.5 wt.%; e.g. 0.05-0.4 wt.%; 0.1-0.3 wt.%; e.g. 0.15-0.25 wt.% e.g. 0.20 wt.%.

[0063] Chelating Agents

[0064] In some embodiments, the compositions of the present disclosure include a chelating molecule, for example ethylenediaminetetraacetic acid (EDTA), disodium EDTA, tetrasodium EDTA, pentasodium penetate, sodium metasilicate and phosphate derivatives, etidronic acid and its derivatives, or galactaric acid. These chelating agents can be added to capture free radicals in solution and prevent oxidation of lipids in the liposomal formula. In some embodiments, the chelating agent(s) are present in the compositions of the present disclosure in an amount of 0.01- 0.5 wt.%; e.g. 0.05-0.20 wt.%; 0.075-0.15 wt.%; e.g. 0.1 wt.%.

[0065] Penetration Enhancers

[0066] In some embodiments, the compositions of the present disclosure include one or more penetration enhancers. Examples of penetration enhancers include, but are not limited to, alcohols, glycols (e.g., diethylene glycol and tetraethylene glycol; diethylene glycol monoethyl ether; PEG-6 Caprylic / Capric glyceride, e.g.. Accanon-CC6), fatty acids (e.g., lauric acid, myristic acid and capric acid), fatty esters, fatty ethers, cyclodextrines, occlusive agents, surface active agents, dimethylaminopropionic acid derivatives, terpenes, sulfoxides, cyclic ethers, amides, and amines. Other examples of penetration enhancers can include sulphoxides (such asdimethyl sulphoxide, DMSO, decylmethal sulfoxide), Azones (e.g., 1 -dodecylazacycloheptan-2- onc, laurocapran, or laurocapram), pyrrolidones (e.g., 2-pyrrolidonc, 2P, N-mcthylpyrrilidonc, N-methyl-2-pyrrolidone, NMP, l-propyl-3-dodecyl-2-pyrrolidone, 1- butyl-3-dodecyl-2- pyrrolidone), alcohols and alkanols (ethanol, or decanol), glycols (e.g., propylene glycol), surfactants (e.g., polyoxyethylene-2-oleyl ether, polyoxy ethylene-2-stearly ether, sodium dodecyl sulfate, SDS, sodium lauryl sulfate, SLS), Oxazolidinones (e.g., 4- decyloxazolidin-2- one), urea, 2-(l-nonyl)-l,3-dioxolane, and terpenes. Additional examples of penetration enhancers can include polyester nanosponges, liposomes, phospholipids, cyclopentadecalactone, pentadecalactone, SNAC, salcaprozate sodium N-[8-(2- hydroxybenzoyl) amino]caprylate, CNAC, 5-CNAC, 8-(N-2-hydroxy-5-chloro-benzyl)-amino- caprylic acid, sodium caprate, glyceryl triglyceride, and peptides. Other examples of penetration enhancers can include longer chain 1,2-glycols, e.g., Arachidyl Glycol (1,2-Eicosanediol, CAS 39825-93-9), Cetyl Glycol (1,2-Dihydroxy hexadecane, 1,2-Hexadecanediol, 1 ,2-Hexadecylene Glycol, 2-Hydroxycetyl Alcohol, CAS 6920-24-7), Hexacosyl Glycol (Hexacosil glycol), Lauryl Glycol (1,2- Dihydroxydodecane, 1 ,2-Dodecanediol, 1,2-Dodecylene Glycol, CAS 1119-87-5), Myristyl Glycol (1,2-Tetradecanediol, CAS 21129-09-9), Octacosanyl Glycol (1,2-Octacosanediol, CAS 97338-11-9), Stearyl Glycol (1,2-Dihydroxyoctadecane, 1,2-Octadecanediol, CAS 20294-76-2), Caprylyl Glycol (Capryl Glycol, 1,2-Dihydroxyoctane, 1,2-Octanediol, 1,2-Octylene Glycol, CAS 1117-86-8), Decylene Glycol ( 1 ,2-Decanediol, CAS 1119-86-4), Pentylene Glycol (1,2- Dihydroxypentane, 1,2-Pentanediol, CAS 5343-92-0), 1,2-Butanediol (1,2-Butylene Glycol, 1,2- Dihydroxybutane, CAS 584-03-2), 1 ,2-Hexanediol (1,2-Dihydroxyhexane, CAS 6920-22-5), C14-18 Glycol (Ethylene Glycol Fatty Acid Ester), C15-18 Glycol (Alkylene (15-18) Glycol, Cetyl Stearyl Vicinal Glycol, Glycols C15-18, CAS 70750-40-2, CAS 92128-52-4), C18-30 Glycol (Ethylene Glycol Fatty Acid Ester), C20-30 Glycol (Alkylene (20-30) Glycol). In some embodiments the penetration enhancer(s) is / are present in the composition in a total amount of 0.10 wt.%-3.00 wt.%; e.g. 0.50 wt.%-2.00 wt.%; e.g. 0.75 wt.%-1.25 wt.%; e.g. 1.00 wt.%; or 0.10 wt.%-7.0 wt.%0; e.g. 0.50 wt.%-5.00 wt.%; e.g. 0.75 wt.%-4.0 wt.%; e.g. 2.6 wt.%. Preferred penetration enhancers include, but are not limited to PEG-6 Caprylic / Capric Glycerides, a polyethylene glycol derivative of a mixture of mono-, di-, and triglycerides of caprylic and capric acids with an average of 6 moles of ethylene oxide sold under the trade nameAcconon® CC-6.; diethylene glycol monoethyl ether; Transcutol; one or more of a carbitol, dicthylcnc glycol monocthyl ether, ethyl carbitol, cthylcarbitol, Transcutol, transcutol HP and transcutol P, and Pentylene Glycol (1,2-Dihydroxypentane, 1,2-Pentanediol, CAS 5343-92-0).

