Use of Itaconate and Its Derivatives / Analogs to Induce Hair Growth

Abstract

Methods of using prodrugs of itaconic acid and 1- and 4-methyl itaconic acid to induce hair growth and treat inflammation or immune activation (adaptive or congenital), skin conditions, or other conditions associated with hair loss are disclosed.

Description

Federally sponsored research or development

[0001] This invention was made with government support under grants AR068280 and AR064297 awarded by the National Institutes of Health. The government has certain rights in this invention. [Background technology]

[0002] Alopecia areata (AA) is a widespread inflammatory cause of hair loss, affecting approximately 2% of the population at some point in their lifetime. A typical AA lesion is a non-scarring, hairless, circular patch on the scalp, which may progress to multiple patches. Extensive forms of AA can progress to complete scalp hair loss, termed alopecia totalis (AT), or complete body hair loss, termed alopecia universalis (AU). AA is associated with other skin and autoimmune diseases, particularly atopic dermatitis (AD), as well as vitiligo, lupus erythematosus, psoriasis, autoimmune thyroid disease, and allergic rhinitis. While AA is not life-threatening, psychological comorbidities are common in patients with AA and can significantly impact their lives. Current treatments for severe cases of AA include broad-spectrum immunosuppressants, which can be associated with significant side effects that preclude long-term use, along with rapid hair loss after treatment completion. The pathogenesis of AA is not yet fully understood, and there is currently no cure for alopecia areata or any treatment that induces and maintains remission. Summary of the Invention

[0003] The presently disclosed subject matter provides the use of prodrugs of itaconic acid and 1- and 4-methyl itaconic acid to induce hair growth and treat inflammatory skin conditions or other conditions associated with hair loss, such as alopecia areata.

[0004] More specifically, in some embodiments, the presently disclosed subject matter is a method of inducing hair growth or treating an inflammatory skin condition or immune activation (adaptive or congenital) or other condition associated with hair loss in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a compound of formula (I): [ka] [During the ceremony: R1 and R2 may be the same or different and are each independently selected from one or more of the following and combinations thereof: (a) -OR3 (R3 is H or C1-C6 straight or branched unsubstituted or substituted alkyl); (b) [ka] (wherein n is an integer selected from 1, 2, 3, and 4; R4 is C1-C6 straight or branched unsubstituted or substituted alkyl or -OR5, where R5 is C1-C6 straight or branched unsubstituted or substituted alkyl); (c) [ka] (wherein m is an integer selected from 1, 2, 3, and 4; p is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20; and R6 is a C1-C6 straight or branched unsubstituted or substituted alkyl); (d) [ka] wherein R7 is selected from: (i) -C(=O)-O-R8 (R8 is a C1-C6 straight or branched unsubstituted or substituted alkyl); (ii) [ka] (wherein R9 is H or C1-C4 straight or branched unsubstituted or substituted alkyl; R 10 is a C1-C6 straight or branched unsubstituted or substituted alkyl; R 11 and R 12 are each independently H or a protecting group; and R 13 is a C1-C6 straight or branched unsubstituted or substituted alkyl; (iii) [ka] wherein q is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; R 11 and R 12 are each independently H or a protecting group; R 14 is H or C1-C4 straight or branched unsubstituted or substituted alkyl; and R 15 is a C1-C6 straight or branched unsubstituted or substituted alkyl; and (iv) [ka] (In the formula, R 16 and R 17 are each independently selected from H, C1-C4 straight or branched unsubstituted or substituted alkyl, and a protecting group; R 18 is aryl); (e) [ka] (In the formula, R 19 is a C1-C4 straight or branched unsubstituted or substituted alkyl; (f) [ka] where u is an integer selected from 1, 2, 3, and 4; R 20 is H or C1-C4 straight or branched unsubstituted or substituted alkyl; and R 21 HA-OR 22 where R 22 is a C1-C6 linear or branched unsubstituted or substituted alkyl or -NR 23 R 24 where R 23 and R 24 are each independently H or C1-C4 straight or branched unsubstituted or substituted alkyl. and a pharmaceutically acceptable salt thereof.

[0005] In yet another aspect, the presently disclosed subject matter provides a method for treating a disease, disorder, or condition associated with inflammation or immune system activation (adaptive or congenital), comprising administering a compound of formula (I) or a pharmaceutical composition thereof to a subject in need of such treatment. Representative diseases, disorders, or conditions associated with inflammation or immune system activation (adaptive or congenital) include, but are not limited to, androgenic alopecia, alopecia areata, other forms of alopecia, UV radiation, wound healing defects, psoriasis, atopic dermatitis, contact dermatitis, rosacea, acne, and autoimmune / autoinflammatory conditions.

[0006] While certain aspects addressed in whole or in part by the subject matter of the present disclosure have been described herein, other aspects will become apparent as the description proceeds when taken in conjunction with the accompanying examples and drawings as best described herein below. [Brief explanation of the drawings]

[0007] The patent or application file contains at least one drawing executed in color. Copies of any color drawing(s) in this patent or patent application publication will be provided by the Office upon request and payment of the necessary fee. Having thus described the presently disclosed subject matter in general terms, reference is now made to the accompanying drawings, which are not necessarily drawn to scale. [Figure 1] FIG. 1 shows the release of itaconic acid (FIG. 1A) or monomethyl itaconate (FIG. 1B) prodrugs after 60 minutes of incubation in skin homogenate. [Figure 2] FIG. 1 shows the release of 4-methylitaconate from its prodrug after 60 minutes of incubation in mouse plasma. [Figure 3] FIG. 1 shows monomethyl itaconate release in mouse plasma after oral administration of 10 mg / kg equivalent in male CD1 mice. [Figure 4] Treatment of human keratinocytes with itaconate prodrugs perturbs the TLR3 / IL6 / STAT3 axis. IS-100-142 most effectively inhibits JAK-STAT signaling by mitigating poly(I:C) effects on downstream gene targets (TLR3, IL6, STAT3, WNT7B). [Figure 5] FIG. 1 shows that 20% dimethyl itaconate induces the transition from telogen, the resting phase of the hair cycle, to anagen, the growth phase of the hair cycle, in mice. [Figure 6] FIG. 1 shows the effect of dimethyl itaconate in blunting the effect of dsRNA (PIC; polyinosinic:polycytidylic acid) on increasing TLR3, IL-6, and STAT3 expression. [Figure 7] FIG. 1 shows that compound 5 induces hair growth in mice. [Figure 8] FIG. 1 shows further evidence that compound 5 induces hair growth in mice. [Figure 9]FIG. 1 shows that compound 5 induces hair growth in mice to a level comparable to that of ruxolitinib and tofacitinib. [Figure 10] FIG. 1 shows mouse skin homogenate stability of representative prodrugs. [Figure 11] FIG. 1 shows the release of active monomethyl itaconate (in skin homogenates) from representative prodrugs. [Figure 12] FIG. 1 shows mouse skin penetration in an alopecia model for IS-100-142 (compound 5) (left panel) and monomethyl itaconate (right panel). [Figure 13] Figure 1 shows ex vivo human skin penetration of IS-100-142 (compound 5). Monomethyl itaconate (MMI) levels in the medium and skin were higher than for intact IS-100-142; medium levels of both analytes were similar to those in the skin. Detailed Description of the Invention

[0008] The presently disclosed subject matter will now be described more fully hereinafter with reference to the accompanying drawings, in which some, but not all, embodiments of the invention are shown. Like numbers indicate like elements throughout. The presently disclosed subject matter may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will satisfy applicable legal requirements. Indeed, many modifications and other embodiments of the presently disclosed subject matter described herein will come to mind to one skilled in the art to which the presently disclosed subject matter pertains having the benefit of the teachings presented in the foregoing descriptions and the associated drawings. Therefore, it is to be understood that the presently disclosed subject matter is not limited to the specific embodiments disclosed, and that modifications and other embodiments are intended to be included within the scope of the appended claims.

[0009] I. Use of Itaconate and Its Derivatives / Analogs to Induce Hair Growth

[0010] The compounds of formula (I) of the present disclosure are useful for inducing hair growth and treating one or more inflammatory skin conditions or other conditions associated with hair loss. In some embodiments, the inflammatory skin conditions or other conditions associated with hair loss include alopecia areata.

[0011] Alopecia areata is a common inflammatory cause of hair loss and is thought to be an autoimmune disease in which T lymphocytes attack hair follicles, causing hair to stop growing and enter the telogen phase, at the end of which the hair falls out.

[0012] There are three subtypes of alopecia areata, named according to severity: (i) alopecia areata, which involves mild patchy hair loss on the scalp; (ii) alopecia totalis, which involves loss of all scalp hair; and (iii) alopecia universalis, which involves loss of hair on the scalp and the entire body. Other types of alopecia areata include, but are not limited to, androgenic alopecia, anagen effluvium, self-induced hair loss, telogen effluvium, and scarring alopecia.

