JAK1 pathway inhibitors for the treatment of vitiligo

Abstract

The present disclosure relates to JAK1 pathway inhibitors and their use in treating vitiligo.

Description

[Technical Field]

[0001] Claim of priority This application claims priority to U.S. Provisional Application No. 63 / 122,574 filed on 8 December 2020 and U.S. Provisional Application No. 63 / 246,688 filed on 21 September 2021, the respective disclosures of which are incorporated herein by reference in their entirety.

[0002] This disclosure relates to JAK1 pathway inhibitors and their use in treating vitiligo. [Background technology]

[0003] Vitiligo is a chronic skin disease characterized by depigmented patches of skin due to autoimmune destruction of melanocytes, and is estimated to affect 0.5–2.0% of the world's population (varying geographically). The prevalence of vitiligo is similar in men and women, and no differences in symptoms based on skin type or race are known. The average age of onset is the mid-20s, but it can occur at any age. The disorder tends to progress over time. 15–25% of vitiligo patients tend to have at least one other autoimmune disorder. Non-segmental vitiligo is the most common subtype (up to 90% of vitiligo patients) and is characterized by symmetrical, bilateral white patches. The white patches are usually found on the hands, feet, and around the mouth / face.

[0004] Psychosocial factors can significantly impact the quality of life for those suffering from vitiligo. The chronic nature of vitiligo and the lack of effective treatments can have detrimental psychosocial effects on patients, affecting their quality of life as with other skin conditions. Involvement in cosmetically sensitive areas such as the face and hands can greatly impact self-esteem, ultimately leading to psychological burden and poor quality of life. Individuals with vitiligo affecting more than 25% of their body surface area may find it difficult to perform everyday tasks such as gardening, shopping, and choosing clothes, and may experience difficulties in social activities such as participating in sports or initiating and maintaining romantic relationships.

[0005] Clinical management aims to prevent depigmentation and induce repigmentation. Currently, there are no drugs approved for the treatment of vitiligo patients. Generally, the first-line treatment consists of off-label use of topical corticosteroids and calcineurin inhibitors. The second-line treatment consists of phototherapy (narrowband UVB, psoralen, UVA) and systemic corticosteroids. Less common treatment methods include surgical transplantation techniques and depigmentation treatments. Therefore, there is a need to develop new treatment methods for vitiligo. This application addresses this need and other needs.

Brief Description of the Drawings

[0006] [Figure 1] It is a graph showing melanocyte proliferation, % confluence vs. time (hours). [Figure 2] It is a graph showing the effect of Compound 1 on JAK1-dependent cytokines using T cell fraction, protein T cell medium (pg / ml) vs. specific JAK1-dependent cytokines. [Figure 3] It is a graph showing the effect of Compound 1 on growth factors and chemokines using T cell fraction, protein T cell medium (pg / ml) vs. IL-8 (A), HGF (B), and LIF (C). [Figure 4] It is a graph showing the effect of Compound 1 on JAK1-dependent cytokines using melanocytes, protein melanocyte medium (pg / ml) vs. specific JAK1-dependent cytokines. [Figure 5] It is a graph showing the effect of Compound 1 on growth factors and chemokines using melanocytes, protein melanocyte medium (pg / ml) vs. IL-8 (A), HGF (B), and LIF (C). [Figure 6] [[ID=--23]]It shows the outline of a Phase 2 randomized double-blind placebo-controlled dose range trial regarding the efficacy and safety of Compound 1. [Figure 7] It is a graph showing melanocyte proliferation, % confluence vs. time (hours). [Figure 8]This graph shows the effect of ruxolitinib on JAK1-dependent cytokines, using T cell compartments and protein T cell medium (pg / ml) versus specific JAK1-dependent cytokines. [Figure 9] This graph shows the effects of ruxolitinib on growth factors and chemokines using T cell compartments, protein T cell medium (pg / ml) versus IL-8 (A), and LIF (B). [Figure 10] This graph shows the effect of ruxolitinib on JAK1-dependent cytokines, using melanocytes, protein melanocyte medium (pg / ml), and specific JAK1-dependent cytokines. [Figure 11] This graph shows the effects of ruxolitinib on growth factors and chemokines using melanocytes, protein melanocyte medium (pg / ml) versus IL-8 (A), and LIF (B). [Overview of the Initiative]

[0007] This specification provides a method for treating vitiligo in a subject requiring treatment, the method comprising administering to the subject a therapeutically effective amount of a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof.