[0067] Further preferred penetration enhancers include Accanon-CC6, diethylene glycol monoethyl ether and Transcutol; e.g. Accanon-CC6 at a concentration of, for example, 0.10-3.00 wt.%; e.g. 0.50-2.00 wt.%; e.g. 0.75-1.25 wt.%; e.g. 1.00 wt.%; diethylene glycol monoethyl ether at a concentration of, for example, 0.10-7.00 wt.%; e.g. 0.50-5.00 wt.%; e.g. 0.75-4.0 wt.%; e.g. 2.6 wt.%; Transcutol at a concentration of 0.10 wt.%-3.00 wt.%; e.g. 0.50 wt.%-2.00 wt.%; e.g. 0.75 wt.%-1.25 wt.%; e.g. 1.00 wt.%; or 0.10 wt.%-7.0 wt.%0; e.g. 0.50 wt.%-5.00 wt.%; e.g. 0.75 wt.%-4.0 wt.%; e.g. 2.6 wt.%; or a combination of two or three thereof.

[0068] In some preferred embodiments, the penetration enhancer comprises or consists of Pentylene Glycol (1,2-Dihydroxypentane, 1,2-Pentanediol, CAS 5343-92-0).

[0069] Fragrances and Surfactants

[0070] In some embodiments, the present compositions can further include one or more fragrance compounds and / or surfactants, particularly where the fragrance compound, or another component as described herein, is not completely soluble in the composition. Suitable surfactants include anionic, cationic, nonionic and amphoteric surfactants. In some embodiments the surfactant is a nonionic surfactant, for example polyoxyethylene-2-oleyl ether, polyoxy ethylene-2-stearly ether, or polysorbate 80, or an anionic surfactant such as sodium dodecyl sulfate (SDS; sodium lauryl sulfate (SLS)).

[0071] In some embodiments, the fragrance compound(s) is present in an amount of 0.01- 0.15 wt%; e.g. 0.02-0.10 wt%; e.g. 0.06 wt%.

[0072] In some embodiments, the surfactant (s) is present in an amount of 0.01-1.0 wt%; e.g. 0.10-0.5 wt%; e.g. 0.20-0.30 wt%; e.g. 0.25 wt%.

[0073] Preservatives

[0074] In some embodiments, the composition can further comprise one or more preservatives, for example an antibacterial / antifungal agent. Suitable preservatives includesodium benzoate, parabens, tetrasodium EDTA (ethylenediaminetetraacetic acid), mcthylisothiazolinonc (MIT) and Quatcmium compounds such as Quatcrnium-15, a combination of phenoxyethanol and ethylhexylglycerin (e.g., EUXYL PE 9010), and a combination of caprylhydroxamic acid, 1 ,2-hexanediol and propanediol, e.g., Spectrastat PHL. In some preferred embodiments, the preservative includes or consists of phenoxyethanol and ethylhexylglycerin, e.g. EUXYL PE 9010. In other preferred embodiments, the preservative includes or consists of caprylhydroxamic acid and 1 ,2-hexanediol and propanediol, e.g., Spectrastat PHL.

[0075] It is understood that, in certain cases, an ingredient may possess multiple functions. For example, the buffer system (e.g., TRIS / TRIS HC1 at basic pH) can also act as an alkalinizing agent, and the chelating agent (e.g. EDTA) and the inorganic sulfur source (e.g. sodium metabisulfite) can each also act as a preservative.

[0076] Formulations

[0077] The components of the topical compositions disclosed herein can be formulated as any one or combination of a time release vehicle, a cream, a solution, an emulsion, a lotion, a serum, an ointment, a spray, an aerosol medium, a capsule, a shampoo, a gel, a foam, a cosmetic, a hair conditioner, a hair care product, a hair styling product (e.g., a styling gel, a styling foam or a hair conditioner), a hair mask, a deodorant, an antiperspirant, a moisturizer, or a shaving cream or gel. In some embodiments, the alkalinizing agents described herein and used in the present methods may be formulated to provide immediate release. In some embodiments, the alkalinizing agents described herein and used in the present methods may be formulated to provide extended release.

[0078] In some embodiments, the present composition provide the alkalinizing agent and / or minoxidil with a penetration enhancer as discussed supra, formulated with a carrier or delivery vehicle optimized for delivery of the alkalinizing agent to the scalp. The alkalinizing agent and / or minoxidil can be released using several different formulations or release methods including time release, creams, solutions, lotions, serums, ointments, sprays, capsules, or otherrelease methods. For instance the alkalinizing agent can be incorporated into a shampoo for utilization during showering. In other embodiments, the alkalinizing agent can be included in ointments or other topical creams that could be applied to the scalp so that it can be slowly absorbed into the skin. In other embodiments, the alkalinizing agent can be included in a liquid spray or aerosol medium to be applied to the scalp. In other embodiments, the alkalinizing agent can be incorporated into capsules or other slow release vehicles that would allow the chemical or agent to be slowly released into the dermis of the scalp. Capsules or vehicles that encapsulate the alkalinizing agent or an alkalinizing agent with a penetration enhancer can include, but are not limited to, liposomes, non-ionic liposomes, niosomes, novasome I, erythromycin-Zn complex, microspheres, nanoparticles, solid lipid nanoparticles, and nanoemulsions. In some embodiments, this can include a gel or foam that is applied to the scalp. It is specifically contemplated that the alkalinizing agent or an alkalinizing agent with a penetration enhancer can be formulated in hair care products such as a shampoo, styling gel, styling foam, hair conditioner, hair serum, a hair mask, etc.

[0079] In some embodiments, the present compositions can be delivered as a slow releasing suspension, liposome, in situ gel-forming system, microsphere, non-aqueous solution / suspension, or implant.

