Active agents that modulate the activity of ion channels for use in regulating hair growth - Patents.com

Abstract

An active agent that modulates the activity of ion channels for use in hair growth regulation. In order to provide a novel agent for actively controlling hair growth in a subject, the present invention proposes an active agent for use in hair growth regulation, particularly for use in the treatment (stimulation or inhibition) of hair growth, which activates, enhances, inactivates, blocks or attenuates the cellular response of the transient receptor potential ion channel TRPM5, or interferes with the expression of said ion channel.Furthermore, the present invention is directed to a composition for use as a cosmetic or pharmaceutical in the treatment of hair growth, which comprises at least one of the above-mentioned active agents and at least one auxiliary agent.Furthermore, a non-therapeutic method for hair growth regulation is disclosed, in which an effective amount of at least one of the above-mentioned active agents is administered to a subject.

Description

[Technical Field]

[0001] The present invention is directed to active agents for use in regulating hair growth, particularly for use in treating hair growth, which modulate the activity of an ion channel (including but not limited to activating, enhancing, inactivating or blocking the ion channel, or attenuating a cellular response induced by the ion channel, or interfering with the expression of the ion channel).

[0002] Furthermore, the present invention is directed to a composition for use as a cosmetic or pharmaceutical in the treatment of hair growth, comprising at least one active agent that modulates the activity of an ion channel (including, but not limited to, activating, enhancing, inactivating or blocking the ion channel, or attenuating a cellular response induced by the ion channel, or interfering with the expression of said ion channel).

[0003] Additionally, the present invention is directed to non-therapeutic methods of modulating hair growth, in which an effective amount of at least one active agent that modulates the activity of an ion channel (including, but not limited to, activating, enhancing, inactivating or blocking the ion channel, or attenuating a cellular response induced by the ion channel, or interfering with the expression of said ion channel) is administered to a subject. [Background technology]

[0004] Hair follicle formation occurs once in a healthy mammal's lifetime, and the number of hair follicles is determined in utero. Hair follicle morphogenesis is the development of the entire hair follicle structure from the epidermis and mesoderm. During postnatal life, hair follicles undergo lifelong cyclical transformations that progress through phases of rapid growth (anagen), degeneration (catagen), and relative quiescence (telogen), as classified by morphological and molecular markers.

[0005] There are many common and rare diseases associated with the hair follicle cycle, such as non-scarring alopecia, cicatricial alopecia, chronic and acute hair shedding, hirsutism or hirsutism.In addition, there are many other conditions in which it is desirable to control unwanted hair on the human body or promote hair growth in areas of the human body, such as those involving drug-induced hair loss (e.g., chemotherapy), radiation-induced hair loss (e.g., radiation therapy), and those involving unwanted hair growth due to drugs (e.g., cyclosporin A or diazoxide). Summary of the Invention [Problem to be solved by the invention]

[0006] There is a continuing need for active agents to actively control hair growth in a subject without undesirable side effects. [Means for solving the problem]

[0007] The present inventors found that the transient receptor potential ion channel TRPM5 is expressed in hair follicles. Specifically, strong cytoplasmic expression was detected in the outer root sheath (ORS), while weaker expression was observed in the inner root sheath (IRS). The dermal sheath (DS) and dermal papilla (DP) did not show TRPM5 expression.

[0008] "Ion channels" are pore-forming proteins located in biological membranes. Through the protein's pore, ions can pass through the membrane down their electrochemical gradient. By opening and closing the pore, ion channels can gate and control the flow of ions across the membrane, thereby regulating the intracellular concentration of the ion. Changes in intracellular ion concentrations affect numerous cellular responses and processes, including, but not limited to, proliferation, differentiation, survival, death, mediator release, immune mechanisms, etc.

[0009] "Transient receptor potential ion channels" (TRP channels) are a group of ion channels located primarily on the plasma membrane, and there are approximately 30 TRP channels, including TRPC, TRPV, TRPM, TRPN, TRPA, TRPP, and TRPML. The ion channels are relatively nonselectively permeable to cations, including sodium, calcium, and magnesium.

[0010] "TRPM5" is the official gene symbol for "transient receptor potential cation channel subfamily M member 5" and identifies the protein encoded by the TRPM5 gene in humans (NCBI Gene ID: 29850; HGNC: 14323; NCBI mRNA sequence: NM_014555.3; NCBI protein sequence: NP_055370.1; Status on June 7, 2020.).