[0008] This specification provides JAK1 pathway inhibitors or pharmaceutically acceptable salts thereof for treating vitiligo in subjects requiring treatment.

[0009] This specification provides the use of a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof for the manufacture of a drug used in the treatment of vitiligo in subjects requiring treatment. [Modes for carrying out the invention]

[0010] In particular, the present invention provides a method for treating vitiligo in subjects requiring treatment, the method comprising administering to the subject a therapeutically effective amount of a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof. In some embodiments, the vitiligo is non-segmental vitiligo.

[0011] In some embodiments, the present invention provides a method for treating vitiligo in a subject, the method comprising administering to the subject a therapeutically effective amount of a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof.

[0012] In some embodiments, JAK1 pathway inhibitors or pharmaceutically acceptable salts thereof are more selective for JAK1 than for JAK2, JAK3, and TYK2.

[0013] In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipirazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharmaceutically acceptable salt thereof.

[0014] In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipirazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamidophosphate.

[0015] In some embodiments, the leukodectic mange is non-segmental leukodectic mange.

[0016] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in doses of approximately 5 mg to approximately 95 mg per day, based on free bases.

[0017] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in doses of approximately 15 mg, approximately 45 mg, 75 mg, or approximately 90 mg per day, based on free base.

[0018] In some embodiments, a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in combination with further therapeutic agents.

[0019] In some embodiments, further therapeutic agents include Janus kinase inhibitors.

[0020] In some embodiments, the Janus kinase inhibitor is ruxolitinib or a pharmaceutically acceptable salt thereof.

[0021] In some embodiments, administration involves administering a JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof together with at least one pharmaceutically acceptable carrier or excipient.

[0022] In some embodiments, vitiligo is non-segmental vitiligo. Non-segmental vitiligo exhibits some symmetry in the location of the depigmented patches. Classes of non-segmental vitiligo include generalized vitiligo, generalized vitiligo, vitiligo of the limbs and face, vitiligo of the mucous membranes, and localized vitiligo. In some embodiments, vitiligo is generalized vitiligo. In some embodiments, vitiligo is generalized vitiligo. In some embodiments, vitiligo is vitiligo of the limbs and face. In some embodiments, vitiligo is mucous membranes. In some embodiments, vitiligo is localized vitiligo. Generalized vitiligo, the most common category, affects about 0.5% of the world's population, with an average age of onset of about 24 years, occurring at roughly the same frequency in men and women. There are no ethnic differences, but the disease is more pronounced in people with darker skin tones, which can be psychologically distressing.

[0023] In some embodiments, vitiligo is segmental vitiligo. Segmental vitiligo differs from non-segmental vitiligo in appearance, cause, and prevalence. Segmental vitiligo tends to affect skin areas associated with the dorsal roots of the spinal cord and is almost always unilateral. Partial vitiligo spreads much more rapidly than non-partial vitiligo, and without treatment, partial vitiligo has a much more stable / static course and is not associated with autoimmune disease.

[0024] In some embodiments, the effectiveness of the treatment methods disclosed herein can be established based on the percentage change from various baseline measurements using various indicators. For example, the total percentage of depigmented body surface area (BSA), including facial area and non-facial area, can be used. BSA can be assessed by the Palmer method as follows: the approximate size of the entire palm surface of the subject (i.e., palm and five fingers) should be considered as 1% BSA, and the approximate size of the subject's thumb should be considered as 0.1% BSA.

[0025] As used herein, the term “face” may generally be defined as the area from the forehead to the original hairline, from the cheeks to the jawline, perpendicular to the jawline, and lateral from the corners of the mouth to the tragus. The area of ​​the “face” does not include the surface area of ​​the lips, scalp, ears, or neck, but it does include the nose and eyelids.

[0026] In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in a reduction of approximately 5%, approximately 10%, approximately 20%, approximately 30%, approximately 40%, 50%, approximately 60%, approximately 70%, approximately 80%, approximately 90%, or approximately 95% of the total body surface area (T-BSA) of depigmentation in a subject.

[0027] In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in a reduction of approximately 5%, approximately 10%, approximately 20%, approximately 30%, approximately 40%, 50%, approximately 60%, approximately 70%, approximately 80%, approximately 90%, or approximately 95% of the depigmented facial surface area (F-BSA) in a subject.

