Topical composition for treating and preventing alopecia and gray hair

Abstract

The present disclosure relates, in certain aspects, to pharmaceutical compositions, such as topical compositions, for use in treating or preventing alopecia and / or hair whitening in a subject. In some aspects, kits comprising the compositions are also provided, as are methods of using the compositions in the treatment of conditions, such as alopecia or gray hair.

Description

[0001] Cross-references

[0002] This application claims priority under PCT Article 8(1) and Rule 4.10 to U.S. Provisional Application No. 63 / 431,126, filed on December 8, 2022, and U.S. Provisional Application No. 63 / 530,854, filed on August 4, 2023, both of which are incorporated herein by reference as if fully set forth herein. Technical Field

[0003] The present disclosure relates in certain aspects to pharmaceutical compositions, such as topical compositions, for treating or preventing hair loss and / or graying in a subject. In some aspects, kits comprising the compositions and methods of using the compositions to treat conditions (e.g., hair loss or graying) are also provided. Background Art

[0004] Pattern hair loss, also known as androgenetic alopecia, affects up to 50% of men and 25% of women over the age of 50 (Vary JC, Med Clin North Am. 2015;99(6):1195–1211). The cause of pattern hair loss is unclear but may be related to oxidative stress, scalp microbiota, and / or hormonal abnormalities. Effective treatments for pattern hair loss, particularly those that are effective and do not cause adverse side effects, are lacking. Therefore, the continued prevalence of pattern hair loss creates an unmet need for the relief and treatment of this condition.

[0005] Incorporation by reference

[0006] Each of the cited patents, publications and non-patent documents is incorporated by reference in its entirety as if each were individually incorporated and as if each were fully set forth herein. However, such citation should not be construed as an admission that the cited reference is from fields similar to or directly applicable to the present invention, nor should the citation be construed as an admission that the document or underlying information is prior art or forms part of the common general knowledge in the art in any jurisdiction. Summary of the Invention

[0007] In order to provide a basic understanding of the present invention, the following is a brief summary of some embodiments of the present invention. This summary is not an extensive overview of the present invention. It is not intended to identify key or important elements of the present invention or to describe the scope of the present invention. Its sole purpose is to present some embodiments and aspects of the present invention in a simplified form as a preface to the more detailed description below.

[0008] In a first aspect, a topical composition for treating or preventing hair loss is provided, comprising: (i) triiodothyronine (T3); (ii) a pharmaceutically acceptable excipient; and (iii) a solvent system.

[0009] In some embodiments, the topical composition comprises about 1 nM to 30 nM T3. In some embodiments, the topical composition comprises about 10 nM T3.

[0010] In some embodiments, the pharmaceutically acceptable excipient is a penetration enhancer, a carrier, a diluent, an emulsifier, a stabilizer, a viscosity modifier, an adhesion modifier, a preservative, an antioxidant, a viscous polymer, a solubilizer, a colorant, a binder, a wetting agent, a surfactant, or a gelling agent. In some embodiments, the pharmaceutically acceptable excipient is hydroxyalkyl cellulose. In some embodiments, the pharmaceutically acceptable excipient is hydroxypropyl cellulose.

[0011] In some embodiments, the topical composition comprises about 1% to 10% (w / v) hydroxypropyl cellulose. In some embodiments, the topical composition comprises about 5% (w / v) hydroxypropyl cellulose.

[0012] In some embodiments, the solvent system comprises alcohol. In some embodiments, the solvent system comprises ethanol or propylene glycol. In some embodiments, the solvent system comprises about 10% to 70% (v / v) ethanol. In some embodiments, the solvent system comprises about 60% (v / v) ethanol. In some embodiments, the solvent system comprises about 30% (v / v) ethanol. In some embodiments, the solvent system comprises about 10% to 90% (v / v) propylene glycol. In some embodiments, the solvent system comprises about 20% (v / v) propylene glycol. In some embodiments, the solvent system comprises about 50% (v / v) propylene glycol. In some embodiments, the solvent system comprises water. In some embodiments, the solvent system comprises about 1% to 30% (v / v) water. In some embodiments, the solvent system comprises about 10% (v / v) water.

[0013] Also provided is a topical composition for treating or preventing hair loss comprising: (i) triiodothyronine (T3); (ii) hydroxypropyl cellulose; and (iii) a solvent system.

[0014] Also provided is a topical composition for treating or preventing hair loss comprising: (i) triiodothyronine (T3); (ii) hydroxypropylcellulose; (iii) ethanol; (iv) propylene glycol; and (v) water.

[0015] Also provided is a topical composition for treating or preventing hair loss, comprising: (i) about 10 nM triiodothyronine (T3); (ii) about 5% (w / v) hydroxypropyl cellulose; (iii) about 60% (v / v) ethanol; (iv) about 20% (v / v) propylene glycol; and (v) about 10% (v / v) water.

[0016] In some embodiments, the topical composition further comprises an additional active agent. In some embodiments, the additional active agent is an amino acid, an antioxidant, an anti-inflammatory agent, an analgesic, a 5-alpha reductase inhibitor, a cannabinoid, an immunosuppressant, an immunostimulant, an anticancer agent, an antiulcer agent, an antihistamine, a terpene, a vitamin, a vasodilator, or a vasoconstrictor. In some embodiments, the additional active agent is rapamycin, finasteride, dutasteride, or minoxidil. In some embodiments, the additional active agent is thyroxine (T4).

[0017] In some embodiments, the topical composition is in lyophilized form.

[0018] In another aspect, there is provided use of the topical composition of any of the disclosed embodiments for treating or preventing hair loss.

[0019] In another aspect, there is provided use of the topical composition of any one of the disclosed embodiments in the preparation of a medicament for treating or preventing hair loss.

[0020] Also provided is a method of treating or preventing hair loss in a subject, comprising administering to the subject the topical composition of any of the disclosed embodiments.

[0021] In some embodiments, the method comprises administering to the subject about 0.1 to 10 mL of the composition per unit dose. In some embodiments, the method comprises administering to the subject about 1 mL of the composition per unit dose. In some embodiments, the method comprises administering to the subject about 1 ng to 10 ng of T3 per unit dose. In some embodiments, the method comprises administering to the subject about 6.5 ng of T3 per unit dose.

[0022] In some embodiments, the composition is administered daily. In some embodiments, the composition is administered every other day. In some embodiments, the composition is administered every other day for several weeks, followed by a prolonged period of no administration. In some embodiments, the composition is administered every other day for two weeks, followed by a prolonged period of no administration. In some embodiments, the prolonged period of no administration is at least two weeks.

[0023] In some embodiments, the hair loss is caused by androgenic alopecia, alopecia areata, alopecia areata persevera, alopecia totalis, alopecia universalis, alopecia areata universalis, serpiginous alopecia, cicatricial alopecia, lichen planus, frontal fibrosing alopecia, central centrifugal cicatricial alopecia (CCCA), traction alopecia, alopecia barbae, or postpartum alopecia.

[0024] In some embodiments, the method results in an increase in hair shaft output. In some embodiments, hair shaft output is increased by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to a baseline value measured prior to administration of the composition.

[0025] In some embodiments, the method results in an extension of the anagen phase. In some embodiments, the anagen phase is extended by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to a baseline value measured before application of the composition.

[0026] In some embodiments, the method results in an increase in FGF7 expression. In some embodiments, the expression of FGF7 is increased by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a baseline value measured before administration of the composition.

[0027] In some embodiments, the method results in increased proliferation of bulge epithelial stem cells. In some embodiments, proliferation of bulge epithelial stem cells is increased by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to a baseline value measured before administration of the composition.

[0028] In some embodiments, the method results in an increase in expression of keratin 15. In some embodiments, the expression of keratin 15 is increased by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to a baseline value measured prior to application of the composition.

[0029] In some embodiments, the method results in a decrease in p-S6 expression. In some embodiments, the expression of p-S6 is decreased by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% compared to a baseline value measured before administration of the composition.

[0030] In another aspect, there is provided use of the topical composition of any of the disclosed embodiments for treating or preventing graying of hair.

[0031] In another aspect, there is provided use of the topical composition of any one of the disclosed embodiments in the preparation of a medicament for treating or preventing graying of hair.

[0032] Also provided is a method of treating or preventing graying of hair in a subject, comprising administering to the subject the topical composition of any disclosed embodiment.

[0033] In some embodiments, the topical composition includes: (i) about 1 nM triiodothyronine (T3); (ii) about 5% (w / v) hydroxypropylcellulose; (iii) about 30% (v / v) ethanol; (iv) about 50% (v / v) propylene glycol; and (v) about 10% (v / v) water.

[0034] In some embodiments, the method results in a reduction in hair depigmentation. In some embodiments, hair depigmentation is reduced by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%. In some embodiments, the method results in hair re-pigmentation.

[0035] The foregoing has summarized and summarized certain relevant features of the present disclosure so that the detailed description of the present invention below can be better understood and the contribution of the present invention to the prior art can be more fully understood. Therefore, the summary of the invention is understood to be a brief and general summary of only some of the objects and embodiments disclosed herein, which is provided solely for the benefit and convenience of the reader and is not intended to limit the scope of the legally required protection of the claims or the scope of equivalents in any way. Additional features of the present invention are described below. It should be understood by those skilled in the art that all disclosed specific compositions and methods are merely exemplary and can be easily used as the basis for modifying or designing other compositions and methods for achieving the same purpose. Such equivalent compositions and methods will be understood to also fall within the scope and spirit of the present invention set forth in the claims. It should also be understood that the titles herein are intended only to facilitate the reader's reading. They should not be interpreted as limiting the present invention in any way. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] In order to further illustrate various aspects of the present invention, a more specific description is presented by reference to certain exemplary embodiments shown in the accompanying drawings. It should be understood that these drawings describe only exemplary embodiments of the present invention and should not be considered as limiting the scope of the present invention. They are provided merely as illustrative illustrations of certain concepts of some embodiments of the present invention. These drawings and the elements described therein are not necessarily drawn to a consistent scale or any proportion. Unless the context otherwise indicates, similar numbers represent similar elements. Therefore, with reference to the accompanying drawings, certain aspects of the present invention are further described and explained as having additional features and details, but still only as examples, in which:

[0037] Figure 1A Shown are the percentages (mean + / - SEM) of hair follicle (HF) growth after 6 days of topical treatment with vehicle formulation A (Formulation A), Formulation A containing T3 (1 nM and 10 nM), or Formulation A containing T4 (1 μM and 10 μM); n = 8 HF from 1 donor (vehicle, T3, and T4) and 24 HF from 3 donors (vehicle and T3+T4 combination); Mann-Whitney test.

[0038] Figure 1B Shown are the percentages (mean + / - SEM) of hair follicle growth after 6 days of topical treatment with vehicle Formulation B (Formulation A), Formulation B containing T3 (1 nM and 10 nM), or Formulation B containing T4 (1 μM and 10 μM); n = 8 HFs from 1 donor (vehicle, T3, and T4) and 24 HFs from 3 donors (vehicle and T3+T4 combination); Mann-Whitney test.

[0039] Figure 2A Shown are 6 mm hair skin biopsies topically treated with vehicle Formulation A (A, B), Formulation A containing 1 nM T3 (C, D), Formulation A containing 10 nM T3 (E, F) for 1 day (A, C, E), and for 6 days (B, D, F).

[0040] Figure 2B Shown are 6 mm hair skin biopsies topically treated with vehicle Formulation B (A, B), Formulation B containing 1 nM T3 (C, D), Formulation B containing 10 nM T3 (E, F) for 1 day (A, C, E), and for 6 days (B, D, F).

[0041] Figure 3A Shown are the percentages of hair follicles in various hair cycle phases following administration of vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing a combination of T3 + T4 (mean + / - SEM); n = 20-35 HFs from 3 donors; Mann-Whitney test.

[0042] Figure 3B Shown are the percentages of hair follicles in various hair cycle phases following administration of vehicle Formulation B, Formulation B containing T3 (1 nM and 10 nM), Formulation B containing T4 (1 μM and 10 μM), and Formulation B containing a combination of T3 + T4 (mean + / - SEM); n = 20-35 HFs from 3 donors; Mann-Whitney test.

[0043] Figure 4AShown are the percentages of hair follicles in various hair cycle phases following administration of Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing a combination of T3+T4 (mean + / - SEM); n = 20-35 HFs from 3 donors; Mann-Whitney test.

[0044] Figure 4B Shown are the percentages of hair follicles in various hair cycle phases following administration of Formulation B, Formulation B containing T3 (1 nM and 10 nM), Formulation B containing T4 (1 μM and 10 μM), and Formulation B containing a combination of T3+T4 (mean + / - SEM); n = 20-35 HFs from 3 donors; Mann-Whitney test.

[0045] Figure 5 Shown are images of Warthin-Starry histochemistry staining (left column), Ki-67 immunofluorescence (middle column), and Casp-3 immunofluorescence (right column) of anagen hair follicles treated with vehicle formulation A, formulation A containing 1 nM T3, and formulation A containing 10 nM T3.

[0046] Figure 6A The Ki-67 expression in the hair matrix after treatment with vehicle formulation A, formulation A containing T3 (1 nM and 10 nM), formulation A containing T4 (1 μM and 10 μM), and formulation A containing a combination of T3 and T4 is shown. + Percentage of cells (mean + / - SEM); n = 17-22 HFs from 2 donors; Mann-Whitney test, *p < 0.05.

[0047] Figure 6B Ki-67 in the hair matrix after treatment with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing a combination of T3 and T4 is shown. + Percentage of cells (mean + / - SEM); n = 18-28 HFs from 2 donors; Mann-Whitney test.

[0048] Figure 7 Confocal images of Ki-67 immunofluorescence signals of anagen hair follicles treated with vehicle formulation A, formulation A containing 1 nM T3, and formulation A containing 10 nM T3 are shown.

[0049] Figure 8A Shown are melanin production (mean + / - SEM) after treatment with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing a combination of T3+T4; n=17-33 HFs from 3 donors; Mann-Whitney test.

[0050] Figure 8B Shown are melanin production (mean + / - SEM) after treatment with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing a combination of T3 + T4; n = 15-29 HFs from 3 donors; Mann-Whitney test.

[0051] Figure 9 Shown are Warthin-Starry histochemical staining of melanin granules within follicles treated with vehicle formulation A, formulation A containing 1 nM T3, and formulation A containing 10 nM T3.

[0052] Figure 10A Shown are gp100 expression levels (mean + / - SEM) in anagen follicles treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing the combination of T3 + T4; n = 13-22 HFs from 13 donors; Student's t-test, *p < 0.05.

[0053] Figure 10B Shown are gp100 expression levels (mean + / - SEM) in anagen follicles treated with vehicle Formulation B, Formulation B containing T3 (1 nM and 10 nM), Formulation B containing T4 (1 μM and 10 μM), and Formulation B containing a combination of T3 + T4; n = 13-22 HFs from 13 donors; Student's t-test, *p < 0.05.

[0054] Figure 11A Shown are MITF expression levels (mean + / - SEM) in anagen follicles treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing a combination of T3+T4; n=13-22 HFs from 13 donors; Student's t-test, *p<0.05.

[0055] Figure 11B Shown are MITF expression levels (mean + / - SEM) in anagen follicles treated with vehicle Formulation B, Formulation B containing T3 (1 nM and 10 nM), Formulation B containing T4 (1 μM and 10 μM), and Formulation B containing a combination of T3+T4; n=13-22 HFs from 13 donors; Student's t-test.

[0056] Figure 12 Shown are confocal images of immunofluorescence signals of gp100 (left column) and MITF (right column) in anagen hair follicles treated with vehicle Formulation A, Formulation A containing 1 nM T3, and Formulation A containing 10 nM T3.

[0057] Figure 13AShown are MTCO1 expression levels (mean + / - SEM) in the hair matrix apex treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing a combination of T3+T4; n=30-35 HFs from 3 donors; Mann-Whitney test, *p<0.05, ***p<0.001.

[0058] Figure 13B Shown are MTCO1 expression levels (mean + / - SEM) in the hair matrix apex treated with vehicle Formulation B, Formulation B containing T3 (1 nM and 10 nM), Formulation B containing T4 (1 μM and 10 μM), and Formulation B containing a combination of T3 + T4; n = 21-30 HFs from 3 donors; Mann-Whitney test, *p < 0.05, **p < 0.01, ***p < 0.001.

[0059] Figure 14A Shown are MTCO1 expression levels (mean + / - SEM) in the outer root sheath (ORS) treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing the combination of T3 + T4; n = 30-35 HFs from 3 donors; Mann-Whitney test, *p < 0.05, **p < 0.01, ***p < 0.001.

[0060] Figure 14B Shown are MTCO1 expression levels (mean + / - SEM) in ORS treated with vehicle Formulation B, Formulation B containing T3 (1 nM and 10 nM), Formulation B containing T4 (1 μM and 10 μM), and Formulation B containing a combination of T3 + T4; n = 21-30 HFs from 3 donors; Mann-Whitney test, *p < 0.05, **p < 0.01, ***p < 0.001.

[0061] Figure 15A Shown are K15 expression levels (mean + / - SEM) in bulge cells treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing a combination of T3 + T4; n = 21-34 HFs from 3 donors; Mann-Whitney test, *p < 0.05, **p < 0.01.

[0062] Figure 15B Shown are K15 expression levels (mean + / - SEM) in bulge cells treated with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing a combination of T3 + T4; n = 21-34 HFs from 3 donors; Mann-Whitney test.

[0063] Figure 16A The results show the effect of K15 in the bulge area treated with vehicle formulation A, formulation A containing T3 (1 nM and 10 nM), formulation A containing T4 (1 μM and 10 μM), and formulation A containing a combination of T3 and T4. + Number of cells (mean + / - SEM); n = 21-34 HFs from 3 donors; Mann-Whitney test.

[0064] Figure 16B The results show the effect of K15 in the bulge area treated with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing a combination of T3 and T4. + Number of cells (mean + / - SEM); n = 21-34 HFs from 3 donors; Mann-Whitney test, *p < 0.05, **p < 0.01.

[0065] Figure 17A Shown are the numbers (mean + / - SEM) of proliferating K15 cells (K15 and Ki67 positive cells) in the bulge area treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing the combination of T3+T4; n=21-34 HFs from 3 donors; Mann-Whitney test.

[0066] Figure 17B Shown are the numbers (mean + / - SEM) of proliferating K15 cells (K15 and Ki67 positive cells) in the bulge region treated with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing the combination of T3 + T4; n = 21-34 HFs from 3 donors; Mann-Whitney test.

[0067] Figure 18A Shown are the numbers (mean + / - SEM) of apoptotic K15 cells (K15 and Cas3 positive cells) in the bulge region treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing the combination of T3+T4; n=21-34 HFs from 3 donors; Mann-Whitney test, *p<0.05, **p<0.01.

[0068] Figure 18BShown are the numbers (mean + / - SEM) of apoptotic K15 cells (K15 and Cas3 positive cells) in the bulge area treated with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing the combination of T3 + T4; n = 21-34 HFs from 3 donors; Mann-Whitney test, *p < 0.05, **p < 0.01.

[0069] Figure 19 Shown are confocal images of immunofluorescence signals for K15 (left column), Ki67 (middle column), and Cas3 (right column) in bulge cells treated with vehicle formulation A, formulation A containing 1 nM T3, and formulation A containing 10 nM T3.

[0070] Figure 20A Shown are the expression levels of IGF-1 in ORS keratinocytes treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing the combination of T3+T4 (mean + / - SEM); n = 18-35 HFs from 3 donors; Mann-Whitney test.

[0071] Figure 20B Shown are the expression levels of IGF-1 in ORS keratinocytes treated with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing the combination of T3+T4 (mean + / - SEM); n = 18-35 HFs from 3 donors; Mann-Whitney test.