[0011] The present inventors found that the pheromones 2-heptanone (hereinafter referred to as "2-Hep") and 2,5-dimethylpyrazine (hereinafter referred to as "DMP"), both of which are agonists of the transient receptor potential ion channel TRPM5, opening the channel, sustain human hair growth ex vivo. Indeed, the number of hair follicles that remained in anagen during 7 days of culture was higher in the pheromone-treated group than in the control group. Furthermore, treatment of hair follicles with 2-Hep resulted in increased intrafollicular expression of AXIN2 (a known hair growth promoter) and suppression of levels of TGFβ2 (a known hair growth inhibitor).

[0012] Conversely, inhibition of TRPM5 with the selective inhibitor triphenylphosphine oxide (TPPO) consistently promoted catagen in human HF organ cultures. Similarly, specific knockdown of TRPM5 by RNA silencing (thereby mimicking inhibition of TRPM5-coupled signaling) reduced cell proliferation in the hair matrix and proximal hair bulb, hallmarks of hair growth inhibition. Furthermore, knockdown of TRPM5 by RNA silencing induced a tendency toward apoptosis. Finally, knockdown of TRPM5 by RNA silencing resulted in suppression of intrafollicular expression of LEF1 and IGF1 (known hair growth promoters) and increased levels of TGFβ2 and SFRP1 (known hair growth inhibitors).

[0013] In addition to the active agents described above, the present inventors have identified several further active agents that act on TRPM5 and are therefore suitable for use in the treatment of hair growth.

[0014] In conclusion, the present invention features an active agent for use in the treatment of hair growth, which activates, enhances, inactivates or blocks TRPM5, or attenuates cellular responses induced by the transient receptor potential ion channel TRPM5, or interferes with the expression of said ion channel.

[0015] According to one option of the present invention, the active agent that activates, enhances, inactivates or blocks the ion channel or attenuates the cellular response induced by the transient receptor potential ion channel TRPM5 is an agonist or antagonist of the ion channel.

[0016] As referred to herein, an "agonist" of the transient receptor potential ion channel TRPM5 is a substance that binds to and activates or potentiates the ion channel, thereby increasing a cellular response associated with TRPM5.

[0017] As referred to herein, an "antagonist" of the transient receptor potential ion channel TRPM5 is a substance that binds to said ion channel and blocks TRPM5, thereby attenuating the cellular response to an agonist rather than activating or enhancing it as an agonist does.

[0018] An "inverse agonist" of the transient receptor potential ion channel TRPM5 is a substance that binds to the ion channel and then induces a cellular response opposite to that seen when an agonist is applied.

[0019] With respect to the transient receptor potential ion channel TRPM5 of the present invention, the term "cellular response" should be primarily understood as a change in ion concentration in a cell resulting from agonist / antagonist / inverse agonist binding to the transient receptor potential ion channel TRPM5. However, this term also encompasses cases where the binding of an agonist / antagonist / inverse agonist induces a cellular response without changing the intracellular ion concentration.

[0020] Thus, agonists of the transient receptor potential ion channel TRPM5 of the present invention activate or enhance the ion channel to produce a cellular response as do endogenous agonists, whereas antagonists of the ion channel of the present invention block or attenuate the ion channel to produce a cellular response that is normally activated / enhanced by the endogenous agonist, and inverse agonists of the present invention modulate the ion channel to produce a cellular response that is opposite to that normally activated / enhanced by the endogenous agonist.

[0021] In those embodiments in which the active agent is an agonist of the transient receptor potential ion channel TRPM5, the active agent is used in the treatment of unwanted hair loss. In those embodiments in which the active agent is an antagonist / inverse agonist of the transient receptor potential ion channel TRPM5, the active agent is used in the treatment of unwanted hair growth.

[0022] In certain embodiments of the present invention, the ion channel agonist / antagonist / inverse agonist used is a TRPM5 agonist selected from the group consisting of dimethylpyrazine, dimethylethylpyrazine, tetramethylpyrazine, 2-heptanone, eugenol, SID2848719 (CAS No. 702636-90-6, SMILES, NC(=O)C1(CCN(CC1)S(=O)(=O)c1ccc2OCCCOc2c1)N1CCCCC1), rutamarin, bergapten, xanthotoxin, isopimpinellin, carbachol, 3-deoxyglucosone, glucagon-like peptide 1, (E)-N-(3,4 dimethoxybenzylidene)-2-naphthalen-1-yl)acetohydrazide, or a combination thereof. Alternatively, a TRPM5 agonist is an aptamer that binds to the TRPM5 ion channel and activates or enhances the ion channel to generate a cellular response.

[0023] As used herein, the term "aptamer" refers to a DNA, RNA or XNA oligonucleotide or peptide molecule that binds to a specific target molecule, such as an ion channel molecule.

