Topical Compositions and Uses Thereof

Abstract

The present invention relates to a compound represented by formula (I) JPEG2025506878000030.jpg4553 (In the formula, R1 is a C1-C3 alkyl, preferably methyl or ethyl, more preferably ethyl; R2 is H, CHO or CH=N-OH, preferably H, CHO or (E)-CH=N-OH, more preferably H; R3 is a C1-C4 alkyl, preferably ethyl or propyl, more preferably n-propyl; R4 is a C1-C6 alkyl, preferably ethyl or propyl, more preferably n-propyl; R5 is SO2NR 13 R 14 and R 13 and R 14 together with the nitrogen atom to which they are attached form a heterocycle, said heterocycle being selected from piperidine and piperazine, said heterocycle being selected from at least one R 15 and R 15 are independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, 15 at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt thereof, The topical composition comprises from about 0.0001% (w / v) to about 0.05% (w / v) of the compound of formula I, preferably compound (1), and the use of said topical composition for treating a disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, preferably a disease or disorder of the skin mediated by PDE5 activity and / or NO-related endothelial dysfunction, in particular for locally treating a skin disease, preferably skin aging or steroid-induced skin atrophy, erectile dysfunction, for preventing or treating hair loss or for promoting hair growth in a subject, preferably a human.

Description

[Technical field]

[0001] The present invention relates to a topical composition comprising a compound of formula I or a pharma- ceutically acceptable salt thereof.The present invention also relates to the use of said topical composition for treating a disease or disorder alleviated by inhibition of PDE5 and / or NO-related endothelial dysfunction in a subject, preferably a human, in particular for topically treating a skin disease, preferably skin aging or steroid-induced skin atrophy, erectile dysfunction, for preventing or treating hair loss, or for promoting hair growth. [Background technology]

[0002] Phosphodiesterases (PDEs) are enzymes that catalyze the hydrolysis and thus the degradation of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), thereby regulating the intracellular levels of second messengers. Inhibition of PDEs leads to an increase in the intracellular concentration of endogenous cAMP / cGMP. Thus, inhibition of PDEs can mediate various physiological mechanisms at different cellular and organ levels. Phosphodiesterase type 5 (PDE5) specifically hydrolyzes cyclic guanylate monophosphate (cGMP) to 5'GMP. Selective inhibition of PDE5 has been validated as a relevant approach, and strategies aimed at promoting the inhibition of PDE5 activity have been applied as therapeutic tools, especially in neurological, vascular and cardiovascular diseases and disorders. Furthermore, the introduction of PDE5 inhibitors has revolutionized the treatment of male erectile dysfunction (Andersson KE, British Journal of Pharmacology (2018) 175: 2554-2565; Das A et al., Pharmacol Ther. (2015) 147: 12-21; Dobhal T et al., Critical Review in Pharmaceutical Sciences (2012) 1 (3): 13-27). The most prominent examples of PDE5 inhibitors are sildenafil, tadalafil and vardenafil, which are described, inter alia, in, for example, WO 99 / 24433, WO 01 / 60825, EP 995'751 and WO 2011 / 075655. Recently, a new class of PDE5 inhibitors with dual pharmacology has been described, which releases NO in addition to its PDE5 inhibition in an additional manner (WO 2018 / 215433).

[0003] Hair shedding is a condition where many hairs fall out abnormally. Although it is often hair loss, abnormal hair loss of beard, eyebrows, pubic hair, armpit hair, etc. is also called hair shedding. Hair is produced in hair follicles located in the dermis layer, which is located above the subcutaneous fat, below the epidermis layer. Hair follicles are located in all parts of the body except the lips, palms, and soles of the feet, and new hair is produced from the hair matrix at the bottom of the follicle. Living cells in the hair matrix multiply and push upward, and as these cells rapidly dry out and die, they are compressed into a dense, hard mass to form the hair shaft. The hair shaft, which is made of dead protein, is covered with a delicate layer of plate-like scales (cuticle).

[0004] The hair growth cycle consists of three stages: the anagen phase, during which hair grows most actively, followed by the catagen phase, during which hair degeneration begins, and the telogen phase, during which hair growth stops. At the end of the telogen phase, the hair falls out and a new cycle begins again as new hair grows from the hair follicle. The anagen phase of eyebrows and eyelashes lasts about one to six months, while the anagen phase of hair lasts about two to six years. In general, at the end of the telogen phase, about 50 to 100 hairs fall out every day.

[0005] The most common type of hair loss in men is male pattern baldness or alopecia, which develops gradually over several years, beginning most notably on the crown of the head and progressing to the front of the head. In women, hair loss occurs in a more scattered form as the hair thins, and often occurs after menopause.

[0006] Much research has been conducted to reduce or treat alopecia, and in particular in the cosmetics or pharmaceutical industry, research has been conducted for many years to develop new substances to promote hair growth or reduce hair loss, leading to the development of drug therapies and hair transplants using female hormones, blood flow enhancers, etc. Currently, the most commonly used drugs for treating hair loss are 2,4-diamino-6-piperidinopyrimidine-3-oxide (also known as "minoxidil" preparations, see U.S. Patent Nos. 4,193,619 and 4,596,812), which are approved by the FDA, and finasteride, a specific inhibitor of type II 5α-reductase.

[0007] Minoxidil preparations are drugs that induce hair growth by increasing blood flow through vasodilatory action and supplying nutrients to hair roots, and are known to be particularly effective in alleviating hair loss symptoms in the forehead, and a drug using it as an active ingredient is sold under the trade name Rogaine (a trade name of Pharmacia & Upjohn Company). Rogaine is known to reduce hair loss by up to 10% and promote hair growth in men suffering from male pattern baldness. Rogaine must be used regularly for a long period of time, and apparently does not have a good effect on hair loss in areas other than the forehead area.

[0008] Finasteride is a drug that contains the active ingredient Propecia (Merck & Co., Inc.) and is known as an oral pill that inhibits the function of type II 5α-reductase and suppresses the conversion of testosterone to dihydrotestosterone (DHT), thereby preventing hair loss. Finasteride requires continuous and regular administration, and some patients experience side effects such as decreased libido and erectile dysfunction. Propecia can only be used by adult men.

[0009] Thus, there remains a medical need for the treatment of diseases or disorders mediated by PDE5 activity and / or NO-related endothelial dysfunction, particularly for preventing or treating hair loss and alopecia, and for promoting hair growth. Summary of the Invention

[0010] The present inventors have surprisingly found that topical treatment with the compound and composition of the present invention shows excellent therapeutic effects not only for male pattern baldness but also for female pattern baldness, improves hair growth in in vivo experiments, and shows a remarkably excellent effect in hair loss induced by chemotherapy used in cancer treatment and the like.In particular, it shows excellent therapeutic effects compared with minoxidil, which is one of the most used and commercially available drugs at present.Furthermore, it has been shown that administration of the compound and composition of the present invention before the actual hair loss can prevent hair loss.In particular, the excellent therapeutic effects obtained by the compound and composition of the present invention require significantly low amounts and concentrations of the compound and composition of the present invention, respectively.

[0011] Thus, in a first aspect, the present invention provides a compound of formula I, preferably an effective amount of a compound of formula I [ka] (In the formula, R1 is a C1-C3 alkyl, preferably methyl or ethyl, more preferably ethyl; R2 is H, CHO or CH=N-OH, preferably H, CHO or (E)-CH=N-OH, more preferably H; R3 is a C1-C4 alkyl, preferably ethyl or propyl, more preferably n-propyl; R4 is a C1-C6 alkyl, preferably ethyl or propyl, more preferably n-propyl; R5 is SO2NR 13 R 14 and R 13 and R 14 together with the nitrogen atom to which they are attached form a heterocycle selected from piperidine and piperazine, and the heterocycle is selected from at least one R 15 is substituted with, and the R 15 is independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, and at least one of R 15at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt thereof, Preferably, a topical composition is provided, wherein the topical composition comprises from about 0.0001% (w / v) to about 0.05% (w / v) of the compound of formula I, preferably compound 1. Thus, preferably, the compound of formula I, preferably compound 1, is present in the topical composition at from about 0.0001% (w / v) to about 0.05% (w / v).

[0012] In another aspect, the present invention provides a compound of formula I [ka] (In the formula, R1 is a C1-C3 alkyl, preferably methyl or ethyl, more preferably ethyl; R2 is H, CHO or CH=N-OH, preferably H, CHO or (E)-CH=N-OH, more preferably H; R3 is a C1-C4 alkyl, preferably ethyl or propyl, more preferably n-propyl; R4 is a C1-C6 alkyl, preferably ethyl or propyl, more preferably n-propyl; R5 is SO2NR 13 R 14 and R 13 and R 14 together with the nitrogen atom to which they are attached form a heterocycle selected from piperidine and piperazine, and the heterocycle is selected from at least one R 15 is substituted with, and the R 15 is independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, and at least one of R 15 at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt thereof, The topical composition provides a topical composition comprising from about 0.0001% (w / v) to about 0.05% (w / v) of the compound of formula I, preferably compound 1. Thus, the compound of formula I, preferably compound 1, is present in the topical composition at from about 0.0001% (w / v) to about 0.05% (w / v).

[0013] In another aspect, the present invention provides a topical composition comprising: (a) Formula I [ka] (In the formula, R1 is a C1-C3 alkyl, preferably methyl or ethyl, more preferably ethyl; R2 is H, CHO or CH=N-OH, preferably H, CHO or (E)-CH=N-OH, more preferably H; R3 is a C1-C4 alkyl, preferably ethyl or propyl, more preferably n-propyl; R4 is a C1-C6 alkyl, preferably ethyl or propyl, more preferably n-propyl; R5 is SO2NR 13 R 14 and R 13 and R 14 together with the nitrogen atom to which they are attached form a heterocycle selected from piperidine and piperazine, and the heterocycle is selected from at least one R 15 is substituted with, and the R 15 is independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, and at least one of R 15 at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt thereof, the topical composition comprising from about 0.0001% (w / v) to about 0.05% (w / v) of the compound of formula I, preferably compound 1, and thus the compound of formula I, preferably compound 1, is present in the topical composition at from about 0.0001% (w / v) to about 0.05% (w / v); (b) a solvent; (c) optionally, one or more other pharma- ceutically acceptable excipients.

[0014] The topical composition of the present invention can be used for the local treatment of skin diseases or disorders mediated or caused by PDE5 activity and / or NO-related endothelial dysfunction. Furthermore, the topical composition of the present invention can be used for the local treatment of skin diseases or disorders mediated or caused by PDE5 activity and / or NO-related endothelial dysfunction. Furthermore, the topical composition of the present invention can be used for the local treatment of skin diseases or disorders mediated or caused by PDE5 activity and / or NO-related endothelial dysfunction, in particular, the topical composition of the present invention can be used for the local treatment of diseases or disorders in a subject, preferably a human, and the locally treating the disease or disorder includes (a) treating a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treating erectile dysfunction; (c) preventing or treating hair loss, wherein the hair loss is preferably alopecia, and more preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA). (iv) increasing the thickness of the skin; (v) promoting angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancing blood flow within the hair follicle.

[0015] Thus, in a further aspect, the present invention provides a topical composition of the present invention for use in a method for locally treating a disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, preferably a disease or disorder of the skin mediated by PDE5 activity and / or NO-related endothelial dysfunction, in a subject, preferably a human. Preferably, the disease or disorder is selected from: (a) treatment of a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treatment of erectile dysfunction; (c) preventing or treating hair loss, preferably alopecia, more preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata, or (d) promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle. Highly preferably, the disease or disorder is preventing or treating hair loss, wherein the hair loss is alopecia, preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata. Highly preferably, the disease or disorder is promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle.