[0072] Figure 21A Shown are TGFβ-2 expression levels (mean + / - SEM) in ORS keratinocytes treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing the combination of T3+T4; n=20-28 HFs from 3 donors; Mann-Whitney test.

[0073] Figure 21B Shown are TGFβ-2 expression levels (mean + / - SEM) in ORS keratinocytes treated with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing the combination of T3 + T4; n = 22-35 HFs from 3 donors; Mann-Whitney test.

[0074] Figure 22 Confocal images of TGFβ-2 immunofluorescence signals in ORS keratinocytes treated with vehicle formulation A, formulation A containing 1 nM T3, and formulation A containing 10 nM T3 are shown.

[0075] Figure 23A Shown are FGF7 expression levels (mean + / - SEM) in ORS treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing a combination of T3+T4; n=24-32 HFs from 3 donors; Student's t-test, **p<0.01, ***p<0.001.

[0076] Figure 23B Shown are the expression levels of FGF7 in ORS treated with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing a combination of T3+T4 (mean + / - SEM); n = 17-33 HFs from 3 donors; Student's t-test.

[0077] Figure 24 Confocal images of FGF7 immunofluorescence signals in ORS keratinocytes treated with vehicle formulation A, formulation A containing 1 nM T3, and formulation A containing 10 nM T3 are shown.

[0078] Figure 25A Shown are pS6 expression levels (mean + / - SEM) in hair matrix keratinocytes treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing a combination of T3+T4; n=24-32 HFs from 3 donors; Mann-Whitney test, **p<0.01.

[0079] Figure 25B Shown are the expression levels of pS6 in hair matrix keratinocytes treated with vehicle Formulation B, Formulation B containing T3 (1 nM and 10 nM), Formulation B containing T4 (1 μM and 10 μM), and Formulation B containing a combination of T3+T4 (mean + / - SEM); n = 22-34 HFs from 3 donors; Mann-Whitney test, *p < 0.05.

[0080] Figure 26A Shown are K85 expression levels (mean + / - SEM) in hair matrix keratinocytes treated with vehicle Formulation A, Formulation A containing T3 (1 nM and 10 nM), Formulation A containing T4 (1 μM and 10 μM), and Formulation A containing a combination of T3 + T4; n = 22-34 HFs from 3 donors; Mann-Whitney test, **p < 0.01.

[0081] Figure 26BShown are K85 expression levels (mean + / - SEM) in hair matrix keratinocytes treated with vehicle Formulation B, Formulation B containing T3 (1 nM and 10 nM), Formulation B containing T4 (1 μM and 10 μM), and Formulation B containing a combination of T3 + T4; n = 22-34 HFs from 3 donors; Mann-Whitney test, **p < 0.01.

[0082] Figure 27 Shown are confocal images of K85 immunofluorescence signals in the cortical pre-hair matrix treated with vehicle formulation A, formulation A containing 10 nM T3.

[0083] Figure 28A The expression of CD31 in the dermis after treatment with vehicle formulation A, formulation A containing T3 (1 nM and 10 nM), formulation A containing T4 (1 μM and 10 μM), and formulation A containing a combination of T3 and T4 is shown. + Number of endothelial cells (mean + / - SEM); n = 22-34 HFs from 3 donors; Mann-Whitney test, *p < 0.05, **p < 0.01.

[0084] Figure 28B The expression of CD31 in the dermis after treatment with vehicle formulation B, formulation B containing T3 (1 nM and 10 nM), formulation B containing T4 (1 μM and 10 μM), and formulation B containing a combination of T3 and T4 is shown. + Number of endothelial cells (mean + / - SEM); n = 22-34 HFs from 3 donors; Mann-Whitney test, *p < 0.05, **p < 0.01.

[0085] Figure 29 Shown are confocal images of CD31 immunofluorescence signals in the dermis treated with vehicle formulation A, formulation A containing 10 nM T3. DETAILED DESCRIPTION

[0086] The above summarizes various aspects and features of certain embodiments, and the following detailed description describes several exemplary embodiments in more detail to enable those skilled in the art to practice these embodiments and realize and use the full scope of the claimed invention. The examples described are provided for illustrative purposes and are not intended to limit the scope of the invention or its applications. It should be understood that those skilled in the art may make many modifications, substitutions, changes and variations to the examples, embodiments, applications and details of the invention described herein without departing from the spirit of the invention or the scope of the invention described in the appended claims. It should also be understood that the titles herein are only used to facilitate the reader's reading. They should not be interpreted as limiting the present invention in any way.

[0087] The scope of the present invention includes all embodiments and formulations thereof, not just the embodiments and formulations thereof explicitly described below, and it should be understood that those skilled in the art may make many modifications, substitutions, changes and variations to the embodiments, applications and details of the invention described herein without departing from the spirit of the invention or the scope of the invention as described in the appended claims.

[0088] A. General Definitions and Terminology

[0089] As used in this specification and the appended claims, the singular forms "a", "an", "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "an active agent" includes reference to a combination of two or more active agents, and reference to "an excipient" includes reference to a combination of two or more excipients. Although the term "one or more" is used, its absence (or replacement by the singular) does not simply indicate the singular, but simply emphasizes the possibility of multiple agents or ingredients in a particular embodiment. The terms "comprising", "including", "for example", and "having" are inclusive rather than exclusive (i.e., other elements may be present in addition to the listed elements). The term "or" means the term "and / or" in this article and can be used interchangeably with the term "and / or" unless the context clearly dictates otherwise.

[0090] Unless otherwise indicated, all numbers representing quantities of ingredients, properties (such as concentrations, reaction conditions), etc. used to describe and claim certain embodiments of the present invention are to be understood as being modified in some cases by the term "about". Thus, in some embodiments, the numerical parameters set forth in the written description and the appended claims are approximate values that may vary depending on the desired properties sought to be obtained in the specific embodiment. In some embodiments, "about" refers to plus or minus five percent (5%) of the stated unit of measure. The term "substantially", when applied to modify a feature or limitation herein, will be understood in the context of the present invention and the knowledge in the art to provide appropriate certainty, such as by measuring the meaning of "substantially" as a term of degree using standards recognized in the art, or by determining a range as recognized by those skilled in the art.

[0091] In some embodiments, numerical parameters should be interpreted according to the number of reported significant figures and by applying conventional rounding techniques. Although the numerical ranges and parameters setting forth the broad scope of some embodiments of the present invention are approximate, the numerical values set forth in the specific examples are reported as accurately as possible. The numerical values given in some embodiments may contain certain errors, which inevitably come from the standard deviation present in their respective test measurements.

[0092] Unless otherwise defined, all technical and scientific terms herein have the meaning commonly understood by one of ordinary skill in the art to which the invention belongs, which may be referred to as "technician" as a shorthand. The following definitions are further provided to help the reader understand the disclosed embodiments; however, it should be understood that these definitions are not intended to limit the scope of the invention, which should be appropriately interpreted and understood by reference to the entire specification (and any ordinary meaning known to one of ordinary skill in the relevant art) in view of the language used in the appended claims. The terms used herein are intended only to describe specific embodiments and are not intended to be limiting.

[0093] In this article, "hair cycle phases" refer to anagen (growth phase), catagen (transition phase) and telogen (resting phase), which are key phases of human hair (Paus et al. J Investigative Derm. 2001; 117(1):3-15).

[0094] As used herein, "melanin" refers to a complex polymer derived from the amino acid tyrosine. Melanin is present in varying degrees in human skin and contributes to the distinctive color of eyes, hair, and skin (Cao et al. J Am Chem Soc. 2021; 143(7): 2622-2637).

[0095] As used herein, "Ki-67" refers to a monoclonal antibody that can be used to determine the number of circulating cells in hair follicles (Baaret al. Acta Derm Venereol. 1992; 72(2): 161-164).

[0096] As used herein, "caspase-3" is a caspase protein that interacts with caspase-8 and caspase-9. Caspases are regulators of programmed cell death, and it is likely that some specific caspases may serve as mediators of the hair growth cycle (Sawaya et al. Eur J Dermatol. 2001; 11(4): 304-308).

[0097] As used herein, "IGF-1" or "insulin-like growth factor 1" refers to a hair anagen-extending growth factor. IGF-1 has a high degree of structural and functional homology to insulin and exhibits anti-apoptotic effects (Ahn et al. Ann Dermatol. 2012; 24(1): 26-31).

[0098] As used herein, "KGF / FGF7" or "keratinocyte growth factor / fibroblast growth factor-7" refers to a hair anagen extension growth factor.

[0099] As used herein, "TGFβ-2" or "transforming growth factor β2" refers to a hair catagen growth-promoting factor (Xu et al. Bone Research. 2018; 6(2): 1-31).

[0100] "MTCOI" or "mitochondrial cytochrome c oxidase subunit I" herein refers to a component of cytochrome c oxidase, which is the last enzyme in the mitochondrial electron transport chain that drives oxidative phosphorylation (Vidali et al. J Invest Dermatol. 2016; 136(10):2003-2012).

[0101] Herein, "Gp100" refers to a well-established and sensitive tracer of melanosome transfer between melanocytes and keratinocytes (Singh et al. Exp Dermatol. 2008; 17(5):418-426).

[0102] As used herein, "MITF" or "melanocyte-inducing transcription factor" refers to a key transcription factor in melanocyte development and differentiation (Levt et al. Trends Mol Med. 2006; 12(9): 406-414).

[0103] Herein, "p-S6" refers to the direct downstream kinase of mTORC1. Increased p-S6 indicates mTORC1 activation, which is associated with aging and hair graying (Suzuki et al. EMBO Reports. 2023; 24: e56574).

[0104] In this article, "K85" is a sensitive marker of hair shaft keratin production (Ramot et al. Br JDermatol. 2013; 169(1): 146-51).

[0105] As used herein, "CD31," also known as "platelet endothelial cell adhesion molecule-1," is a marker of intraepithelial angiogenesis measured by CD31 immunoreactivity and the number of CD31-positive cells. CD31 is a transmembrane cognate receptor expressed by endothelial cells, platelets, granulocytes, macrophages, dendritic cells, T- and B-cells, and natural killer cells (Berg et al. J Cell Sci. 2013; 126(11): 2343-2352).

[0106] As used herein, "treating" or "treatment" of a disease, such as hair loss, a hair loss disorder, or androgenic alopecia, includes any treatment of a disease in a mammal, preferably a human, including: (a) preventing the development of the disease in a patient who may be susceptible to the disease but has not yet been diagnosed with the disease; (b) inhibiting the disease, i.e., arresting its development, including prevention; (c) alleviating the disease, i.e., causing regression of the disease or its clinical symptoms; (d) preventing or alleviating symptoms or pathology caused by or associated with the disease; (e) reducing, decreasing, inhibiting, ameliorating, or preventing the onset, severity, duration, progression, frequency, or probability of one or more symptoms or pathology associated with the disease; and (f) preventing or inhibiting the worsening or progression of symptoms or pathology associated with the disease or comorbid with the disease.

[0107] As used herein, an "effective amount," "therapeutically effective amount," or "pharmacologically effective amount" refers to an amount of an active agent (e.g., triiodothyronine (T3), thyroxine (T4), or a combination thereof) that is non-toxic and sufficient to provide the desired therapeutic effect at a reasonable benefit / risk ratio in any medical treatment. The effective amount will vary depending on the subject, their weight and age, the severity of the symptoms or degree of health benefit sought, the mode of administration, and the like, all of which can be readily determined by one skilled in the art.

[0108] As used herein, "therapeutic effect" or "therapeutic efficacy" refers to a response in a subject (preferably a human) that is judged to be desirable and beneficial following treatment. Thus, these responses will vary depending on the condition being treated, or the improvement in health or function being sought, as well as on the specific components of the present method being considered, but will be readily understood by those skilled in the art.

[0109] As used herein, the terms "subject," "user," "patient," and "individual" are used interchangeably and refer to a human, mammal, or any other animal susceptible to hair loss, a hair loss condition, or androgenic alopecia. Preferably, the subject is a human. The subject can be a human infant, a human child, an adult, or an elderly person. These terms will be understood to include anyone who has an indication for which the methods described herein may be effective, or who may benefit from the present invention. In general, all disclosed methods will be considered effective for all individuals, although individual differences are expected and understood. The disclosed treatment methods can also be modified to treat multiple patients, including couples or families, simultaneously. Therefore, these terms will be understood to also refer to two or more individuals.

[0110] Generally speaking, the terms used herein and the procedures performed are those known in the fields (e.g., biology, biochemistry, dermatology, pharmacology, and medicine) relevant to one or more aspects of the present invention, and are well-known and commonly used in these fields. Standard techniques and procedures are generally performed according to conventional methods in the art.

[0111] Additional definitions and abbreviations are provided elsewhere in this document.

[0112] B. Topical Compositions

[0113] The present disclosure relates in certain aspects to methods for treating or preventing hair loss (e.g., hair loss caused by a hair loss disorder, such as androgenic alopecia) in a subject (e.g., preferably a human). In some aspects, the present disclosure also relates to pharmaceutical (e.g., topical) compositions and kits for use in the methods. In some aspects, useful features of the methods of the present disclosure include curing or alleviating symptoms in a subject suffering from a hair loss disorder (e.g., androgenic alopecia).

[0114] Without being bound by theory, thyroid hormone may stimulate the proliferation and differentiation of stem cells in the hair matrix by downregulating TGFβ-2 (an important hair catagen growth factor) and by upregulating K15 expression (an upregulator of epithelial progenitor cell markers). For example, see Paus et al. The J Clin Endocrinol Metab. 2008; 93(11): 4381–4388; Paus et al. J Invest Dermatol. 2014; 134(1): 33-42; Paus, et al. PLoS One. 2019; 14(3): e0212659; Paus et al. J Investigative Derm. 2016; 136: 1711–1714; Paus et al. Experimental Derm. 2020; 29(9): 910-923; each of which is incorporated herein by reference in its entirety. Because of the complexity of the intracellular responses downstream of free and bound thyroid hormone, other mechanisms of action may exist but are still unknown.

[0115] In one aspect, a topical composition is provided, comprising a therapeutically effective amount of triiodothyronine (T3), thyroxine (T4), or a combination thereof. In some embodiments, the topical composition comprises T3 and T4. In some embodiments, the composition comprises a therapeutically effective amount of T3. In some embodiments, the composition comprises a therapeutically effective amount of T4. In some embodiments, the composition comprises a therapeutically effective amount of T3 and T4. In some embodiments, the composition comprises a combination of a therapeutically acceptable amount of T3 and T4. In some embodiments, the composition is suitable for topical or transdermal administration. In some embodiments, the composition is formulated for topical administration. In some embodiments, the composition is formulated for transdermal administration. In some embodiments, the topical composition comprises a pharmaceutically acceptable excipient.

[0116] In another aspect, a topical composition for treating or preventing hair loss is provided, comprising: (i) T3; (ii) a pharmaceutically acceptable excipient; and (iii) a solvent system. In some embodiments, the topical composition comprises T3 as the sole active ingredient. Thus, a topical composition for treating or preventing hair loss is also provided, comprising: (i) T3 as the sole active ingredient; (ii) a pharmaceutically acceptable excipient; and (iii) a solvent system. Also provided is a topical composition for treating or preventing hair loss consisting essentially of: (i) T3; (ii) a pharmaceutically acceptable excipient; and (iii) a solvent system.

[0117] In another aspect, a topical composition for treating or preventing graying of hair is provided, comprising: (i) T3; (ii) a pharmaceutically acceptable excipient; and (iii) a solvent system. In some embodiments, the topical composition comprises T3 as the sole active ingredient. Thus, a topical composition for treating or preventing graying of hair is also provided, comprising: (i) T3 as the sole active ingredient; (ii) a pharmaceutically acceptable excipient; and (iii) a solvent system. Also provided is a topical composition for treating or preventing graying of hair, consisting essentially of: (i) T3; (ii) a pharmaceutically acceptable excipient; and (iii) a solvent system.

[0118] In some embodiments, the topical compositions of the present disclosure do not include T4. In some embodiments, the topical compositions do not include iodothyronamine (T1a). In some embodiments, the topical compositions do not include thyronamine (TOa). In some embodiments, the topical compositions do not include cardiac glycosides. In some embodiments, the topical compositions do not include sterols. In some embodiments, the compositions do not include phytosterols. In some embodiments, the compositions do not include 13-sitosterol. In some embodiments, the compositions do not include hormones other than thyroid hormones (e.g., T3 or T4). In some embodiments, the topical compositions do not include human growth hormone. In some embodiments, the topical compositions do not include insulin. In some embodiments, the topical compositions do not include estradiol. In some embodiments, the topical compositions do not include an estrogen source. In some embodiments, the topical compositions do not include a dihydrotestosterone blocker. In some embodiments, the topical compositions do not include a progesterone source. In some embodiments, the topical compositions do not include estradiol benzoate. In some embodiments, the topical compositions do not include medroxyprogesterone acetate. In some embodiments, the topical compositions do not include a vasodilator. In some embodiments, the topical composition does not comprise a monoamine oxidase inhibitor. In some embodiments, the topical composition does not comprise an IL-15 polynucleotide, polypeptide, or a compound that is bound to an antibody that specifically recognizes an IL-15 polypeptide or that specifically binds to the α chain of the IL-15 receptor.

[0119] In some embodiments, the composition comprises T3 at a concentration between about 1 nM and 30 nM. In some embodiments, the composition comprises T3 at a concentration of less than about 1 nM, about 1 nM, about 2 nM, about 3 nM, about 4 nM, about 5 nM, about 6 nM, about 7 nM, about 8 nM, about 9 nM, about 10 nM, about 15 nM, about 20 nM, about 25 nM, about 30 nM, or greater than about 30 nM, including ranges between these values. In some embodiments, the composition comprises T3 at a concentration of less than about 1 nM. In some embodiments, the composition comprises T3 at a concentration of about 1 nM. In some embodiments, the composition comprises T3 at a concentration of about 2 nM. In some embodiments, the composition comprises T3 at a concentration of about 3 nM. In some embodiments, the composition comprises T3 at a concentration of about 4 nM. In some embodiments, the composition comprises T3 at a concentration of about 5 nM. In some embodiments, the composition comprises T3 at a concentration of about 6 nM. In some embodiments, the composition comprises T3 at a concentration of about 7 nM. In some embodiments, the composition comprises T3 at a concentration of about 8 nM. In some embodiments, the composition comprises T3 at a concentration of about 9 nM. In some embodiments, the composition comprises T3 at a concentration of about 10 nM. In some embodiments, the composition comprises T3 at a concentration of about 15 nM. In some embodiments, the composition comprises T3 at a concentration of about 20 nM. In some embodiments, the composition comprises T3 at a concentration of about 25 nM. In some embodiments, the composition comprises T3 at a concentration of about 30 nM. In some embodiments, the composition comprises T3 at a concentration greater than about 30 nM.

[0120] In some embodiments, the composition comprises T3 at a concentration between about 1 μM and 15 μM. In some embodiments, the composition comprises T3 at a concentration of less than about 1 μM, about 1 μM, about 2 μM, about 3 μM, about 4 μM, about 5 μM, about 6 μM, about 7 μM, about 8 μM, about 9 μM, about 10 μM, about 11 μM, about 12 μM, about 13 μM, about 14 μM, about 15 μM, or greater than about 15 μM. In some embodiments, the composition comprises T3 at a concentration of about 1 μM. In some embodiments, the composition comprises T3 at a concentration of about 2 μM. In some embodiments, the composition comprises T3 at a concentration of about 3 μM. In some embodiments, the composition comprises T3 at a concentration of about 4 μM. In some embodiments, the composition comprises T3 at a concentration of about 5 μM. In some embodiments, the composition comprises T3 at a concentration of about 6 μM. In some embodiments, the composition comprises T3 at a concentration of about 7 μM. In some embodiments, the composition comprises T3 at a concentration of about 8 μM. In some embodiments, the composition comprises T3 at a concentration of about 9 μM. In some embodiments, the composition comprises T3 at a concentration of about 10 μM. In some embodiments, the composition comprises T3 at a concentration of about 11 μM. In some embodiments, the composition comprises T3 at a concentration of about 12 μM. In some embodiments, the composition comprises T3 at a concentration of about 13 μM. In some embodiments, the composition comprises T3 at a concentration of about 14 μM. In some embodiments, the composition comprises T3 at a concentration of about 15 μM. In some embodiments, the composition comprises T3 at a concentration of greater than about 15 μM.