[0024] In another embodiment of the present invention, the ion channel agonist / antagonist / inverse agonist used is a TRPM5 antagonist / inverse agonist selected from triphenylphosphine oxide, econazole, miconazole, chlorpromazine, or a combination thereof. Alternatively, the TRPM5 antagonist / inverse agonist is an aptamer that binds to the TRPM5 ion channel and inactivates, blocks, or attenuates the ion channel generating a cellular response.

[0025] According to one option of the present invention, an active agent that interferes with the expression of ion channel TRPM5 is used in regulating hair growth.The active agent that interferes with the expression of ion channel TRPM5 is either miRNA, siRNA or ribozyme that targets TRPM5 gene or mRNA corresponding to TRPM5 gene.

[0026] The term "miRNA" refers to microRNAs, small non-coding RNA molecules containing 21-23 nucleotides that function in RNA knockdown and post-transcriptional regulation of gene expression. miRNAs function through base pairing with complementary sequences within mRNA molecules. As a result, these mRNA molecules are knocked down by mRNA truncation, destabilization, and / or inefficient translation.

[0027] The term "siRNA" refers to small interfering RNA, a small, non-coding RNA molecule 20-25 base pairs in length. siRNAs interfere with the expression of specific genes with complementary nucleotide sequences by degrading the mRNA after transcription, thereby preventing translation.

[0028] In the present invention, a gene is "targeted" by an miRNA, siRNA, or ribozyme if the miRNA, siRNA, or ribozyme molecule selectively reduces or inhibits the expression of TRPM5. As used herein, the phrase "selectively reduces or inhibits" refers to an miRNA, siRNA, or ribozyme that affects the expression of TRPM5.

[0029] In certain embodiments of the present invention, the miRNA or siRNA interferes with gene expression of the ion channel TRPM5 by hybridizing to the gene transcript, i.e., TRPM5 mRNA, under stringent conditions, where hybridizing "under stringent conditions" means annealing to the target mRNA region under standard conditions that tend to disfavor hybridization, such as high temperature (e.g., <60°C for 2 hours) and / or low salt content (e.g., 0.1xSSC).

[0030] According to the present invention, one of the above active agents is used in the treatment of hair growth. In certain embodiments of the present invention, the treatment is carried out locally, i.e., in the skin to be treated, on the skin, or on the skin. In some embodiments of the present invention, the treatment is carried out superficially, where the term "topical" refers to a drug treatment that is applied to a specific location on the skin and / or its appendages (e.g., hair). In certain embodiments, superficial application is on the skin, and means that the agonist or antagonist is applied directly to the skin. In other embodiments of the present invention, the treatment is carried out transdermally, where the term "transdermal" refers to a drug treatment that is applied across the stratum corneum to deeper skin layers, for example, by injection using a standard needle or microneedle. In other embodiments of the present invention, the treatment is carried out transappendageally, where the term "transappendageal" refers to a drug treatment that penetrates the skin through skin appendage structures (e.g., hair follicles, sebaceous glands, and sweat glands) to deeper skin layers.

[0031] As used herein, the term "treatment" refers to any action that results in a change in a physical condition. In particular, "treatment of hair growth" refers to any change in the initial hair growth state, such as the undesired presence of hair, the undesired absence of hair, the undesired slow / fast hair growth, chemotherapy-related hair loss, drug-related hair growth, and radiation-related hair loss. In other words, the present invention is directed to any kind of hair growth regulation.

[0032] In a particular embodiment of the present invention, the above-mentioned active agent is used as a cosmetic in the treatment of hair growth. In particular, the cosmetic use is carried out non-therapeutically but to achieve a change in an initial hair growth state, such as an undesired presence of hair, an undesired absence of hair, an undesired slow / fast hair growth, wherein said initial state is not caused by a disease or disorder.

[0033] In those embodiments in which the above-described active agents are used cosmetically, the active agents used should be cosmetically acceptable, where "cosmetically acceptable" means that the active agent should not be toxic or harmful or have other adverse side effects upon application to hair and / or skin.

[0034] In another particular embodiment of the present invention, the above-mentioned active agent is used as a medicament in the superficial treatment of a hair growth disorder, wherein the term "disorder" refers to any functional abnormality or disturbance of normal health and the term "medicine" refers to a substance useful in the cure, treatment or prevention of the disorder condition.

[0035] In some embodiments, the active agent is used as a medicament in the superficial treatment of hair growth disorders selected from the group consisting of nonscarring or non-cicatricial alopecia, such as alopecia areata, telogen effluvium, androgenic alopecia, anagen effluvium, loose anagen syndrome, and female pattern hair loss; scarring or cicatricial alopecia, such as cutaneous lichen planopilaris, frontal fibrosing alopecia, discoid lupus erythematosus, dissecting cellulitis, and folliculitis barbae; and excessive hair growth, such as hypertrichosis and hirsutism.