[0080] In some embodiments, the present compositions may comprise an exfoliating agent to promote abrasion of the surface of the scalp. Examples of the exfoliating agent include (1) inorganic and / or metallic particles such as: boron nitride, in body -centered cubic form (Borazon®); aluminosilicate (e.g. nepheline); zircon; mixed oxides of aluminum such as emery; zinc oxide; aluminum oxides such as aluminas or corundum; titanium oxide; titanium oxide coated mica; carbides, in particular silicon carbide (carborundum); or other metal oxides; metals, and metal alloys such as iron shot, steel shot, and in particular perlite; silicates such as glass, quartz, sand, or vermiculite; calcium carbonate (e.g. Bora-Bora sand or Rose de Brignoles sand) or magnesium carbonate; sodium chloride; pumice stone; amorphous silica; diamond; ceramics, and (2) organic particles such as: fruit stones, in particular' apricot stones, e.g. Scrubami® apricot; wood cellulose, e.g. ground bamboo stem; coconut shell, e.g. coconut exfoliator; polyamides, in particular Nylon-6; sugars; plastic microbeads, e.g. polyethylenes or polypropylenes; ground walnut; ground apricot seed; ground shells, and (3) mixed particlesassociating organic and inorganic compounds, and particles coated in the above compounds. The exfoliating agents may be in the form of microbcads of less than five millimeters in its largest dimension that have an exfoliating effect.

[0081] Any of the aforementioned formulations can be used routinely, e.g., once daily, twice daily, every other day, once a week, monthly, etc., and applied at a predetermined frequency (e.g., once or twice per day, once per 24 hours, once per 12 hours, once per 6 hours, etc.). In some embodiments, the aforementioned formulations are topical solutions or emulsions. Routine use of the alkalinizing agent, with or without a penetration enhancer, would be indicated as an adjuvant therapy for minoxidil in androgenetic alopecia patients or subjects. . It is specifically envisioned that a composition (e.g., a topical solution or emulsion, or a shampoo) of any of the aforementioned compositions of the present disclosure can be used daily, or at any predetermined frequency by a person to increase the effectiveness of the minoxidil in the formulation.

[0082] The compositions and methods of the present disclosure can be administered to patients in a therapeutic setting to treat conditions and disorders as described herein, and can also be administered to subjects in cosmetic setting to provide cosmetic benefits as disclosed herein.

[0083] In cases where encapsulation techniques have been implemented in the above embodied formulations - for example, liposomes, micelles, nanogels, lipid nanoparticles, selenium nanoparticles, bilosomes, dendrimers and / or carbon nanotubes - the stability of the formula will require optimization to prevent degradation. For example, liposomes are vesicle structures created from one (unilamellar) or more bilayers of amphipathic lipids. In such systems lipids are often in direct contact with aqueous solution and arc subject to hydrolysis (the breakdown of lipids from water moisture) and oxidation (degradation of lipids due to the loss of electrons). Similarly, delivery systems using encapsulation techniques are optimized for stratum corneum penetration via the size of the encapsulated structure. For example, structural instabilities such as aggregation or the conversion of unilamellar structures to multilamellar or other liposome structures in solution would decrease the efficacy of payload delivery through skin.

[0084] Many measurement techniques have been described to track the degradation of encapsulated structures for formula stability optimization. For example, subsequent tohydrolysis, free fatty acids are produced and can be detected with UV-visible (UV / Vis) spectroscopy. UV / Vis spectroscopy can also be used to detect peroxidized lipids if oxidative degradation has occurred in a formula. Vesicle structural instabilities can be monitored by probing the hydrodynamic radius of liposomes with the techniques of dynamic light scattering, cryogenic transmission electron microscopy, and small-angle neutron scattering.

[0085] Formulas utilizing encapsulation techniques to increase dermal penetration can be optimized by adjusting pH, temperature, and particle size (extrusion). In one embodiment of the present invention, a liposomal solution is stabilized by adjusting the pH to a range of 7-11. In another embodiment the pH is adjusted to a range of 7.5 to 10. In yet another embodiment, a liposomal solution is stabilized by adjusting the pH to a range of 8.0-8.7. In one embodiment a buffering solution, for example, acetate, citrate, or HEPES is used to stabilize the pH of a liposomal solution. In another embodiment, a Tris buffer is used. In another embodiment, a carbonate buffering or a phosphate buffering system is used. In one embodiment of the present invention, a liposomal solution is stabilized by adjusting the pH to a range of 8.0-9.0 using a Tris buffer.

[0086] Additional Modalities

[0087] In some embodiments, the compositions of the present disclosure are administered with a device designed to increase transdermal penetration. Examples of devices designed to increase transdermal penetration include micro-needle arrays and iontophoretic patches.

[0088] In some embodiments, the compositions of the present disclosure are administered with a topical proton pump agonist, which aids in alkalinizing the intracellular pH of stem cells in the outer root sheath of a hair follicle.

[0089] In some embodiments, the topical application of compositions of the present disclosure containing an alkalinizing agent provides treatment of alopecia and other hair loss disorders by inducing hair follicle stem cell (HFSC) differentiation and hair anagen cycle elongation by raising the extracellular’ pH and / or intracellular' pH of cells in the outer root sheath of the hair follicle.

[0090] In further embodiments, the present disclosure provides methods for increasing hair graft survival and reducing shock hair loss post hair surgery by applying a topical compositionaccording to the present disclosure that will raise the extracellular pH of cells in the outer root sheath of the hair follicle. In some embodiments, the topical composition is applied with a device designed to increase transdermal penetration, for example micro-needle arrays or iontophoretic patches.

[0091] In further embodiments, the present disclosure provides methods for up-regulating the sulfonating capacity of hair bearing skin, hair follicles, and / or keratinocyte cells by applying to a hair follicle cell and / or a keratinocyte cell a composition according to the present disclosure that comprises an agent that increases the state of oxidative stress of a cell, e.g., an alkalinizing agents that increase intracellular pH (alkaline pH range 7.4-11.0), such as an alkaline solution containing a buffer, e.g., TRIS of bicarbonate buffer.

[0092] In one embodiment of the present invention a buffering system for increasing the shelf-life of a liposomal solution is optimized using UV-visible (UV / Vis) spectroscopy measurements over time. In yet another embodiment, samples of optimized liposomal solutions are stored at elevated temperatures to accelerate degradation prior to UV- visible (UV / Vis) spectroscopy measurements over time.