[0121] In some embodiments, the composition comprises T3 at a concentration of about 50 pM, 60 pM, 70 pM, 80 pM, 90 pM, 100 pM, 110 pM, 120 pM, 130 pM, 140 pM, 150 pM, 160 pM, 170 pM, 180 pM, 190 pM, or 200 pM, or a dose therebetween. In some embodiments, the composition comprises T3 at a concentration of about 50 pM. In some embodiments, the composition comprises T3 at a concentration of about 60 pM. In some embodiments, the composition comprises T3 at a concentration of about 70 pM. In some embodiments, the composition comprises T3 at a concentration of about 80 pM. In some embodiments, the composition comprises T3 at a concentration of about 90 pM. In some embodiments, the composition comprises T3 at a concentration of about 100 pM. In some embodiments, the composition comprises T3 at a concentration of about 110 pM. In some embodiments, the composition comprises T3 at a concentration of about 120 pM. In some embodiments, the composition comprises T3 at a concentration of about 130 pM. In some embodiments, the composition comprises T3 at a concentration of about 140 pM. In some embodiments, the composition comprises T3 at a concentration of about 150 pM. In some embodiments, the composition comprises T3 at a concentration of about 160 pM. In some embodiments, the composition comprises T3 at a concentration of about 170 pM. In some embodiments, the composition comprises T3 at a concentration of about 180 pM. In some embodiments, the composition comprises T3 at a concentration of about 190 pM. In some embodiments, the composition comprises T3 at a concentration of about 200 pM.

[0122] In some embodiments, the composition comprises T4 at a concentration between about 1 μM and 30 μM. In some embodiments, the composition comprises T4 at a concentration of less than about 1 μM, about 1 μM, about 2 μM, about 3 μM, about 4 μM, about 5 μM, about 6 μM, about 7 μM, about 8 μM, about 9 μM, about 10 μM, about 15 μM, about 20 μM, about 25 μM, about 30 μM, or greater than about 30 μM, including ranges between these values. In some embodiments, the composition comprises T4 at a concentration of less than about 1 μM. In some embodiments, the composition comprises T4 at a concentration of about 1 μM. In some embodiments, the composition comprises T4 at a concentration of about 2 μM. In some embodiments, the composition comprises T4 at a concentration of about 3 μM. In some embodiments, the composition comprises T4 at a concentration of about 4 μM. In some embodiments, the composition comprises T4 at a concentration of about 5 μM. In some embodiments, the composition comprises T4 at a concentration of about 6 μM. In some embodiments, the composition comprises T4 at a concentration of about 7 μM. In some embodiments, the composition comprises T4 at a concentration of about 8 μM. In some embodiments, the composition comprises T4 at a concentration of about 9 μM. In some embodiments, the composition comprises T4 at a concentration of about 10 μM. In some embodiments, the composition comprises T4 at a concentration of about 15 μM. In some embodiments, the composition comprises T4 at a concentration of about 20 μM. In some embodiments, the composition comprises T4 at a concentration of about 25 μM. In some embodiments, the composition comprises T4 at a concentration of about 30 μM. In some embodiments, the composition comprises T4 at a concentration greater than about 30 μM.

[0123] In some embodiments, the composition comprises T4 at a concentration between about 10 μM and 15000 μM. In some embodiments, the composition comprises T4 at a concentration of less than about 10 μM, about 10 μM, about 20 μM, about 50 μM, about 100 μM, about 250 μM, about 500 μM, about 1000 μM, about 2500 μM, about 5000 μM, about 10000 μM, about 12000 μM, about 15000 μM, or greater than about 15000 μM, including ranges between these values. In some embodiments, the composition comprises T4 at a concentration of less than about 10 μM. In some embodiments, the composition comprises T4 at a concentration of about 10 μM. In some embodiments, the composition comprises T4 at a concentration of about 20 μM. In some embodiments, the composition comprises T4 at a concentration of about 50 μM. In some embodiments, the composition comprises T4 at a concentration of about 100 μM. In some embodiments, the composition comprises T4 at a concentration of about 250 μM. In some embodiments, the composition comprises T4 at a concentration of about 500 μM. In some embodiments, the composition comprises T4 at a concentration of about 1000 μM. In some embodiments, the composition comprises T4 at a concentration of about 2500 μM. In some embodiments, the composition comprises T4 at a concentration of about 5000 μM. In some embodiments, the composition comprises T4 at a concentration of about 10000 μM. In some embodiments, the composition comprises T4 at a concentration of about 12000 μM. In some embodiments, the composition comprises T4 at a concentration of about 15000 μM. In some embodiments, the composition comprises T4 at a concentration of greater than about 15000 μM.

[0124] In some embodiments, the composition comprises T4 at a concentration of about 50 nM, 60 nM, 70 nM, 80 nM, 90 nM, 100 nM, 110 nM, 120 nM, 130 nM, 140 nM, 150 nM, 160 nM, 170 nM, 180 nM, 190 nM, or 200 nM, or a dose therebetween. In some embodiments, the composition comprises T4 at a concentration of about 50 nM. In some embodiments, the composition comprises T4 at a concentration of about 60 nM. In some embodiments, the composition comprises T4 at a concentration of about 70 nM. In some embodiments, the composition comprises T4 at a concentration of about 80 nM. In some embodiments, the composition comprises T4 at a concentration of about 90 nM. In some embodiments, the composition comprises T4 at a concentration of about 100 nM. In some embodiments, the composition comprises T4 at a concentration of about 120 nM. In some embodiments, the composition comprises T4 at a concentration of about 130 nM. In some embodiments, the composition comprises T4 at a concentration of about 140 nM. In some embodiments, the composition comprises T4 at a concentration of about 150 nM. In some embodiments, the composition comprises T4 at a concentration of about 160 nM. In some embodiments, the composition comprises T4 at a concentration of about 170 nM. In some embodiments, the composition comprises T4 at a concentration of about 180 nM. In some embodiments, the composition comprises T4 at a concentration of about 190 nM. In some embodiments, the composition comprises T4 at a concentration of about 200 nM.

[0125] In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of between about 1:100 and about 1:10000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of between about 1:100 and about 1:1000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:100, 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, or 1:1000, including ranges between these values. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of less than about 1:100. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:100. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:200. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:300. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:400. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:500. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:600. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:700. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:800. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:900. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:1000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of greater than about 1:1000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of between about 1:1000 and about 1:10000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:1000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:2000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:3000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:4000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:5000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:6000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:7000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:8000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1:9000.In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio of about 1: 10,000. In some embodiments, the composition comprises T3 and T4 in a T3:T4 ratio greater than about 1:10,000.

[0126] In some embodiments, the composition comprises T3 at a concentration of about 100 pM and T4 at a concentration of about 100 nM. In some embodiments, the composition comprises T3 at a concentration of about 100 pM, T4 at a concentration of about 100 nM, and a therapeutically effective amount of rapamycin. In some embodiments, the composition comprises T3 at a concentration of about 10 nM and T4 at a concentration of about 1 μM. In some embodiments, the composition comprises T3 at a concentration of about 10 nM and T4 at a concentration of about 10 μM. In some embodiments, the composition comprises T3 at a concentration of about 1 nM and T4 at a concentration of about 10 μM.

[0127] In some embodiments, the topical composition comprises:

[0128] a. Triiodothyronine (T3);

[0129] b. Thyroxine (T4); and

[0130] c.Solvent.

[0131] In some embodiments, the topical composition comprises:

[0132] a. Triiodothyronine (T3);

[0133] b. Thyroxine (T4); and

[0134] c. Ethanol.

[0135] In some embodiments, the topical composition comprises:

[0136] a. Triiodothyronine (T3);

[0137] b. Thyroxine (T4); and

[0138] c. Water.

[0139] In some embodiments, the topical composition comprises:

[0140] a. Triiodothyronine (T3);

[0141] b. Thyroxine (T4); and

[0142] c. Propylene glycol.

[0143] In some embodiments, the topical composition comprises:

[0144] a. Triiodothyronine (T3);

[0145] b.Thyroxine (T4);

[0146] c. ethanol; and

[0147] d. Water.

[0148] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0149] b.Thyroxine (T4);

[0150] c. ethanol; and

[0151] d. Propylene glycol.

[0152] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0153] b.Thyroxine (T4);

[0154] c. propylene glycol; and

[0155] d. Water.

[0156] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0157] b.Thyroxine (T4);

[0158] c. ethanol;

[0159] d. propylene glycol; and

[0160] e. Water.

[0161] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0162] b.Thyroxine (T4);

[0163] c. Hydroxypropyl cellulose; and

[0164] d.Solvent.

[0165] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0166] b.Thyroxine (T4);

[0167] c. Hydroxypropyl cellulose; and

[0168] d. Ethanol.

[0169] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0170] b.Thyroxine (T4);

[0171] c. Hydroxypropyl cellulose; and

[0172] d. Water.

[0173] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0174] b.Thyroxine (T4);

[0175] c. Hydroxypropyl cellulose; and

[0176] d. Propylene glycol.

[0177] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0178] b.Thyroxine (T4);

[0179] c. Hydroxypropyl cellulose;

[0180] d. ethanol; and

[0181] e. Water.

[0182] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0183] b.Thyroxine (T4);

[0184] c. Hydroxypropyl cellulose;

[0185] d. ethanol; and

[0186] e. Propylene glycol.

[0187] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0188] b.Thyroxine (T4);

[0189] c. Hydroxypropyl cellulose;

[0190] d. propylene glycol; and

[0191] e. Water.

[0192] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0193] b.Thyroxine (T4);

[0194] c. Hydroxypropyl cellulose;

[0195] d. ethanol;

[0196] e. propylene glycol; and

[0197] f. Water.

[0198] In some embodiments, a topical composition comprises: a. triiodothyronine (T3); and b. a solvent.

[0199] In some embodiments, the topical composition comprises: a. triiodothyronine (T3); and b. ethanol.

[0200] In some embodiments, the topical composition comprises: a. triiodothyronine (T3); and b. water.

[0201] In some embodiments, the topical composition comprises: a. triiodothyronine (T3); and b. propylene glycol.

[0202] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0203] b. ethanol; and

[0204] c. Water.

[0205] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0206] b. ethanol; and

[0207] c. Propylene glycol.

[0208] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0209] b. propylene glycol; and

[0210] c. Water.

[0211] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0212] b. ethanol;

[0213] c. propylene glycol; and

[0214] d. Water.

[0215] In some embodiments, a topical composition comprises: a. triiodothyronine (T3); b. hydroxypropylcellulose; and c. a solvent.

[0216] In some embodiments, the topical composition comprises: a. triiodothyronine (T3); b. hydroxypropylcellulose; and c. ethanol.

[0217] In some embodiments, the topical composition comprises: a. triiodothyronine (T3); b. hydroxypropylcellulose; and c. water.

[0218] In some embodiments, the topical composition comprises: a. triiodothyronine (T3); b. hydroxypropylcellulose; and c. propylene glycol.

[0219] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0220] b. Hydroxypropyl cellulose;

[0221] c. ethanol; and

[0222] d. Water.

[0223] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0224] b. Hydroxypropyl cellulose;

[0225] c. ethanol; and

[0226] d. Propylene glycol.

[0227] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0228] b. Hydroxypropyl cellulose;

[0229] c. propylene glycol; and

[0230] d. Water.

[0231] In some embodiments, the topical composition includes: a. triiodothyronine (T3);

[0232] b. Hydroxypropyl cellulose;

[0233] c. ethanol;

[0234] d. propylene glycol; and

[0235] e. Water.

[0236] In some embodiments, the topical composition comprises: a. thyroxine (T4); and b. a solvent.

[0237] In some embodiments, the topical composition comprises: a. thyroxine (T4); and b. ethanol.

[0238] In some embodiments, the topical composition comprises: a. thyroxine (T4); and b. water.

[0239] In some embodiments, the topical composition includes: a. thyroxine (T4); and b. propylene glycol.

[0240] In some embodiments, the topical composition includes: a. thyroxine (T4);

[0241] b. ethanol; and

[0242] c. Water.

[0243] In some embodiments, the topical composition includes: a. thyroxine (T4);

[0244] b. ethanol; and

[0245] c. Propylene glycol.

[0246] In some embodiments, the topical composition includes: a. thyroxine (T4);

[0247] b. propylene glycol; and

[0248] c. Water.

[0249] In some embodiments, the topical composition includes: a. thyroxine (T4);

[0250] b. ethanol;

[0251] c. propylene glycol; and

[0252] d. Water.

[0253] In some embodiments, the topical composition includes: a. thyroxine (T4);

[0254] b. Hydroxypropyl cellulose; and

[0255] c.Solvent.

[0256] In some embodiments, the topical composition includes: a. thyroxine (T4);

[0257] b. Hydroxypropyl cellulose; and

[0258] c. Ethanol.

[0259] In some embodiments, the topical composition includes: a. thyroxine (T4);

[0260] b. Hydroxypropyl cellulose; and

[0261] c. Water.

[0262] In some embodiments, the topical composition comprises: a.(T3);

[0263] b.Thyroxine (T4);

[0264] c. Hydroxypropyl cellulose; and

[0265] d. Propylene glycol.

[0266] In some embodiments, the topical composition includes: a. thyroxine (T4);

[0267] b. Hydroxypropyl cellulose;

[0268] c. ethanol; and

[0269] d. Water.

[0270] In some embodiments, the topical composition comprises:

[0271] a. Thyroxine (T4);

[0272] b. Hydroxypropyl cellulose;

[0273] c. ethanol; and

[0274] d. Propylene glycol.

[0275] In some embodiments, the topical composition comprises:

[0276] a. Thyroxine (T4);

[0277] b. Hydroxypropyl cellulose;

[0278] c. propylene glycol; and

[0279] d. Water.

[0280] In some embodiments, the topical composition comprises:

[0281] a. Thyroxine (T4);

[0282] b. Hydroxypropyl cellulose;

[0283] c. ethanol;

[0284] d. propylene glycol; and

[0285] e. Water.

[0286] In some embodiments, the composition further comprises a therapeutically effective amount of an additional active compound. In some embodiments, the additional active agent is selected to provide a synergistic effect. In embodiments, "synergism" will be understood to include an increase in efficacy, biological activity, bioaccessibility, bioavailability, or therapeutic effect that is greater than the cumulative contribution of the components acting alone, and / or greater than the contribution of the isolated compounds themselves. There are many methods known to those skilled in the art for determining whether there is synergy of a particular effect, that is, when two or more components are mixed together, whether the effect is greater than the sum of the effects of the individual components when applied alone, thereby producing a "1+1>2" effect, one of which is isobologram analysis (or contour method) (Huang et al. 2019).

[0287] In some embodiments, the additional active agents are selected to provide additional therapeutic effects, such as antioxidant, anti-inflammatory, analgesic, antinociceptive, immunostimulatory, immunosuppressive, anticancer, antiemetic, antiulcer, antihistamine, vasodilatory, and vasoconstrictive effects.

[0288] In some embodiments, the additional active agent is an amino acid, an antioxidant, an anti-inflammatory agent, an analgesic, a 5-alpha reductase inhibitor, a cannabinoid, an immunosuppressant, an immunostimulant, an anticancer agent, an antiulcer agent, an antihistamine, a terpene, a vitamin, a vasodilator, or a vasoconstrictor. These active agents may be in ionic, free base, or salt form, including polymorphs, and may be isomers.

[0289] In some embodiments, the additional active compound is rapamycin. In some embodiments, the additional active compound is rapamycin. In some embodiments, the additional active compound is finasteride. In some embodiments, the additional active compound is dutasteride. In some embodiments, the additional active compound is minoxidil.

[0290] In another aspect, provided is a use of the topical composition of any of the disclosed embodiments for treating hair loss. In some embodiments, the hair loss is caused by androgenic alopecia.

[0291] In another aspect, provided is a use of the topical composition of the disclosed embodiments for treating or preventing hair loss. In some embodiments, the hair loss is caused by androgenic alopecia.

[0292] In some embodiments, the topical compositions can be applied and dosed according to good medical practice, taking into account the method and schedule of administration, previous and concomitant medications and medical supplements, the individual patient's clinical condition and the severity of the underlying disease, the patient's age, sex, weight, and other factors relevant to the physician, as well as knowledge of the specific compound being used. Therefore, dosage levels may vary from patient to patient, from patient to patient over time, and for different compositions and formulations, but should be able to be determined by routine techniques.

[0293] Determining the appropriate dosage should include determining not only a single dose, but also the number and timing of doses, as well as the optimal time of administration.

[0294] The present disclosure also provides a package comprising the disclosed compositions. In some embodiments, the package provides the disclosed compositions in unit dosage form. The package generally includes suitable packaging. The package can include one or more containers, and the container includes any composition described herein. Each component (if there are multiple components) can be packaged in separate containers, or when cross-reactivity and shelf life allow, some components can be combined in one container. The package can be a unit dosage form, a large package (such as a multi-dose package) or a subunit dose.

[0295] For example, suit of the present disclosure can comprise the present composition of sufficient dosage, for individual one section extension time, for example one week, two weeks, three weeks, four weeks, six weeks, eight weeks, three months, four months, five months, seven months, eight months, nine months or longer.Suit may also comprise multiple unit doses and instructions for use, and the package quantity is enough to store at home or in retail outlets.Suit optionally comprises one group of instructions relevant to the assembly using the disclosed method, normally written instructions, or the electronic storage medium (such as magnetic disk or CD) comprising instructions.The instructions contained in suit generally include relevant assembly and information used by individuals thereof.

[0296] In some embodiments, compositions of the present disclosure (whether or not in unit dosage form) can be provided in lyophilized form. Lyophilization, also referred to as freeze drying, is a method commonly used in preserving and stabilizing pharmaceutical compounds and compositions. Lyophilization can obtain a dry and storage-resistant product, which can then be reconstituted before use. The lyophilization of compositions of the present disclosure can provide some advantages. For example, in some embodiments, compared with the same compositions not provided in lyophilized form, the compositions of the present disclosure in lyophilized form have improved stability (for example, improved storage stability) and / or reduce sensitivity to chemical, thermal or biological degradation. In some embodiments, compared with the same compositions not provided in lyophilized form, the compositions of the present disclosure in lyophilized form have reduced weight and / or volume, which can reduce the cost and overall difficulty of composition transportation, storage, distribution and use.

[0297] C. Method

[0298] In one aspect, a method of treating or preventing hair loss in a subject is provided, the method comprising administering to the subject a therapeutically effective amount of a topical composition of the present disclosure. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of T3. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of T4. In some embodiments, the method comprises administering to the subject a therapeutically effective amount of T3 and T4. In some embodiments, the hair loss is caused by androgenetic alopecia.

[0299] In some embodiments, T3, T4, or a combination of T3 and T4 is administered topically. In some embodiments, T3 is administered topically. In some embodiments, T4 is administered topically. In some embodiments, T3 and T4 are administered topically (i.e., a combination is administered topically).

[0300] In some embodiments, T3 and T4 are administered simultaneously. The simultaneous administration of T3 and T4 can be achieved, for example, by administering a composition comprising T3 and T4 (e.g., a topical composition of the present disclosure). In another embodiment, the simultaneous administration of T3 and T4 can be achieved, for example, by administering separate T3 and T4 compositions simultaneously.