[0028] In some embodiments, the area affected by depigmentation due to vitiligo can be assessed using the Vitiligo Area Scoring Index (VASI). The VASI is based on a composite estimate of the involvement rate of vitiligo (%BSA, using the Palmer method) and the degree of depigmentation within the vitiligo lesion. In some embodiments, the degree of depigmentation can be determined and estimated to the closest value of the following percentages: 0%, 10%, 25%, 50%, 75%, 90%, or 100%. 100% depigmentation means no pigment is present; 90% means pigmented patches are present; 75% means the depigmented area exceeds the pigmented area; 50% means the depigmented area and pigmented area are equal; 25% means the pigmented area exceeds the depigmented area; 10% means only depigmented patches are present. 0% depigmentation means no depigmentation is present.

[0029] In some embodiments, the facial vitiligo area scoring index (F-VASI) is derived by multiplying the facial vitiligo involvement rate (%F-BSA) by the degree of depigmentation within the vitiligo lesion and summing the values ​​for all lesions.

[0030] In some embodiments, efficacy can be measured using a whole-body VASI (T-VASI) derived by multiplying the whole-body vitiligo involvement rate (%T-BSA) by the degree of depigmentation in vitiligo lesions in all body parts, and summing the values ​​for all parts (possibly ranging from 0 to 100). For the purpose of T-VASI evaluation, the body is divided into the following six mutually exclusive parts: (1) head / neck (including scalp), (2) hands, (3) upper limbs (including axillae and armpits) (excluding hands), (4) trunk (including genitals, excluding buttocks), (5) lower limbs (including buttocks, excluding feet), and (6) feet.

[0031] In some embodiments, efficacy can be evaluated based on the percentage change from baseline in T-VASI. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of T-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 50% or more in T-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 75% or more in T-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 90% or more in T-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 40% to about 90% or more in T-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 50% to about 95% or more in T-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in a reduction of approximately 5%, approximately 10%, approximately 20%, approximately 30%, approximately 40%, 50%, approximately 60%, approximately 70%, approximately 80%, approximately 90%, or approximately 95% of T-VASI in the subject.

[0032] In some embodiments, effectiveness can be evaluated based on the percentage of subjects who achieved a reduction of approximately 50% or more in T-VASI within a specific period. In some embodiments, effectiveness can be evaluated based on the percentage of subjects who achieved a reduction of approximately 50% or more, approximately 75% or more, or approximately 90% or more in T-VASI within a specific period.

[0033] In some embodiments, efficacy can be evaluated based on the percentage change from baseline in F-VASI. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of F-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 50% or more in F-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 75% or more in F-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 90% or more in F-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 40% to about 90% or more in F-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a reduction of about 50% to about 95% or more in F-VASI in the subject. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in a reduction of approximately 5%, approximately 10%, approximately 20%, approximately 30%, approximately 40%, 50%, approximately 60%, approximately 70%, approximately 80%, approximately 90%, or approximately 95% of F-VASI in the subject.

[0034] The Vitiligo Expansion Score (VES) is a validated measure that represents the overall involvement of vitiligo on the body. VES uses 19 individual anatomical area photographs to assess the degree of vitiligo (%BSA).

[0035] The Vitiligo Expansion Score Plus (VESplus) is a validated measure that represents the overall involvement of vitiligo on the body and assesses the pattern of perifollicular repigmentation. VESplus uses 19 individual area photographs to assess the degree of vitiligo (%BSA) and the pattern of perifollicular pigmentation, each categorized into seven groups (0%, 5%, 10%, 25%, 50%, 75%, 90%).

[0036] In some embodiments, VESplus can be used to evaluate its effectiveness based on improvements in the extent of vitiligo and the pattern of perifollicular repigmentation. Typically, VESplus uses a calculation in which BSA is multiplied by the degree of perifollicular repigmentation. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein, results in an improvement in VESplus in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein, results in a VESplus of about 50% or more in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein, results in a VESplus of about 75% or more in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein, results in a VESplus of about 90% or more in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein, results in a VESplus of about 50% to about 90% or more in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein, results in approximately 5% to approximately 75% or more of VESplus in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein, results in approximately 5% to approximately 90% or more of VESplus in the subject.