[0093] In further embodiments, the present disclosure also provides a method for stabilizing a composition for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response; the method comprising: encapsulating a composition, for example a composition according to the present disclosure, in a solution comprising liposomes, non-ionic liposomes, niosome, novasome I, erythromycin-Zn complex, microspheres, nanoparticles, solid lipid nanoparticles, nanoemulsions, micelles, nanogels, selenium nanoparticles, bilosomes, dendrimers, or carbon nanotubes; wherein the encapsulation increases water solubility of the composition, decreases water solubility of the composition, reduces loss through degradation, or promotes penetration through the stratum comeum.

[0094] Assays

[0095] A colorimetric assay for detecting sulfotransferase in plucked hair samples is described in U.S. Patent No. 8,691,518, which is incorporated by referenc e herein in its entirety.

[0096] As used herein, the terms "prevent" or "prevention" and other derivatives of the words, when used in reference to alopecia, e.g., androgenetic alopecia, refer to a reduced likelihood of alopecia in an individual receiving a given treatment relative to that of a similar individual at risk for alopecia but not receiving that treatment. As such, the terms "prevent" and "prevention" encompass a treatment that results in a lesser degree of alopecia, e.g., androgenetic alopecia, than would be otherwise expected for a given individual. Efficacy for prevention of alopecia, e.g., androgenetic alopecia, can be established through controlled studies, e.g., in which a subject is administered a treatment (e.g., a topical treatment) and another subject is administered a placebo. Under these circumstances, if the subject treated with the topical treatment undergoes less hair loss over time relative to the subject receiving the placebo, e.g., at least 5% less, at least 10% less, at least 15% less, at least 20% less, at least 25% less, at least 30% less, at least 35% less, at least 40% less, at least 45% less, at least 50% less or beyond, the treatment is effective for the prevention of alopecia, e.g., androgenetic alopecia.

[0097] As used herein, the terms "treat," "treatment," or "treating" refer to therapeutic treatments, wherein the object is to reverse, alleviate, ameliorate, inhibit, slow down or stop the progression or severity of a disease or condition, e.g., androgenetic alopecia or other form of alopecia. The term "treating" includes reducing or alleviating at least one adverse effect or symptom of a disease or condition, e.g., androgenetic alopecia or other form of alopecia. Treatment is generally "effective" if one or more symptoms are reduced. Alternatively, treatment is "effective" if the progression of a disease is reduced or halted. That is, "treatment" includes not just the improvement of symptoms, but also a cessation of, or at least slowing of, progress or worsening of symptoms compared to what would be expected in the absence of treatment. Beneficial or desired clinical results include, but are not limited to, alleviation of one or more symptom(s), diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, remission (whether partial or total), and / or decreased mortality. For example, treatment is considered effective if the extent or amount of hair loss is reduced, or the progression of hair loss is slowedor halted. The term "treatment" of a disease also includes providing relief from the symptoms or side-effects of the disease (including palliative treatment).

[0098] As used herein the term "comprising" or "comprises" is used in reference to compositions, methods, etc. refers to component(s) or method steps that are present in the method or composition, yet allows for the composition, method, etc. to also include unspecified elements.

[0099] The term "consisting of' refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the embodiment.

[0100] As used herein the term "consisting essentially of refers to those elements required for a given embodiment. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment.

[0101] As used herein the term "alopecia" refers to all forms of hair loss in men and women including but not limited to traction alopecia, androgenetic alopecia, male pattern baldness (MPB), female pattern hair loss (FPHL), alopecia areata, alopecia universalis, telogen effluvium, chemotherapy induced alopecia, hair shedding, eyebrow hair loss, beard hair loss, hair thinning, etc.. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment.

[0102] As used herein the term "ion carriers" refers to all cellular ion carriers that either increase or decrease the intracellular pH. In the context of this application ion carriers can be referred to as proton pumps. The term permits the presence of elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment.

[0103] The singular terms "a," "an," and "the" include plural referents unless context clearly indicates otherwise. Similarly, the word "or" is intended to include "and" unless the context clearly indicates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. The abbreviation, "e.g." is derived from the Latin exempli gratia, and is used herein to indicate a non-limiting example. Thus, the abbreviation "e.g." is synonymous with the term "for example."

[0104] Various aspects of the technology described measuring the sulfonating ability of a hair bearing skin, hair follicle, and / or kcratinocytc cell. An increase in the sulfonating ability of a hair bearing skin, hair follicle, and / or keratinocyte cell can be interpreted to mean that enzymes or substrates required for this reaction have been increased in concentration, i.e., an increase in substrates will lead to an increase in reaction products (Le Chatelier's principle). For example, increasing the available sulfotransferase will increase the sulfonating ability of a hair bearing skin, hair follicle, and / or keratinocyte cell. Similarly, increasing the available PAPS or PAPSS (which produces PAPS) will increase the sulfonating ability of a hair bearing skin, hair follicle, and / or keratinocyte cell.

[0105] Measurement of the sulfonating ability of a hair bearing skin, hair follicle, and / or keratinocyte cell or a hair follicle can be performed, if necessary, via a colorimetric assay adapted for that purpose. Examples are described in, e.g., Goren A, Shapiro J, Roberts J, McCoy J, Desai N, Zarrab Z, Pietrzak A, Lotti T. Clinical utility and validity of minoxidil response testing in androgenetic alopecia. Dermatol Ther 2015: 28(1): 13-16, ("the Minoxidil Response Test") which is incorporated herein in its entirety by reference. Briefly, plucked anagen hairs are collected from the scalp and inspected visually for an intact bulb. Suitable hairs are trimmed to a length of -1 cm and immersed, bulb first, in 100 pL of an assay solution containing 50 mM phosphate buffer (pH8), 5 mM potassium p-nitrophenyl sulfate, 20 pM adenosine 3', 5'- diphosphate, 100 pM minoxidil and 5 mM MgC12. Hairs are allowed to react with the solution for 24 hours at room temperature. After incubation, hairs are removed and the optical absorbance of the solution at 405 nm is determined with a spectrophotometer (e.g., Shimadzu UV-1700, Kyoto, Japan) using a single scan and 1 cm path length.