[0301] In some embodiments, T3 is administered at a concentration of about 1 nM to 30 nM. In some embodiments, T3 is administered at a concentration of less than about 1 nM, about 1 nM, about 2 nM, about 3 nM, about 4 nM, about 5 nM, about 6 nM, about 7 nM, about 8 nM, about 9 nM, about 10 nM, about 15 nM, about 20 nM, about 25 nM, about 30 nM, or greater than about 30 nM, including ranges between these values. In some embodiments, T3 is administered at a concentration of less than about 1 nM. In some embodiments, T3 is administered at a concentration of about 1 nM. In some embodiments, T3 is administered at a concentration of about 2 nM. In some embodiments, T3 is administered at a concentration of about 3 nM. In some embodiments, T3 is administered at a concentration of about 4 nM. In some embodiments, T3 is administered at a concentration of about 5 nM. In some embodiments, T3 is administered at a concentration of about 6 nM. In some embodiments, T3 is administered at a concentration of about 7 nM. In some embodiments, T3 is administered at a concentration of about 8 nM. In some embodiments, T3 is administered at a concentration of about 9 nM. In some embodiments, T3 is administered at a concentration of about 10 nM. In some embodiments, T3 is administered at a concentration of about 15 nM. In some embodiments, T3 is administered at a concentration of about 20 nM. In some embodiments, T3 is administered at a concentration of about 25 nM. In some embodiments, T3 is administered at a concentration of about 30 nM. In some embodiments, T3 is administered at a concentration greater than about 30 nM.

[0302] In some embodiments, T3 is administered at a concentration between about 1 μM and 15 μM. In some embodiments, T3 is administered at a concentration of less than about 1 μM, about 1 μM, about 2 μM, about 3 μM, about 4 μM, about 5 μM, about 6 μM, about 7 μM, about 8 μM, about 9 μM, about 10 μM, about 11 μM, about 12 μM, about 13 μM, about 14 μM, about 15 μM, or greater than about 15 μM, including ranges between these values. In some embodiments, T3 is administered at a concentration of less than about 1 μM. In some embodiments, T3 is administered at a concentration of about 1 μM. In some embodiments, T3 is administered at a concentration of about 2 μM. In some embodiments, T3 is administered at a concentration of about 3 μM. In some embodiments, T3 is administered at a concentration of about 4 μM. In some embodiments, T3 is administered at a concentration of about 5 μM. In some embodiments, T3 is administered at a concentration of about 6 μM. In some embodiments, T3 is administered at a concentration of about 7 μM. In some embodiments, T3 is administered at a concentration of about 8 μM. In some embodiments, T3 is administered at a concentration of about 9 μM. In some embodiments, T3 is administered at a concentration of about 10 μM. In some embodiments, T3 is administered at a concentration of about 11 μM. In some embodiments, T3 is administered at a concentration of about 12 μM. In some embodiments, T3 is administered at a concentration of about 13 μM. In some embodiments, T3 is administered at a concentration of about 14 μM. In some embodiments, T3 is administered at a concentration of about 15 μM. In some embodiments, T3 is administered at a concentration greater than about 15 μM.

[0303] In some embodiments, T3 is administered at a concentration of about 50 pM, 60 pM, 70 pM, 80 pM, 90 pM, 100 pM, 110 pM, 120 pM, 130 pM, 140 pM, 150 pM, 160 pM, 170 pM, 180 pM, 190 pM, or 200 pM, or a dose between these values. In some embodiments, T3 is administered at a concentration of about 50 pM. In some embodiments, T3 is administered at a concentration of about 60 pM. In some embodiments, T3 is administered at a concentration of about 70 pM. In some embodiments, T3 is administered at a concentration of about 80 pM. In some embodiments, T3 is administered at a concentration of about 90 pM. In some embodiments, T3 is administered at a concentration of about 100 pM. In some embodiments, T3 is administered at a concentration of about 110 pM. In some embodiments, T3 is administered at a concentration of about 120 pM. In some embodiments, T3 is administered at a concentration of about 130 pM. In some embodiments, T3 is administered at a concentration of about 140 pM. In some embodiments, T3 is administered at a concentration of about 150 pM. In some embodiments, T3 is administered at a concentration of about 160 pM. In some embodiments, T3 is administered at a concentration of about 170 pM. In some embodiments, T3 is administered at a concentration of about 180 pM. In some embodiments, T3 is administered at a concentration of about 190 pM. In some embodiments, T3 is administered at a concentration of about 200 pM.

[0304] In some embodiments, T4 is administered at a concentration between about 1 μM and 30 μM. In some embodiments, T4 is administered at a concentration of less than about 1 μM, about 1 μM, about 2 μM, about 3 μM, about 4 μM, about 5 μM, about 6 μM, about 7 μM, about 8 μM, about 9 μM, about 10 μM, about 15 μM, about 20 μM, about 25 μM, about 30 μM, or greater than about 30 μM. In some embodiments, T4 is administered at a concentration of less than about 1 μM. In some embodiments, T4 is administered at a concentration of about 1 μM. In some embodiments, T4 is administered at a concentration of about 2 μM. In some embodiments, T4 is administered at a concentration of about 3 μM. In some embodiments, T4 is administered at a concentration of about 4 μM. In some embodiments, T4 is administered at a concentration of about 5 μM. In some embodiments, T4 is administered at a concentration of about 6 μM. In some embodiments, T4 is administered at a concentration of about 7 μM. In some embodiments, T4 is administered at a concentration of about 8 μM. In some embodiments, T4 is administered at a concentration of about 9 μM. In some embodiments, T4 is administered at a concentration of about 10 μM. In some embodiments, T4 is administered at a concentration of about 15 μM. In some embodiments, T4 is administered at a concentration of about 20 μM. In some embodiments, T4 is administered at a concentration of about 25 μM. In some embodiments, T4 is administered at a concentration of about 30 μM. In some embodiments, T4 is administered at a concentration greater than about 30 μM.

[0305] In some embodiments, T4 is administered at a concentration of about 10 μM to 15,000 μM. In some embodiments, T4 is administered at a concentration of less than about 10 μM, about 10 μM, about 20 μM, about 50 μM, about 100 μM, about 250 μM, about 500 μM, about 1000 μM, about 2500 μM, about 5000 μM, about 10,000 μM, about 12,000 μM, 15,000 μM, or greater than 15,000 μM, including ranges between these values. In some embodiments, T4 is administered at a concentration of less than about 10 μM. In some embodiments, T4 is administered at a concentration of about 10 μM. In some embodiments, T4 is administered at a concentration of about 20 μM. In some embodiments, T4 is administered at a concentration of about 50 μM. In some embodiments, T4 is administered at a concentration of about 100 μM. In some embodiments, T4 is administered at a concentration of about 250 μM. In some embodiments, T4 is administered at a concentration of about 500 μM. In some embodiments, T4 is administered at a concentration of about 1000 μM. In some embodiments, T4 is administered at a concentration of about 2500 μM. In some embodiments, T4 is administered at a concentration of about 5000 μM. In some embodiments, T4 is administered at a concentration of about 10000 μM. In some embodiments, T4 is administered at a concentration of about 12000 μM. In some embodiments, T4 is administered at a concentration of about 15000 μM. In some embodiments, T4 is administered at a concentration greater than about 15000 μM.

[0306] In some embodiments, T4 is administered at a concentration of about 50 nM, 60 nM, 70 nM, 80 nM, 90 nM, 100 nM, 110 nM, 120 nM, 130 nM, 140 nM, 150 nM, 160 nM, 170 nM, 180 nM, 190 nM, or 200 nM, or a dose between these values. In some embodiments, T4 is administered at a concentration of about 50 nM. In some embodiments, T4 is administered at a concentration of about 60 nM. In some embodiments, T4 is administered at a concentration of about 70 nM. In some embodiments, T4 is administered at a concentration of about 80 nM. In some embodiments, T4 is administered at a concentration of about 90 nM. In some embodiments, T4 is administered at a concentration of about 100 nM. In some embodiments, T4 is administered at a concentration of about 120 nM. In some embodiments, T4 is administered at a concentration of about 130 nM. In some embodiments, T4 is administered at a concentration of about 140 nM. In some embodiments, T4 is administered at a concentration of about 150 nM. In some embodiments, T4 is administered at a concentration of about 160 nM. In some embodiments, T4 is administered at a concentration of about 170 nM. In some embodiments, T4 is administered at a concentration of about 180 nM. In some embodiments, T4 is administered at a concentration of about 190 nM. In some embodiments, T4 is administered at a concentration of about 200 nM.

[0307] In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:100 to about 1:10000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:100 to about 1:1000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:100, 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, or 1:1000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of less than about 1:100. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:100. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:200. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:300. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:400. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:500. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:600. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:700. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:800. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:900. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:1000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of greater than about 1:1000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:1000 to about 1:10000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:1000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:2000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:3000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:4000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:5000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:6000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:7000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:8000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1:9000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of about 1: 10,000. In some embodiments, T3 and T4 are administered in a T3:T4 ratio of greater than about 1: 10,000.

[0308] In some embodiments, T3 is administered at a concentration of about 100 pM and T4 is administered at a concentration of about 100 nM. In some embodiments, T3 is administered at a concentration of about 10 nM and T4 is administered at a concentration of about 1 μM. In some embodiments, T3 is administered at a concentration of about 10 nM and T4 is administered at a concentration of about 10 μM. In some embodiments, T3 is administered at a concentration of about 1 nM and T4 is administered at a concentration of about 10 μM.

[0309] In some embodiments, the total unit dose volume of the topical composition of the present disclosure is between about 0.1 mL and 10 mL. In some embodiments, the total unit dose volume is about 0.1 mL, about 0.5 mL, about 1 mL, about 2 mL, about 3 mL, about 4 mL, about 5 mL, about 6 mL, about 7 mL, about 8 mL, about 9 mL, or about 10 mL. In some embodiments, the total unit dose volume is about 0.1 mL. In some embodiments, the total dose volume is about 0.5 mL. In some embodiments, the total unit dose volume is about 1 mL. In some embodiments, the total unit dose volume is about 2 mL. In some embodiments, the total dose volume of the pharmaceutical composition of the present disclosure is about 3 mL. In some embodiments, the total unit dose volume is about 4 mL. In some embodiments, the total unit dose volume is about 5 mL. In some embodiments, the total unit dose volume is about 6 mL. In some embodiments, the total unit dose volume is about 7 mL. In some embodiments, the total unit dose volume is about 8 mL. In some embodiments, the total dose volume of the pharmaceutical composition of the present disclosure is about 9 mL. In some embodiments, the total unit dose volume is about 10 mL.

[0310] In some embodiments, the methods of the present disclosure are performed in accordance with good medical practice, taking into account the method and schedule of administration, previous and concomitant medications and medical supplements, the individual patient's clinical condition and the severity of the underlying disease, the patient's age, sex, weight, and other factors relevant to the physician, as well as knowledge of the specific compound being used. Therefore, dosage levels may vary for different patients, from patient to patient over time, and for different compositions and formulations, but should be able to be determined by routine techniques.

[0311] In some embodiments, the topical composition is applied every other day for one day for a period of time and then not applied for a long time. For example, in some embodiments, the topical composition is applied every other day for one day for a week and then not applied for a long time. In some embodiments, the topical composition is applied every other day for one day for 2 weeks and then not applied for a long time. In some embodiments, the topical composition is applied every other day for one day for 3 weeks and then not applied for a long time. In some embodiments, the topical composition is applied every other day for one day for 4 weeks and then not applied for a long time. In some embodiments, the topical composition is applied every other day for one day for 5 weeks and then not applied for a long time. In some embodiments, the topical composition is applied every other day for one day for 6 weeks and then not applied for a long time.

[0312] In some embodiments, the long period of no administration lasts one day. In some embodiments, the long period of no administration lasts two days. In some embodiments, the long period of no administration lasts three days. In some embodiments, the long period of no administration lasts four days. In some embodiments, the long period of no administration lasts five days. In some embodiments, the long period of no administration lasts six days. In some embodiments, the long period of no administration lasts seven days. In some embodiments, the long period of no administration lasts ten days. In some embodiments, the long period of no administration lasts fourteen days. In some embodiments, the long period of no administration lasts thirty days.

[0313] In some embodiments, the topical composition is applied daily for several consecutive days, followed by a prolonged period of time. For example, in some embodiments, the topical composition is applied daily for 2 consecutive days, followed by a prolonged period of time. In some embodiments, the topical composition is applied daily for 3 consecutive days, followed by a prolonged period of time. In some embodiments, the topical composition is applied daily for 4 consecutive days, followed by a prolonged period of time. In some embodiments, the topical composition is applied daily for 5 consecutive days, followed by a prolonged period of time. In some embodiments, the topical composition is applied daily for 6 consecutive days, followed by a prolonged period of time. In some embodiments, the topical composition is applied daily for 7 consecutive days, followed by a prolonged period of time. In other embodiments, the topical composition is applied for only one day, followed by a prolonged period of time.

[0314] In some embodiments, the long period of no administration lasts one day. In some embodiments, the long period of no administration lasts two days. In some embodiments, the long period of no administration lasts three days. In some embodiments, the long period of no administration lasts four days. In some embodiments, the long period of no administration lasts five days. In some embodiments, the long period of no administration lasts six days. In some embodiments, the long period of no administration lasts seven days. In some embodiments, the long period of no administration lasts ten days. In some embodiments, the long period of no administration lasts fourteen days. In some embodiments, the long period of no administration lasts thirty days.

[0315] In some embodiments, T3, T4, or a combination of T3 and T4 are administered as a topical composition. In some embodiments, the methods of the present disclosure include administering a combination of T3 and T4. In some embodiments, T3 and T4 are in the same composition. In other embodiments, T3 and T4 are in different compositions.

[0316] It should be understood that the frequency or duration of the disclosed methods can be increased or decreased depending on the desired clinical outcome, the state of the pathology or symptoms, any adverse side effects of the treatment or therapy, or the indications of the concomitant medication. In some embodiments, the subject receives the disclosed methods every day for several consecutive days, followed by an extended period of no administration. This is referred to herein as "pulse dosing" or "pulse therapy." In some embodiments, the subject receives the disclosed methods every day for 2 consecutive days, followed by an extended period of no administration. In some embodiments, the subject receives the disclosed methods every day for 3 consecutive days, followed by an extended period of no administration. In some embodiments, the subject receives the disclosed methods every day for 4 consecutive days, followed by an extended period of no administration. In some embodiments, the subject receives the disclosed methods every day for 5 consecutive days, followed by an extended period of no administration. In some embodiments, the subject receives the disclosed methods every day for 6 consecutive days, followed by an extended period of no administration. In some embodiments, the subject receives the disclosed methods every day for 7 consecutive days, followed by an extended period of no administration. In some embodiments, the subject receives the disclosed methods every day for 10 consecutive days, followed by an extended period of no administration. In some embodiments, the subject receives the disclosed methods every day for 14 consecutive days, followed by an extended period of no administration. In some embodiments, the subject receives the disclosed methods for only one day, followed by an extended period of no administration.

[0317] In some embodiments, the long period of no administration lasts one day. In some embodiments, the long period of no administration lasts two days. In some embodiments, the long period of no administration lasts three days. In some embodiments, the long period of no administration lasts four days. In some embodiments, the long period of no administration lasts five days. In some embodiments, the long period of no administration lasts six days. In some embodiments, the long period of no administration lasts seven days. In some embodiments, the long period of no administration lasts ten days. In some embodiments, the long period of no administration lasts fourteen days. In some embodiments, the long period of no administration lasts thirty days.

[0318] In some embodiments, the amount of T3 administered during a single application of a topical composition of the present disclosure (i.e., per unit dose) is between about 0.00001 μg and about 0.1 μg. In some embodiments, the amount of T3 administered per unit dose is between about 0.00001 μg and about 0.001 μg. In some embodiments, the amount of T3 administered per unit dose is between about 0.00001 μg and about 0.01 μg. In some embodiments, the amount of T3 administered per unit dose is between about 0.001 μg and about 0.01 μg. In some embodiments, the amount of T3 administered per unit dose is between about 0.01 μg and about 0.1 μg.

[0319] In some embodiments, the amount of T3 administered per unit dose is between about 1 ng and 20 ng. In some embodiments, the amount of T3 administered per unit dose is between about 1 ng and 10 ng. In some embodiments, the amount of T3 administered per unit dose is between about 5 ng and 10 ng. In some embodiments, the amount of T3 administered per unit dose is about 1 ng, 2 ng, 3 ng, 4 ng, 5 ng, 6 ng, 7 ng, 8 ng, 9 ng, or 10 ng. In some embodiments, the amount of T3 administered per unit dose is about 6.5 ng.

[0320] In some embodiments, the amount of T3 administered per unit dose is between about 0.1 ng and 1 ng. In some embodiments, the amount of T3 administered per unit dose is about 0.1 ng, 0.2 ng, 0.3 ng, 0.4 ng, 0.5 ng, 0.6 ng, 0.7 ng, 0.8 ng, 0.9 ng, or 1.0 ng. In some embodiments, the amount of T3 administered per unit dose is about 0.65 ng.

[0321] In some embodiments, the amount of T4 administered during a single application of a topical composition of the present disclosure (i.e., per unit dose) is from about 0.1 μg to about 100 μg. In some embodiments, the amount of T4 administered per unit dose is from about 0.1 μg to about 1 μg. In some embodiments, the amount of T4 administered per unit dose is from about 0.1 μg to about 10 μg. In some embodiments, the amount of T4 administered per unit dose is from about 1 μg to about 10 μg. In some embodiments, the amount of T4 administered per unit dose is from about 10 μg to about 100 μg.

[0322] In some embodiments, the compositions of the present disclosure are formulated in unit dosage forms, each dosage containing an effective amount of the active ingredient, for example, a dosage as disclosed above. The term "unit dosage form" refers to physically discrete units of a unit dosage suitable for individual consumption, each unit containing a predetermined amount of active substance calculated to produce the desired effect. Unit dosage forms are generally used for ease of administration and uniform dosage. A unit dosage form can contain a single or separate dose or a composition of a unit, subdose, or an appropriate fraction thereof.

[0323] In embodiments where the compositions of the present disclosure are used to produce a desired effect, it will be readily understood that dosage and dosage form may vary depending on the individual's general health, age, sex, and race, bioavailability, potential systemic, regional, or local side effects, any existing conditions or diseases of the individual, and other factors understood by those skilled in the art (e.g., medical or family history).

[0324] Generally, the dosage, frequency, or duration may be increased or decreased depending upon the desired therapeutic result or effect, the desired beneficial result or effect, and / or the specific subjective result or effect desired.

[0325] Those skilled in the art will understand the factors that may influence the dosage, frequency, and timing required to provide an amount sufficient or effective to provide the desired effect, and to achieve this in order to avoid or minimize side effects depending on the type of effect desired.

[0326] Dosage levels may vary from patient to patient, individual to individual over time, and for different compositions and formulations, but should be determined by routine techniques. Determining the appropriate dosage should include determining not only a single dose, but also the number and timing of administration, as well as the preferred time of administration.

[0327] D. Accessories

[0328] In some embodiments, the compositions of the present disclosure are formulated for topical administration (e.g., as topical dosage forms), for example, by using one or more pharmaceutically acceptable excipients. Topical dosage forms include transmucosal and transdermal preparations, such as aerosols, emulsions, sprays, ointments, salves, gels, pastes, lotions, liniments, oils, and creams. Pharmaceutically acceptable excipients include, for example, penetration enhancers, carriers, diluents, emulsifiers, stabilizers, solvents and cosolvents, viscosity modifiers (e.g., thickeners), adhesion modifiers (e.g., tackifiers), preservatives, antioxidants, viscous polymers, solubilizers, colorants, adhesives, wetting agents, surfactants, gelling agents, and other ingredients commonly known to those skilled in the art.