[0036] In those embodiments in which the above-described active agents are used as pharmaceuticals, the active agents used should be pharmaceutically acceptable, where "pharmaceutically acceptable" means that the active agent should not be toxic or harmful or have other adverse side effects upon application to hair and / or skin.

[0037] In some embodiments, at least one of the active substances of the present invention is used as a component of a composition for cosmetic or pharmaceutical use in the superficial treatment of hair growth, said composition further comprising at least one auxiliary selected from the group consisting of carriers, excipients, adjuvants, diluents and disintegrants.

[0038] In certain embodiments of such compositions, the excipient is selected from the group consisting of liposomes, nanoparticles, carboxymethylcellulose, hydroxyethylcellulose, mineral oil, petrolatum, glycerin, polysorbate 80, hydroxyethyl starch, dextran, and polyethylene glycol.

[0039] In the compositions of the present invention, the concentration of the active agent is typically in the range of 10 to 10,000 μM. In some embodiments, the lower limit of the active agent concentration is 30 μM or even 100 μM. In some embodiments, the upper limit is 3,000 μM or 1,000 μM. This results in preferred ranges, such as 30 to 10,000 μM, 30 to 3,000 μM, 10 to 3,000 μM, and 100 to 3,000 μM.

[0040] In certain embodiments, such compositions further comprise at least one other active agent that is effective in the treatment of hair growth.

[0041] In such embodiments, the other active agent is a hair growth inhibitor, e.g., an inhibitor of ornithine decarboxylase (including difluoromethylornithine (DFMO)), an antiandrogen compound, an inhibitor of 5-α-reductase, an inhibitor of the androgen ion channel, an inhibitor of S-adenosylmethionine decarboxylase, an inhibitor of γ-glutamyltranspeptidase, an inhibitor of adenylosuccinate synthetase, an inhibitor of aspartate transcarbamylase, an inhibitor of transglutaminase, an inhibitor of L-asparagine synthetase, pantothenic acid and analogs thereof, a sulfhydryl-reactive compound, an inhibitor of lipoxygenase, an inhibitor of cyclooxygenase, an inhibitor of nitric oxide synthet ... synthetase), inhibitors of ornithine aminotransferase, inhibitors of cysteine ​​synthesis pathway enzymes, inhibitors of protein kinase C, catechin compounds, green tea polyphenols, nonsteroidal antiangiogenic agents, inhibitors of arginase, inhibitors of the metabolic pathway for the conversion of glucose to acetyl-CoA, compounds that inhibit the formation of glycoproteins, proteoglycans and glycosaminoglycans, inhibitors of matrix metalloproteinase, inhibitors of the cholesterol synthesis pathway, inhibitors of DNA topoisomerase, inhibitors of aminoacyl-tRNA synthetase, inhibitors of the hypusine biosynthetic pathway, compounds that activate androgen conjugation, inhibitors of alkaline phosphatase, inhibitors of protein tyrosine kinase, and compounds that increase cellular ceramide levels. Specific examples include cyproterone acetate, progesterone, acivicin, anthglutin, L-alanosine, guanidino-succinic acid, ethacrynic acid, D-pantothenic acid, pantoyl alcohol, gabaculine, canaline, isonicotinic acid, verapamil, phentolamine, pentosan polysulfate, nafoxidine, tripelennamine, octapine, phloretin, argaric acid, simvastatin, atorvastatin, lovastatin, fluvastatin, mevastatin, N G-Methyl-L-arginine, NG-nitro-L-arginine, benzoyl-L-argininamide, L-argininamide, quercetin, apigenin, nordihydroguaiaretic acid acid:NDGA), ketoprofen, naproxen, tolmetin, diclofenac, diflunisal, sulindac, thiosalicylic acid, cysteamine, diethyldithiocarbamic acid, D-penicillamine, N-acetyl-L-cysteine, bathocuproine, enalapril, tamoxifen, cimetidine, mycophenolic acid, tetracycline, doxycycline, minocycline, thioridazine, trifluoperidine, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine, glycyrrhetinic acid, epigallocatechin gallate, epicatechin gallate, epigallocatechin, epicatechin, fusidic acid, and nitroso-acetyl-penicillamine.

[0042] In yet other embodiments, the other active agent is a preventative agent for chemotherapy- or radiation-induced alopecia or hair loss, such as, but not limited to, 4-((cyanoimino((1,2,2-trimethylpropyl)amino)methyl)amino)benzonitrile, epidermal growth factor, fibroblast growth factor, keratinocyte growth factor (FGF7), prostaglandins, cyclin-dependent kinase, p53 inhibitor, caspase-3 inhibitor, N-acyl cysteine, parathyroid hormone antagonist, α-tocopherol, cyclosporine, angiotensin ion channel blocker, and minoxidil.