[0016] In another aspect, the present invention provides a topical composition of the present invention for use in a method of locally treating a disease or disorder in a subject, preferably a human, wherein the disease or disorder is selected from: (a) treating a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treating erectile dysfunction; (c) preventing or treating hair loss, preferably alopecia, more preferably wherein the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata, or (d) promoting hair growth through (i) inducing anagen in hair follicles; (ii) promoting melanogenesis; (iii) increasing the number of hair follicles; (iv) increasing the thickness of the skin; (v) promoting angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancing blood flow within the hair follicle. Highly preferably, the disease or disorder is preventing or treating hair loss, wherein the hair loss is alopecia, preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata. Highly preferably, the disease or disorder is promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle.

[0017] In another aspect, the present invention provides a method for treating a disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, preferably a skin disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, in a subject, preferably a human, comprising topically administering a topical composition of the present invention to the subject, preferably the human.

[0018] In another aspect, the present invention provides a method of treating a disease or disorder in a subject, preferably a human, selected from: (a) treating a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treating erectile dysfunction; (c) preventing or treating hair loss, preferably alopecia, more preferably, wherein the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata, or (d) promoting hair growth through (i) inducing anagen in hair follicles; (ii) promoting melanogenesis; (iii) increasing the number of hair follicles; (iv) increasing skin thickness; (v) promoting angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancing blood flow within the hair follicle, the method comprising topically administering a topical composition of the present invention to the subject, preferably the human. Highly preferably, the disease or disorder is preventing or treating hair loss, wherein the hair loss is alopecia, preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata. Highly preferably, the disease or disorder is promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle.

[0019] In another aspect, the present invention provides a use of a topical composition of the present invention for preparing a medicament for locally treating a disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, preferably a skin disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, in a subject, preferably a human, the method comprising topically administering the topical composition of the present invention to the subject, preferably the human.

[0020] In another aspect, the present invention provides use of a topical composition of the present invention for preparing a medicament for locally treating a disease or disorder in a subject, preferably a human, selected from: (a) treatment of a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treatment of erectile dysfunction; (c) preventing or treating hair loss, preferably hair loss, which is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata, or (d) promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle, said method comprising topically administering a topical composition of the present invention to said subject, preferably said human. Highly preferably, the disease or disorder is preventing or treating hair loss, wherein the hair loss is alopecia, preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata. Highly preferably, the disease or disorder is promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle.

[0021] Further aspects and embodiments of the invention will become apparent as this description continues. [Brief description of the drawings]

[0022] [Figure 1] Figure 1: Mimic diagram of in vivo mouse model experiment to confirm the treatment effect of compound 1 and compound 2 dissolved in PEG-400:EtOH / 7:3 on hair loss. [Figure 2A] Dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 compared with minoxidil for changes in body weight in male mice. [Figure 2B] Hair growth effects of dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 compared to minoxidil in male mice. a, b indicate p<0.05 and p<0.01, respectively, for Compound 1 and minoxidil compared to the vehicle group. [Figure 3A] A set of photographs comparing the anagen induction effects of dose-specific application of Compound 1 dissolved in PEG-400:EtOH / 7:3 versus minoxidil application in male C57BL / 6 mice. [Figure 3B] A set of photographs comparing the melanogenic effect (visual melanogenesis) of dose-specific application of Compound 1 dissolved in PEG-400:EtOH / 7:3 versus minoxidil application in male C57BL / 6 mice. [Figure 4A] Comparison of cross-sectional skin sections in male mice with dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 versus minoxidil application with H&E staining. [Figure 4B] Comparison of hair follicle numbers following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 versus minoxidil application in male mice by H&E staining. [Figure 4C] Comparison of skin thickness following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 versus minoxidil application in male mice using H&E staining. [Figure 5A] Comparison of body weight changes following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 compared to minoxidil application in female mice. [Figure 5B] Comparison of hair growth effects of dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 compared to Minoxidil application in female mice. a, b, c show p<0.05 p<0.01 and p<0.001, respectively, when comparing Compound 1 group with vehicle group. [Figure 6A]A set of photographs comparing the anagen induction effects of dose-specific application of Compound 1 dissolved in PEG-400:EtOH / 7:3 versus minoxidil application in female C57BL / 6 mice. [Figure 6B] A set of photographs comparing the melanogenic effect (visual melanogenesis) of dose-specific application of Compound 1 dissolved in PEG-400:EtOH / 7:3 versus minoxidil application in female C57BL / 6 mice. [Figure 7A] Comparison of transverse skin sections in female mice with dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 versus minoxidil application with H&E staining. [Figure 7B] Comparison of hair follicle numbers following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 versus minoxidil application in female mice by H&E staining. [Figure 7C] Comparison of skin thickness following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 versus minoxidil application in female mice using H&E staining. [Figure 8A] Dose-specific application of Compound 2 in PEG-400:EtOH / 7:3 compared with minoxidil for changes in body weight in female mice. [Figure 8B] Hair growth effects of dose-specific application of Compound 2 in PEG-400:EtOH / 7:3 compared to minoxidil in female mice. a, b, c, d show p<0.05, p<0.01, p<0.001, and p<0.0001, respectively, when comparing Compound 2 group with vehicle, and e shows p<0.05 when comparing 0.02% Compound 2 group with 2% minoxidil. [Figure 9A] A set of photographs comparing the anagen induction effects of dose-specific application of Compound 2 dissolved in PEG-400:EtOH / 7:3 versus minoxidil application in female C57BL / 6 mice. [Figure 9B]A set of photographs comparing the melanogenic effects (visual melanogenesis) of dose-specific application of Compound 2 dissolved in PEG-400:EtOH / 7:3 versus minoxidil application in female C57BL / 6 mice. [Figure 10A] Comparison of transverse skin sections in female mice with dose-specific application of Compound 2 in PEG-400:EtOH / 7:3 versus minoxidil application with H&E staining. [Figure 10B] Comparison of hair follicle numbers following dose-specific application of Compound 2 in PEG-400:EtOH / 7:3 versus minoxidil application in female mice by H&E staining. [Figure 10C] Comparison of skin thickness following dose-specific application of Compound 2 in PEG-400:EtOH / 7:3 versus minoxidil application in female mice using H&E staining. [Figure 11] FIG. 1 is a mimic diagram of an in vivo mouse model experiment to confirm the treatment effect of Compound 1 dissolved in PEG-400:EtOH / 7:3 on chemotherapy-induced hair loss. [Figure 12A] Body weight following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 compared to minoxidil. [Figure 12B] Hair growth effects of dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 compared to Minoxidil application. a, b, c, d show p<0.05, p<0.01, p<0.001, and p<0.0001, respectively, when comparing Compound 1 group to vehicle, and e, f, g, h show p<0.05, p<0.01, p<0.001, and p<0.0001, respectively, when comparing Compound 1 group to 2% Minoxidil group. [Figure 12C] Hair weight following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 compared to minoxidil application. ***, **** and ns indicate p<0.001, p<0.0001 and not significant, respectively, when comparing pre- and post-treatment animals from the same group. [Figure 13A]A set of photographs comparing the anagen induction effect of dose-specific application of Compound 1 dissolved in PEG-400:EtOH / 7:3 versus minoxidil application. [Figure 13B] A set of photographs comparing the melanogenic effect (visual melanogenesis) of dose-specific application of Compound 1 dissolved in PEG-400:EtOH / 7:3 versus minoxidil application. [Figure 14A] Comparison of transverse and longitudinal skin sections in female mice following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 versus minoxidil application with H&E staining. [Figure 14B] Comparison of hair follicle numbers following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 versus minoxidil application in female mice by H&E staining. [Figure 14C] Comparison of skin thickness following dose-specific application of Compound 1 in PEG-400:EtOH / 7:3 versus minoxidil application in female mice using H&E staining. [Figure 15A] Comparison of expression of CD-31 angiogenesis marker following dose-specific application of compound 1 in PEG-400:EtOH / 7:3 with minoxidil. [Figure 15B] Comparison of expression of CD-31 angiogenesis marker following dose-specific application of compound 1 in PEG-400:EtOH / 7:3 with minoxidil. [Figure 15C] Comparison of expression of Ki-67 cell proliferation marker following dose-specific application of compound 1 dissolved in PEG-400:EtOH / 7:3 with minoxidil. [Figure 15D] Comparison of expression of Ki-67 cell proliferation marker following dose-specific application of compound 1 dissolved in PEG-400:EtOH / 7:3 with minoxidil. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. The embodiments, preferred and highly preferred embodiments described and disclosed herein should be applied to all aspects and other embodiments, preferred and highly preferred embodiments, whether or not specifically mentioned again.

[0024] As used herein, the term "about" shall mean + / - 10%. For example, about 50% shall mean 45% to 55%. Preferably, as used herein, the term "about" shall mean + / - 5%. For example, about 50% shall mean 47.5% to 52.5%.

[0025] As used herein, the phrase "number X to number Y" includes number X and number Y. For example, the phrase "0.01 μmol to 50 μmol" refers to 0.01 μmol and 50 μmol, as well as values ​​therebetween. The same applies to the phrase "about number X to about number Y."

[0026] As used herein, the term "% (v / v)" refers to (volume of component / total volume of composition) x 100. As an example, 70% (v / v) PEG 400 refers to 70 ml of PEG 400 per 100 ml of total composition.

[0027] The terms "a" or "an" are used herein to mean "at least one," unless otherwise specified.

[0028] As used herein, the terms "topically" and "topical" refer to the application of the compositions of the present invention to the surface of the skin of a subject, preferably a human, and thus, as used herein, the term "topical administration" refers to administration to the skin of a subject, preferably a human, to prevent or treat hair loss (wherein the hair loss is alopecia, preferably the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata), or to promote hair growth.

[0029] As used herein, the term "topical composition" refers to a composition that is suitable for topical administration and can be applied to the skin of a subject, preferably a human.

[0030] As used herein, the terms "treatment", "treat", "treated" or "treating" refer to prophylaxis and / or therapy. In one embodiment, the terms "treatment", "treat", "treated" or "treating" refer to therapeutic treatment. In another embodiment, the terms "treatment", "treat", "treated" or "treating" refer to prophylactic treatment. Preferably, beneficial or desired clinical results of the treatment include, but are not limited to, alleviation of symptoms, attenuation of the extent of the disease or disorder, a stabilized (i.e., not worsening) state of the disease or disorder, a delay or slowing of the progression of the disease or disorder, an improvement or alleviation of the disease or disorder state.

[0031] As used herein, the term "effective amount" refers to an amount necessary or sufficient to achieve a desired biological effect. Preferably, the term "effective amount" refers to an amount of the compound of formula I of the present invention that (i) treats or prevents a particular disease or disorder, (ii) attenuates, improves or eliminates one or more symptoms of a particular disease or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease or disorder described herein. An effective amount of the compound of formula I of the present invention, or the topical composition or the pharmaceutical composition, is an amount that achieves this selected result, and such an amount can be routinely determined by one of ordinary skill in the art. More preferably, as used herein, the term "effective amount" refers to an amount necessary or sufficient to be effective in activating soluble guanylyl cyclase (sGC) and / or increasing the inhibition of PDE5. The effective amount can vary depending on the particular composition applied and the size of the subject. One of ordinary skill in the art can empirically determine the effective amount of a particular compound of formula I of the present invention, or the topical composition or the pharmaceutical composition of the present invention, without undue experimentation.

[0032] As used herein, the term "mammal" includes, but is not limited to, humans, mice, rats, guinea pigs, monkeys, dogs, cats, horses, cows, pigs, and sheep. As used herein, the term "mammal" preferably refers to humans. As used herein, the term "subject" includes, but is not limited to humans and mammals. As used herein, the term "subject" preferably refers to humans.