[0037] In some embodiments, effectiveness can be assessed based on improvement in the Facial Static Investigator Overall Assessment (FSIGA) score. The severity of facial vitiligo can be assessed using the 5-point FSIGA scale (Table 1). [Table 1]

[0038] In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a subject with no (0) or nearly no (1) FSIGA score. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in an improvement of at least 2 points in the FSIGA score. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a subject with no (0) or nearly no (1) and an improvement of at least 2 points in the FSIGA score.

[0039] In some embodiments, effectiveness can be assessed based on improvements in the Static Investigator Overall Assessment (SIGA) score. The severity of vitiligo can be assessed using the 5-point SIGA scale (Table 2). [Table 2]

[0040] In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a SIGA score of none (0) or nearly none (1) in the subject. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in an improvement of at least 2 points in the SIGA score. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in an improvement of at least 1, 2, or 3 points in the SIGA score. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a SIGA score of none (0) or nearly none (1) and at least 2 points in the subject.

[0041] Vitiligo is characterized by the progressive loss of functional melanocytes and is associated with auditory abnormalities such as sensorineural hearing loss (SNHL), with a prevalence of 4.0–68.8% compared to healthy controls. During embryogenesis, precursor melanocyte cells (melanoblasts) migrate to the dermis and further to the base of the cochlea, which is responsible for hearing high frequencies. Inner ear melanocytes appear to be crucial for cochlear hair cell function and normal hearing. Individuals suffering from vitiligo can undergo a baseline SNHL assessment to evaluate hearing and confirm potential improvements in hearing compared to exposure to a JAK1 inhibitor, e.g., compound 1, throughout treatment. In some embodiments, efficacy can be assessed based on the improvement in the subject's hearing. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in an improvement in the subject's baseline auditory parameters.

[0042] In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement of about 5%, about 10%, about 20%, about 30%, about 40%, 50%, about 60%, about 70%, about 80%, about 90%, or about 95% of the baseline auditory parameters of the subject. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement of about 50% or more of the baseline auditory parameters of the subject.

[0043] In some embodiments, efficacy can be evaluated based on improvement in the subject's response to a color matching question (i.e., patient-reported outcome (PRO)). In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in improvement in the subject's response to a color matching question. In the color matching question, the subject may be shown a current facial image (the subject may use a mirror) for reference, and the subject is asked to answer the following question: "At this stage of treatment, how well does your skin color match the skin color of the facial area treated for vitiligo to that of a normal facial area?" Possible responses: (1) Very good, (2) Very good, (3) Good, (4) Poor, and (5) Very poor. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in improvement of at least 2 points in the color matching question. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in improvement of at least 1, 2, or 3 points in the color matching question.

[0044] In some embodiments, efficacy can be assessed based on the subject's assessment of the PRO, the response to a baseline facial photograph. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in an improvement in the subject's assessment of the PRO to a baseline facial photograph. The Vitiligo Visibility Scale (VNS) is a patient-reported measure of the success of vitiligo treatment and consists of a 5-point scale. A baseline facial photograph can be shown to the subject for reference, and a mirror can be provided for the subject to assess the vitiligo on their face. The subject is asked to answer the following question: "How noticeable are the vitiligo patches now compared to before treatment?" Possible responses are: (1) more noticeable, (2) equally noticeable, (3) slightly less noticeable, (4) considerably less noticeable, (5) no longer noticeable. How noticeable are the vitiligo patches now compared to before treatment? Responses: (1) more noticeable, (2) equally noticeable, (3) slightly less noticeable, (4) considerably less noticeable, (5) no longer noticeable. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a VNS score of 4 or 5.

[0045] In some embodiments, efficacy can be assessed based on the subject's response to facial overall impression change - vitiligo (F-PaGIC-V), improvement in PRO. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement in the subject's response to F-PaGIC-V. F-PaGIC-V is an assessment of improvement. It is a 7-point scale comparing the baseline vitiligo area to the treated area of ​​facial vitiligo in the subject. The subject is shown baseline photographs and current facial images (a mirror may be provided to the subject) for reference. The subject is asked to answer the following question: Since initiating treatment in this study, the facial vitiligo treated with the investigational drug has: (1) greatly improved, (2) considerably improved, (3) slightly improved, (4) no change, (5) slightly worsened, (6) considerably worsened, (7) greatly worsened. In some embodiments, Compound 1 and / or the method of use described herein results in an F-PaGIC-V score of 1 or 2.