[0106] This disclosure uses the terms optical density (OD) and absorbance units (AU) interchangeably .

[0107] Increased intracellular pH is necessary for adult epithelial and embryonic stem cell differentiation. Various aspects of the invention describe the increased or decreased rate of hair follicle stem cells (HFSC), rate of differentiation, and / or proliferation. An increase in intracellular pH (pHi) can be used to increase the rate of hair follicle stem cells (HFSC) rate of differentiation and / or proliferation. Similarly, a decrease in intracellular pH (pHi) can be used todecrease the rate of hair follicle stem cells (HFSC) rate of differentiation and / or proliferation. Tn one aspect of the invention, pHi can be changed by changing the extracellular pH (pHc).

[0108] Efficacy of treatment to treat or prevent androgenetic alopecia can be determined by monitoring the density of hairs on a given area of the subject's body, e.g., a given area of the scalp. If the rate of hair loss is reduced, e.g., by 10% or more following treatment, the treatment is effective for the prevention of androgenetic alopecia. Similarly, if hair density remains the same, the treatment is effective for the prevention of androgenetic alopecia. If the density of hair increases, e.g., by 5% or more, e.g., by 10% or more following treatment, the treatment is also considered effective for the treatment and / or prevention of androgenetic alopecia.

[0109] Efficacy of treatment to treat or prevent androgenetic alopecia can be determined by monitoring global photography. For example, the patient or an expert can assess the treatment response utilizing before and after global photographs.

[0110] As noted above, it is contemplated that all forms of alopecia can benefit from the technology described herein. For example, the technology described herein can be applicable to prevent or treat androgenic alopecia.

[0111] It will be understood that the ranges described above, and throughout this document, are also intended to encompass single values contained within these ranges. For example, for a formulation comprising a particular ingredient in a range between 1-50%, a percentage of 5% or 49% is also intended to be disclosed.

[0112] The various methods and techniques described above provide a number of ways to carry out the invention. Of course, it is to be understood that not necessarily all objectives or advantages described can be achieved in accordance with any particular embodiment described herein. Thus, for example, those skilled in the art will recognize that the methods can be performed in a manner that achieves or optimizes one advantage or group of advantages as taught herein without necessarily achieving other objectives or advantages as taught or suggested herein. A variety of alternatives are mentioned herein. It is to be understood that some embodiments specifically include one, another, or several features, while others specifically exclude one, another, or several features, while still others mitigate a particular feature by inclusion of one, another, or several advantageous features.

[0113] Furthermore, the skilled artisan will recognize the applicability of various features from different embodiments. Similarly, the various elements, features and steps discussed above, as well as other known equivalents for each such element, feature or step, can be employed in various combinations by one of ordinary skill in this art to perform methods in accordance with the principles described herein. Among the various elements, features, and steps some will be specifically included and others specifically excluded in diverse embodiments.

[0114] Although the application has been disclosed in the context of certain embodiments and examples, it will be understood by those skilled in the art that the embodiments of the application extend beyond the specifically disclosed embodiments to other alternative embodiments and / or uses and modifications and equivalents thereof.

[0115] The recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (for example, "such as") provided with respect to certain embodiments herein is intended merely to better illuminate the application and does not pose a limitation on the scope of the application otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the application.

[0116] Certain embodiments of this application are described herein. Variations on those embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. It is contemplated that skilled artisans can employ such variations as appropriate, and the application can be practiced otherwise than specifically described herein. Accordingly, many embodiments of this application include all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the application unless otherwise indicated herein or otherwise clearly contradicted by context.

[0117] All patents, patent applications, publications of patent applications, and other material, such as articles, books, specifications, publications, documents, things, and / or the like, referenced herein are hereby incorporated herein by this reference in their entirety for all purposes, excepting any prosecution file history associated with same, any of same that is inconsistent with or in conflict with the present document, or any of same that can have a limiting affect as to the broadest scope of the claims now or later associated with the present document. By way of example, should there be any inconsistency or conflict between the description, definition, and / or the use of a term associated with any of the incorporated material and that associated with the present document, the description, definition, and / or the use of the term in the present document shall prevail.

[0118] Examples

[0119] Example 1: In-vitro Evaluation of Sulfotransferase Up-Regulation

[0120] The following experiment was conducted to assess the up-regulation of sulfotransferase in hair follicles by Compositions of the present disclosure.

[0121] Method: For each Composition tested (Composition 1.27 and Compositions 2.28- 2.38), 24 hairs are plucked from a human subject.

[0122] Hairs re visually inspected to confirm the presence of the hair bulb. Two hairs are placed in to 12 clean sample tubes (i.e., two hairs per reaction) containing lOOuL of Williams' E media with 0.292g / L L-glutamine. Compositions of the invention are (Composition 1.27 and Compositions 2.28-2.38) are prepared such that when 1 uL of the Composition solution is added to the sample media the final concentration of the alkalinizing agent is 50 to 500 mM and / or the inorganic sulfur source is 0.1-3.0%. 1 uL of DMSO with no Composition is added to control samples. Hairs are allowed to incubate with the Compositions for 24 h at room temperature. All samples are run in triplicate.

[0123] After 24h of incubation hairs are removed from the sample media and washed briefly with clean water. Hairs are trimmed to a length of ~1 cm and immersed, bulb first, in 100 pL of an assay solution containing 50 mM phosphate buffer (pH8), 5 mM potassium p- nitrophenyl sulfate, 20 pM adenosine 3',5'-diphosphate, 100 pM minoxidil and 5 mM MgC H. Hairs are allowed to react with the solution for 24 hours at room temperature. After incubation, hairs areremoved and the optical absorbance of the solution at 405 nm is determined with a spectrophotometer using a single scan and 1 cm path length.

[0124] Results: The Compositions of the present disclosure are expected to induce sulfotransferase activity in plucked hair follicles.