[0033] Thus, in a first aspect, the present invention provides a compound of formula I, preferably an effective amount of a compound of formula I [ka] (In the formula, R1 is a C1-C3 alkyl, preferably methyl or ethyl, more preferably ethyl; R2 is H, CHO or CH=N-OH, preferably H, CHO or (E)-CH=N-OH, more preferably H; R3 is a C1-C4 alkyl, preferably ethyl or propyl, more preferably n-propyl; R4 is a C1-C6 alkyl, preferably ethyl or propyl, more preferably n-propyl; R5 is SO2NR 13 R 14 and R 13 and R 14 together with the nitrogen atom to which they are attached form a heterocycle selected from piperidine and piperazine, and the heterocycle is selected from at least one R 15 is substituted with, and the R 15 is independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, and at least one of R 15 at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt thereof, Preferably, a topical composition is provided, wherein the topical composition comprises from about 0.0001% (w / v) to about 0.05% (w / v) of the compound of formula I, preferably compound 1. Thus, preferably, the compound of formula I, preferably compound 1, is present in the topical composition at from about 0.0001% (w / v) to about 0.05% (w / v).

[0034] In another aspect, the present invention provides a compound of formula I [ka] (In the formula, R1 is a C1-C3 alkyl, preferably methyl or ethyl, more preferably ethyl; R2 is H, CHO or CH=N-OH, preferably H, CHO or (E)-CH=N-OH, more preferably H; R3 is a C1-C4 alkyl, preferably ethyl or propyl, more preferably n-propyl; R4 is a C1-C6 alkyl, preferably ethyl or propyl, more preferably n-propyl; R5 is SO2NR 13 R 14 and R 13 and R 14 together with the nitrogen atom to which they are attached form a heterocycle selected from piperidine and piperazine, and the heterocycle is selected from at least one R 15 is substituted with, and the R 15 is independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, and at least one of R 15 at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt thereof, The topical composition provides a topical composition comprising from about 0.0001% (w / v) to about 0.05% (w / v) of the compound of formula I, preferably compound 1. Thus, the compound of formula I, preferably compound 1, is present in the topical composition at from about 0.0001% (w / v) to about 0.05% (w / v).

[0035] In another aspect, the present invention provides a topical composition comprising: (a) Formula I [ka] (In the formula, R1 is a C1-C3 alkyl, preferably methyl or ethyl, more preferably ethyl; R2 is H, CHO or CH=N-OH, preferably H, CHO or (E)-CH=N-OH, more preferably H; R3 is a C1-C4 alkyl, preferably ethyl or propyl, more preferably n-propyl; R4 is a C1-C6 alkyl, preferably ethyl or propyl, more preferably n-propyl; R5 is SO2NR 13 R 14 and R 13and R 14 together with the nitrogen atom to which they are attached form a heterocycle selected from piperidine and piperazine, and the heterocycle is selected from at least one R 15 is substituted with, and the R 15 is independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, and at least one of R 15 at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt thereof (the topical composition comprises from about 0.0001% (w / v) to about 0.05% (w / v) of the compound of formula I, preferably compound 1, and thus the compound of formula I, preferably compound 1, is present in the topical composition at from about 0.0001% (w / v) to about 0.05% (w / v). (b) a solvent; (c) optionally, one or more other pharma- ceutically acceptable excipients.

[0036] The compound of formula I contained in the topical composition of the present invention includes pharmaceutically acceptable salts of said compound.In particular, as used herein, the term "pharmaceutically acceptable salts" refers to pharmaceutically acceptable organic or inorganic salts of the compound of the present invention, particularly acid addition salts.Exemplary salts include, but are not limited to, salts of physiologically acceptable mineral acids such as hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid, or salts of organic acids such as methanesulfonic acid, p-toluenesulfonic acid, lactic acid, malic acid, tartaric acid, acetic acid, trifluoroacetic acid, citric acid, succinic acid, fumaric acid, maleic acid and salicylic acid.Further examples of pharmacologically acceptable salts of the compound of formula I are alkali metal and alkaline earth metal salts, such as sodium, potassium, lithium, calcium or magnesium salts, ammonium salts or salts of organic bases, such as methylamine, dimethylamine, triethylamine, piperidine, ethylenediamine, lysine, choline hydroxide, meglumine, morpholine or arginine salts. Further examples of pharma- ceutically acceptable salts of the compounds of formula I include the hydrochloride, hydrobromide, sulfate, bisulfate, phosphate, hydrogen phosphate, nitrate, acetate, benzoate, succinate, fumarate, maleate, lactate, citrate, benzenesulfonate, p-toluenesulfonate, and the like.

[0037] In a preferred embodiment, R1 is methyl or ethyl. In a preferred embodiment, R1 is methyl. In a preferred embodiment, R1 is ethyl. In another preferred embodiment, R2 is H. In a further preferred embodiment, R3 is ethyl or n-propyl. In a further preferred embodiment, R3 is ethyl. In a further preferred embodiment, R3 is n-propyl. In another preferred embodiment, R4 is ethyl or n-propyl. In another preferred embodiment, R4 is ethyl. In another preferred embodiment, R4 is n-propyl. In yet a further preferred embodiment, R1 is methyl or ethyl, R2 is H, R3 is ethyl or n-propyl, and R4 is ethyl or n-propyl. In yet a further preferred embodiment, R1 is ethyl, R2 is H, R3 is n-propyl, and R4 is n-propyl.

[0038] In a preferred embodiment, R5 is SO2NR 13 R 14 where R 13 and R 14 together with the nitrogen atom to which they are attached form a heterocycle, the heterocycle being selected from piperidine and piperazine, the heterocycle being selected from at least one R 15 and the R 15 are independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, and at least one of R 15 At least one of R5 contains at least one ONO2 moiety. In a preferred embodiment, the heterocyclic ring is piperidine. In another preferred embodiment, the heterocyclic ring is piperazine. In a preferred embodiment, R5 is SO2NR 13 R 14 where R 13 and R 14 together with the nitrogen atom to which they are attached form a heterocycle selected from piperidine and piperazine, and the heterocycle is selected from six R 15is replaced with 1, 2, 3, 4, or 5 of the R 15 are independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, and at least one of R 15 At least one, preferably 1, 2, 3 or 4 of R5 contains at least one ONO2 moiety. In a preferred embodiment, R5 is SO2NR 13 R 14 where R 13 and R 14 together with the nitrogen atom to which they are attached form a heterocycle selected from piperidine and piperazine, and the heterocycle is selected from one, two, or three R 15 and the R 15 are independently selected from C1 to C6 alkyl optionally substituted with OH or ONO2, and at least one of R 15 At least one, preferably one, two or three of the above comprises at least one ONO2 moiety.

[0039] In a preferred embodiment, the compound of formula I is a compound of formula I* or a compound of formula I**, [ka] wherein R1, R2, R3 and R4 are as defined herein, including all preferred and highly preferred embodiments of R1, R2, R3 and R4, alone or in combination; X is CR 21 or N, R 20 is a C1-C6 alkyl optionally substituted with OH and / or ONO2, R 21 is H or C1-C6 alkyl optionally substituted with OH and / or ONO2, The R 20 and R 21 at least one of which comprises at least one ONO2 moiety; R 22is H, or C1-C6 alkyl optionally substituted with OH and / or ONO2, R 23 and R 24 are each independently a C1-C6 alkyl optionally substituted with OH and / or ONO2, The R 22 , R 23 and R 24 at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt, solvate or hydrate thereof.

[0040] In a preferred embodiment, the compound of formula I is a compound of formula I* [ka] wherein R1, R2, R3 and R4 are as defined herein, including all preferred and highly preferred embodiments of R1, R2, R3 and R4, alone or in combination; X is CR 21 or N, R 20 is a C1-C6 alkyl optionally substituted with OH and / or ONO2, R 21 is H or C1-C6 alkyl optionally substituted with OH and / or ONO2, The R 20 and R 21 at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt, solvate or hydrate thereof.

[0041] In a further preferred embodiment, the R 20 is a C1-C6 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C4 alkyl optionally substituted with OH and / or ONO2, 20 and R 21In a further preferred embodiment, at least one of the R 20 is a C1-C6 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C3 alkyl optionally substituted with OH and / or ONO2, 20 and R 21 In a further preferred embodiment, at least one of the R 20 is a C1-C6 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C2 alkyl optionally substituted with OH and / or ONO2, 20 and R 21 In a further preferred embodiment, at least one of X is CH or N and R 20 is a C1-C6 alkyl optionally substituted with OH and / or ONO2. 20 is a C1-C4 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C4 alkyl optionally substituted with OH and / or ONO2, 20 and R 21 In a further preferred embodiment, at least one of the R 20 is a C1-C3 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C3 alkyl optionally substituted with OH and / or ONO2, 20 and R 21 In a further preferred embodiment, at least one of the R 20 is a C1-C4 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C2 alkyl optionally substituted with OH and / or ONO2, 20 and R 21At least one of the comprises at least one ONO2 moiety.

[0042] In a further preferred embodiment, the R 20 or R 21 In a further preferred embodiment, R 20 Or the R 21 In a further highly preferred embodiment, the R 20 In a further highly preferred embodiment, the R 21 In a further preferred embodiment, R 20 and the R 21 In a further preferred embodiment, the R 20 and the R 21 In a further preferred embodiment, R 20 In a further preferred embodiment, R 20 and the R 21 Each of R contains exactly one ONO2 moiety. 20 and the R 21 Each of R contains exactly one ONO2 moiety. 20 and the R 21 In a further preferred embodiment, the R 20 and the R 21 In a preferred embodiment, R 21 is H.

[0043] In a further preferred embodiment, the R 20 is a C1-C3 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C3 alkyl optionally substituted with OH and / or ONO2, 20 and R 21Each of the R independently contains at least one, preferably one or two, ONO2 moieties. 20 is a C1-C4 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C2 alkyl optionally substituted with OH and / or ONO2, 20 and R 21 Each of the R independently contains at least one, preferably one or two, ONO2 moieties. 20 is a C1-C3 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C3 alkyl optionally substituted with OH and / or ONO2, 20 and R 21 Each of R contains exactly one ONO2 moiety. 20 is a C1-C4 alkyl group optionally substituted with OH and / or ONO2, 21 is H or C1-C2 alkyl optionally substituted with OH and / or ONO2, 20 and R 21 Each of R contains exactly one ONO2 moiety. 20 is a C2-C3 alkyl group optionally substituted with OH and / or ONO2, 21 is H or a C2-C3 alkyl group optionally substituted with OH and / or ONO2, 20 and R 21 Each of the R independently contains at least one, preferably one or two, ONO2 moieties. 20 is a C2-C3 alkyl group optionally substituted with OH and / or ONO2, 21 is H or a C2-C3 alkyl group optionally substituted with OH and / or ONO2, 20 and R 21 Each of R contains exactly one ONO2 moiety. 20is selected from CH2ONO2, CH2CH2ONO2, CH(OH)CH2ONO2, CH2CH2CH2ONO2, CH(ONO2)CH2OH, CH(ONO2)CH2ONO2, C(OH)(CH2ONO2)CH2ONO2, C(OH)(CH2CH2ONO2)CH2CH2ONO2, and the R 21 is selected from H, CH2ONO2, CH2CH2ONO2, CH(OH)CH2ONO2, CH2CH2CH2ONO2, CH(ONO2)CH2OH, CH(ONO2)CH2ONO2. In a further highly preferred embodiment, the R 20 and the R 21 is CH2CH2ONO2.

[0044] In a preferred embodiment, the compound of formula I is a compound of formula I**: [ka] wherein R1, R2, R3 and R4 are as defined herein, including all preferred and highly preferred embodiments of R1, R2, R3 and R4, alone or in combination; R 22 is H or C1-C6 alkyl optionally substituted with OH and / or ONO2, R 23 and R 24 are each independently a C1-C6 alkyl optionally substituted with OH and / or ONO2, The R 22 , R 23 , and R 24 at least one of which contains at least one ONO2 moiety or a pharma- ceutically acceptable salt, solvate or hydrate thereof.