[0046] In some embodiments, efficacy can be assessed based on the subject's response to overall patient impression change—vitiligo (T-PaGIC-V), improvement in PRO. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement in the subject's response to T-PaGIC-V. T-PaGIC-V is an assessment of improvement. It is a 7-point scale comparing baseline vitiligo area to the treated area of ​​generalized vitiligo in the subject. Subjects are asked to answer the following question: Since initiating treatment in this study, generalized vitiligo treated with the investigational drug has: (1) greatly improved, (2) considerably improved, (3) slightly improved, (4) no change, (5) slightly worsened, (6) considerably worsened, (7) greatly worsened.

[0047] In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in a T-PaGIC-V score of 1 or 2.

[0048] In some embodiments, efficacy can be assessed based on improvement in the subject's self-reported vitiligo severity score (SA-VES), PRO. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in an improvement in the subject's response to SA-VES from baseline. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the method of use described herein results in an improvement of approximately 5%, approximately 10%, approximately 20%, approximately 30%, approximately 40%, 50%, approximately 60%, approximately 70%, approximately 80%, approximately 90%, or approximately 95% of the subject's response to SA-VES from baseline. The self-reported vitiligo severity score (SA-VES) is a validated tool in which a patient can select the extent of the disease (shown in images) across 12 anatomical areas.

[0049] In some embodiments, efficacy can be assessed based on improvement in the subject's quality of life (VitiQoL), PRO. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement in the subject's response to VitiQoL from baseline. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement of approximately 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% of the subject's response to VitiQoL from baseline. VitiQoL is a 15-item quality of life assessment in which subjects rate various aspects of their condition over the past month on a 7-point scale (from "never" to "always").

[0050] In some embodiments, efficacy can be assessed based on improvement in the subject's Dermatological Quality of Life Index (DLQI), PRO. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement in the subject's response to DLQI from baseline. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement of about 5%, about 10%, about 20%, about 30%, about 40%, 50%, about 60%, about 70%, about 80%, about 90%, or about 95% of the subject's response to DLQI from baseline. The DLQI is a 10-question validated questionnaire to measure how much a skin problem has affected a subject over the past 7 days. Subjects can answer the questionnaire on the scale of (1) very, (2) quite, (3) a little, or (4) none at all. The questionnaire is analyzed under the following six headings: Symptoms and Emotions (Questions 1 and 2); Daily Activities (Questions 3 and 4); Leisure (Questions 5 and 6); Workplace and School (Question 7); Relationships (Questions 8 and 9); and Treatment (Question 10).

[0051] In some embodiments, efficacy can be assessed based on improvement, PRO, in the subject's Hospital Anxiety and Depression Scale (HADS). In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement in the subject's response to the HADS from baseline. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement of approximately 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% in the subject's response to the HADS from baseline. The HADS is a 14-item questionnaire that assesses the level of anxiety and depression a person is currently experiencing. There are seven questions to measure anxiety and seven questions to measure depression, with four possible responses for each question (responses are scored as 0, 1, 2, or 3). Separate scores are calculated for anxiety and depression.

[0052] In some embodiments, efficacy can be evaluated based on the subject's improvement in the WHO-5, PRO. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement in the subject's response to the WHO-5 from baseline. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the method of use described herein results in an improvement of about 5%, about 10%, about 20%, about 30%, about 40%, 50%, about 60%, about 70%, about 80%, about 90%, or about 95% of the subject's response to the WHO-5 from baseline. The WHO-5 is a validated, self-administered, five-item questionnaire designed to assess mental health status over the past two weeks and can be used as an outcome measure for desired and undesirable effects of treatment. The questionnaire consists of five statements, which respondents rate according to the following scale: 0 = never; 1 = sometimes; 2 = less than half; 3 = more than half; 4 = almost; 5 = always.

[0053] The raw score is calculated by summing the values ​​of five responses on a scale of 0 to 25. 0 represents the worst possible quality of life, and 25 represents the best possible quality of life. A score below 13 indicates low happiness.