[0329] Pharmaceutical preparations can be prepared into liquid suspensions or solutions by using sterile liquids, such as, but not limited to, oils, water, alcohols, and pharmaceutically suitable surfactants, suspending agents, and emulsifiers. Suspensions can include oils. Such oils include peanut oil, sesame oil, cottonseed oil, corn oil, and olive oil. Suitable oils also include carrier oils, such as MCT and long-chain triglyceride (LCT) oils. Suspension formulations can also contain fatty acid esters, such as ethyl oleate, isopropyl myristate, fatty acid glycerides, and acetylated fatty acid glycerides. Suspension formulations can include alcohols (such as ethanol, isopropyl alcohol, hexadecanol), glycerol, and propylene glycol. Ethers (such as polyethylene glycol), petroleum hydrocarbons (such as mineral oil and petrolatum), and water can also be used in suspension formulations. Therefore, suspensions can include aqueous liquids or non-aqueous liquids, oil-in-water liquid emulsions, or water-in-oil emulsions.

[0330] "Pharmaceutically acceptable" for an excipient or other ingredient means that the ingredient is generally safe and, within the scope of sound medical judgment, suitable for use in contact with human and other animal cells without undue toxicity, irritation, allergic response, or complications, and commensurate with a reasonable risk / benefit ratio. In some embodiments, "pharmaceutically acceptable" means that the particular ingredient is approved by the FDA for topical use in cosmetics.

[0331] In some embodiments, the composition includes a penetration enhancer. Without being bound by theory, the feature of a penetration enhancer is generally that it increases the ability of biological barriers (e.g., scalp) permeability. In some embodiments, a penetration enhancer is included in the composition to increase the bioavailability of an active agent (e.g., T3, T4, and / or any additional active ingredient) by improving the ability of the active agent to diffuse into skin tissue. Penetration enhancers include, for example, fatty acids and oils, such as castor oil, coconut oil, medium-chain triglycerides (MCT), jojoba oil, sunflower oil, argan oil, almond oil, olive oil, mineral oil, petrolatum, cocoa butter, shea butter, or other esters, triglycerides, or functional derivatives thereof. In some embodiments, the penetration enhancer is 1,2-lauryl ether, aprotinin, azone, benzalkonium chloride, benzalkonium bromide, cetylpyridinium chloride, cetyltrimethylammonium, cyclodextrin, dextran sulfate, ethylene glycol, lauric acid, lauric acid, propylene, lysophosphatidylcholine, menthol, phosphatidylcholine, polyethylene oxide, polysorbate 80, sodium EDTA, chitosan, sodium glycocholate, sodium deoxyglycocholate, sodium lauryl sulfate, sodium salicylate, sodium taurocholate, dimethyl sulfoxide, or a combination thereof. In some embodiments, the penetration enhancer is selected from a lower alcohol having a carbon chain length of 1 to 5, sodium glycocholate, sodium deoxycholate, sodium taurocholate, sodium glycodeoxycholate, sodium taurodeoxycholate, oleic acid, capric acid, lauric acid, lecithin, myristic acid, palmitic acid, lysophosphatidylcholine, phosphatidylcholine, azone, cyclodextrin, sodium lauryl sulfate, polyoxyethylene-9-lauryl ether, polyoxyethylene-20-hexadecyl ether, benzalkonium chloride, cetylpyridinium chloride, vitamin E, TPGS, caprylocaproyl polyoxyglycerol, stearoyl macrogol glyceride, propylene glycol dicaprate, or a mixture thereof. In some embodiments, the compositions of the present disclosure may include a penetration enhancer at a concentration of about 0.01%, about 0.02%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, based on the weight or volume of the composition. , about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 75%, about 75% and about 80%.

[0332] In some embodiments, the matrix may further include a conditioning agent that prevents dry skin and hair in combination with an active agent (e.g., T3, T4, and / or any additional active ingredients). Representative conditioning agents may include, but are not limited to, glycerol, propylene glycol, alpha hydroxy acids, urea, lactic acid, oils, lanolin, and silicones and their derivatives. In some embodiments, the conditioning agent is physically and chemically compatible with the essential components of the composition and does not excessively compromise the stability, aesthetics, or performance of the product. In some embodiments, the concentration of the conditioning agent in the composition is sufficient to provide the desired conditioning benefits, which is apparent to those of ordinary skill in the art. The concentration can vary depending on the conditioning agent, the desired conditioning performance, the average size of the conditioning agent particles, the type and concentration of other components, and other similar factors.

[0333] In some embodiments, the composition comprises a carrier. The carrier can be designed to provide controlled release characteristics, improved circulation time and better transdermal permeability. In some embodiments, the carrier is a hydrophobic drug carrier. The hydrophobic drug carrier can have the advantages of slow sustained release and can adhere well to biological surfaces. The hydrophobic drug carrier can have slow (i.e., extended) release kinetics, or can be configured to have fast or immediate release characteristics. New technologies include developing hydrophilic coatings on hydrophobic nanoparticles to improve their delivery on tissue surfaces while maintaining sustained release characteristics. These include polyethylene glycol and chitosan coatings (see de la Fuente, et al. Nanomedicine 2008; 3: 845–857). Various pharmaceutically acceptable carriers can be used, including but not limited to aqueous media such as water, saline, glycine, hyaluronic acid, etc.; solid carriers such as starch, magnesium stearate, mannitol, sodium saccharin, talc, cellulose, glucose, sucrose, lactose, trehalose, magnesium carbonate, etc.; solvents; dispersion media; coatings; antibacterial and antifungal agents; isotonic and absorption delaying agents; or any other inactive ingredients. The choice of a pharmacologically acceptable carrier depends on the mode of administration. Non-limiting examples of specific uses of such pharmaceutical carriers can be found in Pharmaceutical Dosage Forms and Drug Delivery Systems (Howard C. Ansel et al., eds., Lippincott Williams & Wilkins Publishers, 7th ed. 1999); Remington: The Science and Practice of Pharmacy (Alfonso R. Gennaro ed., Lippincott, Williams & Wilkins, 20th ed. 2000); Goodman & Gilman's The Pharmacological Basis of Therapeutics (Joel G. Hardman et al., eds., McGraw-Hill Professional, 10th ed. 2001); Handbook of Pharmaceutical Excipients (Raymond C. Rowe et al., APhA Publications, 4th edition 2003).In some embodiments, the compositions of the present disclosure may include a carrier at a concentration of about 0.01%, about 0.02%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about About 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 75%, about 75% and about 80%.

[0334] In some embodiments, the composition comprises an emulsifier. The emulsifier can be an anionic, cationic or neutral emulsifier. In certain embodiments, the emulsifier is an anionic emulsifier, selected from alkyl sulfate, aralkyl sulfate, alkyl ethoxy ether sulfate, alkylaryl sulfonate, alkyl succinate, alkyl sulfosuccinate, N-alkanoyl sarcosinate, isethionate, N-acyl taurate, sodium lauryl sulfate, sodium lauryl ether sulfate, sodium oleyl succinate, sodium dodecylbenzenesulfonate and sodium lauryl sarcosinate. Exemplary nonionic or neutral emulsifiers include sorbitan esters, ethoxylated sorbitan esters, ethoxylated alkyl ethers, ethoxylated fatty acid ethers, fatty alcohols, ethoxylated fatty alcohols and glycerol and fatty acid esters. In certain embodiments, the emulsifier is synthetic or natural polymers. In certain embodiments, the emulsifier comprises silicon. In certain embodiments, the emulsifier is a silicone, such as dimethicone, phenyl trimethicone, PEG dimethicone, PPG dimethicone, etc. In some embodiments, the compositions of the present disclosure may include an emulsifier at a concentration of about 0.01%, about 0.02%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about About 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 75%, about 75% and about 80%.

[0335] In some embodiments, the compositions comprise an antidandruff agent or other ingredients typically used on the scalp or hair, including antimicrobial agents, if desired, in amounts typically found useful for topical application. One of ordinary skill in the art can readily determine the type and amount of antidandruff agent to be used in the formulations of the present invention.

[0336] In some embodiments, the composition comprises an antioxidant. The antioxidant can be an amino acid (e.g., glycine, histidine, tyrosine, tryptophan) and its derivatives, an imidazole (e.g., urocanic acid) and its derivatives, a peptide (e.g., D,L-carnosine, D-carnosine, L-carnosine and its derivatives, such as anserine), a carotenoid, a carotene (e.g., β-carotene, lycopene) and its derivatives, a chlorogenic acid and its derivatives, a lipoic acid and its derivatives (e.g., dihydrolipoic acid), aurothioglucose, propylthiouracil, and other thiols (e.g., thioredoxin, glutathione, cysteine, cystine, cystamine, and its glycosyl, N-acetylcholine). yl, methyl, ethyl, propyl, amyl, butyl and lauryl, palmitoyl, oleyl, gamma-linolenic, cholesteryl and glyceryl esters) and their salts, dilauryl thiodipropionate, distearyl thiodipropionate, thiodipropionic acid and its derivatives (esters, ethers, peptides, lipids, nucleotides, nucleosides and salts), and sulfoximine compounds (e.g., buthionine sulfoximine, homocysteine sulfoximine, buthionine sulfone, pentathionine sulfoximine, hexathionine sulfoximine and heptathionine sulfoximine), present at very low tolerated doses (e.g., pmol / kg to μmol / kg) , in addition, (metal) chelating agents (e.g. α-hydroxy fatty acids, palmitic acid, phytic acid, lactoferrin), α-hydroxy acids (e.g. citric acid, lactic acid, malic acid), humic acid, gallic acid, bile extract, bilirubin, biliverdin, EDTA and its derivatives, unsaturated fatty acids and their derivatives (e.g. γ-linolenic acid, linoleic acid, oleic acid), folic acid and its derivatives, ubiquinone and ubiquinol and their derivatives, vitamin C and its derivatives (e.g. sodium ascorbate, ascorbyl palmitate, magnesium ascorbyl phosphate, ascorbyl acetate), tocopherol and its derivatives (vitamin benzoyl peroxide, ferulic acid, furanyl, furfurylidene glucitol, carnosine, butylated hydroxytoluene, butylated hydroxyanisole, nordihydroguaiaretic acid, nordihydroguaiaretic acid, trihydroxybutanone, uric acid and its derivatives, mannose and its derivatives, zinc and its derivatives (such as ZnO, ZnSO4), selenium and its derivatives (such as selenium methionine), stilbene compounds and their derivatives (such as stilbene oxide, trans-stilbene oxide).

[0337] In some embodiments, the composition comprises vitamins, which may be riboflavin (vitamin B2), niacinamide (vitamin B3), pantothenic acid (vitamin B5), pyridoxine (vitamin B6), or biotin (vitamin B7).

[0338] In some embodiments, the composition comprises a conventional thickener. Conventional thickeners can be cross-linked polyacrylic acid and derivatives thereof, polysaccharides and derivatives thereof, such as xanthan gum, agar, alginate or filler, cellulose derivatives (such as carboxymethyl cellulose or hydroxycarboxymethyl cellulose), fatty alcohols, monoglycerides and fatty acids, polyvinyl alcohol and polyvinyl pyrrolidone. Preferably, nonionic thickeners are used.

[0339] In some embodiments, the composition comprises cosmetic and / or dermocosmetic active substances. Cosmetic and / or dermocosmetic active substances may be color-imparting active substances, skin and hair coloring compositions, dyeing compositions, tanning compositions, bleaching agents, keratin-hardening substances, antimicrobial active substances, light filter active substances, protective active substances, substances with hyperemic activity, substances with keratosoftening and keratosculpting activity, antidandruff active substances, anti-inflammatory agents, substances with keratinizing activity, antioxidant active substances or active substances acting as free radical scavengers, skin moisturizing substances or skin humectants, lipid replenishing active substances, substances with anti-erythematous or antiallergic activity, branched-chain fatty acids (e.g. 18-methylarachidic acid), and mixtures thereof.

[0340] In some embodiments, the composition comprises a fragrance oil. Natural fragrances are extracts of flowers (lily, lavender, rose, jasmine, orange blossom, ylang-ylang), stems and leaves (geranium, patchouli, orange leaf), fruits (fennel, coriander, caraway, juniper), peels (bergamot, lemon, orange), roots (nutmeg, angelica, celery, cardamom, costus, iris, vine), wood (pine, sandalwood, guava, cedar, rosewood), herbs and grasses (tarragon, lemongrass, sage, thyme), needles and branches (spruce, fir, pine, dwarf tree), resins and balsams (galbanum, elemi, benzoin, myrrh, frankincense, red myrrh). Typical synthetic fragrance compounds include esters, ethers, aldehydes, ketones, alcohols, and hydrocarbons. Low-volatility essential oils commonly used as fragrance components are also suitable for use as perfume oils, such as sage oil, chamomile oil, clove oil, lemon balm oil, peppermint oil, cinnamon leaf oil, lime tree flower oil, juniper oil, vetiver oil, frankincense oil, geranium, lavender oil, bergamot oil, dihydromyrcenol, lilyral, lyral, citronellol, phenylethyl alcohol, α-hexylcinnamaldehyde, geraniol, benzylacetone, cyclocinnamaldehyde, linalool, ambergris, ambroxan, indole, methyl dihydrojasmonate, sandelice, lemon oil, citrus oil, orange oil, geranium ester, cyclovertal, lavender oil, musk sage oil, G39 damascenone, bourbon geranium oil, cyclohexyl salicylate, methyl cedar ketone ( Coeur), ambergris Musk NP), evemyl, iraldein gamma, phenylacetic acid, geranyl acetate, benzyl acetate, rose oxide, romillate, irotyl and floramat.

[0341] In some embodiments, the topical composition comprises hydroxyalkyl cellulose. When hydroxyalkyl cellulose is included as an adjuvant, it can have multiple functions. For example, hydroxyalkyl cellulose can serve as any one of a penetration enhancer, carrier, emulsifier, stabilizer, viscosity modifier, adhesion modifier, antioxidant, viscous polymer, solubilizer, adhesive, wetting agent and / or gelling agent. In some embodiments, the topical composition comprises hydroxymethyl cellulose. In some embodiments, the topical composition comprises hydroxyethyl cellulose. In some embodiments, the topical composition comprises hydroxypropyl cellulose.

[0342] In some embodiments, the composition comprises a solvent and an optional cosolvent. Any solvent and cosolvent can be collectively referred to as a "solvent system". Without being bound by theory, the selected solvent system can affect the stability, bioavailability and overall efficacy of the composition. In some embodiments, the solvent system can dissolve or solubilize the active ingredient and any adjuvant included at a desired concentration, and should be stable and compatible with the components in the composition (e.g., T3, T4, any additional activating agent and any other adjuvant). In some embodiments, wherein the solvent system comprises more than one solvent, the ratio of the cosolvent is optimized, thereby, for example, increasing the permeability or bioavailability of the active ingredient. Preferred solvent systems are also safe and nontoxic for human use. In some embodiments, when selecting a solvent to be included in the solvent system of the present disclosure, potential adverse effects, such as irritation or allergic reactions, are considered and minimized. The solvents that may be included in the compositions of the present disclosure may include, but are not limited to, water, ethanol, polyols (e.g., glycerol), 1,3-butylene glycol, propylene glycol, hexylene glycol, propylene glycol, ethylene glycol, diethylene glycol, dipropylene glycol, diglycerol, sorbitol, other sugars that are liquid at room temperature, water-soluble alkoxylated nonionic polymers (e.g., polyethylene glycol), and combinations thereof. The solvents may be present in the composition alone or in combination (if more than one solvent is included) in an amount of about 0.1% to about 95% by weight (based on the total weight of the solvents in the composition divided by the total weight of the composition).

[0343] In some embodiments, the solvent system is an aqueous solvent system. In some embodiments, the solvent system includes water. In some embodiments, the composition comprises about 0.1% (v / v) to 90% (v / v) water. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 0.1% (v / v) to 1% (v / v) water. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 0.1% (v / v) to 10% (v / v) water. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 1% (v / v) to 90% (v / v) water. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 1% (v / v) to 50% (v / v) water. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 1% (v / v) to 30% (v / v) water. In some embodiments, the solvent system includes ethanol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 5% (v / v) to 50% (v / v) water. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 5% (v / v) to 30% (v / v) water. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 5% (v / v) to 20% (v / v) water. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 10% (v / v) to 20% (v / v) water. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 10% (v / v) to 20% (v / v) water. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 10% (v / v) water. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 15% (v / v) water. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 20% (v / v) water.

[0344] In some embodiments, the solvent system comprises ethanol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 60% (v / v) ethanol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 10% (v / v) to 70% (v / v) ethanol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 10% (v / v) to 60% (v / v) ethanol. In some embodiments, the solvent system includes ethanol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 15% (v / v) to 55% (v / v) ethanol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 20% (v / v) to 50% (v / v) ethanol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 20% (v / v) to 40% (v / v) ethanol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 25% (v / v) to 35% (v / v) ethanol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 25% (v / v) ethanol. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 30% (v / v) ethanol. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 35% (v / v) ethanol.

[0345] In some embodiments, the solvent system comprises propylene glycol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 20% (v / v) propylene glycol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 10% (v / v) to 90% (v / v) propylene glycol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 20% (v / v) to 80% (v / v) propylene glycol. In some embodiments, the solvent system includes propylene glycol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 30% (v / v) to 70% (v / v) propylene glycol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 40% (v / v) to 60% (v / v) propylene glycol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 45% (v / v) to 55% (v / v) propylene glycol. In some embodiments, the pharmaceutical composition of the present disclosure comprises about 45% (v / v) propylene glycol. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 50% (v / v) propylene glycol. In some embodiments, the pharmaceutical compositions of the present disclosure comprise about 55% (v / v) propylene glycol.

[0346] In some embodiments, the pharmaceutical compositions of the present disclosure include a viscosity modifier. In some embodiments, the viscosity modifier is a thickener. Common thickeners include, but are not limited to, acrylates, carbomers, cellulose matrices, silicones, carrageenans, gums, resins, polysaccharides, and high melting point waxes and oils, such as beeswax, coconut oil, palm oil, soybean oil, stearic acid, rapeseed, cocoa butter, shea butter, gums, rosin, resins, paraffin, and vaseline. In some embodiments, the viscosity modifier is a carbohydrate. Exemplary carbohydrates include monosaccharides, disaccharides, oligosaccharides, and polysaccharides. Exemplary polysaccharides include cellulose, methylcellulose, hydroxypropyl methylcellulose, chitin, galactoarabinan, polygalactose, and polyarabinan. Exemplary glycerides include hydroxystearic acid monoglyceride, hydroxystearic acid diglyceride, isostearic acid monoglyceride, isostearic acid diglyceride, oleic acid monoglyceride, oleic acid diglyceride, ricinoleic acid monoglyceride, ricinoleic acid diglyceride, linoleic acid monoglyceride, linoleic acid diglyceride, linolenic acid monoglyceride, linolenic acid diglyceride, erucic acid monoglyceride, erucic acid diglyceride, tartaric acid monoglyceride, tartaric acid diglyceride, citric acid monoglyceride, citric acid diglyceride, malic acid monoglyceride, malic acid monoglyceride, malic acid diglyceride, and mixtures thereof. In some embodiments, the viscosity modifier is a polymer. Polymer can be natural or synthetic polymers. Natural polymers include polysaccharides, nucleic acids and proteins. Synthetic polymers include polyesters, polyureas, polycarbonates, polyvinyl alcohols, polyamides, polyethers, polyesters, polyamines, polytyrosines, polyanhydrides, polyphosphazenes, polyacrylamides, polyacrylates, polymethacrylates, polyvinylpyrrolidone etc. Exemplary thickening agents include alginate derivatives, pre-neutralized carbomer 430, hydrophilic silica, polysaccharides, xanthan gum, guar gum, agar, carboxymethyl cellulose, hydroxyethyl cellulose, polyacrylates, polyacrylamides, polyvinyl pyrrolidone, and salts. In some embodiments, the compositions of the present disclosure may include a viscosity modifier at a concentration of about 0.01%, about 0.02%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 75%, about 75% and about 80%.