[0045] In a further preferred embodiment, the R 22 is H or C1-C4 alkyl optionally substituted with OH and / or ONO2, and the R 23 and R 24are each independently a C1-C4 alkyl group optionally substituted with OH and / or ONO2. 22 is H or C1-C2 alkyl, preferably H or methyl, and the R 23 and R 24 are each independently a C1-C3 alkyl group optionally substituted with OH and / or ONO2. 22 is H or methyl, and the R 23 and R 24 are each independently a C1-C2 alkyl group optionally substituted with OH and / or ONO2. 22 is H or methyl, and the R 23 and R 24 are each independently a C2-C3 alkyl group optionally substituted with OH and / or ONO2. 22 is methyl, and the R 23 and R 24 are each independently a C1-C2 alkyl group optionally substituted with OH and / or ONO2. 22 is methyl, and the R 23 and R 24 are each independently a C2 to C3 alkyl optionally substituted with OH and / or ONO2.

[0046] In a further preferred embodiment, the R 22 , R 23 and R 24 In a further preferred embodiment, the R 22 , R 23 and R 24 In a further preferred embodiment, R 23 and the R 24 In a further preferred embodiment, R 23 and the R 24 contains exactly one ONO2 moiety.

[0047] In a further preferred embodiment, the compound of formula I is [ka] , and [ka] is selected from.

[0048] In a further preferred embodiment, the compound of formula I is (1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl)dinitrate (1); 2-(4-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperazin-1-yl)ethyl nitrate (2); 2-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)ethyl nitrate (3); 3-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)propyl nitrate (4); (R)-1-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)ethane-1,2-diyl dinitrate (5); (S)-1-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)ethane-1,2-diyl dinitrate (6); ((2R,6S)-4-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)-1-methylpiperazine-2,6-diyl)bis(ethane-2,1-diyl) dinitrate (7); ((2S,6S)-4-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)-1-methylpiperazine-2,6-diyl)bis(ethane-2,1-diyl) dinitrate (8); 3-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)-3-hydroxypentane-1,5-diyl dinitrate (9); and The compound is selected from 2-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)-2-hydroxypropane-1,3-diyl dinitrate (10).

[0049] In a further preferred embodiment, the compound of formula I is compound 1 or compound 2. [ka]

[0050] In a further highly preferred embodiment, the compound of formula I is compound 1. [ka]

[0051] In a preferred embodiment, the compound of formula I, preferably compound 1, is present in the topical composition at about 0.0001% (w / v) to about 0.02% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I, preferably compound 1, is present in the topical composition at about 0.0002% (w / v) to about 0.02% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I, preferably compound 1, is present in the topical composition at about 0.0002% (w / v) to about 0.01% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the topical composition comprises about 0.0002% (w / v) to about 0.02% (w / v) of the compound of formula I, preferably compound 1. In a preferred embodiment, the topical composition comprises from about 0.0002% (w / v) to about 0.01% (w / v) of the compound of formula I, preferably compound 1.

[0052] In a preferred embodiment, the compound of formula I, preferably compound 1, is present in the topical composition at about 0.0002% (w / v) to about 0.008% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I, preferably compound 1, is present in the topical composition at about 0.0002% (w / v) to about 0.006% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the topical composition comprises about 0.0002% (w / v) to about 0.008% (w / v) of the compound of formula I, preferably compound 1. In a preferred embodiment, the topical composition comprises about 0.0002% (w / v) to about 0.006% (w / v) of the compound of formula I, preferably compound 1.

[0053] In a preferred embodiment, the compound of formula I, preferably compound 1, is present in the topical composition at about 0.0002% (w / v) to about 0.005% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I, preferably compound 1, is present in the topical composition at about 0.0002% (w / v) to about 0.004% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the topical composition comprises about 0.0002% (w / v) to about 0.005% (w / v) of the compound of formula I, preferably compound 1. In a preferred embodiment, the topical composition comprises about 0.0002% (w / v) to about 0.004% (w / v) of the compound of formula I, preferably compound 1.

[0054] In a preferred embodiment, the compound of formula I is compound 1, and compound 1 is present in the topical composition at about 0.0001% (w / v) to about 0.02% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I is compound 1, and compound 1 is present in the topical composition at about 0.0002% (w / v) to about 0.02% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I is compound 1, and compound 1 is present in the topical composition at about 0.0002% (w / v) to about 0.01% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at about 0.0002% (w / v) to about 0.02% (w / v). In a preferred embodiment, the compound of formula I is Compound 1, and the topical composition comprises Compound 1 in an amount of from about 0.0002% (w / v) to about 0.01% (w / v).

[0055] In a preferred embodiment, the compound of formula I is compound 1, and compound 1 is present in the topical composition at about 0.0002% (w / v) to about 0.008% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I is compound 1, and compound 1 is present in the topical composition at about 0.0002% (w / v) to about 0.006% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises about 0.0002% (w / v) to about 0.008% (w / v) of compound 1. In a preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises about 0.0002% (w / v) to about 0.006% (w / v) of compound 1.

[0056] In a preferred embodiment, the compound of formula I is compound 1, and compound 1 is present in the topical composition at about 0.0002% (w / v) to about 0.005% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I is compound 1, and compound 1 is present in the topical composition at about 0.0002% (w / v) to about 0.004% (w / v) based on the total volume of the topical composition. In a preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises about 0.0002% (w / v) to about 0.005% (w / v) of compound 1. In a preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises about 0.0002% (w / v) to about 0.004% (w / v) of compound 1.

[0057] In a more preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, in a concentration of about 1.5 μM to about 300 μM, preferably about 3 μM to about 200 μM, more preferably about 5 μM to about 100 μM.

[0058] In a more preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 1.5 μM to about 300 μM, preferably about 3 μM to about 200 μM, and more preferably about 5 μM to about 100 μM.

[0059] In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 5 μM, preferably about 6 μM to about 75 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 5 μM, preferably about 6 μM to about 60 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 5 μM, preferably about 6 μM to about 50 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 5 μM, preferably about 6 μM to about 45 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 5 μM, preferably about 6 μM to about 40 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 6 μM to about 40 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 6 μM to about 38 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 6 μM to about 35 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 6 μM to about 8 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 25 μM to about 35 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 6 μM. In a further preferred embodiment, the topical composition comprises the compound of formula I, preferably compound 1, at a concentration of about 30 μM.

[0060] In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 5 μM, preferably about 6 μM to about 75 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 5 μM, preferably about 6 μM to about 60 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 5 μM, preferably about 6 μM to about 50 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 5 μM, preferably about 6 μM to about 45 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 5 μM, preferably about 6 μM to about 40 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 6 μM to about 40 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 6 μM to about 38 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 6 μM to about 35 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 6 μM to about 8 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 25 μM to about 35 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 6 μM. In a further preferred embodiment, the compound of formula I is compound 1, and the topical composition comprises compound 1 at a concentration of about 30 μM.

[0061] In some embodiments, the topical compositions can be formulated into different dosage forms, such as solutions, suspensions, creams, ointments, lotions, pastes, emulsions, foams and gels.

[0062] In a preferred embodiment, the topical composition is formulated as a solution or gel. In a preferred embodiment, the topical composition is a liquid topical composition. In a further preferred embodiment, the topical composition is a liquid topical composition and the liquid topical composition is formulated as a solution. In a further preferred embodiment, the topical composition is a liquid topical composition and the liquid topical composition is formulated as a gel.

[0063] In a preferred embodiment, the topical composition comprises at least one solvent selected from the group consisting of PEG 200, PEG 240, PEG 300, PEG 350, PEG 400, PEG 540, ethanol, 2-(2-ethoxyethoxy)ethanol (transcutol), glycerin, propylene glycol, polypropylene glycol, poloxamer 101, poloxamer 182, poloxamer 188, poloxamer 237, poloxamer 331, poloxamer 338, poloxamer 407, water, and mixtures thereof. In a preferred embodiment, the topical composition comprises at least one solvent selected from the group consisting of PEG 400, PEG 350, PEG 240, ethanol, 2-(2-ethoxyethoxy)ethanol (transcutol), glycerin, propylene glycol polypropylene glycol, poloxamer 407, poloxamer 188, and water. As used herein, the term "Transcutol" refers to 2-(2-ethoxyethoxy)ethanol.

[0064] In a preferred embodiment, the topical composition comprises a combination of solvents, the combination of solvents comprising a solvent selected from the group consisting of PEG 200, PEG 240, PEG 300, PEG 350, PEG 400, PEG 540, ethanol, 2-(2-ethoxyethoxy)ethanol (Transcutol), glycerin, propylene glycol, polypropylene glycol, poloxamer 101, poloxamer 182, poloxamer 188, poloxamer 237, poloxamer 331, poloxamer 338, poloxamer 407, and water. In a preferred embodiment, the topical composition comprises a combination of solvents, the combination of solvents comprising a solvent selected from the group consisting of PEG 400, PEG 350, PEG 240, ethanol, 2-(2-ethoxyethoxy)ethanol (Transcutol), glycerin, propylene glycol polypropylene glycol, poloxamer 407, poloxamer 188, and water.

[0065] In a preferred embodiment, the topical composition comprises a solvent combination, the solvent combination comprising PEG 400 and ethanol. In a preferred embodiment, the topical composition comprises a solvent combination, the solvent combination comprising, and preferably consisting of, PEG 400 and ethanol, the amount of PEG 400 being about 60% to about 75% (vol / vol), preferably about 65% to about 75% (vol / vol), more preferably about 68% to about 72% (vol / vol), based on the total volume of the topical composition, and the amount of ethanol being about 25% to about 40% (vol / vol), preferably about 25% to about 35% (vol / vol), more preferably about 28% to about 32% (vol / vol), based on the total volume of the topical composition.

[0066] In a preferred embodiment, the topical composition includes a solvent combination consisting of PEG 400 and ethanol, the amount of PEG 400 is about 60% to about 75% (vol / vol), preferably about 65% to about 75% (vol / vol), more preferably about 68% to about 72% (vol / vol) based on the total volume of the topical composition, and the amount of ethanol is about 25% to about 40% (vol / vol), preferably about 25% to about 35% (vol / vol), more preferably about 28% to about 32% (vol / vol) based on the total volume of the topical composition.

[0067] In a preferred embodiment, the topical composition comprises a solvent combination consisting of PEG 400 and ethanol, the amount of PEG 400 being about 68% to about 72% (vol / vol) based on the total volume of the topical composition, and the amount of ethanol being about 28% to about 32% (vol / vol) based on the total volume of the topical composition.

[0068] In a preferred embodiment, the topical composition comprises a solvent combination consisting of PEG 400 and ethanol, the amount of PEG 400 being about 70% (vol / vol) based on the total volume of the topical composition, and the amount of ethanol being about 30% (vol / vol) based on the total volume of the topical composition.

[0069] In a preferred embodiment, the topical composition comprises a solvent, which is a combination of PEG 400 and ethanol, the ratio of PEG 400 to ethanol (vol / vol) being about 60:40 to 75:25, preferably about 65:35 to 75:25, more preferably about 68:32 to 72:28, again more preferably about 70:30.

[0070] In a preferred embodiment, the topical composition comprises a combination of solvents, the combination of solvents comprising, and preferably consisting of, PEG400, ethanol, transcutol, and water, the amount of PEG400 is about 5% to about 15% (vol / vol), preferably about 8% to about 12% (vol / vol), based on the total volume of the topical composition, the amount of ethanol is about 20% to about 40% (vol / vol), preferably about 25% to about 35% (vol / vol), based on the total volume of the topical composition, the amount of transcutol is about 5% to about 15% (vol / vol), preferably about 8% to about 12% (vol / vol), based on the total volume of the topical composition, and the amount of water is about 30% to about 70% (vol / vol), preferably about 40% to about 60%, more preferably about 45% to about 55% (vol / vol), based on the total volume of the topical composition.