[0054] In some embodiments, efficacy can be assessed based on improvement in the Treatment Satisfaction Questionnaire (TSQM) for the medication in question. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the use described herein results in an improvement in the subject's response to the TSQM from baseline. In some embodiments, a JAK1 inhibitor, e.g., Compound 1, and / or the use described herein results in an improvement of approximately 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% in the subject's response to the TSQM from baseline. The TSQM is a validated nine-item questionnaire that measures the subject's satisfaction with treatment using the past two to three weeks, or the recall period since the last use of treatment. A seven-point scale is used for each question in the questionnaire.

[0055] In some embodiments, efficacy can be evaluated based on improvement in the subject's EQ-5D-5L questionnaire. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the use described herein results in an improvement in the subject's response to the EQ-5D-5L questionnaire from baseline. In some embodiments, a JAK1 inhibitor, e.g., compound 1, and / or the use described herein results in an improvement of approximately 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95% of the subject's response to the EQ-5D-5L questionnaire from baseline. The EQ-5D-5L questionnaire is a standardized and validated means for use as a measure of health outcomes. The EQ-5D-5L questionnaire provides data for use in economic models and analyses, such as in the development of healthcare businesses and quality-adjusted life years (QALYs). The EQ-5D-5L questionnaire consists of two sections: the EQ-5D Description System and the EQVAS. The descriptive system consists of five aspects: mobility, self-care, usual activity, pain / discomfort, and anxiety / depression.

[0056] Each dimension has five levels: Level 1 is "No problem," Level 2 is "Mild problem," Level 3 is "Moderate problem," Level 4 is "Severe problem," and Level 5 is "Extreme problem." This part of the EQ-5D-5L questionnaire provides descriptive profiles that can be used to generate health status profiles. For example, a person with "Health Status 12345" has no problems with mobility, mild problems with self-care (laundry and dressing), moderate problems with performing normal activities, severe pain or discomfort, or extreme anxiety or depression. Each health status may be assigned a summary index score based on the weight of social preferences for that health status. These weights, also called utility, are often used to calculate QALYs for use in health economic analysis. Health status index scores generally range from less than 0 (0 is a health status equivalent to death, negative values ​​represent a health status worse than death) to 1 (value of perfect health), with higher scores indicating greater health utility. Health status preferences can vary from country / region to country, as they often reflect national or regional values. The EQ VAS records the subject's self-reported health status using a visual analog scale (0-100), with endpoints assigned as "best possible health" (100 score) and "worst possible health" (0 score).

[0057] The methods described herein utilize JAK1 pathway inhibitors, particularly JAK1 selective inhibitors. JAK1 selective inhibitors are compounds that preferentially inhibit JAK1 activity over other Janus kinases. JAK1 plays a central role in the signaling pathways of many cytokines and growth factors that, in dysregulation, can lead to or contribute to disease states. For example, levels of IL-6 (a pro-inflammatory cytokine) are elevated in subjects with vitiligo (Singh, et al., Indian J Dermatology, 2012 Jan-Feb;57(1):12-14). In other autoimmune diseases and cancers, elevated systemic levels of inflammatory cytokines that activate JAK1 may also contribute to the disease and / or associated symptoms. Therefore, patients with autoimmune diseases such as vitiligo may benefit from JAK1 inhibition. Selective JAK1 inhibitors may be beneficial while avoiding unnecessary and potentially unwanted effects caused by inhibiting other JAK kinases.

[0058] In some embodiments, JAK1 pathway inhibitors or pharmaceutically acceptable salts thereof are selective to JAK1 over JAK2, JAK3, and TYK2 (i.e., JAK1 selective inhibitors). For example, the compounds described herein, or their pharmaceutically acceptable salts, preferentially inhibit JAK1 over one or more JAK2, JAK3, and TYK2. In some embodiments, the compounds preferentially inhibit JAK1 over JAK2 (e.g., JAK2 / JAK1 IC2). 50 Ratio > 1). In some embodiments, the compound or salt is about 10 times more selective to JAK1 than to JAK2. In some embodiments, the compound or salt IC2 at 1 mM ATP. 50 Calculations based on measurements show that JAK2 is approximately 3 times, 5 times, 10 times, 15 times, or 20 times more selective than JAK1 (see, for example, Example A).