[0347] For example, in some embodiments, the pharmaceutical compositions of the present disclosure include hydroxypropyl cellulose as a viscosity modifier. However, it should be understood that when included in the composition, hydroxypropyl cellulose or another disclosed excipient can play a variety of functions, as described elsewhere herein. In some embodiments, the pharmaceutical compositions of the present disclosure include about 1% (w / v) to about 10% (w / v) hydroxypropyl cellulose. In some embodiments, the pharmaceutical compositions of the present disclosure include about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9% or about 10% (w / v) hydroxypropyl cellulose.

[0348] In some embodiments, the pharmaceutical composition of the present disclosure comprises an adhesion regulator. In some embodiments, the pharmaceutical composition of the present disclosure comprises an adhesive polymer. The adhesive polymer has physicochemical properties that allow for prolonged binding to the tissue surface. In some embodiments, the inclusion of an adhesive polymer in the composition increases the time for the active agent to contact and diffuse through the barrier (e.g., scalp). In some embodiments, the adhesive polymer is chitosan, gelatin, guar gum, lectin, sodium alginate, soluble starch, tragacanth gum, xanthan gum, deacetylated gum, polyacrylic acid, polyvinyl alcohol, hydroxypropyl methylcellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, sodium carboxymethyl cellulose, thiomer, polycarbophil, hyaluronic acid, dermatan sulfate, or a combination thereof. In some embodiments, the adhesion regulator is a tackifier. Common tackifiers include, but are not limited to, gums, resins (natural or modified), carbomers, or other natural or synthetic polymers. In some embodiments, the compositions of the present disclosure may include an adhesion modulator at a concentration of about 0.01%, about 0.02%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 75%, about 75% and about 80%.

[0349] In some embodiments, the present pharmaceutical composition comprises a preservative. Preservatives can be used to inhibit microbial growth or increase the stability of the composition, thereby extending the shelf life of the composition. Suitable preservatives are known in the art and include EDTA, EGTA, benzalkonium chloride or benzoic acid or benzoate (e.g., sodium benzoate), vitamin A, vitamin C (ascorbic acid), citric acid, vitamin E, and tocopherol.

[0350] In some embodiments, pharmaceutical composition of the present disclosure comprises antioxidant.Not bound by theory, antioxidant can usually delay or suppress the oxidative decomposition of the component (such as activating agent, such as T3, T4 or any other active ingredient) of the disclosed composition, thereby can improve the stability of the disclosed composition and extend its shelf life.In some embodiments, antioxidant is alpha-tocopherol, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, methionine, citric acid, ascorbic acid, sodium ascorbate, sodium thiosulfate, sodium bisulfite, sodium pyrosulfite, ascorbyl palmitate, thioglycerol, propyl gallate, cysteine or its combination.In some embodiments, antioxidant is cyclodextrin, D-alpha-tocopherol, rosmarinic acid or its combination. In some embodiments, the compositions of the present disclosure may include an antioxidant at a concentration of about 0.01%, about 0.02%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 4%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 75%, about 75% and about 80%.

[0351] In some embodiments, the composition may include one or more vitamins. Any vitamin with properties such as nourishing hair, inhibiting hair loss and / or promoting hair growth can be used. Representative suitable vitamins may include, but are not limited to, essential B vitamins, such as thiamine, riboflavin, niacin, vitamin B6, folic acid, vitamin B12, biotin, and pantothenic acid. In some embodiments, the concentration of the vitamin in the composition is sufficient to provide the desired benefits for treating hair loss and / or promoting hair growth while remaining compatible with the disclosed composition (e.g., active agent, such as T3, T4, or any other active ingredient). The concentration can vary depending on the type and concentration of the selected vitamin, the desired effect, and other components, as well as other similar factors. Representatively, the composition may include 1 mg to 200 mg of vitamins; in some embodiments, 50 mg to 250 mg of vitamins.

[0352] In some embodiments, the pharmaceutical composition of the present disclosure includes a solubilizing agent. Without being bound by theory, the solubilizing agent typically forms a complex with the active ingredient, and the complex can have physicochemical properties different from those of a single active ingredient. The properties of the complex can increase the solubility of T3, T4, or any other active ingredient in the composition. In some embodiments, the solubilizing agent is a water-soluble organic solvent, a nonionic surfactant, a water-insoluble lipid, an organic liquid, a cyclodextrin, or a phospholipid. In some embodiments, the solubilizing agent is a water-soluble enhancer. In some embodiments, the water-soluble enhancer is polyethylene glycol 300, polyethylene glycol 400, ethanol, propylene glycol, xanthan gum, glycerol, N-methyl-2-pyrrolidone, dimethylacetamide, dimethyl sulfoxide, or a combination thereof. In some embodiments, the solubilizing agent is propylene glycol. In some embodiments, the solubilizing agent is xanthan gum. In some embodiments, the solubilizing agent is a nonionic surfactant. In some embodiments, the nonionic surfactant is Cremophor EL, Cremophor RH 40, Cremophor RH 60, d-tocopheryl polyethylene glycol 1000 succinate, polysorbate 20, polysorbate 80, polyethylene glycol-15-hydroxystearate (Solutol HS15), sorbitan monooleate, poloxamer 407, oleic acid macrogolglycerides (Labrafil) M-1944CS, Labrafil M-2125CS, caprylic capric macrogolglycerides (Labrasol), lauric macrogolglycerides (Gellucire) 44 / 14, caprylic capric macrogolglycerides (Softigen) 767, fatty acid monoesters and fatty acid diesters of PEG 300, 400 or 1750. In some embodiments, the solubilizing agent is an organic liquid. In some embodiments, the organic liquid is beeswax, d-α-tocopherol, oleic acid, or a medium-chain mono- or diglyceride. In some embodiments, the solubilizer is a cyclodextrin. In some embodiments, the solubilizer is α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxypropyl-β-cyclodextrin, and sulfobutyl ether-β-cyclodextrin. In some embodiments, the solubilizer is α-cyclodextrin. In some embodiments, the solubilizer is β-cyclodextrin. In some embodiments, the solubilizer is γ-cyclodextrin. In some embodiments, the solubilizer is hydroxypropyl-β-cyclodextrin. In some embodiments, the solubilizer is sulfobutyl ether-β-cyclodextrin. In some embodiments, the solubilizer is a phospholipid. In some embodiments, the phospholipid is hydrogenated soy phosphatidylcholine, distearoylphosphatidylglycerol, L-α-dimyristoylphosphatidylcholine, or L-α-dimyristoylphosphatidylglycerol. In some embodiments, the solubilizer is lecithin.In some embodiments, the compositions of the present disclosure may include a solubilizing agent at a concentration of about 0.01%, about 0.02%, about 0.05%, about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about 49%, about 50%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%, about 56%, about 57%, about 58%, about 59%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about 75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%, about 82%, about 83%, about 8 %, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about 23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, about 70%, about 75%, about 75%, and about 80%.

[0353] In some embodiments, the present pharmaceutical composition comprises coloring agent.Suitable coloring agent and / or dye and / or pigment can include but not limited to following color, for example white, black, yellow, blue, green, pink, red, orange, purple, indigo, brown and combination thereof, pigment such as Timica Extra Large Sparkle, titanium dioxide and chromium oxide green, ultramarine blue and pink and iron oxide.Coloring agent and / or dye and / or pigment can be present in the present composition (by the gross weight of coloring agent and / or dye and / or pigment in the composition divided by the gross weight of composition) with the amount of about 0.01 % by weight to about 5 % by weight alone or all (if comprising more than one coloring agent).Coloring agent can be present in the present composition (by the gross weight of coloring agent in the composition divided by the gross weight of composition) with the amount of about 0.01 % by weight to about 5 % by weight alone or all (if comprising more than one coloring agent).

[0354] In some embodiments, pharmaceutical composition of the present disclosure comprises adhesive.Suitable adhesive includes but is not limited to polyvinylpyrrolidone (PVP), marine colloids (marine colloids), carboxyvinyl polymer, starch, cellulose polymer (such as hydroxyethyl cellulose, carboxymethyl cellulose (carmellose), hypromellose, hydroxyethyl propyl cellulose, hydroxybutyl methyl cellulose and its salt (such as sodium carboxymethyl cellulose)), natural gum (such as karaya gum, xanthan gum, carrageenan, gellan gum, locust bean gum, gum arabic and tragacanth), chitosan, colloidal magnesium aluminum silicate and colloidal silicon dioxide.Adhesive can be present in the disclosed composition alone or entirely (if comprising more than one adhesive) with the amount of about 0.01 weight % to about 5 weight % (by the gross weight of adhesive in composition divided by the gross weight of composition).

[0355] In some embodiments, the pharmaceutical compositions of the present disclosure include a wetting agent. The wetting agent, such as a low molecular weight polyethylene glycol (e.g., PEG6-PEG12), can be present in the composition alone or in combination (if more than one wetting agent is included) in an amount of up to about 10 wt %, up to about 5 wt %, up to about 3 wt %, up to about 1 wt %, or up to about 0.1 wt % (based on the total weight of the wetting agent in the composition divided by the total weight of the composition).

[0356] In some embodiments, pharmaceutical compositions of the present disclosure include surfactants. The surfactant that can be included in the composition can be anionic, nonionic or amphoteric compounds. Suitable examples of anionic surfactants are one or more higher alkyl sulfates (such as potassium or sodium lauryl sulfate), higher fatty acid monoglyceride monosulfate (such as the salt of the monosulfated monoglyceride of hydrogenated coconut oil fatty acid), alkyl sulfonates (such as sodium dodecylbenzenesulfonate), higher fatty acid sulfoacetate, the higher fatty acid ester of 1,2 dihydroxypropane sulfonate. The example of water-soluble nonionic surfactants is the condensation product of ethylene oxide and various hydrogenated compounds with it being reactive and having a long hydrophobic chain (such as an aliphatic chain of about 12-20 carbon atoms), the condensation product containing hydrophilic polyoxyethylene moiety, such as the condensation product of polyethylene oxide with fatty acid, fatty alcohol, fatty amide and other fat moieties, and with the condensation product of propylene oxide and polypropylene oxide, such as Pluronic materials, such as Pluronic F127. Exemplary suitable alkyl polyglycoside (APG) surfactants that can be used in the composition may include APG C8-C10, APG C10-C16, decyl glucoside, coco glucoside, anionic APG carboxylates, sodium lauryl glucose carboxylate, lauryl glucoside, D-pyranose (oligomer, C10-16 glycoside, carboxymethyl ether, sodium salt), C12-C16 fatty alcohol glycosides, and combinations thereof. Exemplary APG surfactants that can be used may have 2000NUP / MB, LGC Sorb, 1200N UP / MB and The surfactants, alone or collectively (if more than one surfactant is included), may be present in the composition in an amount of from about 0.01% to about 10% by weight (based on the total weight of surfactants in the composition divided by the total weight of the composition).

[0357] In some embodiments, the pharmaceutical composition of the present disclosure includes a gelling agent. Exemplary gelling agents that can be used in the compositions of the present disclosure may include pectin, starch, and gelatin forms derived from animals or plants (e.g., pork gelatin). The pectin in the composition may include, for example, high methoxyl pectin, low methoxyl pectin, or a combination thereof. In some embodiments, pectin is amidated pectin. In other embodiments, pectin is non-amidated pectin. In certain embodiments, pectin is a combination of amidated pectin and non-amidated pectin. The gelatin in the composition may include type A gelatin, type B gelatin, hide gelatin or skin gelatin (e.g., cowhide, pigskin), and / or bone gelatin (e.g., cow bone, pig bone), used alone or in combination. The gelling agent may be present in the composition in an amount of about 0.1% by weight to about 20% by weight (by the total weight of the gelling agent in the composition divided by the total weight of the composition) alone or in combination. In some embodiments, the composition does not include a gelling agent.

[0358] Depending on the unit dose volume and total volume, the composition can be provided as a final packaged product, for example, in a bottle or any other suitable container. In some embodiments, the bottle is a dropper bottle, a fine mist spray bottle, a pump bottle, a glass bottle, or a plastic bottle. In some embodiments, the bottle is a dropper bottle. In some embodiments, the bottle is a fine mist spray bottle. In some embodiments, the bottle is a pump bottle. In some embodiments, the bottle is a glass bottle. In some embodiments, the bottle is a plastic bottle. In some embodiments, the bottle is between about 15mL and 90mL. In some embodiments, the bottle is about 15mL (i.e., about 0.5 ounces). In some embodiments, the bottle is about 20mL. In some embodiments, the bottle is about 30mL (i.e., about 1 ounce). In some embodiments, the bottle is about 40mL. In some embodiments, the bottle is about 50mL. In some embodiments, the bottle is about 60mL (i.e., about 2 ounces). In some embodiments, the bottle is about 70mL. In some embodiments, the bottle is about 80mL. In some embodiments, the bottle is about 90mL (i.e., about 3 ounces). In some embodiments, the bottle is greater than 90mL.

[0359] In some embodiments, when included in a composition, the excipients disclosed herein can perform more than one function. For example, hydroxypropyl cellulose can be used to change the viscosity of the composition while also affecting the adhesive properties of the composition, or to stabilize the active agent in the composition, promote emulsification, or other functions described herein.

[0360] Those of ordinary skill in the art will appreciate that the selection of suitable excipients for the disclosed compositions may depend on a variety of factors. Relevant factors for selecting suitable excipients include, for example, the compatibility of the excipient with the active agent (e.g., T3, T4, any additional active agent), the desired active agent permeation kinetics, processing parameters, biocompatibility, and user preference.

[0361] E. Exemplary Features of the Disclosed Compositions and Methods

[0362] In some embodiments, the compositions and methods of the present disclosure produce fewer adverse effects (e.g., side effects) than comparable methods or compositions (e.g., standard of care for specific hair loss conditions, such as androgenic alopecia). Without wishing to be bound by theory, it is proposed that thyroid hormones (e.g., T3, T4) may be absorbed primarily through the epidermis and dermis before entering the bloodstream. This would localize potential side effects and potentially mitigate them through pulsed therapy, as described in various embodiments herein.

[0363] In some embodiments, the hair loss is caused by alopecia. In some embodiments, the hair loss is caused by androgenic alopecia, alopecia areata, alopecia areata perseverant, alopecia totalis, alopecia universalis, alopecia areata universalis, alopecia serpiginosa, cicatricial alopecia, lichen planus, frontal fibrosing alopecia, central centrifugal cicatricial alopecia (CCCA), traction alopecia, alopecia barbae, or postpartum alopecia. In some embodiments, the hair loss is caused by androgenic alopecia. In some embodiments, the hair loss is male pattern hair loss. In some embodiments, the hair loss is female pattern hair loss.

[0364] In some embodiments, the compositions or methods of the present disclosure are particularly suitable for use with subjects having certain characteristics (e.g., biomarkers) indicating a high likelihood of treatment success or a low risk of treatment. For example, in some embodiments, subjects whose thyroid hormone levels are only below a threshold value (e.g., the level of T3, T4) or other biomarkers receive the methods or compositions of the present disclosure. In some embodiments, prior to treatment with the methods or compositions of the present disclosure, subjects undergo a blood test.

[0365] The present invention also provides kits for practicing the disclosed methods, comprising one or more thyroid hormones described herein (e.g., T3, T4, or a combination thereof), or pharmaceutical compositions comprising one or more thyroid hormones described herein (e.g., T3, T4, or a combination thereof).

[0366] In some embodiments, administration of a composition of the present disclosure increases hair shaft yield. Hair shaft yield refers to the number of hairs produced by a hair follicle. In some embodiments, the composition increases hair shaft yield by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0367] In some embodiments, administration of a composition of the present disclosure extends the anagen phase of hair growth. The anagen phase is the first of three hair growth phases, during which hair follicles actively produce hair. In some embodiments, the composition extends the anagen phase by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0368] In some embodiments, administration of the disclosed compositions increases FGF7 expression. FGF7 is a growth factor that promotes hair growth by regulating the onset and extension of the anagen phase. In some embodiments, the compositions increase FGF7 expression by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0369] In some embodiments, administration of the disclosed compositions increases the proliferation of bulge epithelial stem cells. Bulb epithelial stem cells are located in the bulge region of the hair follicle, have a high proliferation capacity, and have the multipotency to regenerate keratinocytes, sebaceous glands, and epidermal tissue. In some embodiments, the compositions increase the proliferation of bulge stem cells by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0370] In some embodiments, administration of a composition of the present disclosure increases keratin 15 expression. Keratin 15 is a widely used biomarker for bulge epithelial stem cells. An increase in keratin 15 indicates an increased presence of bulge epithelial stem cells. In some embodiments, the composition increases keratin 15 expression by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0371] In some embodiments, administration of the disclosed compositions reduces the expression of p-S6. p-S6 is a direct downstream kinase of the mechanistic target of rapamycin complex 1 (mTORC1), which is associated with aging and increased graying of hair. A decrease in p-S6 expression indicates downregulation of the pathway. In some embodiments, the composition reduces the expression of p-S6 by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0372] In some embodiments, administration of the compositions of the present disclosure can reduce the rate of graying (or depigmentation) of hair in a subject. Graying / depigmentation may be caused by dysfunction of differentiated melanocytes in the pigment units of hair follicles. Graying / depigmentation is usually temporarily reversible, and pigmentation can begin in hair follicles that have previously stopped pigmentation. Once the hair follicle melanocyte stem cells are exhausted, graying / depigmentation becomes irreversible. Without being bound by theory, the topical compositions of the present disclosure downregulate mTORC1, a protein complex that negatively regulates human hair follicle growth and pigmentation (Suzuki T et al. EMBO Rep. 2023; 24(7): e56574). Without being bound by theory, downregulation of mTORC1 by the topical compositions promotes hair growth and pigmentation.

[0373] In some embodiments, the compositions of the present disclosure reduce depigmentation by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%. In some embodiments, the compositions increase re-pigmentation by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0374] In some embodiments, the compositions of the present disclosure prevent hair follicles from initiating depigmentation. In some embodiments, the compositions prevent 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the hair follicles in a subject from initiating depigmentation. In some embodiments, the compositions increase the amount of pigment-producing hair follicles by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0375] F. Examples

[0376] The following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

[0377] Example 1: Human skin organ culture study

[0378] Purpose: The study described herein was designed to test the effects of applying the disclosed topical compositions to organotypic human scalp skin. Previous work (e.g., Paus, et al. J. Clin. Endocrinol. Metab. 2008;93(11):4381–4388) tested the application of thyroid hormones to hair follicles extracted from the scalp by injecting the cultures themselves with solutions. In this study, T4 increased the proliferation of hair matrix keratinocytes and prolonged the growth phase of the hair follicle cycle, while T3 reduced apoptosis of hair matrix keratinocytes.

[0379] This study was conducted by applying the topical composition of the present disclosure to the scalp skin.

[0380] method:

[0381] Skin biopsy: Skin biopsies (4 mm) containing terminal hair follicles were prepared and placed at the air-liquid interface in William's E medium without serum supplementation (Samra et al. Int J Mol Sci. 2023; 24(2); Gherardini et al. Int J Cosmet Sci. 2019; 41(2): 164-182) and incubated at 37°C in a humidified atmosphere of 5% CO2. 35, 43 After 24 hours of equilibration, the skin biopsies were topically treated with 2 μL of a viscous formulation (containing ethanol, hydroxypropylcellulose, propylene glycol, and laboratory-prepared purified water). This formulation enhances the permeability of the test agent and is very viscous, thereby preventing the test compound from escaping from the skin surface into the medium containing vehicle medium or T3 (1 nM or 10 nM) or T4 (1 μM or 10 μM) or their combination (T3 10 nM + T4 10 μM).