[0071] In a preferred embodiment, the topical composition comprises a combination of solvents, the combination of solvents being PEG400, ethanol, transcutol, and water, the amount of PEG400 being about 5% to about 15% (vol / vol), preferably about 8% to about 12% (vol / vol), based on the total volume of the topical composition, the amount of ethanol being about 20% to about 40% (vol / vol), preferably about 25% to about 35% (vol / vol), based on the total volume of the topical composition, the amount of transcutol being about 5% to about 15% (vol / vol), preferably about 8% to about 12% (vol / vol), based on the total volume of the topical composition, and the amount of water being about 30% to about 70% (vol / vol), preferably about 40% to about 60%, more preferably about 45% to about 55% (vol / vol), based on the total volume of the topical composition.

[0072] In a preferred embodiment, the topical composition comprises a solvent, which is a combination of PEG400, ethanol, transcutol, and water, in a ratio (vol / vol) of about 5:20:5:30 to 15:40:15:70, preferably about 8:25:8:40 to about 12:35:12:60, and more preferably about 10:30:10:50.

[0073] In a preferred embodiment, the solvent is present in a concentration of about 90% w / w to about 99% w / w, preferably about 95% w / w to about 99% w / w, and more preferably about 97% w / w to about 98% w / w, based on the total weight of the topical composition.

[0074] In a preferred embodiment, the topical composition optionally includes one or more other pharma- ceutically acceptable excipients. In a preferred embodiment, the topical composition includes one or more other pharma- ceutically acceptable excipients. In a preferred embodiment, the pharma-ceutically acceptable excipients are selected from the group consisting of thickeners, stabilizers, antioxidants, chelating agents, oily materials, emulsifiers, penetration enhancers, pH adjusters, preservatives, antibacterial agents, opacifiers, fragrances, colorants, gelling agents, moisturizers, surfactants, and combinations thereof. In one embodiment, the pharma-ceutically acceptable excipients are selected from the group consisting of thickeners, stabilizers, antioxidants, pH adjusters, preservatives, antibacterial agents, and combinations thereof.

[0075] Pharmaceutically acceptable excipients used in the topical compositions of the present invention can act in more than one way, for example, a thickening agent can also function as a gelling agent, and a solubilizing agent can also function as a solvent.

[0076] In a preferred embodiment, the topical composition comprises one or more other pharma- ceutically acceptable excipients, one of which is a thickening agent. In a preferred embodiment, the topical composition comprises a thickening agent. Thickening agents are known to those skilled in the art and include cellulose derivatives, polyvinylpyrrolidone, carbomer polymers, carbomer derivatives, maltodextrin, polydextrose, dextrates, carboxypolymethylene, polyvinyl alcohol, poloxamers, and mixtures thereof.

[0077] In a preferred embodiment, the topical composition comprises one or more pharma- ceutically acceptable excipients selected from cellulose derivatives, polyvinylpyrrolidone, carbomer polymers, carbomer derivatives, maltodextrin, polydextrose, dextrates, carboxypolymethylene, polyvinyl alcohol, poloxamers, or mixtures thereof.

[0078] In a preferred embodiment, the topical composition comprises one or more other pharma- ceutically acceptable excipients, and one of the pharma-ceutically acceptable excipients is an antioxidant.In a preferred embodiment, the topical composition comprises an antioxidant.Antioxidants are known to those skilled in the art, and include, for example, butylated hydroxyanisole, butylated hydroxytoluene, vitamin C, vitamin E, vitamin A, lutein, lycopene, retinyl palmitate, potassium metabisulfite, sodium metabisulfite, sodium thiosulfate pentahydrate, 3,4-dihydroxybenzoic acid, propyl gallate, α-lipoic acid, ascorbyl palmitate, sodium pyrosulfite, ubiquinone, selenium, or combinations thereof.

[0079] In a preferred embodiment, the topical composition comprises one or more pharma- ceutically acceptable excipients selected from the group consisting of butylated hydroxyanisole, butylated hydroxytoluene, vitamin C, vitamin E, vitamin A, lutein, lycopene, retinyl palmitate, potassium metabisulfite, sodium metabisulfite, sodium thiosulfate pentahydrate, 3,4-dihydroxybenzoic acid, propyl gallate, alpha-lipoic acid, ascorbyl palmitate, sodium metabisulfite, ubiquinone, selenium, or combinations thereof.

[0080] In a preferred embodiment, the topical composition comprises one or more other pharma- ceutically acceptable excipients, one of which is a preservative.In a preferred embodiment, the topical composition comprises a preservative.Preservatives are known to those skilled in the art and include, for example, benzyl alcohol, benzoic acid, phenol, m-cresol, methylparaben, propylparaben, or combinations thereof.

[0081] In a preferred embodiment, the topical composition comprises one or more pharma- ceutically acceptable excipients selected from benzyl alcohol, benzoic acid, phenol, m-cresol, methylparaben, propylparaben, or combinations thereof.

[0082] In a preferred embodiment, the topical composition comprises one or more pharma- ceutically acceptable excipients selected from PEG-20 hydrogenated castor oil, PEG-35 hydrogenated castor oil, PEG-40 hydrogenated castor oil (polyoxyl 40 hydrogenated castor oil), PEG-60 hydrogenated castor oil, PEG-100 hydrogenated castor oil, PEG-1000 hydrogenated castor oil, PEG-2000 hydrogenated castor oil, PEG-4000 hydrogenated castor oil, PEG-6000 hydrogenated castor oil, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, and polysorbate 100.

[0083] In a preferred embodiment, the topical composition comprises PEG-20 hydrogenated castor oil, PEG-35 hydrogenated castor oil, PEG-40 hydrogenated castor oil (Polyoxyl 40 hydrogenated castor oil), PEG-60 hydrogenated castor oil, PEG-100 hydrogenated castor oil, PEG-1000 hydrogenated castor oil, PEG-2000 hydrogenated castor oil, PEG-4000 hydrogenated castor oil, PEG-6000 hydrogenated castor oil, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, polysorbate 100, cellulose derivatives, polyvinylpyrrolidone, carbomer polymers, carbomer derivatives, maltodextrin, polydextrose, dextrates, carboxypoly The composition may include one or more pharma- ceutically acceptable excipients selected from methylene, polyvinyl alcohol, poloxamer, butylated hydroxyanisole, butylated hydroxytoluene, vitamin C, vitamin E, vitamin A, lutein, lycopene, retinyl palmitate, potassium metabisulfite, sodium metabisulfite, sodium thiosulfate pentahydrate, 3,4-dihydroxybenzoic acid, propyl gallate, alpha-lipoic acid, ascorbyl palmitate, sodium pyrosulfite, ubiquinone, selenium, benzyl alcohol, benzoic acid, phenol, m-cresol, methylparaben, propylparaben, or combinations or mixtures thereof.

[0084] In a preferred embodiment, the compound of formula I is compound 1, [ka] The topical composition comprises compound 1 at a concentration of about 1.5 μM to about 300 μM, preferably about 3 μM to about 200 μM, and more preferably about 5 μM to about 100 μM, the topical composition comprises a solvent combination, the solvent combination comprising, and preferably consisting of, PEG400 and ethanol, the amount of PEG400 being about 60% to about 75% (vol / vol), preferably about 65% to about 75% (vol / vol), more preferably about 68% to about 72% (vol / vol), based on the total volume of the topical composition, and the amount of ethanol being about 25% to about 40% (vol / vol), preferably about 25% to about 35% (vol / vol), more preferably about 28% to about 32% (vol / vol), based on the total volume of the topical composition.

[0085] In a preferred embodiment, the compound of formula I is compound 1, [ka] The topical composition comprises compound 1 at a concentration of about 1.5 μM to about 300 μM, preferably about 3 μM to about 200 μM, and more preferably about 5 μM to about 100 μM; The topical composition comprises a combination of solvents, the combination of solvents comprising, and preferably consisting of, PEG400, ethanol, transcutol, and water, the amount of PEG400 being about 5% to about 15% (vol / vol), preferably about 8% to about 12% (vol / vol), based on the total volume of the topical composition, the amount of ethanol being about 20% to about 40% (vol / vol), preferably about 25% to about 35% (vol / vol), based on the total volume of the topical composition, the amount of transcutol being about 5% to about 15% (vol / vol), preferably about 8% to about 12% (vol / vol), based on the total volume of the topical composition, and the amount of water being about 30% to about 70% (vol / vol), preferably about 40% to about 60%, more preferably about 45% to about 55% (vol / vol), based on the total volume of the topical composition.

[0086] In a preferred embodiment, the compound of formula I is compound 1, [ka] The topical composition comprises Compound 1 at a concentration of about 5 μM to about 75 μM, preferably about 5 μM to about 60 μM, and more preferably about 5 μM to about 50 μM; the topical composition comprises a solvent combination consisting of PEG 400 and ethanol, the amount of PEG 400 being about 68% to about 72% (vol / vol) based on the total volume of the topical composition, and the amount of ethanol being about 28% to about 32% (vol / vol) based on the total volume of the topical composition.

[0087] In a preferred embodiment, the compound of formula I is compound 1, [ka] The topical composition contains compound 1 at a concentration of about 1.5 μM to about 300 μM, preferably about 3 μM to about 200 μM, and more preferably about 5 μM to about 100 μM, and the topical composition contains a solvent, which is a combination of PEG400, ethanol, transcutol, and water, and the ratio (vol / vol) of PEG400:ethanol:transcutol:water is about 5:20:5:30 to 15:40:15:70, preferably about 8:25:8:40 to about 12:35:12:60, and more preferably about 10:30:10:50.

[0088] In a preferred embodiment, the compound of formula I is compound 1, [ka] The topical composition comprises compound 1 at a concentration of about 5 μM to about 40 μM, preferably about 5 μM to about 38 μM, and more preferably about 5 μM to about 35 μM, and the topical composition comprises a solvent, the solvent being a combination of PEG 400 and ethanol, the ratio of PEG 400 to ethanol (vol / vol) being about 60:40 to 75:25, preferably about 65:35 to 75:25, more preferably about 68:32 to 72:28, and again more preferably about 70:30.

[0089] The topical composition of the present invention can be used for the local treatment of skin diseases or disorders mediated or caused by PDE5 activity and / or NO-related endothelial dysfunction. Furthermore, the topical composition of the present invention can be used for the local treatment of skin diseases or disorders mediated or caused by PDE5 activity and / or NO-related endothelial dysfunction. Furthermore, the topical composition of the present invention can be used for the local treatment of skin diseases or disorders mediated or caused by PDE5 activity and / or NO-related endothelial dysfunction, in particular, the topical composition of the present invention can be used for the local treatment of diseases or disorders in a subject, preferably a human, and the locally treating the disease or disorder includes (a) treating a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treating erectile dysfunction; (c) preventing or treating hair loss, wherein the hair loss is preferably alopecia, and more preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA). (iv) increasing the thickness of the skin; (v) promoting angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancing blood flow within the hair follicle.

[0090] Thus, in a further aspect, the present invention provides a topical composition of the present invention for use in a method for locally treating a disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, preferably a disease or disorder of the skin mediated by PDE5 activity and / or NO-related endothelial dysfunction, in a subject, preferably a human. Preferably, the disease or disorder is selected from: (a) treatment of a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treatment of erectile dysfunction; (c) preventing or treating hair loss, preferably alopecia, more preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata, or (d) promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle. Highly preferably, the disease or disorder is preventing or treating hair loss, wherein the hair loss is alopecia, preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata. Highly preferably, the disease or disorder is promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle.

[0091] In another aspect, the present invention provides a topical composition of the present invention for use in a method of locally treating a disease or disorder in a subject, preferably a human, wherein the disease or disorder is selected from: (a) treating a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treating erectile dysfunction; (c) preventing or treating hair loss, preferably alopecia, more preferably wherein the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata, or (d) promoting hair growth through (i) inducing anagen in hair follicles; (ii) promoting melanogenesis; (iii) increasing the number of hair follicles; (iv) increasing the thickness of the skin; (v) promoting angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancing blood flow within the hair follicle. Highly preferably, the disease or disorder is preventing or treating hair loss, wherein the hair loss is alopecia, preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata. Highly preferably, the disease or disorder is promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle.