[0013] Male pattern baldness includes male pattern baldness and female pattern baldness, accounting for 95% of all hair loss. Anagen effluvium is sudden hair loss that occurs as a result of exposure to chemicals or radiation, such as the alopecia that occurs during certain types of chemotherapy or radiation treatment, or as a result of exposure to toxic chemicals such as thallium and arsenic. In most cases, hair growth returns to normal once the treatment is completed or the toxin exposure is eliminated. Self-induced hair loss can be caused consciously or unconsciously. The two main types of self-induced hair loss are trichotillomania and traction alopecia. Trichotillomania is self-induced hair loss caused by persistent pulling or plucking of hair. Traction alopecia is usually caused by continuous and excessive pulling of hair from various types of hair styling, resulting in gradual hair loss that can be permanent. Telogen effluvium is hair loss associated with sudden or severe stress, resulting in the appearance of a general thinning of the scalp. Sudden or stressful events can cause hair follicles to prematurely stop growing and enter the resting phase. Other causes of telogen phase effluvium include thyroid insufficiency (hypothyroidism or hyperthyroidism, which occur when the thyroid gland produces too little or too much of the thyroid hormone thyroxine, respectively); diabetes; anemia; and the autoimmune disease systemic lupus erythematosus. Cicatricial alopecia occurs as a result of inflammation of hair follicles due to infection. Cicatricial alopecia can be caused by discoid lupus erythematosus, a diffuse connective tissue disease; lichen planus, an inflammatory disease that primarily affects the skin and mucous membranes; pseudopelade of brocq, a rare cicatricial alopecia with no potential for regrowth; congenital atrophy of the scalp, a rare disorder that often manifests as small, vesicular atrophic areas present at birth, usually in the midline of the scalp; or congenital atrichia. Other types of hair loss include syphilitic alopecia, a secondary manifestation of syphilis; scleroderma; and tinea capitis (ringworm).

[0014] More specifically, the subject matter of the present disclosure is a method of inducing hair growth or treating an inflammatory skin condition or other condition associated with hair loss in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a compound of formula (I): [ka] [During the ceremony: R1 and R2 may be the same or different and are each independently selected from one or more of the following and combinations thereof: (a) -OR3 (R3 is H or C1-C6 straight or branched unsubstituted or substituted alkyl); (b) [ka] (wherein n is an integer selected from 1, 2, 3, and 4; R4 is C1-C6 straight or branched unsubstituted or substituted alkyl or -OR5, where R5 is C1-C6 straight or branched unsubstituted or substituted alkyl); (c) [ka] (wherein m is an integer selected from 1, 2, 3, and 4; p is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20; and R6 is a C1-C6 straight or branched unsubstituted or substituted alkyl); (d) [ka] wherein R7 is selected from: (i) -C(=O)-O-R8 (R8 is a C1-C6 straight or branched unsubstituted or substituted alkyl); (ii) [ka] (wherein R9 is H or C1-C4 straight or branched unsubstituted or substituted alkyl; R 10 is a C1-C6 straight or branched unsubstituted or substituted alkyl; R 11 and R 12 are each independently H or a protecting group; and R 13 is a C1-C6 straight or branched unsubstituted or substituted alkyl; (iii) [ka] wherein q is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; R 11 and R 12 are each independently H or a protecting group; R 14 is H or C1-C4 straight or branched unsubstituted or substituted alkyl; and R 15 is a C1-C6 straight or branched unsubstituted or substituted alkyl; and (iv) [ka] (In the formula, R 16 and R 17 are each independently selected from H, C1-C4 straight or branched unsubstituted or substituted alkyl, and a protecting group; R 18 is aryl); (e) [ka] (In the formula, R 19 is a C1-C4 straight or branched unsubstituted or substituted alkyl; (f) [ka] where u is an integer selected from 1, 2, 3, and 4; R20 is H or C1-C4 straight or branched unsubstituted or substituted alkyl; and R 21 HA-OR 22 where R 22 is a C1-C6 linear or branched unsubstituted or substituted alkyl or -NR 23 R 24 where R 23 and R 24 are each independently H or C1-C4 straight or branched unsubstituted or substituted alkyl). and a pharmaceutically acceptable salt thereof.

[0015] In certain embodiments of the disclosed methods, (a) R1 is -OR3 and R2 is [ka] wherein R7 is selected from [ka] Selected from; (b) R1 is [ka] and R2 is [ka] wherein R7 is selected from [ka] Selected from; (c) R1 is [ka] and R2 is [ka] wherein R7 is selected from [ka] Selected from; (d) R1 is [ka] and R2 is [ka] wherein R7 is selected from [ka] Selected from; (e) R1 is [ka] and R2 is [ka] wherein R7 is selected from [ka] Selected from; (f) R1 is [ka] wherein R7 is —C(═O)—O—R8 and R2 is [ka] wherein R7 is selected from [ka] Selected from; (g) R1 is [ka] wherein R7 is [ka] and R2 is [ka] wherein R7 is selected from [ka] Selected from; (h) R1 is [ka] wherein R7 is [ka] and R2 is [ka] wherein R7 is selected from [ka] selected from; and (i) R1 is [ka] wherein R7 is [ka] and R2 is [ka] wherein R7 is selected from [ka] Selected from; and pharmaceutically acceptable salts thereof.

[0016] In a more particular embodiment of the disclosed method, (ai) R1 is -OH and R2 is [ka] wherein R7 is —C(═O)—O—R8; (a-ii) R1 is -OR3 and R2 is [ka] wherein R7 is selected from [ka] Selected from; (b) R1 is [ka] and R2 is [ka] Selected from; (c) R1 is [ka] and R2 is -OR3; (d) R1 is [ka] and R2 is [ka] Selected from; (e) R1 is [ka] wherein R7 is -C(=O)-O-R8 and R2 is -OR3; (f) R1 is [ka] wherein R7 is [ka] and R2 is -OR3; (g) R1 is [ka] wherein R7 is [ka] and R2 is -OR3; (h) R1 is [ka] wherein R7 is [ka] and R2 is -OR3; and (i) R1 is [ka] and R2 is -OR3.

[0017] In some embodiments of the disclosed methods, R3, R4, R5, R6, R8, R9, R 10 , R 13 , R 15 and R 22may each independently be a C1, C2, C3, C4, C5 or C6 straight or branched unsubstituted or substituted alkyl selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, iso-butyl, t-butyl, pentyl, isopentyl, neopentyl, n-hexyl, 2-methylpentane, 3-methylpentane, 2,2-dimethylbutane and 2,3-dimethylbutane.

[0018] In some embodiments of the disclosed methods, R, R 14 , R 16 , R 17 , R 19 , R 20 , R 23 and R 24 may each independently be a C1, C2, C3 or C4 straight or branched unsubstituted or substituted alkyl selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, iso-butyl and t-butyl.

[0019] Representative substituents include, but are not limited to, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate, cyano, mercapto, and alkylthio.

[0020] In certain embodiments of the disclosed methods, the protecting group is selected from tert-butoxycarbonyl (boc), carbobenzyloxy (Cbz), p-methoxybenzylcarbonyl (Moz or MeOZ), 9-fluorenylmethyloxycarbonyl (Fmoc) group, benzyl (Bn), p-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DMPM), tosyl (Ts), Troc (trichloroethyl chloroformate), (4-nitrophenyl)sulfonyl (Nosyl), and nitrophenylsulfenyl (Nps). In certain embodiments, the protecting group is tert-butoxycarbonyl (boc).

[0021] In some embodiments of the disclosed methods, R 18 is selected from phenyl.

[0022] In certain embodiments of the disclosed methods, (ai) R1 is -OH and R2 is [ka] Selected from; (a-ii) R1 is selected from -OCH3, -OCH(CH3)2 and -OC(CH3)3, and R2 is [ka] Selected from; (b) R1 is [ka] and R2 is [ka] is; (c) R1 is [ka] and R2 is -OCH3(12,13); (d) R1 is [ka] and R2 is [ka] Selected from; (e) R1 is [ka] and R2 is -OCH3(14); (f) R1 is [ka] and R2 is -OCH3(17,19); (g) R1 is [ka] and R2 is -OCH3(15,16); (h) R1 is [ka] and R2 is -OCH3(18,20); and (i) R1 is [ka] and R2 is -OCH3(3,4).

[0023] In certain embodiments of the disclosed methods, the compound of formula (I) is [ka] [ka] is selected from.

[0024] Representative compounds of formula (I) and the active agents released by each respective prodrug are provided in Table 1.

[0025] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4]

[0026] In some embodiments, the methods of the present disclosure further include a pharmaceutical composition comprising a compound of formula (I) and a pharmaceutically acceptable excipient.

[0027] In certain embodiments of the disclosed methods, the inflammatory skin condition or other condition associated with hair loss comprises alopecia areata. In certain embodiments, the alopecia areata comprises patchy alopecia areata. In other embodiments, the alopecia areata comprises alopecia totalis. In yet other embodiments, the alopecia areata comprises alopecia universalis.

[0028] In some embodiments, the method for inducing hair growth induces a transition from the telogen phase to the anagen phase of the hair growth cycle. Briefly, the three stages of hair growth are anagen, catagen, and telogen. Anagen is known as the growth phase, during which hair physically grows. Catagen is known as the transition phase. Telogen is known as the resting phase, during which the follicle is dormant.

[0029] In certain embodiments, the method further comprises increasing the number of hair follicles present in the subject in the anagen phase, hi certain embodiments, the subject has a reduced number of hair follicles in the anagen phase compared to a control subject prior to administration of a therapeutically effective amount of a compound of Formula (I).

[0030] In other embodiments, the subject suffers from or is at risk of developing alopecia from an underlying health disorder or therapeutic treatment. In certain embodiments, the underlying health disorder is selected from acquired immune deficiency (AIDS), hypothyroidism, hyperthyroidism, lupus erythematosus, diabetes, anemia, syphilis, zinc deficiency, trichotillomania, Cushing's syndrome, systemic lupus erythematosus, infection, discoid lupus erythematosus, lichen planus, pseudopelade of brocq, congenital skin defects, congenital atrichosis, scleroderma, and tinea capitis (ringworm).