[0125] Example 2 - In vivo study

[0126] A study is conducted on thirty human subjects (30 men) with low sulfotransferase activity as determined by colorimetric enzymatic test described by Goren el al. Subjects are recruited based on an enzymatic test result of OD<0.4 The subjects are randomized to an active group and a placebo group in a 1:1 ratio. All subjects apply a composition according to Composition 1.27 or 2.28-2.38 once daily. The placebo group applies a vehicle composition lacking the alkalinizing agent and / or inorganic sulfur source. At the end of 1 week the average sulfotransferase activity of all subjects is determined, and is expected to increase almost 3 fold. The average sulfotransferase activity is also expected to remain high for a follow-up period of 6 months. At 4 months, the average increase in hair counts in the active group is expected to be over 100% higher than the placebo. In addition, 60% of the subjects in the active group are expected to respond to minoxidil compared to 0% of the placebo group. This will demonstrate that embodiments of the method and compositions disclosed herein can convert androgenetic alopecia patients who are non-responders to minoxidil into responders.

[0127] Example 3 - in vivo study

[0128] A study is conducted on thirty human subjects (30 women) with low sulfotransferase activity as determined by colorimetric enzymatic test described by Goren et al. Subjects are recruited based on an enzymatic test result of OD<0.4 The subjects are randomized to an active group and a placebo group in a 1:1 ratio. All subjects apply a composition according to Composition 1.27 or 2.28-2.38 once daily. The active group applies a vehicle composition lacking the alkalinizing agent and / or inorganic sulfur source. At the end of 1 week the average sulfotransferase activity of all subjects is determines, and is expected to increase almost 3 fold. The average sulfotransferase activity is also expected to remain high for a follow-up period of 6 months. At 6 months, the average increase in hair counts in the active group is expected to beover 100 % higher than the placebo. In addition, 60% of the subjects in the active group are expected to respond to minoxidil compared to 0% of the placebo group. This demonstrates that embodiments of the method and compositions disclosed herein can convert androgenetic alopecia patients who are non-responders to minoxidil into responders.

Claims

What is claimed is:

1. A composition for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising:(i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person;(ii) an alkalinizing agent;(iii) liposomes encapsulating at least some of the alkalinizing agent;(iv) an antioxidant configured to prevent oxidative degradation of the liposomes; and(v) a buffer system configured to increase dermal penetration of the liposomes, and / or stabilize the liposome solution; and(vi) a solvent; and(vii) minoxidil or a salt thereof; wherein the alkalinizing agent and the buffer system are the same, and / or comprise the same components.

2. The composition according to claim 1, wherein the agent configured to increase in the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is an inorganic sulfur source, e.g. cysteine, L-cysteine, hydrogen sulfide, elemental sulfur, sulfite, thiosulfate, polythionates, magnesium sulfate, sodium sulfate or sodium metabisulfite; for example sodium metabisulfite.

3. The composition according to claim 1 or claim 2, wherein the alkalinizing agent comprises a pH regulating agent, for example a buffer system selected from bicarbonate, acetate, citrate, Tris, or HEPES; for example TRIS / TRIS HC1 buffer at a pH above 7; for example from 7-10; for example from 7-9; for example from 8 to 9 of from 8 to 9.

4. The composition according to any one of the preceding claims, wherein the buffer system is selected from carbonate, c.g. sodium bicarbonatc / sodium carbonate; acetate, c.g., acetic acid / sodium acetate; citrate, e.g. mono-, di- or trisodium citrate; TRIS, e.g. TRIS / TRIS HC1; or HEPES.

5. The composition according to any one of the preceding claims, wherein the buffer system comprises TRIS / TRIS HC1 at a pH of from 8 to 9.

6. The composition according to any one of the preceding claims, wherein the alkalinizing agent and the buffer system both comprise TRIS, e.g. TRIS / TRIS HC1.

5. The composition according to any one of the preceding claims, wherein the antioxidant is selected from ascorbic acid, monothioglycerol, potassium metabisulfite, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium sulfite, sodium thiosulfate, tocopherol, sodium metabisulfite, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), ascorbyl palmitate, propyl gallate, acai oil, alpha lipoic acid, green tea extract, retinal, vitamin C, coenzyme Q10 (Co Q-10), isoflavones, polyphenols, curcumin, turmeric, pomegranate, rosemary extract, glutathione, selenium, zinc, a chelating molecule, ethylenediaminetetraacetic acid (EDTA), disodium EDTA, tetrasodium EDTA, pentasodium penetrate, sodium metasilicate and phosphate derivatives, etidronic acid and / or its derivatives, and / or galactaric acid; for example tocopherol, e.g. tocopherol acetate.

6. The composition according to any one of the preceding claims further comprising a penetration enhancer; for example diethylene glycol monoethyl ether or a polyethylene glycol derivative of a mixture of mono-, di-, and triglycerides of caprylic and capric acids with an average of 6 moles of ethylene oxide; or longer chain 1,2-glycols, e.g., Pentylene Glycol.

7. The composition according to any one of the preceding claims further comprising a chelating agent; for example tetrasodium EDTA.

8. The composition according to any one of the preceding claims further comprising one or more of: a preservative, for example an antibacterial / antifungal agt, e.g. EUXYL PE 9010 and / or Spectrastat PHL; a fragrance compound; a surfactant, e.g., polysorbate 80; and glycerol.

9. The composition according to claim 1, wherein: the agent configured to increase in the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is an inorganic sulfur source, e.g. sodium metabisulfite; the alkalinizing agent and the buffer system each comprise TRIS / TRIS HO; the antioxidant comprises tocopherol; e.g. tocopherol acetate; the solvent is water; and the liposomes comprise phosphatidylcholine.

10. The composition according to claim 9, further comprising: a chelating agent that is tetrasodium EDTA; an antibacterial / antifungal agt., e.g. EUXYL PE 9010; a fragrance compound; a surfactant, e.g., polysorbate 80; and glycerol.