[0092] In another aspect, the present invention provides a method for treating a disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, preferably a skin disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, in a subject, preferably a human, comprising topically administering a topical composition of the present invention to the subject, preferably the human.

[0093] In another aspect, the present invention provides a method of treating a disease or disorder in a subject, preferably a human, selected from: (a) treating a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treating erectile dysfunction; (c) preventing or treating hair loss, preferably alopecia, more preferably, wherein the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata, or (d) promoting hair growth through (i) inducing anagen in hair follicles; (ii) promoting melanogenesis; (iii) increasing the number of hair follicles; (iv) increasing skin thickness; (v) promoting angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancing blood flow within the hair follicle, the method comprising topically administering a topical composition of the present invention to the subject, preferably the human. Highly preferably, the disease or disorder is preventing or treating hair loss, wherein the hair loss is alopecia, preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata. Highly preferably, the disease or disorder is promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle.

[0094] In another aspect, the present invention provides a use of a topical composition of the present invention for preparing a medicament for locally treating a disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, preferably a skin disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, in a subject, preferably a human, the method comprising topically administering the topical composition of the present invention to the subject, preferably the human.

[0095] In another aspect, the present invention provides use of a topical composition of the present invention for preparing a medicament for locally treating a disease or disorder in a subject, preferably a human, selected from: (a) treatment of a skin disease, preferably skin aging or steroid-induced skin atrophy; (b) treatment of erectile dysfunction; (c) preventing or treating hair loss, preferably hair loss, which is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata, or (d) promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle, said method comprising topically administering a topical composition of the present invention to said subject, preferably said human. Highly preferably, the disease or disorder is preventing or treating hair loss, wherein the hair loss is alopecia, preferably, the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata. Highly preferably, the disease or disorder is promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle.

[0096] In a further aspect, the present invention relates to a pharmaceutical composition for preventing or treating hair loss or promoting hair growth, comprising a compound of formula I, its derivative or a pharma- ceutically acceptable salt thereof as an active ingredient, preferably, said compound of formula I is compound 1 or compound 2, more preferably, said compound of formula I is compound 1.

[0097] In a preferred embodiment, the disease or disorder is the prevention or treatment of hair loss. In a preferred embodiment, the disease or disorder is the prevention or treatment of hair loss, and the hair loss is alopecia. In a preferred embodiment, the disease or disorder is the prevention or treatment of hair loss, and the hair loss is alopecia, and the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata. In a preferred embodiment, the alopecia is androgenetic alopecia (AGA). In a preferred embodiment, the alopecia is chemotherapy-induced alopecia (CIA). In a preferred embodiment, the alopecia is alopecia areata.

[0098] In a preferred embodiment, the disease or disorder is promoting hair growth through (i) induction of anagen in hair follicles; (ii) promotion of melanogenesis; (iii) increase in the number of hair follicles; (iv) increase in skin thickness; (v) promotion of angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) enhancement of blood flow within the hair follicle.

[0099] example Synthesis of compounds of formula I: Compounds of formula I and their synthesis are disclosed in WO 2018 / 215433, the disclosure of which is incorporated herein by reference in its entirety. In particular, Examples 7, 14, 20, 55, 57, 103, 106 and 107 of WO 2018 / 215433 contain detailed descriptions of the preparation of preferred compounds of formula I.

[0100] Materials: The following materials were used to prepare the exemplary compositions described herein, all of which are known to those of skill in the art and are described in more detail in the Handbook of Pharmaceutical Excipients, co-published by the American Pharmaceutical Association and The Pharmaceutical Society of Great Britain (eds RC Rowe, PJ Sheskey and ME Quinn; Pharmaceutical Press, London, 2009): Transcutol (diethylene glycol monoethyl ether [IUPAC: 2-(2-ethoxyethoxy)ethanol], 99% purity or higher, Sigma Aldrich); Kolliphor RH40 (Sigma Aldrich, polyoxyl 40 hydrogenated castor oil); PEG-400 (C 2n H 4n+2 O n+1 ;n=8.2~9.1;VWR);Ethanol BioUltra (Sigma Aldrich).

[0101] Analytical HPLC: HPLC-MS was used to analyze the exemplary compositions described herein. The content of Compound 1 in the compositions was within the prescribed tolerance range.

[0102] Instrument: AGILENT 1100, MS SCIEC API 4000 Q-TRAP mass spectrometer Analytical conditions: Column: Zorbax XBD-C18 column, 4.6 × 50 mm, 3.5 μm. Column temperature: 40 °C, Mobile phase A: water / acetonitrile (95 / 5: v / v) + 0.1% formic acid in water, Mobile phase B: water / acetonitrile (5 / 95: v / v) + 0.1% formic acid in water, Flow rate: 0.4 mL / min, Injection volume: 20 μL (single injection), Detection: Wavelength 250 nm.

[0103] [Table 0]

[0104] All exemplary compositions were clear solutions upon visual inspection.

[0105] Example 1 Preparation of the Composition Vehicle A. Preparation of Vehicle Formulation A For 100 ml of vehicle composition A, 70 ml of PEG-400 was mixed with 30 ml of ethanol and the solution was stirred for 10 minutes.

[0106] B. Preparation of Vehicle Composition B For 100 ml of vehicle composition B, 10 ml of PEG-400 was mixed with 10 ml of Transcutol and 30 ml of ethanol and the solution was stirred for 10 minutes. 0.130 ml of Kolliphor was added to the solution and stirred at room temperature for 24 hours. After 24 hours, 50 ml of water was added and stirred for 15 minutes.

[0107] Example 2 Preparation of the Compositions of the Invention A. Preparation of Topical Composition A1 For 100 ml of topical composition A1, 70 ml of PEG-400 was mixed with 30 ml of ethanol, and the solution was stirred for 10 minutes. 0.2 mg (0.3 μmol) of compound 1 was added to the solution, and the solution was stirred at room temperature for 24 hours.

[0108] B. Preparation of Topical Composition A2 For 100 ml of topical composition A2, 70 ml of PEG-400 was mixed with 30 ml of ethanol and the solution was stirred for 10 minutes. 2 mg (3 μmol) of compound 1 was added to the solution and stirred at room temperature for 24 hours.

[0109] C. Preparation of Topical Composition A3 For 100 ml of topical composition A3, 70 ml of PEG-400 was mixed with 30 ml of ethanol and the solution was stirred for 10 minutes. 20 mg (30 μmol) of compound 1 was added to the solution and stirred at room temperature for 24 hours.

[0110] D. Preparation of Topical Composition A4 For 100 ml of topical composition A4, 70 ml of PEG-400 was mixed with 30 ml of ethanol and the solution was stirred for 10 minutes. 0.4 mg (0.6 μmol) of Compound 1 was added to the solution and stirred at room temperature for 24 hours.

[0111] E. Preparation of Topical Composition A5 For 100 ml of topical composition A5, 70 ml of PEG-400 was mixed with 30 ml of ethanol and the solution was stirred for 10 minutes. 2 mg (3.5 μmol) of compound 2 was added to the solution and stirred at room temperature for 24 hours.

[0112] F. Preparation of Topical Composition A6 For 100 ml of topical composition A3, 70 ml of PEG-400 was mixed with 30 ml of ethanol and the solution was stirred for 10 minutes. 20 mg (35 μmol) of compound 2 was added to the solution and stirred at room temperature for 24 hours.

[0113] G. Preparation of Topical Composition B1 For 100ml of vehicle composition B1, 10ml of PEG-400 was mixed with 10ml of Transcutol and 30ml of ethanol, and the solution was stirred for 10 minutes. 0.2mg (0.3μmol) of Compound 1 and 0.130ml of Kolliphor were added to the solution, and the solution was stirred at room temperature for 24 hours. After 24 hours, 50ml of water was added, and the solution was stirred for 15 minutes.

[0114] Example 3 Topical dermal administration of Compound 1 and its effect on hair growth in a C57BL / 6 mouse model of alopecia in vivo The back hair of C57BL / 6 mice is known to have a time-synchronized hair cycle. From about 18-21 days (2-3 weeks) and 47-95 days (6-14 weeks) of age, the back hair is in telogen / quiescent phase (visually pink skin) [Lee BH, Lee JS, Kim YC. Hair growth-promoting effects of lavender oil in C57BL / 6 mice. Toxicol Res. (2016) 32(2): 103-108; Choi HI, Kang BM, Jang J, Hwang ST, Kwon O. Novel effect of sildenafil on hair growth. Biochem Biophys Res Commun. (2018) 505(3): 685-691]. Thus, C57BL / 6 mice are an established model for studying hair growth promotion.

[0115] animal Male and female C57BL / 6 mice aged 6–9 weeks and in a stable telogen phase of the hair growth cycle were used (obtained from Vivo Bio Tech Ltd, Telangana, India). Details of the different treatment groups are given in Table 1.

[0116] [Table 1]

[0117] Compound treatment application 50 μL of clear solutions of compound 1 at concentrations of 0.02%, 0.002%, and 0.0002% (w / v) in the corresponding vehicle (PEG-400:EtOH / 7:3) (PEG-400: Central Drug House Ltd., Delhi, India; Ethanol: Changshu Hongsheng fine chemicals Co. Ltd., China) and positive control, minoxidil (Chemieliva, China) at 2% solution (females) or 5% solution (males) (w / v) in the corresponding vehicle (PEG-400:EtOH / 7:3) were topically applied to the shaved area on the back of the mice. The solutions were applied over a distance of approximately 2 cm. 2The treatment regimen was applied twice daily for 5 days / week for a total of 4 weeks.

[0118] Induction of anagen hair growth in C57BL / 6 mice The animals selected on day 1 weighed 19.8–20.2 g, with an average of 20.05 g ± 0.17 g. 2 ) was clipped using electric clippers without touching the animal's skin 0 days before treatment with Compound 1, and the hair was collected. Treatment was monitored following visual analysis for the onset of anagen, changes in skin color, and the appearance of hair growth. After the end of the experiment, the dorsal skin of all animals was removed and visual melanin formation was reported. Skin sections were then stored in 10% formalin solution for further histopathological analysis.

[0119] The animals were observed for 30 days and hair growth scores were recorded daily. The criteria for scoring hair growth were as follows: no hair growth, pink skin - score 0; skin color changed from pink to gray / light gray with no visible hair growth - score 0.5, onset of anagen (skin color changed from gray / light gray to dark gray / black with no visible hair growth) - score 1; sparse hair growth - score 1.5; diffuse short hair growth - score 2, moderate hair growth - score 2.5 and dense normal hair coat - score 3 (Lee BH, Lee JS, Kim YC. Hair growth-promoting effects of lavender oil in C57BL / 6 mice. Toxicol Res. (2016) 32(2): 103-108).

[0120] Photographs of the animals were captured before the start of the study and throughout the course of the study. To maintain the integrity of the study, animals that showed sudden hair regrowth in random areas of the dorsal shaved skin were considered outliers and were not included in the experiment.

[0121] Histology and immunohistochemistry After the experiment was completed, all animals were sacrificed by euthanasia (Choi HI, Kang BM, Jang J, Hwang ST, Kwon O. Novel effect of sildenafil on hair growth. Biochem Biophys Res Commun. (2018) 505(3):685-691). The dorsal skin of each animal was then removed by peeling. The peeled skin was spread and visually checked for induction of melanogenesis, indicated by blackening of the back of the skin. The skin samples were spread evenly on pre-labeled glass slides and kept in 10% formalin until further use. The skin samples were then embedded in paraffin and sectioned at 5 microns using a microtome, and corresponding slides were prepared for each animal. The resulting slides were used for hematoxylin & eosin (H&E) staining, and the total number of hair follicles in the dermis and subcutaneous tissue was determined for each animal in the micrograph data of the cross-sections and longitudinal sections. Skin thickness was measured using Image J 1.44 software.