[0059] In some embodiments, the JAK1 pathway inhibitor is one of the compounds listed in Table 3, or a pharmaceutically acceptable salt thereof. The compounds in Table 3 are selective JAK1 inhibitors (more selective than JAK2, JAK3, and TYK2). IC obtained by the method of Example A with 1 mM ATP. 50 The values ​​are shown in Table 3.

[0060] The compounds in Table 3 include, for example, U.S. Patent Publication No. 2011 / 0224190 filed on March 9, 2011, U.S. Patent Publication No. 2014 / 0343030 filed on May 16, 2014, U.S. Patent Publication No. 2014 / 0121198 filed on October 31, 2013, U.S. Patent Publication No. 2010 / 0298334 filed on May 21, 2010, U.S. Patent Publication No. 2011 / 0059951 filed on August 31, 2010, and U.S. Patent Publication No. 224190 filed on November 18, 2011. These can be prepared by the synthesis procedures described in U.S. Patent Publication No. 012 / 0149681, U.S. Patent Publication No. 2012 / 0149682 filed November 18, 2011, U.S. Patent Publication No. 2013 / 0018034 filed June 19, 2012, U.S. Patent Publication No. 2013 / 0045963 filed August 17, 2012, and U.S. Patent Publication No. 2014 / 0005166 filed May 17, 2013, each of which is incorporated herein by reference in whole. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] [Table 3-8] [Table 3-9] [Table 3-10] [Table 3-11]

[0061] In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide (compound 1) or a pharmaceutically acceptable salt thereof. In some embodiments, the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazole-1-yl)azetidine-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide phosphate. Compound 1 and its salts can be prepared, for example, by the procedure described in U.S. Patent No. 9,382,231, filed May 16, 2014 (see, for example, Example 7), which is incorporated herein by reference in its entirety.

[0062] In some embodiments, the JAK1 pathway inhibitor is U.S. Patent Publication No. 2011 / 0224190 filed March 9, 2011, U.S. Patent Publication No. 2014 / 0343030 filed May 16, 2014, U.S. Patent Publication No. 2014 / 0121198 filed October 31, 2013, U.S. Patent Publication No. 2010 / 0298334 filed May 21, 2010, U.S. Patent Publication No. 2011 / 0059951 filed August 31, 2010, and U.S. Patent Publication No. A selection of compounds or pharmaceutically acceptable salts thereof described in U.S. Patent Publication No. 2012 / 0149681, U.S. Patent Publication No. 2012 / 0149682 filed November 18, 2011, U.S. Patent Publication No. 2013 / 0018034 filed June 19, 2012, U.S. Patent Publication No. 2013 / 0045963 filed August 17, 2012, and U.S. Patent Publication No. 2014 / 0005166 filed May 17, 2013, is used, each of which is incorporated herein by reference in whole.

[0063] In some embodiments, the JAK1 pathway inhibitor is a compound of formula I. [ka] or a pharmaceutically acceptable salt thereof, in the formula, X is N or CH, L is C(=O) or C(=O)NH, A is phenyl, pyridinyl, or pyrimidinyl, each of which is one or two independently selected R 1 It is arbitrarily substituted in the base, and further Each R 1 These are independently fluoromethyl or trifluoromethyl.

[0064] In some embodiments, the compound of formula I is {1-{1-[3-fluoro-2-(trifluoromethyl)isonicotinoyl]piperidine-4-yl}-3[4-(7H-pyrrolo[2,3-d]pyrimidine-4-yl)-1H-pyrazole-1-yl]azetidine-3-yl}acetonitrile, or a pharmaceutically acceptable salt thereof.

[0065] In some embodiments, the compound of formula I is 4-{3-(cyanomethyl)-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]azetidin-1-yl}-N-[4-fluoro-2-(trifluoromethyl)phenyl]piperidine-1-carboxamide or a pharmaceutically acceptable salt thereof.

[0066] In some embodiments, the compound of formula I is [3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(1-{[2-(trifluoromethyl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)azetidin-3-yl]acetonitrile or a pharmaceutically acceptable salt thereof.