[0043] Generally, the composition of the present invention can be used in any formulation suitable for treating hair growth.In a specific embodiment of the present invention, the composition is formulated in the form of ointment, lotion, cream, shampoo, gel, spray, plaster or sustained release plaster.In another specific embodiment of the present invention, the composition is formulated in the form of solution.The solution can be applied by microneedle device or before ultrasonic treatment, electrical stimulation.

[0044] As mentioned above, the inventive use of the above active agents can also be carried out non-therapeutically, where non-therapeutic refers to treatment that is not directed to curing, treating or preventing the disorder condition (see above).

[0045] Accordingly, the present invention is further directed to non-therapeutic methods of regulating hair growth, in which an effective amount of at least one of the above-mentioned active agents is administered to a subject.

[0046] Non-therapeutic methods also include embodiments in which the above-mentioned active agents are administered simultaneously, sequentially, or separately to the subject to be treated together with at least one other active agent useful in the treatment of hair growth (see above).

[0047] The following examples illustrate some of the features of specific embodiments of the present invention. However, those skilled in the art will understand that these embodiments are merely illustrative and are not intended to limit the scope of the present invention to the precise features or combinations of features of the example embodiments.

[0048] In describing the examples, reference is made to the following figures: [Brief explanation of the drawings]

[0049] [Figure 1] FIG. 1 is a photograph showing the results of immunofluorescence analysis of TRPM5 expression in human skin. [Figure 2] FIG. 2 is a photograph showing the results of immunofluorescence analysis of TRPM5 expression in human skin. [Figure 3] FIG. 3 is a graph showing that TRPM5 protein expression is significantly reduced in TRMP5-depleted human hair follicles. [Figure 4] FIG. 4 is a graph showing reduced cell proliferation (Ki67) in siTRPM5-treated hair follicles. [Figure 5] FIG. 5 is a graph showing increased apoptosis (TUNEL) in siTRPM5-treated hair follicles. [Figure 6]FIG. 6 is a graph showing that TRPM5 siRNA reduces the expression levels of LEF1 and IGF1 in human hair follicles. [Figure 7] FIG. 7 is a graph showing that TRPM5 siRNA increases the expression of TGFB2 and SFRP1 in human hair follicles. [Figure 8] FIG. 8 is a graph showing that 2,5-dimethylpyrazine (DMP) extends the anagen phase in human hair follicle organ cultures. [Figure 9] FIG. 9 is a graph showing that 2-heptanone (2-Hep) extends the anagen phase in human hair follicle organ cultures. [Figure 10] FIG. 10 is a graph showing that expression of the catagen-promoting gene TGFB2 was downregulated in 2-Hep-treated hair follicles, while the Wnt target gene AXIN2 was upregulated. [Figure 11] FIG. 11 is a graph showing that triphenylphosphine oxide (TPPO) promotes catagen in human hair follicle organ cultures. [Figure 12] FIG. 12 is a graph showing that the expression levels of IGF1 and FGF7 genes were significantly down-regulated in TPPO-treated hair follicles, while TGFB2 transcripts were up-regulated. [Example]

[0050] example 1. Immunofluorescence analysis of TRPM5 expression in human skin Unfixed frozen sections were dried and then fixed in acetone at -20°C. Subsequently, slides were washed in phosphate-buffered saline (PBS). Sample sections were pretreated with goat serum. Anti-TRPM5 was added to the sample sections and incubated. Slides were washed in PBS, and goat anti-rabbit A488 antibody was added to the sample sections and incubated. Slides were washed in PBS, and sample sections were incubated with DAPI. Slides were washed again in PBS and mounted with a coverslip using Southernbiotech Fluoromount.

[0051] Figures 1 and 2 show the results of immunofluorescence analysis of TRPM5 expression in human skin using the above protocol. From Figure 1, it can be seen that TRPM5 protein expression was primarily detected in the basal layer of the epidermis (arrowhead). From Figure 2, it can be seen that strong TRPM5 expression was detected in the outer root sheath (arrowhead), with weaker expression in the hair matrix (arrow), while no expression was detected in the hair follicle mesenchyme (asterisk).