11. The composition according to claim 1, having the following composition:

12. The composition according to any preceding claim, wherein the minoxidil is present as a salt, c.g., minoxidil tromethamine, minoxidil carbonate, minoxidil acetate, or minoxidil citrate.

13. The composition according to any preceding claim, wherein the minoxidil is present in the liposomes.

14. The composition according to any preceding claim, wherein the composition is a mixture, an emulsion, or a solution.

15. The composition according to any preceding claim, wherein the minoxidil is added to the other ingredients of the composition at the point of use to form the composition.

16. The composition according to any preceding claim, wherein the composition is configured to be administered by applying the composition at a predetermined frequency.

17. The composition according to any preceding claim, wherein the composition is formulated as a cosmetic product.

18. The composition according to any preceding claim, wherein the composition is formulated as any one or combination of: a time release vehicle, a cream, a solution, an emulsion, a lotion, a serum, an ointment, a spray, an aerosol medium, a capsule, a shampoo, a gel, a foam, a cosmetic, a hair conditioner, a hair care product, a hair styling product (e.g., a styling gel, a styling foam or a hair conditioner), a hair mask, a deodorant, an antiperspirant, a moisturizer, or a shaving cream or gel.

19. A Composition for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase,converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising:(i) minoxidil;(ii) an alkalinizing agent; and(iii) a penetration enhancer.

20. The composition according to claim 19, wherein the alkalinizing agent comprises a pH regulating agent that maintains the pH of the Composition at a pH of 7 or greater; for example at a pH of from 7-10; for example at a pH of from 7-9; for example at a pH of from 8-9.

21. The composition according to claim 20, wherein the pH regulating agent comprises or consists of a buffer system selected from bicarbonate, acetate, citrate, TRIS, or HEPES; for example TRIS / TRIS HC1 buffer at a pH of from 7 to 9; preferably from 8 to 9.

22. The composition according to any of claims 19-21, wherein the penetration enhancer is selected from diethylene glycol monoethyl ether, and a polyethylene glycol derivative of a mixture of mono-, di-, and triglycerides of caprylic and capric acids with an average of 6 moles of ethylene oxide; longer chain 1,2-glycols, e.g., Arachidyl Glycol (1,2- Eicosanediol, CAS 39825-93-9), Cetyl Glycol (1,2-Dihydroxyhexadecane, 1,2- Hexadecanediol, 1 ,2-Hexadecylene Glycol, 2-Hydroxycetyl Alcohol, CAS 6920-24-7), Hexacosyl Glycol (Hexacosil glycol), Lauryl Glycol (1,2-Dihydroxydodecane, 1,2- Dodecanediol, 1,2-Dodecylene Glycol, CAS 1119-87-5), Myristyl Glycol (1,2- Tetradecanediol, CAS 21129-09-9), Octacosanyl Glycol (1,2-Octacosanediol, CAS 97338-11-9), Stearyl Glycol (1,2-Dihydroxyoctadecane, 1,2-Octadecanediol, CAS 20294- 76-2), Caprylyl Glycol (Capryl Glycol, 1 ,2-Dihydroxyoctane, 1,2-Octanediol, 1,2- Octylene Glycol, CAS 1117-86-8), Decylene Glycol (1,2-Decanediol, CAS 1119-86-4), Pentylene Glycol (1,2-Dihydroxypentane, 1,2-Pentanediol, CAS 5343-92-0), 1,2-Butanediol (1,2-Butylene Glycol, 1,2-Dihydroxybutane, CAS 584-03-2), 1 ,2-Hexanediol (1,2-Dihydroxyhexane, CAS 6920-22-5), C14-18 Glycol (Ethylene Glycol Fatty AcidEster), Cl 5-18 Glycol (Alkylene (15-18) Glycol, Cetyl Stearyl Vicinal Glycol, Glycols C15-18, CAS 70750-40-2, CAS 92128-52-4), C18-3O Glycol (Ethylene Glycol Fatty Acid Ester), C20-30 Glycol (Alkylene (20-30) Glycol).

23. The composition according to any of claims 19 et seq., wherein the composition further comprises an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells; for example wherein the agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells is an inorganic sulfur source; for example wherein the inorganic sulfur source is selected from cysteine, L-cysteine, hydrogen sulfide, elemental sulfur, sulfite, thiosulfate, poly thionates, magnesium sulfate, sodium sulfate or sodium metabisulfite; for example sodium metabisulfite.

24. The composition according to any of claims 19 et seq., wherein the composition further comprises an antioxidant; for example wherein the antioxidant is selected from ascorbic acid, monothioglycerol, potassium metabisulfite, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium sulfite, sodium thiosulfate, tocopherols or ester derivatives thereof such as tocopherol acetate, sodium metabisulfite, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), ascorbyl palmitate, propyl gallate, acai oil, alpha lipoic acid, green tea extract, retinal, vitamin C, coenzyme Q10 (Co Q-10), isoflavones, polyphenols, curcumin, turmeric, pomegranate, rosemary extract, glutathione, selenium, zinc, a chelating molecule, ethylenediaminetetraacetic acid (EDTA), disodium EDTA, tetrasodium EDTA, pentasodium penetrate, sodium metasilicate and phosphate derivatives, etidronic acid and / or its derivatives, and / or galactaric acid; tocopherols, e.g. tocopherol acetate; for example wherein the antioxidant comprises tocopherol acetate.

25. The composition according to any of claims 19 et seq., wherein the composition further comprises a chelating agent; for example tetrasodium EDTA.

26. The composition according to any of claims 19 et seq., wherein the composition further comprises a preservative; for example wherein the preservative is an antibacterial and / or antifungal agent; for example wherein the preservative is EUXYL PE 9010 and / or Spectrastat PHL.

27. The composition according to any of claims 19 et seq., wherein the composition further comprises a surfactant; for example wherein the surfactant comprises polysorbate 80.

28. The composition according to any of claims 19 et seq., wherein the composition further comprises a solvent; for example wherein the solvent comprises or consists of water.