[0031] In more particular embodiments, the underlying health disorder comprises a hair loss-related disorder or condition, hi particular embodiments, the hair loss-related disorder or condition is selected from androgenic alopecia, alopecia areata, anagen effluvium, self-induced hair loss, telogen effluvium, and scarring alopecia.

[0032] In some embodiments, the therapeutic treatment comprises chemotherapy or radiation therapy.

[0033] In yet another embodiment, the presently disclosed subject matter provides a method for treating a disease, disorder, or condition associated with inflammation or immune system activation (adaptive or congenital), comprising administering a compound of formula (I) or a pharmaceutical composition thereof to a subject in need thereof. Representative diseases, disorders, or conditions associated with inflammation or immune system activation (adaptive or congenital) include, but are not limited to, androgenic alopecia, alopecia areata, other forms of alopecia, UV radiation, wound healing defects, psoriasis, atopic dermatitis, contact dermatitis, rosacea, acne, and autoimmune / autoinflammatory conditions.

[0034] Generally, as used herein, the term "treating" can include reversing, alleviating, inhibiting the progression of, preventing, or reducing the likelihood of the disease, disorder, or condition to which such term applies, or one or more symptomatic manifestations of such disease, disorder, or condition. Prevention refers to preventing a disease, disorder, condition, or its symptoms or manifestations, or worsening of its severity, from occurring. Thus, the compounds of the present disclosure can be administered prophylactically to prevent or reduce the occurrence or recurrence of a disease, disorder, or condition.

[0035] The "subject" treated by the methods of the present disclosure in many embodiments thereof is preferably a human subject, although it will be understood that the methods described herein are effective with respect to all vertebrate species intended to be encompassed by the term "subject." Thus, a "subject" can include a human subject for medical purposes, such as treatment of an existing condition or disease or prophylactic treatment to prevent the onset of a condition or disease, or an animal subject for medical, veterinary, or developmental purposes. Suitable animal subjects include, but are not limited to, primates (e.g., humans, monkeys, apes, etc.); bovines (e.g., cattle, bulls, etc.); ovines (e.g., sheep, etc.); caprines (e.g., goats, etc.); porcines (e.g., pigs, boars, etc.); equines (e.g., horses, donkeys, zebras, etc.); felines (e.g., wild cats, domestic cats, etc.); canines (e.g., roosters, etc.); lagomorphs (e.g., rabbits, hares, etc.); and rodents (e.g., mice, rats, etc.). The animal may be a transgenic animal. In some embodiments, the subject is a human, including, but not limited to, a fetus, a newborn, an infant, a juvenile, and an adult subject. Furthermore, a "subject" can include a patient suffering from or suspected of suffering from a condition or disease. Thus, the terms "subject" and "patient" are used interchangeably herein. The term "subject" also refers to an organism, tissue, cell, or collection of cells from a subject.

[0036] Generally, the "effective amount" of an active agent or drug delivery device refers to the amount required to induce a desired biological response.As understood by those skilled in the art, the effective amount of an agent or device can vary depending on factors such as the desired biological endpoint, the agent to be delivered, the composition of the pharmaceutical composition, the target tissue, etc.

[0037] The term "combination" is used in its broadest sense and refers to the administration of at least two agents to a subject, more specifically, a compound of Formula (I) and at least one beta-lactam antibiotic, and optionally one or more antibacterial agents. More specifically, the term "in combination" refers to the simultaneous administration of two (or more) active agents, for example, for the treatment of a single disease state. As used herein, the active agents may be administered in combination in a single dosage form, simultaneously in separate dosage forms, or in separate dosage forms that are administered alternately or sequentially on the same or different days. In one embodiment of the presently disclosed subject matter, the active agents are administered in combination in a single dosage form. In another embodiment, the active agents are administered in separate dosage forms (e.g., where it is desirable to vary the amount of one but not the other). The single dosage form may contain additional active agents for the treatment of a disease state.

[0038] Additionally, the compounds of Formula (I) described herein can be administered alone or in combination with one or more antibacterial agents in combination with adjuvants that enhance the stability of the compounds of Formula (I), and in certain embodiments can facilitate administration of pharmaceutical compositions containing them, provide increased dissolution or dispersion, enhance inhibitory activity, provide adjunctive therapy with other active ingredients, etc. Advantageously, such combination therapies utilize lower dosages of conventional therapies, thus avoiding toxic and adverse side effects that can occur when those agents are used as monotherapies.

[0039] The timing of administering the compound of formula (I) and at least one additional therapeutic agent can be varied, as long as the beneficial effect of the combination of these agents is achieved.Therefore, the term "in combination" refers to administering the compound of formula (I) and at least one additional therapeutic agent simultaneously, sequentially, or any combination thereof.Therefore, the subject who is administered the combination of the compound of formula (I) and at least one additional therapeutic agent can receive the compound of formula (I) and at least one additional therapeutic agent simultaneously (i.e., simultaneously) or at different times (i.e., sequentially (in any order), on the same day or on different days), as long as the combined effect of both agents is achieved in the subject.

[0040] When administered sequentially, the agents can be administered within 1 minute, 5 minutes, 10 minutes, 30 minutes, 60 minutes, 120 minutes, 180 minutes, 240 minutes, or more of each other. In other embodiments, the sequentially administered agents can be administered within 1 day, 5 days, 10 days, 15 days, 20 days, or more of each other. When the compound of Formula (I) and at least one additional therapeutic agent are administered simultaneously, they can be administered to the subject as separate pharmaceutical compositions, each containing either the compound of Formula (I) or the at least one additional therapeutic agent, or they can be administered to the subject as a single pharmaceutical composition containing both agents.

[0041] When administered in combination, the effective concentration of each agent to elicit a particular biological response may be lower than the effective concentration of each agent when administered alone, thereby allowing for a reduction in the dosage of one or more agents relative to the dosage required when the agent is administered alone. The effects of multiple agents may be, but need not be, additive or synergistic. Agents may be administered multiple times.

[0042] In some embodiments, two or more agents may have a synergistic effect when administered in combination. As used herein, the terms "synergistic," "synergistic," "synergistically," and derivatives thereof, such as in "synergistic effect" or "synergistic combination" or "synergistic composition," refer to a situation in which the biological activity of the combination of a compound of Formula (I) and at least one additional therapeutic agent is greater than the sum of the biological activities of each agent when administered individually.

[0043] Synergy can be expressed as a "synergy index (SI)," which can generally be determined by the method described by F.C. Kull et al., Applied Microbiology 9, 538 (1961), from a ratio determined by the following formula: Q a / Q A +Q b / Q B = Synergy Index (SI) During the ceremony: Q A is the concentration of component A acting alone to produce the endpoint relative to component A; Q a is the concentration of component A in the mixture, which produces the endpoint; Q B is the concentration of component B acting alone to produce the endpoint relative to component B; and Q b is the concentration of component B in the mixture, which produces the endpoint.

[0044] In general, Q a / Q A and Q b / Q BIf the sum is greater than 1, antagonism is indicated. If the sum is equal to 1, additivity is indicated. If the sum is less than 1, synergy is indicated. The lower the SI, the greater the synergistic effect exhibited by that particular mixture. Thus, a "synergistic combination" has greater activity than would be expected based on the activity observed when the individual components are used alone. Furthermore, a "synergistically effective amount" of a component refers to the amount of that component needed to elicit a synergistic effect, for example, with another therapeutic agent present in the composition.

[0045] In another embodiment, the presently disclosed subject matter provides pharmaceutical compositions comprising a compound of Formula (I), alone or in combination with one or more additional therapeutic agents in admixture with a pharmaceutically acceptable excipient. Those skilled in the art will recognize that pharmaceutical compositions also include pharmaceutically acceptable salts of the compounds described above. Pharmaceutically acceptable salts are generally well known to those skilled in the art and include salts of active compounds prepared using relatively non-toxic acids or bases, depending on the particular substituent moieties found on the compounds described herein. When a compound of the present disclosure contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired base, either neat or in a suitable inert solvent, or by ion exchange, thereby replacing one basic counterion (base) in the ionic complex with another. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, or magnesium salts, or similar salts thereof.

[0046] When a compound of the present disclosure contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such a compound with a sufficient amount of the desired acid, either neat or in a suitable inert solvent, or by ion exchange, thereby replacing one acidic counterion (acid) in the ionic complex with another. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphate, dihydrogenphosphate, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphorous acid, as well as salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, methanesulfonic acid, and the like. Also included are salts of amino acids, such as arginine salts, and salts of organic acids, such as glucuronic acid and galacturonic acid (see, e.g., Berge et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 1977, 66, pp. 1-19). Certain compounds of the present disclosure contain both basic and acidic functional groups, allowing the compounds to be converted into either base or acid addition salts.

[0047] Thus, pharmaceutically acceptable salts suitable for use with the presently disclosed subject matter include, but are not limited to, acetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calcium edetate, camsylate, carbonate, citrate, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthalene, hydroxybenzoates ... Examples of suitable pharmaceutically acceptable salts include hydroxynaphthoate, iodide, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, mucate, napsylate, nitrate, pamoate (embonate), pantothenate, phosphate / diphosphate, polygalacturonate, salicylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, or teoclate. Other pharmaceutically acceptable salts can be found, for example, in Remington: The Science and Practice of Pharmacy (20th Edition), Lippincott, Williams & Wilkins (2000).

[0048] For therapeutic and / or diagnostic applications, the compounds of the present disclosure can be formulated for a variety of modes of administration, including systemic administration and topical or localized administration. Techniques and formulations can generally be found in Remington: The Science and Practice of Pharmacy (20th ed.) Lippincott, Williams & Wilkins (2000).