[0382] The skin surface was cleaned before each topical treatment, and the culture medium was changed on days 1, 3, and 5. Two biopsy samples from each experimental group were placed in cryogenic matrix and snap-frozen in liquid nitrogen for further analysis on day 6.

[0383] Snap-frozen samples were sliced using a cryostat (Leica) and 7 μM sections were collected. Serial sections of each full-length hair follicle were collected and stored on slides at -80°C until further analysis.

[0384] Immunochemistry: To assess proliferative and apoptotic keratinocytes in the hair matrix, frozen sections were fixed with 4% paraformaldehyde in PBS and preincubated in 10% goat serum, followed by incubation with mouse anti-Ki-67 antibody (1:800; Cell Signaling Technology) and rabbit anti-human cleaved caspase-3 antibody (1:400; Cell Signaling Technology) overnight at 4°C. After washing three times in PBS, sections were incubated with corresponding secondary antibodies (goat anti-rabbit IgG-Alexa Fluor 488 antibody and goat anti-mouse IgG-Alexa Fluor 555 antibody (1:400; Life Technologies)) at 37°C for 45 minutes.

[0385] IGF-1: To assess IGF-1 protein expression, 58, 40 tissue cryosections were fixed in acetone and incubated with rabbit anti-human IGF-1 antibody (1:250; Novus Biological) overnight at 4°C. Secondary antibody incubation was performed with goat anti-rabbit IgG-Alexa Fluor 555 antibody (1:400; Life Technologies) at 37°C for 45 minutes.

[0386] TGFβ-2: To assess TGFβ-2 protein expression, 58, 40 tissue cryosections were fixed in acetone and incubated with rabbit anti-human TGF-β2 antibody (1:400; Proteintech) overnight at 4°C. Secondary antibody incubation was performed with goat anti-rabbit IgG-Alexa Fluor 488 antibody (1:400; Life Technologies) at 37°C for 45 minutes.

[0387] K15: To evaluate K15 protein expression, tissue cryosections were fixed in 4% paraformaldehyde in PBS and preincubated in 10% goat serum, followed by incubation with mouse anti-human CK15-FITC-conjugated antibody (1:200; Novus biological) at 37°C for 2 h.

[0388] K85: To assess K85 protein expression, tissue cryosections were fixed in acetone and incubated with guinea pig anti-human CK85 antibody (1:1000; Progen) overnight at 4°C. Secondary antibody incubation was performed with goat anti-guinea pig Alexa Fluor 594 antibody (1:400; Life Technologies) at 37°C for 45 minutes.

[0389] gp100 / MITF: To assess gp100 / MITF protein expression, tissue cryosections were fixed in methanol:acetone (1:1), permeabilized, and blocked in TBS containing 10% goat serum + 0.3% Triton X-100. The sections were then incubated with mouse anti-MITF antibody (1:50; Abcam) and rabbit anti-human NKI-beteb (gp100) antibody (1:100; Abcam) overnight at 4°C. After three washes in TBS, the sections were incubated with the corresponding secondary antibodies: goat anti-mouse IgG-FITC antibody (1:400; Jackson immunoresearch) and goat anti-rabbit IgG-Alexa Fluor 555 antibody (1:400; Life Technologies) for 45 minutes at 37°C. An additional amplification step was performed with anti-FITC Alexa Fluor 488 antibody (1:400; Life Technologies) for 30 minutes at 37°C.

[0390] MTCOI: To assess MTCOI protein expression, tissue cryosections were fixed in 4% paraformaldehyde, permeabilized, and blocked in 10% goat serum + 0.3% Triton X-100 in PBS. Sections were then incubated with rabbit anti-human MTCO1 antibody (1:50; Abcam) overnight at 4°C. After three washes in PBS, sections were incubated with goat anti-rabbit IgG-FITC antibody (1:400; Jackson Immunoresearch) for 45 minutes at 37°C. An additional amplification step was performed with anti-FITC Alexa Fluor 488 antibody (1:400; Life Technologies) for 30 minutes at 37°C.

[0391] FGF7: To assess FGF7 protein expression, tissue cryosections were fixed in acetone and incubated with rabbit anti-KGF / FGF7 antibody (1:100; Novus Biological) overnight at 4°C. Secondary antibody incubation was performed with goat anti-rabbit IgG-Alexa Fluor 488 antibody (1:400; Life Technologies) at room temperature for 45 minutes.

[0392] p-S6: To assess p-S6 protein expression, tissue cryosections were fixed in 4% paraformaldehyde and incubated with rabbit anti-phospho-S6 ribosomal protein antibody (1:200; Cell Signaling Technology) overnight at 4°C. Secondary antibody incubation was performed with goat anti-rabbit IgG-Alexa Fluor 488 antibody (1:400; Life Technologies) for 45 minutes at room temperature.

[0393] CD31: To assess CD31 protein expression, tissue cryosections were fixed in acetone and incubated with mouse anti-human CD31 antibody (1:50; Agilent) overnight at 4°C. Secondary antibody incubation was performed with goat anti-mouse IgG-Alexa Fluor 555 (1:400; Life Technologies) for 45 minutes at room temperature.

[0394] Omission of the primary antibody was used as a negative control, and the cells were counterstained with 4',6-diamidino-2-phenylindole (DAPI) / Fluoromount fluorescent mounting medium (EMS) to visualize the cell nuclei and then mounted.

[0395] Quantitative (immuno)histoporesistive and image culture:Evaluations were performed using a Keyence-Biozero 9100 microscope (Keyence Corporation, Japan) and associated software (Biozero-II Analyzer; Keyence Corporation, Japan) at an original magnification of 100× or 200×. Different target areas, namely the dermis (for CD31 quantity and expression), the hair bulge (for K15 expression), the hair bulb (for Ki-67 / Casp-3, Warthin-Starry, FGF7, p-S6, MTCOI, K85, gp100 / MITF), and the proximal outer root sheath (ORS) (for IGF-1 and TGFβ-2), were imaged at 100× or 200× magnification. Melanin content, K15, FGF7, p-S6, MTCOI, K85, gp100 / MITF, IGF-1, and TGFβ-2 were quantified in specific reference areas by measuring relative staining intensity in ImageJ. The percentages of proliferative and apoptotic hair matrix (HM) keratinocytes (Ki-67+ or Casp-3+ cells among DAPI+ cells) were quantified in the hair bulb.

[0396] For hair cycle staging, HFs were microscopically assessed using Warthin-Starry histochemistry and Ki-67 / Caspase-3 immunostaining as previously described (Haslam et al. J Invest Dermatol 2018; Oh et al. J Invest Dermatol 2016; Alam et al. Br J Dermatol. 2020). The percentage of hair follicles in each hair cycle stage (anagen VI, early catagen, mid-catagen, or late catagen) was calculated.

[0397] Statistical analysis: All data are presented as mean ± SEM or mean ± SEM of fold change relative to vehicle. Data were analyzed for Gaussian distribution using the D'Agostino and Pearson omnibus normality test. Significant differences were analyzed using the unpaired Student's t-test for parametric data or the Mann–Whitney test for nonparametric data. Results for each test group were compared using GraphPad Prism 9 (GraphPad Software). A p value < 0.05 was considered statistically significant.

[0398] Vector preparation and experimental design: The two selected carrier formulations for topical administration of T3 and T4 peptide hormone compositions were designed based on the potential to facilitate penetration and transfollicular absorption of thyroid hormones, containing only FDA-approved and cosmetically approved topical ingredients.

[0399] Turbidity tests were performed to confirm satisfactory solubility and miscibility of all components. The turbidity tests showed that T3 and T4 were completely dissolved at the highest concentration tested (T3 = 10 nM, T4 = 10 μM).

[0400] Preparation A Preparation B Ethanol 30% v / v Ethanol 60% v / v Hydroxypropyl cellulose 5% w / v Hydroxypropyl cellulose 5% w / v Propylene glycol 50% v / v Propylene glycol 20% v / v Purified water, diluted to final volume Purified water, diluted to final volume

[0401] The concentrations of T3 and T4 in formulation A were varied as follows:

[0402]

[0403] The experiments in this example were conducted according to the following schedule:

[0404]

[0405] Human scalp skin was obtained from five donors:

[0406] Donor 1: Scalp skin, male, African American (T3 and T4 administered separately)

[0407] Donor 2: Scalp skin, male, African American (T3 and T4 administered separately)

[0408] Donor 3: Scalp skin, female, Spanish (combination of T3 and T4)

[0409] Donor 4: Scalp skin, female, Jamaican (combination of T3 and T4)

[0410] Donor 5: Scalp skin, male, Mediterranean (T3 and T4 administered alone and in combination)

[0411] Skin punch samples were collected from the temporal scalp skin on day 0. The punch biopsies were then placed at the air-liquid interface of culture medium under 5% CO2. After previously removing any residue from previous applications, 2 μL of treatment solution (loaded with T3 or T4) was applied on days 1, 3, and 5. On day 6, the skin biopsies were embedded in cryo-matrix and snap-frozen in liquid nitrogen prior to quantitative (immuno)histomorphometry.

[0412] result:

[0413] Hair shaft produces: like Figure 1A As shown, topical application of Formulation A containing T3 (1 nM and 10 nM) and T4 (1 μM) tends to promote hair shaft production. Figure 1B As shown, topical application of Formulation B containing T4 (10 μM) and the combination of T3+T4 promoted hair growth, whereas Formulation B containing T3 (10 nM) reduced hair growth. The results are expressed as the percentage of hair follicles generated from treatment day 1 to treatment day 6. Figure 2A and Figure 2B Representative photographs of skin punch samples are shown for trials of Formulation A and Formulation B. Measurements for Formulation A containing T3 and T4 are from one donor, as samples from two donors were damaged and could not be analyzed.

[0414] Duration of anagen phase: Figure 3A 、 4A and Figure 5 The results in [ 001 ] showed that formulation A containing T3 (1 nM and 10 nM) and T4 10 μM prolonged the anagen phase of the hair follicle cycle, potentially promoting hair growth in isolated microdissected scalp follicles after the addition of thyroid hormone to the culture medium (simulating systemic application). Figure 5 The Ki-67 and Casp-3 immunofluorescence signals in the anagen phase reflect the extent of hair follicles in the catagen phase. Ki-67 is a biomarker of hair matrix keratinocyte proliferation, while Casp-3 is a biomarker of apoptosis. In formulation A, the greatest increase in the percentage of hair follicles with extended anagen phase cycles was observed after administration of T4 (10 μM). Formulation A containing the combination of T3 + T4 resulted in a decrease in the percentage of hair follicles in the catagen phase. Figure 3B and Figure 4B As shown, T3 (10 nM) and T4 (10 μM) prolonged the anagen phase duration, and the T3+T4 combination did not affect the anagen phase in Formulation B. The results are expressed as mean % ± SEM.

[0415] Hair follicles in the telogen phase have the lowest levels of bioactivity (Lin et al. Front Cell Dev Biol. 2022;10:899095). These results suggest that topical application of thyroid hormone may reduce AGA-related hair shaft shedding (telogen effluvium). This is new evidence that topical thyroid hormone treatment extends the duration of the telogen phase of hair on the human scalp.

[0416] Therefore, topical administration of both thyroid hormones may: 1) inhibit telogen effluvium in AGA patients; and 2) extend the window of time during which miniaturized vellus hairs can reconvert into terminal follicles (only in anagen). In Formulation A, the combination of 1 nM T3 and 10 μM T4 had the opposite effect of either administered alone, suggesting that this combination may induce premature catagen.

[0417] Hair matrix keratinocyte proliferation: like Figure 6A and Figure 6B As shown, in both Formulation A and Formulation B, 10 μM topical T4 and the combination of T3+T4 reduced hair matrix keratinocyte proliferation on human scalp. Figure 6A The results confirmed Figure 4AThe T3+T4 combination inhibited the growth phase of hair observed in the study. T3 at two concentrations and 1 μM T4 had no significant effect on the proliferation of hair matrix keratinocytes. + The percentage of cells is expressed.

[0418] The results for Formulation A are also shown in Figure 7 Among them, Ki-67 + , Caspase-3, and DAPI were used for cell detection. Similar results were observed in Preparation B. Caspase-3 was not detected in any group. + cells, which is within expectations for anagen follicles in healthy scalp skin.

[0419] Melanin production: Figure 8A and Figure 8B The results presented in the present study indicate that in both Formulations A and B, when only well-pigmented anagen follicles were evaluated, thyroid hormone did not further stimulate melanin production. In Formulation A, T3 (10 nM) and the combination of T3 and T4 showed a slight, non-significant increase in melanin content in the evaluated follicles. This suggests that in intact scalp skin, topical application of thyroid hormone has no significant effect on melanin production. Figure 9 The results showed that no pigmentary abnormalities (melanin aggregation or ectopic melanin granules) indicating hair follicle damage were observed, indicating that the hair follicles tolerated the topical thyroid hormone well.

[0420] Hair follicle pigment unit (gp100 and MITF expression):When T3, T4, or a combination of T3 and T4 were added to serum-free culture medium from microdissected, organotypic human scalp hair follicles at anagen stage VI, quantitative Masson-Fontana histochemistry was used to assess hair follicle pigmentation. The biomarker gp100 distinguishes premelanosomes produced by melanocytes. It is used as a sensitive tracer of melanosome transfer between melanocytes and keratinocytes and is a marker of hair follicle pigment unit activity. MITF was upregulated under 1 nM T3 and the combination. MITF is a key transcription factor in melanocyte development and differentiation. MITF regulates the expression of numerous pigmentation genes to promote melanocyte differentiation. Although pigment production itself was not stimulated, the hair follicle pigment unit appeared to be activated, as indicated by stronger gp100 immunoreactivity and longer gp100+ melanocyte dendrites. For more information on gp100 expression and melanosomes / melanin granules, see, for example, Singh et al. Exp. Dermatol. 2008; 17(5): 418-426. These results may indicate a partial hair follicle "rejuvenation" effect of topical T3 and T4 administration. A partial "rejuvenation" effect may be beneficial in the AGA model because AGA-affected hair follicles express some molecular markers of premature aging.

[0421] Figure 10A and Figure 11A The results in [ 0015 ] showed that in formulation A, topical T3 (1 nM and 10 nM) and the T3 + T4 combination significantly activated hair follicle pigment units as assessed by gp100 and MITF immunoreactivity. Figure 10B and Figure 11B The results in [ 001 ] showed that in formulation B, the T3 + T4 combination significantly reduced hair follicle pigment unit activity, while T3 and T4 administered alone tended to increase activity. Figure 12 It was shown that the immunoexpression of gp100 and MITF in the pigment units of hair follicles was increased by administration of Formulation A containing the T3 composition.

[0422] MTCO1 expression: MTCO1 is a component of cytochrome c oxidase, the final enzyme in the mitochondrial electron transport chain that drives oxidative phosphorylation. Therefore, MTCO1 is a useful screening parameter for detecting mitochondrial activity and its excess.

[0423] Figure 13A and Figure 14AThe results presented here show no evidence that the thyroid hormone in Formulation A overstimulated mitochondrial activity, which could increase reactive oxygen species and lead to oxidative damage. In contrast, Formulation A containing 10 μM T4 and the T3+T4 combination decreased MTCO1 expression, suggesting that their use may have a counter-regulatory effect in reducing mitochondrial activity in response to long-term T4 administration. This suggests a lower risk of oxidative damage caused by topical thyroid hormone. Figure 13B The results shown in Figure 3 showed that in formulation B, T3 (1 nM and 10 nM) and the combination of T3+T4 significantly reduced MTCO1 expression in the hair follicle epithelium. Figure 14B The T4 (10 μM) composition showed the same effect in ORS. As in Formulation A, the composition in Formulation B also showed no signs of excessive stimulation of mitochondrial activity by thyroid hormone.

[0424] Hair follicle epithelial stem cell proliferation: Figure 15A It was shown that in formulation A, T3 (10 nM) and T4 (1 μM) increased K15 expression in the bulge region. Figure 15B The results showed that in Formulation B, the T4 composition tended to increase K15 expression in the bulge region, but not significantly. Higher K15 expression tends to correlate with improved epithelial stem cell function, so these results suggest that topical thyroid hormone activates hair follicle epithelial stem cells. Therefore, activation of hair follicle epithelial stem cells can inhibit hair follicle miniaturization and promote its reversal. Figure 16A The thyroid hormone in preparation A showed the effect of K15 + There was no effect on cell proliferation, but Figure 16B The T4 composition in Formulation B significantly increased K15 + Cell proliferation. K15 + Cell proliferation may be desirable because it counteracts the miniaturization of hair follicles in AGA, but undesirably, it may also lead to K15 + Cell differentiation into K6 / K16 + ORS keratinocytes, which may chronically deplete the bulge stem cell niche (Tiede et al. Eur. J. Cell Biol. 2007; 86(7):355-376).

[0425] In addition, if Figure 17A and Figure 18A As shown by the percentage of cells expressing K15 and Ki67 in the 3D images, in formulation A, T3 (10 nM) and T4 (1 μM) tend to stimulate the proliferation of hair follicle stem cells. However, the administration of T3 (10 nM), T4 (10 μM) and the combination of T3 + T4 may also promote stem cell apoptosis. Figure 17B As shown in Figure 2, T3 in formulation B tends to stimulate the proliferation of hair follicle epithelial stem cells. Figure 18BAs shown, T3 (1 nM) composition B may also promote apoptosis. Increased K15+ cell proliferation and increased apoptosis in the bulge area may indicate a homeostatic mechanism. For example, excess stem cells induced by T3 proliferation may undergo apoptosis through the process of stem cell niche shaping. Figure 19 It is shown that Formulation A treatment with T3 (10 nM) increased Ki67 immunofluorescence compared to the vehicle group.

[0426] IGF-1 expression: Figure 20A The results showed that T4 (1 μM) and the combination of T3+T4 tended to increase IGF-1 protein expression in ORS keratinocytes in formulation A. Although the difference was not significant compared with the vehicle group, this trend suggests that upregulation of IGF-1 may be a mechanism for its anagen extension effect. Figure 20B It was shown that in formulation B, the T3+T4 combination significantly increased the expression of IGF-1 in ORS keratinocytes.

[0427] TGFβ-2 expression: Figure 21A The results showed that T3, T4, or the combination of T3 and T4 had no effect on TGFβ-2 protein expression in ORS keratinocytes in Formulation A. This is a promising result because TGFβ-2 promotes the onset of catagen, which limits hair growth. Figure 21B It was shown that the T4 (1 μM) composition significantly increased TGFβ-2 protein expression in ORS keratinocytes in Formulation B. Stimulating the production of TGFβ-2, a key physiological catagen growth factor, is undesirable for hair loss management. Figure 22 No difference in TGFβ-2 immunofluorescence was shown for vehicle and T3 treatment in Formulation A.

[0428] FGF7 expression: Figure 23A It was shown that in formulation A, T3 (1 nm) and T4 (1 μM) alone as well as the T3+T4 combination significantly increased the expression of FGF7 protein in ORS keratinocytes. Figure 23B The results showed that in formulation B, T3 (1 nm and 10 nm) and T4 (1 μM and 10 μM) tended to increase the expression of FGF7 protein in ORS keratinocytes, but not significantly. FGF7 is an important hair growth-promoting factor, and its inhibition will prevent hair growth. Figure 24 It was shown that FGF7 immunofluorescence was significantly increased in T3 (1 nM) treatment compared to vehicle alone in Formulation A, and there was a trend toward increased FGF7 immunofluorescence in T3 (10 nM).