29. The composition according to any of claims 19 et seq., wherein the composition further comprises glycerin.

30. Any of the foregoing Compositions 19 et seq., wherein the composition further comprises a fragrance compound.

31. The composition according to any of claims 19 et seq., wherein the composition further comprises one or more of: a preservative, for example an antibacterial / antifungal agt., e.g. EUXYL PE 9010 and / or Spectrastat PHL; a fragrance compound; a surfactant, e.g., polysorbate 80; and glycerol.

32. The composition according to claim 20, wherein: the alkalinizing agent comprises TRIS / TRIS HC1; and the penetration enhancer is selected from longer chain 1,2-glycols.

33. The composition according to claim 32, wherein the composition further comprises: an antioxidant that comprises tocopherol; e.g. tocopherol acetate; and optionally, an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells; for example an inorganic sulfur source; for example sodium metabisulfite.

34. The composition according to claim 33, wherein the composition further comprises one or more of: a chelating agent that is tetrasodium EDTA; an antibacterial / antifungal agt., e.g. EUXYL PE 9010; a surfactant, e.g., polysorbate 80; glycerol; and a fragrance compound.

35. The composition according to claim 20, wherein the Composition comprises the following components:

36. The composition according to claim 35, further comprising one or more of the following components:

37. The composition according to claim 36, further comprising one or more of the following components:

38. The composition according to any of claims 19 et seq., wherein the minoxidil is present as a salt, e.g., minoxidil tromethamine, minoxidil carbonate, minoxidil acetate, or minoxidil citrate.

39. The composition according to any of claims 19-37, wherein the minoxidil is present in non-salt form.

40. The composition according to any of claims 19 et seq., wherein the composition is a mixture, an emulsion, or a solution.

41. The composition according to any of claims 19 et seq., wherein the composition comprises liposomes encapsulating at least some of the alkalinizing agent.

42. The composition according to claim 41 , wherein the liposomes comprise or consist of phosphatidylcholine or lecithin.

43. The composition according to claim 41, wherein the liposomes comprise or consist of phosphatidylcholine.

44. The composition according to any of claims 41-43, further comprising the following component:

45. The composition according to any of claims 41 et seq., wherein the minoxidil is present in the liposomes.

46. The composition according to any of claims 41 et seq., wherein the minoxidil is present outside of the liposomes.

47. The composition according to any of claims 41 et seq., wherein the minoxidil is present both inside and outside of the liposomes.

48. The composition according to any of claims 19 et seq., wherein the composition further comprises a buffer system configured to increase dermal penetration of the alkalinizing agent and / or the liposomes, and / or stabilize the liposome in the solution or emulsion.

49. The composition according to any of claims 19 et seq., wherein the minoxidil is added to the other ingredients of the composition at the point of use to form the composition.

50. The composition according to any of claims 19 et seq., wherein the composition is configured to be administered by applying the composition at a predetermined frequency.

51. The composition according to any of claims 19 ct scq., wherein the composition is formulated as a cosmetic product.

52. The composition according to any of claims 19 et seq., wherein the composition is formulated as any one or combination of a time release vehicle, a cream, a solution, an emulsion, a lotion, a serum, an ointment, a spray, an aerosol medium, a capsule, a shampoo, a gel, a foam, a cosmetic, a hair conditioner, a hair care product, a hair styling product (e.g., a styling gel, a styling foam or a hair conditioner), a hair mask, a deodorant, an antiperspirant, a moisturizer, or a shaving cream or gel.

53. The composition according to any of claims 19 et seq., wherein the minoxidil is present in a concentration of 2-15%.

54. The composition according to any of claims 19 et seq., wherein the composition is a therapeutic product.

55. A kit for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, the kit comprising a composition and a dispenser, implant, or pill; wherein the composition comprises a composition according to any of claims 1- 55.

56. The kit of claim 55, wherein the composition is formulated as any one or combination of: a time release vehicle, a cream, a solution, an emulsion, a lotion, a serum, an ointment, a spray, an aerosol medium, a capsule, a shampoo, a gel, a foam, a cosmetic, a hair conditioner, a hair care product, a hair styling product (e.g., a styling gel, a styling foam ora hair conditioner), a hair mask, a deodorant, an antiperspirant, a moisturizer, or a shaving cream or gel.

57. The kit of claim 56, wherein the composition is a composition according to any of claims 11 and 35-44.

58. A method for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising administering a composition according to any of claims 1-54 to a patient or subject in need thereof.

59. A method for preparing a composition for increasing the sulfonating capacity of a hair follicle cell and / or keratinocyte cell, inducing sulfotransferase in a hair follicle, inducing expression of sulfotransferase, increasing sulfotransferase activity, upregulating sulfotransferase, converting non-responders to minoxidil responders, and / or improving minoxidil response, comprising:(i) an agent configured to increase the sulfonating ability of hair bearing skin, hair follicles, and / or keratinocyte cells in a person;(ii) an alkalinizing agent;(iii) liposomes encapsulating at least some of the alkalinizing agent;(iv) an antioxidant configured to prevent oxidative degradation of the liposomes; and(v) a buffer system configured to increase dermal penetration of the liposomes, and / or stabilize the liposome solution; and(vi) a solvent; and(vii) minoxidil; optionally, a penetration enhancer; optionally, a chelating agent;wherein the liposomes comprise phosphatidylcholine and / or lecithin; and optionally wherein the alkalinizing agent and the buffer system arc the same, or comprise the same components; the method comprising: a) encapsulating at least some of each of one or more of the agent configured to increase sulfonating ability; the alkalinizing agent; the antioxidant; the penetration enhancer (if present) and the chelating agent (if present) in liposomes to form a liposome solution; and b) adding a preservative, and optionally a fragrance and optionally a surfactant to the liposome solution; c) adding minoxidil wherein the liposomes comprise phosphatidylcholine and / or lecithin.

60. The method of claim 22, wherein the product of the method is a composition according to any of claims 1-54.

61. The method of claim 22, wherein the product of the method is a composition according to any of claims 11 and 35-44.

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