[0049] Depending on the specific condition to be treated, such agents can be formulated into liquid or solid dosage forms and administered systemically or locally. Agents can be delivered, for example, in time-release or sustained-release forms, as known to those skilled in the art. Formulation and administration techniques can be found in Remington: The Science and Practice of Pharmacy (20th ed.) Lippincott, Williams & Wilkins (2000). Suitable routes include oral, buccal, inhalation spray, sublingual, rectal, transdermal, vaginal, transmucosal, nasal or intestinal administration; intramuscular, subcutaneous, intramedullary injection, and parenteral administration such as intrathecal, direct intraventricular, intravenous, intraarticular, intrasternal, intrasynovial, intrahepatic, intracerebral, intraperitoneal, intranasal or intraocular injection, or other modes of administration.

[0050] For injection, the agent of the present disclosure can be formulated and diluted in a physiologically compatible buffer solution, such as aqueous solution, for example, Hank's solution, Ringer's solution, or physiological saline buffer.For such transmucosal administration, a penetrant appropriate to the barrier to be permeated is used in the formulation.Such penetrants are generally known in the art.

[0051] The use of pharmaceutically acceptable inert carriers to formulate the compounds disclosed herein into dosages suitable for systemic administration for the implementation of the present disclosure is within the scope of the present disclosure. By appropriate selection of the carrier and appropriate manufacturing method, the compositions of the present disclosure, particularly those formulated as solutions, can be administered parenterally, such as by intravenous injection. These compounds can be easily formulated into dosages suitable for oral administration using pharmaceutically acceptable carriers well known in the art. Such carriers allow the compounds of the present disclosure to be formulated as tablets, pills, capsules, liquids, gels, syrups, slurries, suspensions, etc. for oral ingestion by the subject (e.g., patient) to be treated.

[0052] For nasal or inhalation delivery, the agents of the present disclosure may also be formulated by methods known to those skilled in the art and may include, but are not limited to, solubilizing, diluent, or dispersing agents such as saline; preservatives such as benzyl alcohol; absorption enhancers; and fluorocarbons.

[0053] Pharmaceutical compositions suitable for use in the present disclosure include compositions containing the active ingredient in an amount effective to achieve its intended purpose. Determination of an effective amount is within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein. In general, compounds according to the present disclosure are effective over a wide dosage range. For example, in treating adult humans, 0.01-1000 mg, 0.5-100 mg, 1-50 mg / day, and 5-40 mg / day are exemplary dosages that may be used. A non-limiting dosage is 10-30 mg / day. The exact dosage will depend on the route of administration, the dosage form of the compound, the subject being treated, the subject's body weight, the bioavailability of the compound, the absorption, distribution, metabolism, and excretion (ADME) toxicity of the compound, and the preference and experience of the attending physician.

[0054] In addition to the active ingredient, these pharmaceutical compositions may contain suitable pharmaceutically acceptable carriers, including excipients and auxiliaries that facilitate the processing of the active compound into pharmaceutically usable preparations. Preparations formulated for oral administration may be in the form of tablets, dragees, capsules, or solutions.

[0055] Oral pharmaceutical preparations can be obtained by combining the active compound with a solid excipient, optionally grinding the resulting mixture, optionally adding suitable auxiliaries, and then processing the granular mixture to obtain tablets or dragee cores.Suitable excipients are, in particular, sugars including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose (CMC), and / or fillers such as polyvinylpyrrolidone (PVP: povidone).If desired, disintegrants such as cross-linked polyvinylpyrrolidone, agar, alginic acid or its salts (such as sodium alginate) can be added.

[0056] Sugar-coated tablet core is provided with suitable coating.For this purpose, can use concentrated sugar solution, lacquer solution and suitable organic solvent or solvent mixture, which can optionally contain gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol (PEG) and / or titanium dioxide.For identification or to characterize different combinations of dosage of active compound, dyes or pigments can be added to tablet or sugar-coated tablet coating.

[0057] Orally usable pharmaceutical preparations include push-fit capsules made of gelatin and soft, sealed capsules made of gelatin, containing plasticizers such as glycerol or sorbitol. Push-fit capsules can contain active ingredients mixed with fillers such as lactose, binders such as starch, and / or lubricants such as talc or magnesium stearate, and optionally stabilizers. In soft capsules, the active compound can be dissolved or suspended in a suitable liquid such as fatty oils, liquid paraffin, or liquid polyethylene glycol (PEG). Additionally, stabilizers may be added.

[0058] In yet other embodiments, the compounds of formula (I) of the present disclosure may be formulated into viscous lotions, creams, ointments, suspensions, pastes, gels, oils, sprays, or aerosols and administered topically. Such viscous lotions, creams, or ointments may be water-based or may further contain an oil (water-in-oil or oil-in-water), such as liquid paraffin or vegetable oils (e.g., arachis oil or castor oil), or a solvent, and may contain one or more other ingredients, including, but not limited to, penetration enhancers (e.g., ethanol and propylene glycol), humectants (including, but not limited to, glycerin and / or glycerol), thickeners and / or gelling agents (including, but not limited to, soft paraffin, aluminum stearate, cetostearyl alcohol, polyethylene glycol, wool fat, beeswax, carboxypolymethylene and cellulose derivatives, and / or glyceryl monostearate), and / or non-ionic emulsifiers, stabilizers, dispersants, and suspending agents. By way of example, a typical liquid formulation may include about 10% to about 60% water, about 10% to about 70% ethanol, about 5% to about 10% propylene glycol, and about 2% to about 5% humectant.

[0059] II. Definition

[0060] Although specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation. 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 this presently described subject matter belongs.

[0061] While the following terms relating to compounds of formula (I) are believed to be well understood by those of skill in the art, the following definitions are provided to facilitate discussion of the presently disclosed subject matter. These definitions are intended to supplement and explain, but not preclude, definitions that would be apparent to one of skill in the art upon review of this disclosure.

[0062] As used herein, the terms substituted (whether preceded by the term "optionally" or not) and substituent refer to the ability to change one functional group on a molecule to another, provided that the valences of all atoms are maintained, as understood by one of ordinary skill in the art. When more than one position in any given structure can be substituted with more than one substituent selected from a particular group, the substituents can be the same or different at all positions. Substituents can also be further substituted (e.g., an aryl group substituent can have another substituent outside it, e.g., another aryl group that is further substituted at one or more positions).

[0063] Where substituents or linking groups are designated by conventional chemical formulas written from left to right, they equally encompass the chemically identical substituents that result from writing the structure from right to left, e.g., -CHO- is equivalent to -OCH-, -C(=O)O- is equivalent to -OC(=O)-, and -OC(=O)NR- is equivalent to -NRC(=O)O-.

[0064] When the term "independently selected" is used, the referenced substituents (e.g., R groups such as groups R1, R2, or variables such as "m", "n", etc.) can be the same or different. For example, R1 and R2 can both be substituted alkyl, or R1 can be hydrogen and R2 can be substituted alkyl, etc.

[0065] As used herein with respect to a group of substituents, the terms "a," "an," or "a(n)" mean at least one. For example, if a compound is substituted with "an" alkyl or aryl, then the compound is optionally substituted with at least one alkyl and / or at least one aryl. Furthermore, if a moiety is substituted with an R substituent, the group may be referred to as "R-substituted." If a moiety is R-substituted, then the moiety is substituted with at least one R substituent, and each R substituent is optionally different.

[0066] A named "R" or group generally has the structure recognized in the art as corresponding to the named group, unless otherwise specified herein. For purposes of illustration, certain representative "R" groups, as described above, are defined below.

[0067] The description of the compounds of the present disclosure is limited by the principles of chemical bonding known to those skilled in the art.Therefore, when a group can be substituted with one or more of a large number of substituents, such substitutions are selected to comply with the principles of chemical bonding and to give compounds that are not inherently unstable and / or compounds that are known to those skilled in the art to be likely to be unstable under ambient conditions, such as aqueous, neutral, and some known physiological conditions.For example, heterocycloalkyl or heteroaryl can be bonded to the rest of the molecule through a ring heteroatom according to the principles of chemical bonding known to those skilled in the art, thereby avoiding inherently unstable compounds.

[0068] Unless expressly defined otherwise, a "substituent" as used herein includes a functional group selected from one or more of the following moieties, as defined herein:

[0069] The term hydrocarbon, as used herein, refers to any chemical group containing hydrogen and carbon. The hydrocarbon may be substituted or unsubstituted. As known to those skilled in the art, when substitutions are made, all valencies must be satisfied. The hydrocarbon may be unsaturated, saturated, branched, unbranched, cyclic, polycyclic, or heterocyclic. Exemplary hydrocarbons are further defined herein below and include, for example, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, allyl, vinyl, n-butyl, tert-butyl, ethynyl, cyclohexyl, and the like.

[0070] The term “alkyl,” by itself or as part of another substituent, means, unless otherwise stated, a straight-chain (i.e., unbranched) or branched-chain, acyclic or cyclic hydrocarbon group, or combinations thereof, having a specified number of carbon atoms (i.e., C 1-10 means 1 to 10 carbons, including 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 carbons, and may be fully saturated, monounsaturated, or polyunsaturated, and may include divalent and polyvalent groups. In certain embodiments, the term "alkyl" refers to C groups including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20 carbons. 1-20 (C1 and C 20 "Alkenyl" means a hydrocarbon radical that is linear (i.e., "straight chain"), branched, or cyclic, saturated or at least partially and optionally fully unsaturated (i.e., alkenyl and alkynyl), derived from a hydrocarbon moiety containing 1 to 20 carbon atoms by the removal of one hydrogen atom.