[0429] p-S6 expression: Figure 25A It was shown that in formulation A, the T3+T4 combination significantly increased p-S6 in hair matrix keratinocytes. Figure 25B The study showed that in Formulation B, the T3 (10 nM) combination significantly reduced pS6 in hair matrix keratinocytes, while T4 (10 μM) and the T3 + T4 combination increased pS6. pS6 is a direct downstream kinase of mTORC1, so an increase in p-S6 indicates increased mTORC1 activity. mOTRC1 activity is associated with aging and hair graying. In Formulation A, the T3 and T4 combination did not increase p-S6, indicating that they do not stimulate this hair graying mechanism. In Formulation B, T3 (10 nM) downregulated mTORC1, thereby slowing hair graying, a desirable property for hair loss management.

[0430] K85 expression: Figure 26A It was shown that in formulation A, T4 (10 μM) and the combination of T3+T4 significantly downregulated the expression of keratin 85 (K85) in the precortical hair matrix. Figure 26B The results showed that in Formulation B, the T3 + T4 combination significantly reduced K85 expression in the pre-cortical hair matrix, while T3 (1 nM) and T4 (1 μM) increased K85 expression. K85 is a sensitive marker of keratin production in the hair shaft, so its downregulation indicates a decrease in keratin production. The T3 formulation did not significantly reduce K85; in fact, T3 (10 nM) tended to increase K85 production. Figure 27 Shown are K85 immunofluorescence signals in cortical pre-hair matrix cells for both T3 treatments compared to vehicle alone in Formulation A.

[0431] CD31 + Express: Figure 28A It was shown that in formulation A, T3 (1 nM) and T4 (10 μM) alone significantly increased CD31 + The number of endothelial cells. Figure 28B It was shown that in formulation B, T3 (1 nM) and T4 (10 μM) significantly increased the expression of CD31 in the dermis. + The number of endothelial cells.CD31 in the dermis + The increase in cell numbers suggests that thyroid hormone stimulates angiogenesis, which could lead to increased perfusion of hair follicles. If angiogenesis does occur, T3 / T4 could also enhance VEGF-A secretion from hair follicles, thereby promoting hair follicle and / or skin recovery. Figure 29 CD31 immunofluorescence signals for both T3 treatments compared to vehicle alone in Formulation A are shown.

[0432] discuss:These results demonstrate that topical thyroid hormones (T3 and T4) in both carrier formulations (Formulation A and Formulation B) extend the anagen phase. Administration of the disclosed topical compositions inhibits telogen effluvium in AGA patients and prolongs the window during which miniaturized vellus hairs can reconvert into terminal follicles. This anagen-extending effect may be primarily mediated by stimulation of FGF-7 and / or IGF-1 expression.

[0433] The significant downregulation of MTCO1 expression in HM and ORS keratinocytes may represent a decrease in mitochondrial activity. In the case of AGA, this decrease in MTCO1 expression may be beneficial because MTCO1 has been reported to be upregulated in DP fibroblasts of scalp skin affected by hair loss (Chew et al. Exp Dermatol. 2022; 31(6): 906-917). The topical compositions of the present disclosure are well tolerated in HF (no pigmentary abnormalities) and do not overstimulate HF mitochondria (low risk of oxidative damage).

[0434] In the hair bulb during anagen, some biomarkers did change, indicating that application of the topical composition successfully affected the hair bulb, demonstrating adequate penetration of T3 and T4. Within the time window tested, the topical composition stimulated HF pigment units but did not stimulate melanin production itself. Further studies may include determining whether this indicates that the composition partially rejuvenates HF. In addition to the reduction in p-S6 expression, this pro-pigmentation effect may indicate that long-term application has an anti-graying effect. In addition, reduced mTORC1 activity may also play an anti-aging role and may extend the anagen phase (Suzuki et al. EMBO Reports. 2023; 24: e56574).

[0435] This example further demonstrates that the topical compositions disclosed herein can activate bulge epithelial stem cells. This may inhibit HF miniaturization and promote its reversal. In fact, as observed in the test topical compositions containing T3 / T4 combination formulation A and the composition containing 1 nM T3 formulation B, treatment first induced the proliferation of K15+ cells and then stimulated their long-term apoptosis, and the composition contained (Tiede S et al. Eur J Cell Biol. 2010; 89(10): 769-777). However, the total number of K15+ cells did not decrease. This effect may also represent a physiological mechanism by which human scalp HF stabilizes the size of the bulge area through apoptosis induction, thereby potentially limiting tumor formation. The topical composition also stimulated CD31+ cells, which may reflect an increase in angiogenesis. This not only potentially increases the size of HF, but also may exert an anti-aging effect (Keren A et al. Sci Adv. 2022; 8(25): eabm6756).

[0436] Furthermore, a topical composition containing T3 (1 nM) Formulation A is promising; further work may include follow-up studies on scalp skin containing at least 20-25% gray or white HF to evaluate the effects on hair loss and graying. In addition, follow-up analyses could be performed to expand the panel of aging biomarkers, hair shaft keratins, mitochondrial markers (i.e., PGC1α, TFAM, porins), HF pigmentation, in situ tyrosinase activity, α-MSH, and other stem cell markers including CD34, CD200, and K6.

[0437]

[0438]

[0439] =No significant change relative to the vector;↑=increase relative to the vector;↓=decrease relative to the vector;

[0440] *p<0.05; **p<0.01; ***p<0.001.

[0441] Example 2: Clinical Study Design for Evaluating the Efficacy of the Topical Compositions of the Present Disclosure

[0442] Purpose: The efficacy of the topical compositions and methods of the present disclosure for treating or preventing hair loss is measured.

[0443] qualifications:All subjects will undergo a medical history and physical examination. A treatment group will be established, which will include individuals with hair loss (e.g., due to androgenic alopecia or other similar conditions). Individuals with hair loss will be diagnosed according to diagnostic techniques known to those skilled in the art, including various potential causes of hair loss. The treatment group will take the topical composition disclosed herein. A control group will also be established. The control group will take a topical composition without an active agent, such as a carrier formulation. Subjects will be 18 years of age or older. Subjects may withdraw from the study at any time for any reason. Subjects who are unwilling to participate in the study, or subjects with medical conditions that are not suitable for treatment with the disclosed methods or combinations will also be excluded from the study.

[0444] Overview of Study Design: The treatment group received a topical composition comprising (i) 10 nM triiodothyronine (T3); (ii) 5% (w / v) hydroxypropylcellulose; (iii) 60% (v / v) ethanol; (iv) 20% (v / v) propylene glycol; and (v) 10% (v / v) water. The topical composition was applied every other day for two consecutive weeks, followed by one week of no application. The control group received a vehicle formulation comprising (i) 5% (w / v) hydroxypropylcellulose; (ii) 60% (v / v) ethanol; (iii) 20% (v / v) propylene glycol; and (iv) 10% (v / v) water. The control group received the vehicle formulation according to the same treatment cycle as the treatment group. At the end of each treatment cycle, overall hair growth was assessed using techniques known to those skilled in the art (e.g., for measuring hair shaft output). Subjects will also undergo laboratory testing for hair growth biomarkers, including melanin production and protein expression (IGF-1, FGF7, TGFβ-2, MITF, p-S6, K85, CD31, gp100, MTCO1, and K15); these can be measured according to the techniques described herein and known in the art.

[0445] result: Relative to the control group, subjects in the treated group are expected to show increased hair growth (e.g., increased hair shaft production) and / or decreased hair loss. Subjects in the treated group are also expected to show improvement in one or more biomarkers indicative of successful treatment (e.g., extended anagen phase, increased FGF7 expression, increased proliferation of bulge epithelial stem cells, increased keratin 15 expression, or decreased p-S6 expression). Topical application of Formulation B containing T3 (10 nM) induces extended anagen phase, increased expression of IGF-1 and FGF-7, and significantly reduces the expression of p-S6 and MTCO1 (see Example 1). This clinical trial will perform scalp skin biopsies before and at the end of treatment to enable systematic comparison of the resulting short-term preclinical data with the in vivo long-term topical application data regarding the long-term response of K15+ eHFSCs, hair shaft mass, anti-aging, and mitochondrial effects.

[0446] Example 3: Individual Administration of a Topical Composition of the Disclosure for Treating Hair Loss

[0447] This example describes exemplary therapeutic regimens for treating or preventing hair loss in an individual in need thereof.

[0448] Patient 1: Patient 1 was diagnosed with androgenic alopecia and suffered from hair loss. Patient 1 self-applied a topical composition comprising (i) 10 nM triiodothyronine (T3); (ii) 5% (w / v) hydroxypropylcellulose; (iii) 60% (v / v) ethanol; (iv) 20% (v / v) propylene glycol; and (v) 10% (v / v) water. The topical composition was applied in a treatment cycle of two weeks of every other day application followed by one week of no application. After one treatment cycle, Patient 1 experienced increased hair growth, e.g., increased hair shaft production. Patient 1 also demonstrated improvement in one or more biomarkers indicative of successful treatment (e.g., prolonged anagen phase, increased FGF7 expression, increased proliferation of bulge epithelial stem cells, increased keratin 15 expression, or decreased p-S6 expression).

[0449] Patient 2: Patient 2 was diagnosed with alopecia areata and experienced hair loss. Patient 2 self-applied a topical composition comprising (i) 10 nM triiodothyronine (T3); (ii) 5% (w / v) hydroxypropylcellulose; (iii) 60% (v / v) ethanol; (iv) 20% (v / v) propylene glycol; and (v) 10% (v / v) water. The topical composition was applied in treatment cycles of two weeks of every other day application followed by one week of no application. After four treatment cycles, Patient 2 experienced a reduction in the rate of hair loss. Patient 2 also demonstrated improvements in one or more biomarkers indicative of successful treatment (e.g., prolonged anagen, increased FGF7 expression, increased proliferation of bulge epithelial stem cells, increased keratin 15 expression, or decreased p-S6 expression).

[0450] Patient 3: Patient 3 was diagnosed with and experienced alopecia totalis. Patient 3 self-applied a topical composition comprising (i) 10 nM triiodothyronine (T3); (ii) 5% (w / v) hydroxypropylcellulose; (iii) 60% (v / v) ethanol; (iv) 20% (v / v) propylene glycol; and (v) 10% (v / v) water. The topical composition was applied in treatment cycles of three consecutive daily applications followed by one week of no application. After two treatment cycles, Patient 3 experienced increased hair growth, e.g., increased hair shaft production. Patient 3 also demonstrated improvement in one or more biomarkers indicative of successful treatment (e.g., prolonged anagen phase, increased FGF7 expression, increased proliferation of bulge epithelial stem cells, increased keratin 15 expression, or decreased p-S6 expression).

[0451] Patient 4: Patient 4 was diagnosed with persistent alopecia areata and experienced hair loss. Patient 4 self-applied a topical composition comprising (i) 10 nM triiodothyronine (T3); (ii) 5% (w / v) hydroxypropylcellulose; (iii) 30% (v / v) ethanol; (iv) 50% (v / v) propylene glycol; and (v) 10% (v / v) water. The topical composition was applied in a treatment cycle of two weeks of every other day application followed by one week of no application. After three treatment cycles, Patient 4 experienced a decrease in the rate of hair loss. Patient 4 also demonstrated improvement in one or more biomarkers indicative of successful treatment (e.g., prolonged anagen phase, increased FGF7 expression, increased proliferation of bulge epithelial stem cells, increased keratin 15 expression, or decreased p-S6 expression).

[0452] Example 4: Individual administration of a topical composition of the present disclosure for treating graying hair

[0453] This example describes exemplary therapeutic regimens for treating or preventing graying of hair in an individual in need thereof.

[0454] Patient 5: Patient 5 was experiencing graying of the hair. Patient 5 self-applied a topical composition containing (i) 1 nM triiodothyronine (T3); (ii) 5% (w / v) hydroxypropylcellulose; (iii) 30% (v / v) ethanol; (iv) 50% (v / v) propylene glycol; and (v) 10% (v / v) water. The topical composition was applied in a treatment cycle of two weeks of every other day application followed by one week of no application. Beginning after one treatment cycle, Patient 5 experienced gradual re-pigmentation of the hair.

[0455] Patient 6: Patient 6 was experiencing hair graying. Patient 6 self-applied a topical composition comprising (i) 1 nM triiodothyronine (T3); (ii) 5% (w / v) hydroxypropylcellulose; (iii) 30% (v / v) ethanol; (iv) 50% (v / v) propylene glycol; and (v) 10% (v / v) water. The topical composition was applied in a treatment cycle of two weeks of every other day application followed by one week of no application. Beginning after one treatment cycle, Patient 6 experienced a decrease in the rate of hair graying (e.g., decreased hair pigmentation).

[0456] For the purpose of explanation, the foregoing description uses specific terms to provide a comprehensive understanding of the present invention. However, it will be apparent to those skilled in the art that specific details are not required to implement the present invention. Therefore, for the purpose of illustration and description, the foregoing description of specific embodiments of the present invention is presented. It is not intended to be exhaustive or to limit the present invention to the precise compositions, formulations, methods, etc. disclosed; in view of the above teachings, there may be many modifications and variations. These embodiments are selected and described in order to better explain the principles of the present invention and its practical application through the illustration of specific examples, thereby enabling other technical personnel in the art to better utilize the present invention and various embodiments, and when the specific uses envisioned exceed the specific examples disclosed, the various embodiments may have various modifications suitable for these specific uses. Therefore, the scope of the present invention will be limited only by the appended claims and their equivalents.

Claims

1. A topical composition for treating or preventing hair loss, comprising: i. Triiodothyronine (T3); ii. pharmaceutically acceptable excipients; and iii. Solvent system.

2. The topical composition of claim 1 comprising about 1 nM to 30 nM T3.

3. The topical composition of claim 2, comprising about 10 nM T3.

4. The topical composition of claim 1, wherein the pharmaceutically acceptable excipient is a penetration enhancer, a carrier, a diluent, an emulsifier, a stabilizer, a viscosity modifier, an adhesion modifier, a preservative, an antioxidant, a viscous polymer, a solubilizer, a colorant, a binder, a wetting agent, a surfactant, or a gelling agent.

5. The topical composition of claim 1, wherein the pharmaceutically acceptable excipient is hydroxyalkylcellulose.

6. The topical composition of claim 1, wherein the pharmaceutically acceptable excipient is hydroxypropyl cellulose.

7. The topical composition of claim 6 comprising about 1 w / v% to 10 w / v% hydroxypropylcellulose.

8. The topical composition of claim 7 comprising about 5 w / v% hydroxypropylcellulose.

9. The topical composition of claim 1, wherein the solvent system comprises an alcohol.

10. The topical composition of claim 1, wherein the solvent system comprises ethanol or propylene glycol.

11. The topical composition of claim 1 , wherein the solvent system comprises about 10 v / v% to 70 v / v% ethanol.

12. The topical composition of claim 11, wherein the solvent system comprises about 60 v / v% ethanol.

13. The topical composition of claim 11, wherein the solvent system comprises about 30 v / v% ethanol.

14. The topical composition of claim 1, wherein the solvent system comprises about 10 v / v% to 90 v / v% propylene glycol.

15. The topical composition of claim 14, wherein the solvent system comprises about 20 v / v% propylene glycol.

16. The topical composition of claim 14, wherein the solvent system comprises about 50 v / v% propylene glycol.

17. The topical composition of claim 1, wherein the solvent system comprises water.

18. The topical composition of claim 17, wherein the solvent system comprises about 1 v / v% to 30 v / v% water.

19. The topical composition of claim 17, wherein the solvent system comprises about 10 v / v% water.

20. A topical composition for treating or preventing hair loss comprising: i. Triiodothyronine (T3); ii. Hydroxypropyl cellulose; and iii. Solvent system.

21. A topical composition for treating or preventing hair loss comprising: i. Triiodothyronine (T3); ii. Hydroxypropyl cellulose; iii. ethanol; iv. propylene glycol; and v.water.

22. A topical composition for treating or preventing hair loss comprising: i. about 10 nM triiodothyronine (T3); ii. about 5 w / v% hydroxypropyl cellulose; iii. about 60 v / v% ethanol; iv. about 20 v / v% propylene glycol; and v. About 10 v / v% water.

23. The topical composition of claim 1, further comprising an additional active agent.

24. The topical composition of claim 23, wherein the additional active agent is an amino acid, an antioxidant, an anti-inflammatory agent, an analgesic, a 5-alpha reductase inhibitor, a cannabinoid, an immunosuppressant, an immunostimulant, an anticancer agent, an antiulcer agent, an antihistamine, a terpene, a vitamin, a vasodilator, or a vasoconstrictor.

25. The topical composition of claim 23, wherein the additional active agent is rapamycin, finasteride, dutasteride, or minoxidil.

26. The topical composition of claim 23, wherein the additional active agent is thyroxine (T4).

27. The topical composition of claim 1 in lyophilized form.

28. Use of the topical composition of any one of claims 1 to 27 for treating or preventing hair loss.

29. Use of the topical composition of any one of claims 1 to 27 in the preparation of a medicament for treating or preventing hair loss.

30. A method of treating or preventing hair loss in a subject, comprising administering to the subject the topical composition of any one of claims 1-27.

31. The method of claim 30, comprising administering to the subject about 0.1 mL to 10 mL of the composition per unit dose.

32. The method of claim 31 , comprising administering to the subject about 1 mL of the composition per unit dose.

33. The method of claim 30, comprising administering to the subject about 1 ng to 10 ng of T3 per unit dose.

34. The method of claim 33, comprising administering to the subject about 6.5 ng of T3 per unit dose.

35. The method of claim 30, wherein the composition is administered daily.

36. The method of claim 30, wherein the composition is administered every other day.

37. The method of claim 30, wherein the composition is administered every other day for two consecutive weeks followed by a prolonged period of no administration.

38. The method of claim 37, wherein the composition is administered every other day for two consecutive weeks followed by a prolonged period of no administration.

39. The method of claim 37 or 38, wherein the long period of no administration is at least two weeks.

40. The method of claim 30, wherein the hair loss is caused by androgenic alopecia, alopecia areata, alopecia areata persevera, alopecia totalis, alopecia universalis, alopecia areata universalis, serpiginous alopecia, cicatricial alopecia, lichen planus, frontal fibrosing alopecia, central centrifugal cicatricial alopecia (CCCA), traction alopecia, alopecia barbae, or postpartum alopecia.

41. The method of claim 30, which results in increased hair shaft production.

42. The method of claim 41, wherein hair shaft output is increased by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a baseline value measured prior to application of the composition.

43. The method of claim 30, which results in extension of the anagen phase of hair.

44. The method of claim 43, wherein the anagen phase is extended by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a baseline value measured before administration of the composition.

45. The method of claim 30, which results in increased expression of FGF7.

46. The method of claim 45, wherein the expression of FGF7 is increased by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a baseline value measured before administration of the composition.

47. The method of claim 30, which results in increased proliferation of bulge epithelial stem cells.

48. The method of claim 47, wherein bulge epithelial stem cell proliferation is increased by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a baseline value measured before administration of the composition.

49. The method of claim 30, which results in increased keratin 15 expression.

50. The method of claim 49, wherein the expression of keratin 15 is increased by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a baseline value measured before application of the composition.

51. The method of claim 30, which results in decreased p-S6 expression.

52. The method of claim 49, wherein the expression of p-S6 is reduced by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100% compared to a baseline value measured before administration of the composition.

53. Use of the topical composition of any one of claims 1 to 27 for treating or preventing graying of hair.

54. Use of the topical composition of any one of claims 1 to 27 in the preparation of a medicament for treating or preventing graying of hair.

55. A method of treating or preventing graying of hair in a subject, comprising administering to the subject the topical composition of any one of claims 1-27.

56. The method of claim 55, wherein the topical composition comprises: i. approximately 1 nM triiodothyronine (T3); ii. about 5 w / v% hydroxypropyl cellulose; iii. about 30 v / v% ethanol; iv. about 50 v / v% propylene glycol; and v. About 10 v / v% water.

57. The method of claim 55, which results in reduced hair depigmentation.

58. The method of claim 57, wherein hair depigmentation is reduced by 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%.

59. The method of claim 55, which results in hair repigmentation.

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