[0052] 2. Inactivation of TRPM5 by RNAi knockdown To analyze the effects of TRPM5 inactivation by RNAi knockdown, human hair follicles were transfected with 1 μM TRPM5-targeting Accell siRNA (siTRPM5) and scrambled siRNA (siScr). After 24 h, transfected hair follicles were either snap-frozen in liquid nitrogen for immunofluorescence analysis or harvested in RNA extraction buffer for RNA isolation. TRPM5 protein expression was detected as described above. As for TRPM5, Ki67 protein expression was detected using a mouse anti-human Ki67 primary antibody and a goat anti-mouse A568 secondary antibody. For TUNEL immunolabeling, hair follicle cryosections were fixed in formalin / ethanol / acetic acid and labeled with digoxigenin-deoxyUTP (ApopTag Fluorescein In Situ Apoptosis detection kit; Millipore) in the presence of terminal deoxynucleotidyl transferase according to the manufacturer's protocol. TUNEL+ cells were visualized with anti-digoxigenin FITC-conjugated antibody (ApopTag kit). Cells positive for Ki-67 or TUNEL were counted per matrix and ORS and normalized to the total number of nuclei (DAPI+).

[0053] Total RNA was isolated using the PicoPure RNA Isolation Kit (Applied BioSystems) according to the manufacturer's protocol. After isolation, 100 ng of total RNA was reverse transcribed into cDNA using the Tetro cDNA synthesis kit (Bioline). Quantitative PCR was performed using the StepOne Plus real-time PCR system (Applied Biosystems) with Taqman fast Advanced master mix and Taqman probes for LEF1, IGF1, TGFB2, and SFRP1 (Applied Biosystems). The amounts of the above transcripts were normalized to the expression of a reference gene (GAPDH) using the ΔΔCt method. Results were performed in triplicate for each experiment.

[0054] Figures 3 to 6 show the results of siRNA targeting TRPM5 as described above (mean + / - sd, Mann Whitney U test, * p<0.05, ** p<0.001).

[0055] Figure 3 shows that TRPM5 protein expression is significantly reduced in TRPM5-depleted human hair follicles. Figures 4 and 5 show decreased cell proliferation (Ki67) and increased apoptosis (TUNEL) in siTRPM5-treated hair follicles. Figures 6 and 7 show that TRPM5 siRNA reduces the expression levels of LEF1 and IGF1 (Figure 6), while increasing the expression of TGFB2 and SFRP1 in human hair follicles (Figure 7).

[0056] 3. Activation of TRPM5 using 2,5-dimethylpyrazine (DMP) Human anagen hair follicles were isolated from the skin of men undergoing hair transplantation with written informed consent. Microdissected hair follicles were cut at the level of the dermal-subcutaneous junction and equilibrated in Williams' E medium (WEM) (Life Technologies) supplemented with 2 mM L-glutamine (Life Technologies), 10 ng / ml hydrocortisone, 10 μg / ml insulin, and antibiotics (all from Siga-Aldrich). After 24 hours, growing, undamaged anagen follicles were selected for treatment with DMP. Human hair follicles were cultured in the presence of DMP (12.5 μM). The culture medium was changed every two days with the addition of DMP. Hair follicles were photographed, and the percentage of follicles in different phases of the hair cycle was calculated on days 3 and 6 of treatment and compared with vehicle.

[0057] The graph shown in Figure 8 shows that the TRPM5 agonist 2,5-dimethylpyrazine (DMP) prolongs anagen in human hair follicle organ cultures compared to vehicle (Veh).

[0058] 4. Activation of TRPM5 with 2-heptanone (2-Hep) Human anagen hair follicles were isolated from the skin of males undergoing hair transplantation with written informed consent. Microdissected hair follicles were cut at the level of the dermal-subcutaneous junction and equilibrated in Williams' E medium (WEM) (Life Technologies) supplemented with 2 mM L-glutamine (Life Technologies), 10 ng / ml hydrocortisone, 10 μg / ml insulin, and antibiotics (all from Siga-Aldrich). After 24 hours, growing, undamaged anagen hair follicles were selected for treatment with 2-Hep (12.5 μM). The culture medium was changed every two days with the addition of 2-Hep. Hair follicles were photographed, and the percentage of follicles in different phases of the hair cycle was calculated on days 3 and 6 of treatment and compared with vehicle.

[0059] The graph shown in Figure 9 indicates that the TRPM5 agonist 2-heptanone (2-Hep) prolongs the anagen phase in human hair follicle organ cultures.

[0060] For gene expression analysis, human hair follicles were microdissected as described above and treated with 2-Hep (12.5 μM) for 6 hours. After treatment, total RNA was isolated and processed for cDNA synthesis as described above. Quantitative PCR was performed using the Taman assay with probes for TGFB2 and AXIN2 (Applied Biosystems).

[0061] FIG. 10 is a graph showing that expression of the catagen-promoting gene TGFB2 was downregulated in 2-Hep-treated hair follicles compared to vehicle (Veh), while the Wnt target gene AXIN2 was upregulated (mean + / - sd, Mann Whitney U test). * p<0.05).