[0071] Representative saturated hydrocarbon groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, isopentyl, neopentyl, n-hexyl, sec-hexyl, n-heptyl, n-octyl, n-decyl, n-undecyl, dodecyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and homologs and isomers thereof.

[0072] "Branched" refers to an alkyl group in which a lower alkyl group, such as methyl, ethyl, or propyl, is attached to a linear alkyl chain. "Lower alkyl" refers to an alkyl group having 1 to about 8 carbon atoms (e.g., 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms) (i.e., C 1-8 "Higher alkyl" refers to an alkyl group having from about 10 to about 20 carbon atoms (e.g., 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms). In certain embodiments, "alkyl" refers to, in particular, C 1-8 In other embodiments, "alkyl" refers to, inter alia, C 1-8 Refers to branched chain alkyl.

[0073] An alkyl group can be optionally substituted with one or more alkyl group substituents ("substituted alkyl"), which can be the same or different. The term "alkyl group substituent" includes, but is not limited to, alkyl, substituted alkyl, halo, arylamino, acyl, hydroxyl, aryloxy, alkoxyl, alkylthio, arylthio, aralkyloxy, aralkylthio, carboxyl, alkoxycarbonyl, oxo, and cycloalkyl. One or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms can be optionally inserted along the alkyl chain, where the nitrogen substituent is hydrogen, lower alkyl (also referred to herein as "alkylaminoalkyl"), or aryl.

[0074] Thus, as used herein, the term "substituted alkyl" includes alkyl groups, as described herein, in which one or more atoms or functional groups of the alkyl group are replaced with another atom or functional group, such as, for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate, cyano, and mercapto.

[0075] The term "heteroalkyl," by itself or in combination with other terms, means, unless otherwise specified, a stable linear or branched chain or cyclic hydrocarbon radical having 1 to 20 carbon atoms or heteroatoms, or 3 to 10 carbon atoms or heteroatoms, or combinations thereof, consisting of at least one carbon atom and at least one heteroatom selected from the group consisting of O, N, P, Si, and S, wherein the nitrogen, phosphorus, and sulfur atoms may optionally be oxidized, and the nitrogen heteroatom may optionally be quaternized. The heteroatom(s) O, N, P, and S, as well as Si, may be placed at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. Examples include, but are not limited to, -CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2-S(O)-CH3, -CH2-CH2-S(O)-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=N-OCH3, -CH=CH-N(CH3)-CH3, O-CH3, -O-CH2-CH3, and -CN. Up to two or three heteroatoms may be consecutive, such as, for example, -CH2-NH-OCH3 and -CH2-O-Si(CH3)3.

[0076] As noted above, heteroalkyl groups, as used herein, include groups that are attached to the remainder of the molecule via a heteroatom, such as -C(O)NR', ​​-NR'R'', -OR', -SR, -S(O)R, and / or -S(O2)R'. When "heteroalkyl" is recited followed by a recitation of a specific heteroalkyl group, such as NR'R, it is understood that the terms heteroalkyl and NR'R'' are not redundant or mutually exclusive. Rather, the specific heteroalkyl group is recited to add clarity. Thus, as used herein, the term "heteroalkyl" should not be interpreted as excluding specific heteroalkyl groups, such as NR'R''.

[0077] "Cyclic" and "cycloalkyl" refer to a non-aromatic monocyclic or polycyclic ring system of about 3 to about 10 carbon atoms, e.g., 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Cycloalkyl groups can optionally be partially unsaturated. Cycloalkyl groups can also be optionally substituted with alkyl group substituents, oxo, and / or alkylene, as defined herein. One or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms can be optionally inserted along the cyclic alkyl chain, where the nitrogen substituent is hydrogen, unsubstituted alkyl, substituted alkyl, aryl, or substituted aryl, thereby providing a heterocyclic group. Representative monocyclic cycloalkyl rings include cyclopentyl, cyclohexyl, and cycloheptyl. Polycyclic cycloalkyl rings include adamantyl, octahydronaphthyl, decalin, camphor, camphane, and noradamantyl, as well as fused ring systems such as dihydro- and tetrahydronaphthalene.

[0078] The terms "cycloalkyl" and "heterocycloalkyl," by themselves or in combination with other terms, represent, unless otherwise stated, cyclic versions of "alkyl" and "heteroalkyl," respectively. Additionally, for heterocycloalkyl, a heteroatom can occupy the position at which the heterocycle is attached to the remainder of the molecule. Examples of cycloalkyl include, but are not limited to, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, and the like. Examples of heterocycloalkyl include, but are not limited to, 1-(1,2,5,6-tetrahydropyridyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothien-2-yl, tetrahydrothien-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. The terms "cycloalkylene" and "heterocycloalkylene" refer to the divalent derivatives of cycloalkyl and heterocycloalkyl, respectively.

[0079] Unsaturated hydrocarbons have one or more double or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher homologs and isomers. Alkyl groups limited to hydrocarbon groups are referred to as "homoalkyl."

[0080] More specifically, the term "alkenyl" as used herein refers to a C alkyl group having at least one carbon-carbon double bond formed by the removal of one hydrogen molecule. 2-20 (C2 and C 20 Alkenyl refers to a monovalent group derived from a straight-chain or branched hydrocarbon moiety (including, for example, ethenyl (i.e., vinyl), propenyl, butenyl, 1-methyl-2-buten-1-yl, pentenyl, hexenyl, octenyl, allenyl, and butadienyl.

[0081] The term "cycloalkenyl" as used herein refers to a cyclic hydrocarbon containing at least one carbon-carbon double bond. Examples of cycloalkenyl groups include cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadiene, cyclohexenyl, 1,3-cyclohexadiene, cycloheptenyl, cycloheptatrienyl, and cyclooctenyl.

[0082] As used herein, the term "alkynyl" refers to a straight or branched chain C alkyl group of a designed number of carbon atoms containing at least one carbon-carbon triple bond. 2-20 It refers to a monovalent group derived from a hydrocarbon. Examples of "alkynyl" include ethynyl, 2-propynyl (propargyl), 1-propynyl, pentynyl, hexynyl, heptynyl, and the like.

[0083] The term "alkylene," by itself or as part of another substituent, refers to a straight-chain or branched divalent aliphatic hydrocarbon group derived from an alkyl group having 1 to about 20 carbon atoms, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms. Alkylene groups can be straight-chain, branched, or cyclic. Alkylene groups can also be optionally unsaturated and / or substituted with one or more "alkyl group substituents." Optionally inserted along the alkylene group can be one or more oxygen, sulfur, or substituted or unsubstituted nitrogen atoms (also referred to herein as "alkylaminoalkyl"), where the nitrogen substituents are alkyl as previously described. Exemplary alkylene groups include methylene (-CH-); ethylene (-CH-CH-); propylene (-(CH-); cyclohexylene (-CH-); cyclohexylene (-CH-). 10 -);-CH=CH-CH=CH-;-CH=CH-CH2-;-CH2CH2CH2CH2-, -CH2CH=CHCH2-, -CH2CsCCH2-, -CH2CH2CH(CH2CH2CH3)CH2-, -(CH2) q -N(R)-(CH2) r-, where each of q and r is independently an integer from 0 to about 20 (e.g., 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20), and R is hydrogen or lower alkyl; methylenedioxyl (-O-CH2-O-); and ethylenedioxyl (-O-(CH2)2-O-). Alkylene groups can have from about 2 to about 3 carbon atoms and can further have from 6 to 20 carbon atoms. Typically, alkyl (or alkylene) groups have from 1 to 24 carbon atoms, with those groups having 10 or fewer carbon atoms being some embodiments of the present disclosure. A "lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, generally having 8 or fewer carbon atoms.

[0084] The term "heteroalkylene" by itself or as part of another substituent means a divalent group derived from heteroalkyl, exemplified by, but not limited to, -CH-CH-S-CH-CH- and -CH-S-CH-CH-NH-CH-. For heteroalkylene groups, heteroatoms can also occupy either or both of the chain termini (e.g., alkyleneoxo, alkylenedioxo, alkyleneamino, alkylenediamino, etc.). Additionally, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied by the direction in which the formula of the linking group is written. For example, the formula -C(O)OR'- represents both -C(O)OR'- and -R'OC(O)-.

[0085] The term "aryl," unless otherwise specified, means an aromatic hydrocarbon substituent which may be a single ring or multiple rings (e.g., 1 to 3 rings) fused or covalently linked together. The term "heteroaryl" refers to an aryl group (or ring) containing 1 to 4 heteroatoms (in each separate ring in the case of multiple rings) selected from N, O, and S, where the nitrogen and sulfur atoms are optionally oxidized and the nitrogen atom(s) are optionally quaternized. A heteroaryl group can be attached to the remainder of the molecule through a carbon or heteroatom. Non-limiting examples of aryl and heteroaryl groups include phenyl, 1-naphthyl, 2-naphthyl, 4-biphenyl, 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl, pyrazinyl, 2-oxazolyl, 4-oxazolyl, 2-phenyl-4-oxazolyl, 5-oxazolyl, 3-isoxazolyl, 4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl, 2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrimidyl, 4-pyrimidyl.

[0023] Examples of aryl and heteroaryl ring systems include 5-benzothiazolyl, purinyl, 2-benzimidazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl, 5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of the above aryl and heteroaryl ring systems are selected from the group of acceptable substituents described below. The terms "arylene" and "heteroarylene" refer to the divalent forms of aryl and heteroaryl, respectively.