[0122] statistical analysis Statistical analysis was performed using GraphPad Prism 9.0.0 software. Statistical significance was determined by one-way or two-way ANOVA followed by Tukey's multiple comparison test. Results are presented as mean + / - SEM from the number of animals per group. Statistical significance was considered as P<0.05.

[0123] result Male mice: All animals were observed to change skin color from pink (telogen) to gray / black (anagen) and to develop new hair regrowth during the study. No adverse effects were observed with respect to the general health or body weight of male animals treated with Compound 1 compared to the vehicle group (Figure 2A). From day 5, hair growth was significantly higher in the 0.002% Compound 1 treatment group compared to the vehicle (Figure 2B). Furthermore, the 0.002% Compound 1 treatment group showed a significant increase in hair growth from day 15 compared to the 5% minoxidil treatment group. No anagen was observed in the vehicle group throughout the study (Figure 2B).

[0124] Mice treated with 0.002% Compound 1 were found to exhibit a skin color change from pink to light gray on day 10 after hair growth induction, indicating a transition from telogen to anagen. Hair growth was also significantly higher in the 0.002% Compound 1 treatment group than in the vehicle group on day 22 (Figure 3A). At the end of the study, skin biopsies were collected and observed for signs of visual melanogenesis at the compound application site. Treatment with 0.002% Compound 1 resulted in the appearance of melanogenesis in the exfoliated skin of 2 / 5 animals. No visual melanogenesis was observed with the vehicle (Figure 3B).

[0125] The thickness of mouse skin changes according to the hair cycle, being maximum in anagen and thinning when it reaches telogen. Histological examination showed more anagen hair follicles in the compound 1 treatment group (Figure 4A). Compound 1 caused a significantly greater number of hair follicles to appear in the subcutaneous tissue layer of the skin and total hair follicle number when compared to the control group (Figure 4B). Furthermore, skin thickness increased after 0.002% compound 1 treatment when compared to the vehicle group (Figure 4C).

[0126] Female mice: During the study, all animals were observed to change skin color from pink (telogen) to gray / black (anagen) and the appearance of new hair regrowth. No adverse effects were observed with respect to the general health or weight of treated animals compared to the vehicle group (Figure 5A). From day 26, hair growth was significantly higher in the 0.002% Compound 1 treatment group compared to the vehicle group (Figure 5B). Furthermore, on day 30, the maximum hair growth score was observed in mice treated with 0.002% Compound 1, followed by the low dose treatment group and the minoxidil group (Figure 5B). No anagen was observed in the vehicle group throughout the 30-day study (Figure 5B). It was found that 0.002% Compound 1-treated mice showed a skin color change from pink to light gray on day 10 after hair growth induction, indicating a transition from telogen to anagen. On day 24, the area of ​​black skin was larger in Compound 1-treated mice compared to the control group (Figure 6A). At the end of the study, skin biopsies were collected and observed for signs of visual melanogenesis at the compound application sites. Treatment with 0.0002% and 0.002% Compound 1 resulted in the appearance of melanogenesis in the exfoliated skin of 2 / 5 mice, respectively. No visual melanogenesis was observed in both the vehicle and positive control groups (Figure 6B).

[0127] Histological examination showed more anagen hair follicles in the compound 1 0.0002% and 0.002% treatment groups compared to vehicle and 2% minoxidil (Figure 7A). Compound 1 caused a significantly greater number of hair follicles to appear in the subcutaneous tissue layer of the skin and a greater total number of hair follicles compared to the control group (Figure 7B). Furthermore, skin thickness increased after compound 1 treatment compared to vehicle and 2% minoxidil groups (Figure 7C).

[0128] Example 4 Topical dermal administration of compound 2 of the present invention and its effect on hair growth in a C57BL / 6 mouse model of alopecia in vivo The experimental set-up and analysis were carried out as described in Example 1.

[0129] [Table 2]

[0130] Compound treatment application 50 μL of a clear solution of compound 2 at concentrations of 0.02%, 0.002% (w / v) in the corresponding vehicle (PEG-400:EtOH / 7:3) (PEG-400: Central Drug House Ltd., Delhi, India; Ethanol: Changshu Hongsheng fine chemicals Co. Ltd., China) and positive control, minoxidil (Chemieliva, China) at 2% solution (females) or 5% solution (males) (w / v) in the corresponding vehicle (PEG-400:EtOH / 7:3) were topically applied to the shaved area on the back of the mice. The solutions were applied over a 2 cm area. 2 The treatment regimen was applied twice daily for 5 days / week for a total of 4 weeks.

[0131] result Female mice: All animals were observed to change skin color from pink (telogen) to gray / black (anagen) and the appearance of new hair regrowth during the study. No adverse effects were observed with respect to the general health or weight of treated animals compared to the vehicle group (Figure 8A). From day 23 onwards, hair growth was significantly higher in treatment group 2 compared to vehicle. Furthermore, treatment group 2 showed a significant increase in hair growth on days 25, 29 and 30 compared to 2% minoxidil. Furthermore, on day 30, maximum hair growth scores were observed in mice in treatment group 2, followed by the low dose treatment group and the minoxidil group (Figure 8B). No anagen was observed in the vehicle group throughout the 30-day study (Figure 8B). Treatment group 2 mice were found to show a skin color change from pink to light gray on day 10 after hair growth induction, indicating a transition from telogen to anagen. Also, in treatment group 3, hair growth was significantly higher than the vehicle group on days 29 and 30. On day 30, the area of ​​black skin was larger in Compound 2-treated mice compared to the control group (Figure 9A). At the end of the study, skin biopsies were collected and observed for visual signs of melanogenesis at the compound application site. Treatment with 0.002% and 0.02% Compound 2 resulted in the appearance of melanogenesis in the exfoliated skin of 1 / 5 animals, respectively. No visual melanogenesis was observed in both the vehicle group and the positive control group (Figure 9B).

[0132] The thickness of mouse skin changes based on the hair cycle, being maximum in anagen and thinning when telogen is reached. By comparing the skin thickness of the experimental and control groups, it was possible to evaluate anagen induction. Histological examination showed more anagen hair follicles in the compound 2 treatment group compared to the control group (Figure 10A). Compound 2 treatment resulted in the appearance of a significantly greater number of hair follicles in the subcutaneous tissue layer of the skin and total hair follicle number when compared to the control group (Figure 10B). Furthermore, the thickness of the skin was increased after 0.02% compound 2 treatment when compared to the vehicle and 2% minoxidil groups (Figure 10C).

[0133] Example 5 Topical dermal administration of the compounds of the present invention and their effect on an in vivo model of prevention of chemotherapy-induced alopecia (CIA) animal Female C57BL / 6 mice aged 6–9 weeks (Vivo Bio Tech Ltd, Telangana, India) in a stable telogen phase of the hair growth cycle were used. Details of the different treatment groups are shown in Table 3.

[0134] [Table 3]

[0135] Compound treatment application 100 μL of clear solutions of compound 1 at concentrations of 0.0004%, 0.002% (w / v) in the corresponding vehicle (PEG-400:EtOH / 7:3) (PEG-400: Central Drug House Ltd., Delhi, India; Ethanol: Changshu Hongsheng fine chemicals Co. Ltd., China) and positive control, minoxidil (Chemieliva, China) at a 2% solution (w / v) in the corresponding vehicle (PEG-400:EtOH / 7:3) were topically applied to the bald area on the back of female mice. The solutions were applied over a 30 cm area of ​​approximately 2 The treatment regimen consisted of 7 days prior to cyclophosphamide injection, followed by 2 injections per day for 3 weeks.

[0136] A C57BL / 6 mouse model of chemotherapy-induced alopecia (CIA) Animals (females) on day 1 ranged in weight from 18.5 to 19.3 g, with an average of 18.8 g ± 0.2. Animals were denuded by gently clipping the hair from their backs using electric clippers. The next day, animals with clear skin were depilated using a depilatory cream. To evaluate the preventive potential of Compound 1 in chemotherapy-induced alopecia mouse model, animals were treated topically with Compound 1 at 0.0004% and 0.002%, Minoxidil at 2% and vehicle on the dorsal depilated skin from day 1 onwards in a twice daily application regimen until day 10, when all animals had reached anagen stage. No application of Compound 1 or vehicle was performed on day 11 after depilation. This was considered as the washout period. On day 12 after depilation, animals were intraperitoneally injected with cyclophosphamide (CYP-HiMedia Laboratories, Mumbai, India) at a dose of 150 mg / kg body weight. The animals were given a 1-day washout period before resuming treatment with Compound 1. The treatment regimen was resumed with Compound 1 at 0.0004% and 0.002%, minoxidil at 2% and vehicle in each group for an additional 3 weeks. All animals were observed from day 1 (start of the experiment) to day 30 (end of the experiment and sacrifice of the animals) and hair growth scores were recorded daily. The criteria for scoring hair growth are as follows: no hair growth, pink skin - score 0; skin color changes from pink to gray / light gray with no visible hair growth - score 0.5; onset of anagen (skin color changes from gray / light gray to dark gray / black with no visible hair growth) - score 1; sparse hair growth - score 1.5; diffuse short hair growth - score 2, moderate hair growth - score 2.5 and dense normal coat - score 3 (Lee BH, Lee JS, Kim YC. Hair growth-promoting effects of lavender oil in C57BL / 6 mice. Toxicol Res. (2016) 32(2): 103-108). Photographs of the animals were captured before and during the study. Photographs of the animals were captured before and during the study.

[0137] Histology and immunohistochemistry After the experiment was completed, all animals were sacrificed by euthanasia (Choi HI, Kang BM, Jang J, Hwang ST, Kwon O. Novel effect of sildenafil on hair growth. Biochem Biophys Res Commun. (2018) 505(3):685-691). The dorsal skin of each animal was then removed by peeling. The peeled skin was spread and visually checked for induction of melanogenesis, indicated by blackening of the back of the skin. The skin samples were spread evenly on pre-labeled glass slides and kept in 10% formalin until further use. The skin samples were then embedded in paraffin and sectioned at 5 microns using a microtome, and corresponding slides were prepared for each animal. The resulting slides were used for hematoxylin & eosin (H&E) staining, and the total number of hair follicles in the dermis and subcutaneous tissue was determined for each animal in the micrograph data of the cross-sections and longitudinal sections. Skin thickness was measured using Image J 1.44 software. Immunohistochemical analysis was performed using CD-31 antibody (anti-CD31, endothelial cells, JC / 70A, XBioGenex) and Ki-67 antibody (rabbit anti-Ki-67, ZytoMED Systems) to observe perifollicular vascularization and proliferation index in hair follicles, respectively. Semiquantitative grading was applied. For CD-31, scoring was based on the number of capillaries near the base of the hair follicle. A score of 1 was assigned to skin sections in which one or two capillaries at the base of the hair follicle were observed, while more than two capillaries near the base of the hair follicle were given a score of 2. For the Ki-67 marker, the presence of positive Ki-67 cells in the outer root sheath cells of the hair follicle was scored 1 or 2 depending on the intensity of the staining.

[0138] result To study the preventive hair growth-promoting ability of Compound 1 in a cyclophosphamide-induced hair loss mouse model, C57BL / 6 female mice in stable telogen phase were used. This 30-day study included washout periods before and after CYP administration (Figure 11A). In each group, animals were observed for the onset of hair loss after CYP administration. No signs of behavioral changes, reactions to treatment, or adverse effects related to general health were observed during the course of the study. Additionally, no adverse effects on body weight were observed in any group (Figure 12A).