[0062] 5. Inactivation of TRPM5 using triphenylphosphine oxide (TPPO) Human anagen hair follicles were isolated from the skin of men undergoing hair transplantation with written informed consent. Microdissected hair follicles were cut at the level of the dermal-subcutaneous junction and equilibrated in Williams' E medium (WEM) (Life Technologies) supplemented with 2 mM L-glutamine (Life Technologies), 10 ng / ml hydrocortisone, 10 μg / ml insulin, and antibiotics (all from Siga-Aldrich). After 24 hours, growing, undamaged anagen follicles were selected for treatment. Human hair follicles were cultured in the presence of TPPO (150 μM). The culture medium was changed every two days with the addition of TPPO. Hair follicles were photographed, and the percentage of follicles in different phases of the hair cycle was calculated on days 3 and 6 of treatment and compared with vehicle.

[0063] The graph shown in Figure 11 shows that the TRPM5 antagonist triphenylphosphine oxide (TPPO) promotes catagen in human hair follicle organ cultures compared to vehicle (Veh).

[0064] For gene expression analysis, human hair follicles were microdissected as described above and treated with TPPO (150 μM) for 6 hours. After treatment, total RNA was isolated and processed for cDNA synthesis as described above. Quantitative PCR was performed using the Taman assay with probes for IGF1, FGF7, and TGFB2 (Applied Biosystems).

[0065] FIG. 12 is a graph showing that the expression levels of IGF1 and FGF7 genes were significantly down-regulated in TPPO-treated hair follicles compared to vehicle (Veh), while TGFB2 transcripts were up-regulated (mean + / - SEM, Mann Whitney U test). * p<0.05). The present application relates to the invention described in the claims, but may also include the following as other aspects. 1. An active agent for use in the treatment of hair growth, which activates, enhances, inactivates, blocks or attenuates the cellular response of the transient receptor potential ion channel TRPM5 or interferes with the expression of said ion channel. 2. An active agent for use in the treatment of hair growth according to claim 1, wherein said active agent is an agonist of the transient receptor potential ion channel TRPM5 for use in the treatment of unwanted hair loss. 3. An active agent for use in the treatment of hair growth according to claim 1, wherein said active agent is an antagonist / inverse agonist of the transient receptor potential ion channel TRPM5 for use in the treatment of unwanted hair growth. 4. The activator is a) any one of the TRPM5 activating agonists dimethylpyrazine, dimethylethylpyrazine, tetramethylpyrazine, 2-heptanone, eugenol, SID2848719 (CAS number 702636-90-6), rutamarin, bergapten, xanthotoxin, isopimpinellin, carbachol, 3-deoxyglucosone, glucagon-like peptide 1, (E)-N-(3,4-dimethoxybenzylidene)-2-naphthalen-1-yl)acetohydrazide, or a combination thereof, or an aptamer that binds to TRPM5 and activates or enhances the ion channel to generate a cellular response; or b) TRPM5 inactivation antagonists / inverse agonists, such as triphenylphosphine oxide, econazole, miconazole, chlorpromazine, or a combination thereof, or an aptamer that binds to TRPM5 and inactivates, blocks, or attenuates the ion channel to generate a cellular response; 4. An active agent for use in the treatment of hair growth according to any one of 1 to 3 above. 5. The active agent for use in the treatment of hair growth according to claim 1, wherein said active agent is a miRNA, siRNA or ribozyme that targets TRPM5. 6. An active agent according to any one of 1 to 5 above for use as a cosmetic in the treatment of hair growth. 7. An active agent according to any one of 1 to 5 above for use as a pharmaceutical in the treatment of hair growth disorders. 8. The active agent for use as a medicine according to claim 7, wherein the hair growth disorder to be treated is non-scarring (or non-cicatricial) alopecia, such as alopecia areata, telogen effluvium, androgenic alopecia, anagen effluvium, anagen effluvium syndrome and female pattern hair loss; scarring (or cicatricial) alopecia, such as cutaneous lichen planus, frontal fibrosing alopecia, discoid lupus erythematosus, dissecting cellulitis, and folliculitis barbae; and excessive hair growth, such as hypertrichosis and hirsutism. 9. A composition for cosmetic or pharmaceutical use in the treatment of hair growth, comprising at least one active agent according to any one of 1 to 8 above, and at least one auxiliary selected from the group consisting of carriers, excipients, adjuvants, diluents and disintegrants. 10. A composition for cosmetic or pharmaceutical use in the treatment of hair growth according to claim 9, wherein said adjuvants are selected from the group consisting of liposomes, nanoparticles, carboxymethylcellulose, hydroxyethylcellulose, mineral oil, petrolatum, glycerin, polysorbate 80, hydroxyethyl starch, dextran and polyethylene glycol. 11. A composition for cosmetic or pharmaceutical use in the treatment of hair growth according to 9 or 10 above, further comprising at least one other active agent effective in the treatment of hair growth. 12. A composition for cosmetic or pharmaceutical use in the treatment of hair growth according to any one of claims 9 to 11 above, wherein the composition is formulated in the form of an ointment, lotion, cream, shampoo, hair conditioner, gel, solution, spray, plaster or sustained release plaster. 13. A non-therapeutic method for regulating hair growth, wherein an effective amount of at least one active agent that activates, enhances, inactivates, blocks or attenuates the cellular response of the transient receptor potential ion channel TRPM5 or interferes with the expression of said ion channel is administered to a subject. 14. The activator is a) any one of the TRPM5 activating agonists dimethylpyrazine, dimethylethylpyrazine, tetramethylpyrazine, 2-heptanone, eugenol, SID2848719 (CAS number 702636-90-6), rutamarin, bergapten, xanthotoxin, isopimpinellin, carbachol, 3-deoxyglucosone, glucagon-like peptide 1, (E)-N-(3,4-dimethoxybenzylidene)-2-naphthalen-1-yl)acetohydrazide, or a combination thereof, or an aptamer that binds to TRPM5 and activates or enhances the ion channel to generate a cellular response; or b) TRPM5 inactivation antagonists / inverse agonists, such as triphenylphosphine oxide, econazole, miconazole, chlorpromazine, or a combination thereof, or an aptamer that binds to TRPM5 and inactivates, blocks, or attenuates the ion channel to generate a cellular response; 14. A non-therapeutic method of regulating hair growth according to claim 13. 15. A non-therapeutic method for regulating hair growth according to claim 13 or 14, wherein the active agent is a miRNA, siRNA or ribozyme that targets the TRPM5 gene or that targets the mRNA corresponding to the TRPM5 gene.