[0086] For brevity, the term "aryl" when used in combination with other terms (e.g., aryloxy, arylthioxy, arylalkyl) includes both aryl and heteroaryl rings as defined above. Thus, the terms "arylalkyl" and "heteroarylalkyl" are intended to include groups in which an aryl or heteroaryl group is bonded to an alkyl group (e.g., benzyl, phenethyl, pyridylmethyl, furylmethyl, etc.), such as an alkyl group in which a carbon atom (e.g., methylene group) is replaced with an oxygen atom or the like (e.g., phenoxymethyl, 2-pyridyloxymethyl, 3-(1-naphthyloxy)propyl, etc.). However, the term "haloalkyl" as used herein is intended to cover only aryl substituted with one or more halogens.

[0087] When a heteroalkyl, heterocycloalkyl, or heteroaryl contains a specific number of members (eg, "3 to 7 members"), the term "member" refers to a carbon or heteroatom.

[0088] Additionally, as used herein, the following formula: [ka] The structure generally represented by the formula: refers to aliphatic and / or aromatic ring structures, such as, but not limited to, 3-carbon, 4-carbon, 5-carbon, 6-carbon, 7-carbon, etc. ring structures, including saturated, partially saturated, and unsaturated ring structures containing substituent R groups, where the substituent R groups may be present or absent, and if present, one or more R groups may each be substituted on one or more available carbon atoms of the ring structure. The presence or absence of R groups and the number of R groups is determined by the value of the variable "n," which is generally an integer having a value ranging from 0 to the number of carbon atoms on the ring available for substitution. When multiple R groups are present, each R group is substituted on an available carbon of the ring structure, rather than on another R group. For example, the above structure, where n is 0-2, may be substituted, but is not limited to, the following: [ka] The compound group includes:

[0089] A dashed line representing a bond in a cyclic structure indicates that the bond may or may not be present within the ring, i.e., the dashed line representing a bond in a cyclic structure indicates that the cyclic structure is selected from the group consisting of saturated cyclic structures, partially saturated cyclic structures, and unsaturated cyclic structures.

[0090] symbol: [ka] indicates the point of attachment of a moiety to the rest of the molecule.

[0091] When a named atom of an aromatic ring or heteroaromatic ring is defined to be "absent," the named atom is replaced with a direct bond.

[0092] As used herein, the term "acyl" refers to an organic acid group in which the -OH of the carboxyl group is replaced with another substituent and has the general formula -RC(=O)-, where R is an alkyl, alkenyl, alkynyl, aryl, carbocyclic, heterocyclic, or aromatic heterocyclic group as defined herein. Thus, the term "acyl" specifically includes arylacyl groups such as 2-(furan-2-yl)acetyl)-, 2-phenylacetyl groups, etc. Specific examples of acyl groups include acetyl and benzoyl. Acyl groups are also intended to include amides, -RC(=O)NR', ​​esters, -RC(=O)OR', ketones, -RC(=O)R', and aldehydes, -RC(=O)H.

[0093] The terms “alkoxyl” or “alkoxy” are used interchangeably herein and refer to a saturated (i.e., alkyl-O—) or unsaturated (i.e., alkenyl-O— and alkynyl-O—) group attached to the parent molecular moiety through an oxygen atom, where the terms “alkyl,” “alkenyl,” and “alkynyl” are as defined above and include, for example, methoxyl, ethoxyl, propoxyl, isopropoxyl, n-butoxyl, sec-butoxyl, tert-butoxyl, n-pentoxyl, neopentoxyl, n-hexoxyl, and the like. 1-20 (C1 and C 20 The oxo-carbon chain may be linear, branched or cyclic, saturated or unsaturated, including oxo-hydrocarbon chains.

[0094] The term "alkoxyalkyl," as used herein, refers to an alkyl-O-alkyl ether, for example, a methoxyethyl or an ethoxymethyl group.

[0095] "Aryloxyl" refers to an aryl-O- group, where the aryl group is as defined above, including substituted aryl. As used herein, the term "aryloxyl" can refer to phenyloxyl or hexyloxyl, and phenyloxyl or hexyloxyl substituted with alkyl, substituted alkyl, halo, or alkoxyl.

[0096] "Aralkyl" refers to an aryl-alkyl- group where the aryl and alkyl are as previously described, including substituted aryl and alkyl. Exemplary aralkyl groups include benzyl, phenylethyl, and naphthylmethyl.

[0097] "Aralkyloxyl" refers to an aralkyl-O- group, where the aralkyl group is as previously described. An exemplary aralkyloxyl group is benzyloxyl, i.e., C6H5-CH2-O-. The aralkyloxyl group can be optionally substituted.

[0098] "Alkoxycarbonyl" refers to an alkyl-OC(=O)- group. Exemplary alkoxycarbonyl groups include methoxycarbonyl, ethoxycarbonyl, butyloxycarbonyl, and tert-butyloxycarbonyl.

[0099] "Aryloxycarbonyl" refers to the group aryl-OC(=O)-. Exemplary aryloxycarbonyl groups include phenoxy- and naphthoxy-carbonyl.

[0100] "Aralkoxycarbonyl" refers to the group aralkyl-OC(=O)-. An exemplary aralkoxycarbonyl group is benzyloxycarbonyl.

[0101] "Carbamoyl" refers to an amide group of the formula -C(=O)NH2. "Alkylcarbamoyl" refers to the group R'RN-C(=O)- where one of R and R' is hydrogen and the other of R and R' is alkyl and / or substituted alkyl as defined above. "Dialkylcarbamoyl" refers to the group R'RN-C(=O)- where each of R and R' is independently alkyl and / or substituted alkyl as defined above.

[0102] The term carbonyldioxyl, as used herein, refers to a carbonate group of formula -OC(=O)-OR.

[0103] "Acyloxyl" refers to an acyl-O- group where acyl is as previously defined.

[0104] The term "amino" refers to the -NH group and also to nitrogen-containing groups known in the art derived from ammonia by replacing one or more hydrogen radicals with organic radicals. For example, the terms "acylamino" and "alkylamino" refer to specific N-substituted organic radicals with acyl and alkyl substituents, respectively.

[0105] As used herein, "aminoalkyl" refers to an amino group covalently attached to an alkylene linker. More specifically, as used herein, the terms alkylamino, dialkylamino, and trialkylamino refer to one, two, or three alkyl groups, as defined above, respectively, attached to the parent molecular moiety through a nitrogen atom. The term alkylamino refers to a group having the structure -NHR', where R' is an alkyl group as defined above; while the term dialkylamino refers to a group having the structure -NR'R'', where R' and R'' are each independently selected from the group consisting of alkyl groups. The term trialkylamino refers to a group having the structure -NR'R''R''', where R', R'', and R''' are each independently selected from the group consisting of alkyl groups. Furthermore, R', R'', and / or R''', taken together, optionally, form -(CH2) k -, where k is an integer from 2 to 6. Examples include, but are not limited to, methylamino, dimethylamino, ethylamino, diethylamino, diethylaminocarbonyl, methylethylamino, isopropylamino, piperidino, trimethylamino, and propylamino.

[0106] An amino group is -NR'R'', where R' and R'' are typically selected from hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted heteroalkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl.

[0107] The terms alkylthioether and thioalkoxyl refer to saturated (i.e., alkyl-S—) or unsaturated (i.e., alkenyl-S— and alkynyl-S—) groups attached to the parent molecular moiety through a sulfur atom. Examples of thioalkoxyl moieties include, but are not limited to, methylthio, ethylthio, propylthio, isopropylthio, n-butylthio, and the like.

[0108] "Acylamino" refers to an acyl-NH- group in which acyl is as previously described. "Aroylamino" refers to an aroyl-NH- group in which aroyl is as previously described.

[0109] The term "carbonyl" refers to the group -C(=O)- and can include aldehyde groups represented by the general formula RC(=O)H.

[0110] The term "carboxyl" refers to a -COOH group. Such a group is also referred to herein as a "carboxylic acid" moiety.

[0111] The term "cyano" refers to the group --C.ident.N.

[0112] As used herein, the terms "halo," "halide," or "halogen" refer to fluoro, chloro, bromo, and iodo groups. Additionally, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl. For example, "halo(C 1-4 The term "alkyl" is meant to include, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0113] The term "hydroxyl" refers to an --OH group.

[0114] The term "hydroxyalkyl" refers to an alkyl group substituted with an --OH group.

[0115] The term "mercapto" refers to the group --SH.

[0116] The term "oxo" as used herein means an oxygen atom that is double bonded to a carbon atom or another element.

[0117] The term "nitro" refers to the group --NO.sub.2.

[0118] The term "thio" refers to a compound as described herein above, in which a carbon or oxygen atom has been replaced with a sulfur atom.

[0119] The term "sulfate" refers to the -SO4 group.

[0120] The term thiohydroxyl or thiol, as used herein, refers to a group of formula -SH.

[0121] More specifically, the term "sulfide" refers to a compound having a group of formula -SR.

[0122] The term "sulfone" refers to a compound having a sulfonyl group -S(O2)R.

[0123] The term "sulfoxide" refers to a compound with a sulfinyl group -S(O)R.

[0124] The term ureido refers to a urea group of formula -NH-CO-NH2.

[0125] Throughout this specification and the claims, a given chemical formula or name is intended to encompass all tautomers, homologs, optical isomers and stereoisomers, as well as racemic mixtures, where such isomers and mixtures exist.