[0139] Skin color of C57BL / 6 mice is pink during telogen phase and darkens when anagen phase begins. Prior to CYP administration, topical application of Compound 1 (0.0004% and 0.002%) resulted in a faster skin color change (telogen to anagen transition) in the majority of animals when compared to the minoxidil-treated group by the end of the first week of application. Similar observations were recorded in animals treated with 2% minoxidil when compared to the vehicle control group. On day 12 (the day of CYP injection), the average hair growth score for all groups was 1. After CYP administration on day 12, animals treated with 0.0004% Compound 1 had minor signs of alopecia and animals treated with 0.002% Compound 1 had no signs of alopecia. In general, no reduction in average hair growth was observed in the Compound 1-treated groups (Figure 12B). Hair samples of all animals were collected prior to the start of treatment. On the day of animal sacrifice, samples of treated animals with hair growth scores of 2-3 were collected for hair weight analysis. From animals where hair regrowth was observed and hair weight was measured, approximately 100 hairs were selected from pre- and post-treatment hair samples. Treatment with 0.0004% and 0.002% TOP-M119 resulted in a significant increase in hair weight in 7 / 7 animals compared to the respective pre-treated hairs of the same animals (Figure 12C). It was also observed that topical application of Compound 1 resulted in a faster increase in hair growth from day 6 / 7 onwards compared to the minoxidil and vehicle groups. Application of Compound 1 resulted in complete hair regrowth in all 7 / 7 animals (hair growth score of 3). Furthermore, a slightly faster increase in hair growth was observed in animals treated with 0.002% when compared to 0.0004% Compound 1 (Figure 13A). Treatment with 2% minoxidil resulted in a faster achievement of hair growth compared to vehicle. However, complete hair regrowth was observed in only 3 / 7 animals (hair regrowth score of 3), while 4 / 7 animals showed moderate hair regrowth (hair regrowth score of 2–2.5).In the vehicle-treated group, 2 / 7 animals achieved complete hair regrowth, 4 / 7 animals showed preliminary to moderate hair regrowth (hair regrowth score of 2–2.5), and 1 animal showed no hair regrowth (Figure 13A).

[0140] In addition, skin biopsies were collected at the end of the study and analyzed for visual melanin formation at the compound application site. Treatment with 0.0004% and 0.002% Compound 1 resulted in the appearance of melanin formation in the stripped skin of 6 / 7 and 7 / 7 mice, respectively. Visual melanin formation was also observed in the stripped skin of 7 / 7 mice treated with 2% minoxidil and 3 / 7 animals in the vehicle group (Figure 13B).

[0141] H&E staining showed that the number of hair follicles was significantly increased in the compound 1-treated group and total hair follicle number in the subcutaneous layer of the skin when compared with the vehicle group (Figure 14A and B).No significant difference was observed when the compound 1-treated group was compared with the 2% minoxidil-treated group.

[0142] The longitudinal sections of the skin (Figure 14A right panel) were analyzed for the extent of hair follicle depth in treated animals with test compound-induced anagen or hair regrowth. The depth and development of the long hair shaft of the hair papilla and hair follicle were observed in most animals treated with 0.0004% and 0.002% compound 1 and 2% minoxidil. Skin thickness showed a slight increase after treatment with 0.0004% and 0.002% compound 1 and 2% minoxidil compared to the vehicle group (Figure 14C).

[0143] To investigate whether Compound 1 promotes perifollicular angiogenesis and affects cell proliferation, immunohistochemistry analysis using CD-31 and Ki-67 antibodies was performed and semiquantitative grading was reported. An increase in the number of blood vessels near the periphery of the hair follicle was observed after treatment with 0.0004% and 0.002% Compound 1, with mean scores of 1.4 and 1.6, respectively, indicating perifollicular angiogenesis. Animals treated with 2% minoxidil also showed angiogenesis near the hair follicle region with a mean score of 1.4, while the vehicle group had a mean score of 0.8 (Figures 15A and B).

[0144] The presence of Ki-67 positive cells near the root sheath of the hair follicle was higher in both Compound 1 treatments (0.0004% and 0.002%) compared to the vehicle group, with average scores of 1.4 and 1.7, respectively. Treatment with 2% minoxidil also resulted in Ki-67 positive cells near the hair follicle region, with a score of 1.1. Vehicle control animals were found to have a weak ring of Ki-67 positive cells near the root sheath of the hair follicle, with a score of 0.4 (Figures 15C and D).

[0145] Example 6 In vitro safety pharmacology of compound 1 In vitro pharmacological profiling is increasingly being used early in the drug discovery process to identify undesirable off-target activity profiles that, if discovered after a drug has been approved, may hinder or stop the development of a candidate drug, or even lead to market withdrawal (Bowes N, et al., Nature Reviews Drug Discovery 11(12):909-22). Therefore, the off-target selectivity of compound 1 at 100 nM was tested in (radioactive) ligand binding assays and enzyme assays against a set of 44 targets (SafetyScreen44™, Eurofins Cerep, France). Compound 1 (100 nM) showed no significant response at all 44 targets, as defined by ≦50% inhibition or stimulation at any of the targets.

[0146] [Table 4]

Claims

1. Formula I 【Chemistry 1】 (In the formula, R １ is C １ ~C ３ alkyl, preferably methyl or ethyl, more preferably ethyl; R ２ is H, CHO or CH═N—OH, preferably H, CHO or (E)—CH═N—OH, more preferably H, R ３ is C １ ~C ４ alkyl, preferably ethyl or propyl, more preferably n-propyl; R ４ is C １ ~C ６ alkyl, preferably ethyl or propyl, more preferably n-propyl; R ５ is SO ２ NR １３ R １４ and R １３ and R １４ together with the nitrogen atom to which they are attached form a heterocycle, said heterocycle being selected from piperidine and piperazine, said heterocycle containing at least one R １５ and R １５ are independently OH, ONO ２ C optionally substituted with １ ~C ６ alkyl, and said R １５ At least one of the ２ (including parts) or a pharmaceutically acceptable salt thereof, A topical composition comprising from about 0.0001% (w / v) to about 0.05% (w / v) of the compound of formula I, preferably Compound 1.

2. R １ is methyl or ethyl, and R ２ is H and R ３ is ethyl or n-propyl, and R ４ The topical composition of claim 1 , wherein is ethyl or n-propyl.

3. R １ is ethyl, and R ２ is H and R ３ is n-propyl, and R ４ The topical composition of claim 1 , wherein is n-propyl.

4. wherein said compound of formula I is 【Chemistry 2】 , and 【Transformation 3】 The topical composition of claim 1 , wherein the compound is selected from the group consisting of:

5. wherein said compound of formula I is (1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidine-4,4-diyl)bis(ethane-2,1-diyl)dinitrate (1); 2-(4-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperazin-1-yl)ethyl nitrate (2); 2-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)ethyl nitrate (3); 3-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)propyl nitrate (4); (R)-1-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)ethane-1,2-diyl dinitrate (5); (S)-1-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)ethane-1,2-diyl dinitrate (6); ((2R,6S)-4-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)-1-methylpiperazine-2,6-diyl)bis(ethane-2,1-diyl)dinitrate (7); ((2S,6S)-4-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)-1-methylpiperazine-2,6-diyl)bis(ethane-2,1-diyl)dinitrate (8); 3-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)-3-hydroxypentane-1,5-diyl dinitrate (9); and 2-(1-((3-(5-ethyl-4-oxo-7-propyl-4,5-dihydro-3H-pyrrolo[3,2-d]pyrimidin-2-yl)-4-propoxyphenyl)sulfonyl)piperidin-4-yl)-2-hydroxypropane-1,3-diyl dinitrate (10).

6. 2. The topical composition of claim 1, wherein said compound of formula I is compound 1 or compound 2, preferably said compound of formula I is compound 1. 【Chemistry 4】

7. 2. The composition of claim 1, wherein the compound of formula I, preferably compound 1, is present in the topical composition at about 0.0001% (w / v) to about 0.02% (w / v), preferably about 0.0002% (w / v) to about 0.02% (w / v), more preferably about 0.0002% (w / v) to about 0.01% (w / v).

8. 2. The composition of claim 1, wherein the compound of formula I, preferably compound 1, is present in the topical composition at about 0.0002% (w / v) to about 0.008% (w / v), preferably about 0.0002% (w / v) to about 0.006% (w / v), again more preferably about 0.0002% (w / v) to about 0.005% (w / v), again more preferably about 0.0002% (w / v) to about 0.004% (w / v).

9. 10. The topical composition of claim 1, wherein the topical composition comprises at least one solvent, and the solvent is selected from the group consisting of PEG 400, PEG 350, PEG 240, ethanol, 2-(2-ethoxyethoxy)ethanol, glycerin, propylene glycol, polypropylene glycol, poloxamer 407, poloxamer 188, water, and mixtures thereof.

10. 10. The topical composition of claim 1, wherein the topical composition comprises a solvent combination, the solvent combination comprising PEG 400 and ethanol.

11. The compound of formula I is Compound 1 【Transformation 5】 and the topical composition comprises Compound 1 at a concentration of about 1.5 μM to about 300 μM, preferably about 3 μM to about 200 μM, and more preferably about 5 μM to about 100 μM; 10. The topical composition of claim 1, wherein the topical composition comprises a solvent, the solvent being a combination of PEG 400 and ethanol, wherein the ratio of the PEG 400 to ethanol (vol / vol) is about 60:40 to 75:25, preferably about 65:35 to 75:25, more preferably about 68:32 to 72:28, again more preferably about 70:30; or the solvent being a combination of PEG 400, ethanol, 2-(2-ethoxyethoxy)ethanol, and water, wherein the ratio of PEG 400:ethanol:2-(2-ethoxyethoxy)ethanol:water (vol / vol) is about 5:20:5:30 to 15:40:15:70, preferably about 8:25:8:40 to about 12:35:12:60, more preferably about 10:30:10:

50.

12. 12. A topical composition according to any one of claims 1 to 11 for use in a method for topically treating a disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, preferably a skin disease or disorder mediated by PDE5 activity and / or NO-related endothelial dysfunction, in a subject, preferably a human.

13. said locally treating said disease or disorder comprising: (a) the treatment of skin disorders, preferably skin aging or steroid-induced skin atrophy; (b) treatment of erectile dysfunction; (c) preventing or treating hair loss; or (d) promoting hair growth, (i) inducing the anagen phase of hair follicles; (ii) promoting melanogenesis; (iii) increasing the number of hair follicles; (iv) increasing skin thickness; (v) promoting angiogenesis; (vi) growing cells in the outer root sheath; and / or (vii) promoting hair growth by increasing blood flow within the hair follicle.

12. A topical composition according to any one of claims 1 to 11 for use in a method for topically treating a disease or disorder in a subject, preferably a human, selected from:

14. 13. The topical composition for use according to claim 12, wherein said topically treating said disease or disorder is preventing or treating hair loss, wherein said hair loss is alopecia, preferably wherein said alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata.

15. A topical composition for use according to claim 13, wherein the topically treating the disease or disorder is preventing or treating hair loss, wherein the hair loss is alopecia, preferably wherein the alopecia is androgenetic alopecia (AGA) or chemotherapy-induced alopecia (CIA) or alopecia areata.

16. The locally treating the disease or disorder comprises: (i) induction of anagen in hair follicles; (ii) promoting melanogenesis; (iii) an increase in the number of hair follicles; (iv) increased skin thickness; (v) promoting angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) increasing blood flow within the hair follicle 13. The topical composition for use according to claim 12, wherein the use is to promote hair growth through

17. The localized treatment of the disease or disorder comprises: (i) induction of anagen in hair follicles; (ii) promoting melanogenesis; (iii) an increase in the number of hair follicles; (iv) increased skin thickness; (v) promoting angiogenesis; (vi) proliferation of cells in the outer root sheath; and / or (vii) increasing blood flow within the hair follicle 14. The topical composition for use according to claim 13, wherein the use is to promote hair growth through

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