Claims

1. 1. An active agent for use in the treatment of hair growth, which activates, enhances, inactivates, blocks or attenuates the cellular response of the transient receptor potential ion channel TRPM5 or interferes with the expression of said ion channel; a) any one of dimethylpyrazine and 2-heptanone, which are TRPM5 activating agonists, or a combination thereof; or b) triphenylphosphine oxide, a TRPM5 inactivation antagonist / inverse agonist; The activator.

2. 10. The active agent according to claim 1 for cosmetic use in the treatment of hair growth.

3. 3. An active agent according to claim 1 or 2 for use as a medicine in the treatment of hair growth disorders.

4. 4. The active agent for use as a medicament according to claim 3, wherein the hair growth disorders to be treated are non-scarring or non-cicatricial alopecia, such as alopecia areata, telogen effluvium, androgenic alopecia, anagen effluvium, anagen effluvium syndrome and female pattern hair loss; scarring or cicatricial alopecia, such as cutaneous lichen planus, frontal fibrosing alopecia, discoid lupus erythematosus, dissecting cellulitis and folliculitis barbae; and excessive hair growth, such as hypertrichosis and hirsutism.

5. A composition for cosmetic or pharmaceutical use in the treatment of hair growth, comprising at least one active agent according to any one of claims 1 to 4, and at least one auxiliary selected from the group consisting of carriers, excipients, adjuvants, diluents and disintegrants.

6. 6. A composition for cosmetic or pharmaceutical use in the treatment of hair growth according to claim 5, wherein said adjuvants are selected from the group consisting of liposomes, nanoparticles, carboxymethylcellulose, hydroxyethylcellulose, mineral oil, petrolatum, glycerin, polysorbate 80, hydroxyethyl starch, dextran and polyethylene glycol.

7. 7. A composition for cosmetic or pharmaceutical use in the treatment of hair growth according to claim 5 or 6, further comprising at least one other active agent effective in the treatment of hair growth.

8. 8. A composition for cosmetic or pharmaceutical use in the treatment of hair growth according to any one of claims 5 to 7, wherein the composition is formulated in the form of an ointment, lotion, cream, shampoo, hair conditioner, gel, solution, spray, plaster or sustained release plaster.

9. A non-therapeutic method of regulating hair growth, comprising administering to a subject an effective amount of at least one active agent that activates, enhances, inactivates, blocks, or attenuates a cellular response of, or interferes with the expression of, the transient receptor potential ion channel TRPM5; The activator is a) any one of dimethylpyrazine and 2-heptanone, which are TRPM5 activating agonists, or a combination thereof; or b) triphenylphosphine oxide, a TRPM5 inactivation antagonist / inverse agonist; The non-therapeutic method.

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