[0126] Certain compounds of the present disclosure may have asymmetric carbon atoms (optical or chiral centers) or double bonds; enantiomers, racemates, diastereomers, tautomers, geometric isomers, stereoisomeric forms that may be defined in terms of absolute stereochemistry as (R)- or (S)-, or for amino acids as D- or L-, and individual isomers are encompassed within the scope of the present disclosure. The present disclosure is intended to include compounds in racemic, scalaminic, and optically pure form. Optically active (R)- and (S)-, or D- and L-isomers, may be prepared using chiral synthesis or chiral reagents or resolved using conventional techniques. When a compound described herein contains an olefinic bond or other center of geometric asymmetry, unless otherwise specified, the compound is intended to include both E- and Z-geometric isomers.

[0127] Unless otherwise stated, structures depicted herein are meant to include all stereochemical forms of the structure, i.e., both R and S configurations of each asymmetric center. Accordingly, single stereochemical isomers as well as enantiomeric and diastereomeric mixtures of the present compounds are within the scope of the disclosure.

[0128] It will be apparent to one of ordinary skill in the art that certain compounds of the present disclosure may exist in tautomeric forms, and all such tautomeric forms of the compounds are within the scope of the present disclosure. As used herein, the term "tautomer" refers to one of two or more structural isomers that exist in equilibrium and are readily converted from one isomeric form to another.

[0129] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds in which a hydrogen is replaced by deuterium or tritium, or a carbon is replaced by an isotopically enriched atom. 13 C- or 14 Compounds having this structure substituted with C-enriched carbons are within the scope of this disclosure.

[0130] The compounds of the present disclosure may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may contain unnatural proportions of atomic isotopes, such as tritium ( 3 H), iodine-125( 125 I) or carbon-14 ( 14 C). All isotopic variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure.

[0131] Certain compounds of the present disclosure can exist in solvated form, including hydrated form, as well as nonsolvated form.In general, solvated form is equivalent to nonsolvated form and is included in the scope of the present disclosure.Certain compounds of the present disclosure can exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the use contemplated by the present disclosure and are intended to be within the scope of the present disclosure.

[0132] The term "protecting group" refers to a chemical moiety that blocks some or all reactive moieties in a compound, preventing them from participating in a chemical reaction until the protecting group is removed, such as those listed and described in T.W. Greene, P.G.M. Buts, Protective Groups in Organic Synthesis, 3rd ed., John Wiley & Sons (1999). When different protecting groups are employed, it may be advantageous for each (different) protecting group to be removable by different means. Protecting groups that are cleaved under completely different reaction conditions allow for differential removal of such protecting groups. For example, protecting groups can be removed by acid, base, and hydrogenolysis. Groups such as trityl, dimethoxytrityl, acetal, and tert-butyldimethylsilyl are acid-labile and can be used to protect carboxy and hydroxy reactive moieties in the presence of amino groups protected with the hydrogenolysis-removable Cbz group and the base-labile Fmoc group. Carboxylic acid and hydroxy reactive moieties can be blocked with base labile groups such as, but not limited to, methyl, ethyl, and acetyl, in the presence of acid labile groups such as tert-butyl carbamate or amines blocked with acid and base stable but hydrolytically removable carbamates.

[0133] Carboxylic acid and hydroxy reactive moieties can also be blocked with hydrolytically removable protecting groups such as benzyl groups, and amine groups capable of hydrogen bonding with acids can be blocked with base-labile groups such as Fmoc. Carboxylic acid reactive moieties can be blocked with oxidatively removable protecting groups such as 2,4-dimethoxybenzyl, and coexisting amino groups can be blocked with fluorine-labile silyl carbamates.

[0134] Allyl blocking groups are useful in the presence of acid- and base-protecting groups because the former are stable and can be subsequently removed by metal or π-acid catalysts. For example, allyl-blocked carboxylic acids can be deprotected by palladium(O)-catalyzed reactions in the presence of acid-labile t-butyl carbamate or base-labile acetate amine protecting groups. Yet another form of protecting group is a resin to which a compound or intermediate can be attached. As long as the residue is attached to the resin, its functional group is blocked and cannot react. Once released from the resin, the functional group becomes reactive.

[0135] Typical blocking / protecting groups include, but are not limited to, the following moieties: p-methoxybenzylcarbonyl (Moz or MeOZ), 3,4-dimethoxybenzyl (DMPM), Troc (trichloroethyl chloroformate), (4-nitrophenyl)sulfonyl (Nosyl), and nitrophenylsulfenyl (Nps), and [ka]

[0136] Following long-standing patent law practice, when used in this application, including the claims, the terms "a," "an," and "the" refer to "one or more." Thus, for example, a reference to "a subject" includes a plurality of subjects unless the context clearly indicates the contrary (e.g., a plurality of subjects).

[0137] Throughout this specification and the claims, the terms "comprise," "comprises," and "comprising" are used in a non-exclusive sense unless the context requires otherwise. Similarly, the term "include" and grammatical variations thereof are intended to be open-ended, such that the recitation of items in a list does not exclude other similar items that may be substituted for or added to the listed items.

[0138] For purposes of this specification and the appended claims, unless otherwise indicated, all numerical values ​​expressing quantities, sizes, dimensions, ratios, shapes, formulations, parameters, proportions, quantities, properties, and other numerical values ​​used in the specification and claims are to be understood as being modified in all instances by the term "about," even if the term "about" does not explicitly appear in conjunction with that value, amount, or range. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and appended claims need not be exact but may be approximate and / or increased or decreased as desired, reflecting tolerances, conversion factors, rounding off, measurement error, and the like, as well as other factors known to those of ordinary skill in the art, depending upon the desired properties sought to be obtained by the presently disclosed subject matter. For example, the term "about" when referring to a value can be meant to encompass variations from the stated amount of, in some embodiments, ±100%, in some embodiments, ±50%, in some embodiments, ±20%, in some embodiments, ±10%, in some embodiments, ±5%, in some embodiments, ±1%, in some embodiments, ±0.5%, and in some embodiments, ±0.1%, as such variations are appropriate for practicing the disclosed methods or employing the disclosed compositions.

[0139] Furthermore, when used in connection with one or more numerical values ​​or numerical ranges, the term "about" should be understood to refer to all such numerical values, including all numerical values ​​within the range, modifying that range by extending the boundaries above and below the stated numerical values. The recitation of numerical ranges by endpoints also includes all numbers subsumed within that range, for example, whole integers including fractions thereof (e.g., recitation of 1 to 5 includes 1, 2, 3, 4, 5, and fractions thereof, e.g., 1.5, 2.25, 3.75, 4.1, etc.), and any range within that range. [Example]

[0140] The following examples are included to provide guidance for those skilled in the art to practice representative embodiments of the presently disclosed subject matter. In light of this disclosure and the general state of the art, those skilled in the art will recognize that the following examples are intended to be illustrative only, and that numerous changes, modifications, and variations can be adopted without departing from the scope of the presently disclosed subject matter. The synthetic descriptions and specific examples provided below are for illustrative purposes only and are not to be construed as limiting the scope of making the compounds of the present disclosure by other methods.

[0141] Example 1 Stability of the prodrug in mouse skin homogenate and plasma The stability of mouse skin homogenates was evaluated using washed mouse skin samples diluted 1:10 in 0.1 M potassium phosphate buffer and homogenized using a tissue homogenizer, as well as CD1 mouse plasma. To assess the stability of intact prodrugs, 1 mL aliquots of skin homogenate or plasma were spiked with the prodrug to a final assay concentration of 20 μM. The spiked skin homogenate and plasma samples were incubated for 1 h at 37°C in an orbital shaker, after which the reaction was quenched in triplicate with three volumes of acetonitrile containing an internal standard (IS; losartan: 0.5 μM). The samples were vortex-mixed for 30 s and centrifuged at 10,000 × g for 10 min at 4°C. 50 μL of the supernatant was diluted with 50 μL of water and transferred to a 250 μL polypropylene vial sealed with a Teflon cap. Released itaconic acid or methyl itaconate was measured by liquid chromatography with tandem mass spectrometry (LC-MS / MS).

[0142] Example 2 Pharmacokinetics in mice Male CD1 mice (25-30 g) were obtained from Harlan and maintained on a 12-hour light-dark cycle with free access to food and water. IS-100-142 (compound 5), MK939 (compound 14), and MK941 (compound 16) were administered monomethyl itaconate at 10 mg / kg molar equivalent by oral gavage. Blood samples were collected at 0.25 and 1 h post-dose (n = 3 per time point). Mice were euthanized with carbon dioxide at the designated time points post-dose, and blood samples (approximately 0.8 mL) were collected by cardiac puncture into heparinized microtubes. Blood samples were centrifuged at 3,000 × g for 10 min at 4 °C. Plasma samples (approximately 300 μL) were collected in polypropylene tubes and stored at -80 °C until bioanalysis. Naive mouse plasma spiked with monomethyl itaconate was used to prepare calibration standards. Monomethyl itaconate standards and samples were extracted from plasma by one-step protein precipitation using methanol (100% v / v) containing the internal standard methyl succinate (5 μM). Aliquots (100 μL) of the supernatant were diluted with water (100 μL), transferred to 250 μL polypropylene vials sealed with Teflon caps, and analyzed by LC-MS / MS.

[0143] Example 3 Treatment of human keratinocytes with itaconate prodrugs Neonatal human epidermal keratinocytes (NHEKs) isolated from neonatal foreskin were seeded at a density of 100,000 cells per well and maintained in KGM supplemented with growth factors (KGM-GOLD Bullet Kit, #192060). The prodrug was reconstituted in DMSO. NHEKs were pretreated with either vehicle (0.1% DMSO) or the prodrug. Two days later, NHEKs were treated with 50 μg / μl poly(I:C) for 24 hours.

[0144] Example 4 RNA isolation and quantitative real-time PCR Total RNA was isolated and purified from cultured NHEKs using the RNeasy Mini Kit (Qiagen, Valencia, CA, #74106). After assessing RNA purity and concentration using a NanoDrop 2000 UV-Vis spectrophotometer, the RNA was converted to cDNA using a reverse transcription kit and random hexamer primers (Applied Biosystems, #4368814). mRNA expression was determined by qRT-PCR using gene-specific fluorophore-based TaqMan probes and a universal master mix (Applied Biosystems, #4366072). qRT-PCR reactions were multiplexed with target and reference gene (RPLP0) probes. Relative mRNA fold changes were then quantified using the ΔΔCt method.

[0145] Example 5 mouse Wild-type (C57BL6 / J) adult mice, all approximately 8.5 weeks old, were obtained from Jackson Labs (#00664). All animals were housed in an AAALAC (Authority for Accreditation of Laboratory Animal Care International)-accredited facility at Johns Hopkins University. All experimental procedures were approved by the Johns Hopkins Institutional Animal Care and Use Committee under protocol #MO17M298.

[0146] Example 6 Itaconate Prodrug Treatment Adult C57BL6 / J mice were treated during the refractory period of hair telogen to allow for the resumption of hair growth (anagen). To determine anagen reentry, mice were visually observed for thickening and darkening of the skin. The dorsum of each mouse was shaved, and the left half of the skin was topically treated with vehicle (DMSO) or a prodrug (20% IS-100-142 (Compound 5)), while the right half was left untreated. The treatment regimen was administered once every other day for a total of two treatment cycles.

[0147] Example 7 Itaconate and derivatives / analogs for inducing hair growth In one aspect, the subject matter of the present disclosure relates to compounds that may be able to promote hair growth.To this end, the ability of dimethyl itaconate to induce the transition from telogen, the resting phase of the hair cycle, to anagen, the growing phase of the hair cycle, was tested in wild-type mice.As shown in Figure 5, 20% dimethyl itaconate strongly induced this transition.

[0148] We next tested whether itaconate derivatives, such as itaconate prodrugs, possess this same ability. To screen compounds, we evaluated those that behaved in vitro with properties similar to itaconate. We first tested the molecular effect of dimethyl itaconate (DI), which blunts the effects of dsRNA (PIC; polyinosinic acid:polycytidylic acid) in increasing the expression of TLR3, IL-6, and STAT3. As shown in Figure 6, DI potently inhibits these markers.

[0149] Next, we tested the ability of the synthesized itaconate prodrugs to behave similarly in in vitro assays. Referring now to Figure 4, compound 5 from Table 1 also potently inhibited the same dsRNA response as DI. The method was as described above, but with dimethyl itaconate substituted for the described prodrug.

[0150] Finally, we tested the ability of Compound 5 to induce hair growth, as observed with DI. This was indeed the case, as shown in Figure 7. Here, mice were treated as above, except that the left side was treated with Compound 5 or DMSO and the right side was left untreated. The small amount of hair growth in the DMSO-treated mice was due to the initiation of the natural anagen cycle.

[0151] These results demonstrate the ability of itaconate and its derivatives / analogs to stimulate hair growth, such as the transition from telogen (resting phase) to anagen (growth phase) of the hair cycle. This approach should stimulate hair growth in many human conditions. These include male-pattern or female-pattern hair loss, in which follicles predominate in the resting phase, as well as telogen phase alopecia, in which the hair cycle prematurely and synchronously enters telogen. This method should also be useful for treating alopecia areata, in which localized areas of the scalp also synchronously enter telogen. This treatment is not only useful locally as shown here, but is also expected to act systemically.

[0152] Example 8 Compound (5) induces hair growth in mice All wild-type (C57BL6 / J) adult mice were approximately 8.5 weeks old and obtained from Jackson Labs. Animals were fed a standard mouse diet. Results from the mouse model demonstrated that Compound 5 promoted the transition from telogen (resting phase) to anagen (anagen phase) in hair follicles (Figure 8). Adult C57BL6 / J mice were treated during the refractory period of hair telogen to allow for the resumption of hair growth (anagen). To determine anagen reentry, mice were visually observed for thickening and darkening of the skin. Mice were shaved on the dorsum, and the right half of the skin was topically treated with vehicle (DMSO), dimethyl itaconate (20%), or 1% Compound 5, 2% Compound 5, or 5% Compound 5, while the left half was left untreated. The treatment regimen was performed once every other day for a total of two or three treatments, with similar results.

[0153] Example 9 Compound (5) induces hair growth in mice at levels comparable to those of ruxolitinib and tofacitinib Referring now to Figure 9, Compound 5 induces hair growth in mice at levels comparable to those of ruxolitinib and tofacitinib, two potent JAK inhibitors known to be successful in treating alopecia areata. Mice were treated as described in Example 8, but were also treated with ruxolitinib and tofacitinib, i.e., 3% ruxolitinib, 3% tofacitinib, and 3% Compound 5, each diluted in DMSO, as shown in Figure 9.

[0154] Example 10 In Vitro Data - Mouse Skin Homogenate Stability Skin samples were diluted 10-fold with 0.1 M potassium phosphate buffer and homogenized using a Polytron and probe sonicator. To assess the stability of intact prodrugs over time, aliquots were prepared and spiked with prodrugs to a final assay concentration of 20 μM. The spiked samples were incubated at 37°C for 1 hour in an orbital shaker, after which the reaction was quenched with three volumes of acetonitrile containing an internal standard (IS; losartan: 0.5 μM). The samples were vortex-mixed and centrifuged at 16,000 g for 5 minutes at 4°C. 50 μL of the supernatant was diluted with 50 μL of water and transferred to a 250 μL polypropylene vial sealed with a Teflon cap. Prodrug disappearance was monitored over time by liquid chromatography-mass spectrometry (LC-MS). Figure 10 shows the mouse skin homogenate stability of representative prodrugs. Figure 11 shows the release of active monomethyl itaconate from a representative prodrug (into skin homogenate).

[0155] Example 11 In Vitro Data - Mouse Skin Penetration Referring now to Figure 12, the mouse skin penetration of IS-100-142 (compound 5) (left panel) and monomethyl itaconate (right panel) in a model of alopecia is shown. After topical application in DMSO (dosage: 5% w / v), both intact and monomethyl itaconate levels are higher in skin than in plasma.

[0156] References

[0157] All publications, patent applications, patents, and other references mentioned in this specification are indicative of the level of ordinary skill in the art to which the subject matter of this disclosure pertains. All publications, patent applications, patents, and other references are incorporated herein by reference to the same extent as if each individual publication, patent application, patent, and other reference was specifically and individually indicated to be incorporated by reference. Although many patent applications, patents, and other documents are referenced herein, it will be understood that such reference does not constitute an admission that any of these documents form part of the general knowledge in the art.

[0158] PCT Patent Application Publication WO2017142855 (Artyomov et al., Immunomodulatory Agents and Methods of Use Thereof, Published August 24, 2017) PCT Patent Application Publication No. WO2019036509 (Artyomov et al., Methods and Compositions for the Treatment of Diseases Associated with Cancer, Inflammation, or Immune Response, Published February 21, 2019) O'Neill, LAJ and Artyomov, MN, Itaconate: the poster child of metabolic reprogramming in macrophage function, Nature Reviews:Immunology, 19, pp. 273-281 (2019)

[0159] Although the foregoing subject matter has been described in some detail by way of illustration and example for clarity of understanding, it will be understood by those skilled in the art that certain changes and modifications may be practiced within the scope of the appended claims.

Claims

1. Use of compounds of formula (I) and pharmaceutically acceptable salts thereof in the manufacture of a pharmaceutical composition for inducing hair growth in a subject: 【Chemical 1】 [In the ceremony: R ２ Ha-OC １－６ is a straight-chain or branched saturated alkyl; R １ teeth 【Chemistry 2】 where n is 1 and R ４ Ha-OC ３ is a straight-chain or branched saturated alkyl; R ７ teeth 【Chemistry 3】 is; R ８ is C １－６ is a straight-chain or branched saturated alkyl; R ９ is H; R １０ is C １－６ is a straight-chain or branched saturated alkyl; R １１ is H and R １２ is H or a protecting group; R １３ is C １－６ is a straight-chain or branched saturated alkyl; R １４ is H; R １５ is methyl; q is an integer selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10; and R １６ is H and R １７ is H or a protecting group, r is 1, and R １８ is phenyl.

2. Use according to claim 1, wherein R １ teeth 【Chemistry 4】 and R ２ Ha-OCH ３ (18, 20).

3. The compound of formula (I) is 【Chemistry 5】 The use according to claim 1, wherein the compound is selected from the group consisting of:

4. A pharmaceutical composition for inducing hair growth in a subject, comprising a compound of formula (I) and a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient: 【Chemistry 6】 wherein R 1 and R 2 are as defined in claim 1.

5. 2. The use of claim 1, wherein the subject has alopecia areata, androgenic alopecia, anagen effluvium, self-induced hair loss, telogen effluvium, or scarring alopecia.

6. The use according to claim 5, wherein the alopecia areata includes patchy alopecia areata.

7. The use according to claim 5, wherein the alopecia areata includes alopecia totalis.

8. The use according to claim 5, wherein the alopecia areata includes alopecia universalis.

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