[0067] In some embodiments, the JAK1 pathway inhibitor is a compound of formula II

Chemical formula

[0068] In some embodiments, the JAK1 pathway inhibitor is a compound of formula III. [ka] Alternatively, a pharmaceutically acceptable salt thereof, in the formula, Cy 4 CN, OH, F, Cl, C 1-3 Alkyl, C 1-3 Haloalkyl, cyano-C 1-3 Alkyl, HO-C 1-3 Alkyl, amino, C 1-3 Alkylamino and di(C) 1-3 A tetrahydro-2H-pyran ring optionally substituted with one or two groups independently selected from alkyl)amino, where this C 1-3 Alkyl and di(C) 1-3 Alkyl)amino is F, Cl, C 1-3 Alkylaminosulfonyl and C 1-3 Optionally substituted with one, two, or three substituents independently selected from the alkylsulfonyl, R 12 These are -CH2-OH, -CH(CH3)-OH, or -CH2-NHSO2CH3.

[0069] In some embodiments, the compound of formula III is ((2R,5S)-5-{2-[(1R)-1-hydroxyethyl]-1H-imidazo[4,5-d]thieno[3,2-b]pyridine-1-yl}tetrahydro-2H-pyran-2-yl)acetonitrile, or a pharmaceutically acceptable salt thereof.

[0070] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in amounts of approximately 10 mg to approximately 100 mg per day, based on free bases. Therefore, in some embodiments, a selective JAK1 pathway inhibitor is administered in amounts of approximately 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, or 100 mg per day, based on free bases. In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in amounts of approximately 1 mg to approximately 100 mg per day, based on free bases. In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in amounts of approximately 10 mg to approximately 80 mg per day, based on free bases. In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered at a dose of approximately 90 mg per day on a free base basis. In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered at a dose of approximately 75 mg per day on a free base basis. In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered at a dose of approximately 45 mg per day on a free base basis. In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered at a dose of approximately 15 mg per day on a free base basis.

[0071] The term "approximately" means "about" (for example, about 10% plus or minus the given value).

[0072] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered in one or more sustained-release formulations, each containing the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof.

[0073] In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered orally.

[0074] The embodiments described herein are intended to be combined in any preferred combination, as if they were multiple dependent claims (for example, embodiments relating to selective JAK1 pathway inhibitors and their doses, embodiments relating to any salt form of the compounds disclosed herein, embodiments relating to individual types of cytokine-related diseases or disorders, and embodiments relating to compositions and / or administrations that can be combined in any combination).

[0075] All possible combinations are not shown separately in this specification for the sake of brevity.

[0076] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). All stereoisomers, such as enantiomers and diastereomers, are intended unless otherwise specified. Compounds containing asymmetrically substituted carbon atoms may be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically inert starting materials are well known in the art, for example, by resolution or stereoselective synthesis of racemic mixtures. Many geometric isomers, such as olefins and C=N double bonds, may also be present in the compounds described herein, and all such stable isomers are intended in the present invention. Cis and trans geometric isomers in the compounds of the present invention are described and may be isolated as mixtures of isomers or as separated isomers.

[0077] In some embodiments, the compound has an (R) configuration. In some embodiments, the compound has an (S) configuration.

[0078] The separation of racemic mixtures of compounds can be carried out by any of the many methods known in the art. An exemplary method is fractional recrystallization using chiral splitting acids, which are optically active salt-forming organic acids. Suitable resolving agents for fractional recrystallization are optically active acids such as D and L-type tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or various optically active camphorsulfonic acids such as β-camphorsulfonic acid. Other suitable resolving agents for fractional crystallization include stereoisomerically pure forms of α-methylbenzylamine (e.g., S and R-type, or diastereomerically pure forms), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, and 1,2-diaminocyclohexane.

[0079] The decomposition of racemic mixtures can also be carried out by elution on a column packed with an optically active decomposition agent (e.g., dinitrobenzoylphenylglycine). Suitable elution solvent compositions can be determined by those skilled in the art.

[0080] The compounds described herein also include tautomeristic forms. Tautomeristic forms are obtained by the exchange of adjacent double and single bonds and the simultaneous transfer of protons. Tautomeristic forms include prototropic tautomers, which are protonated states of isomers having the same empirical formula and total charge. Exemplary prototropic tautomers include ketone-enol pairs, amide-imoid acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which protons can occupy two or more positions in a heterocyclic system, such as 1H- and 3H-imidazoles, 1H-, 2H-, and 4H-1,2,4-triazoles, 1H- and 2H-isoindoles, and 1H- and 2H-pyrazoles. Tautomeristic forms may be in equilibrium or sterically fixed to one form by appropriate substitution.

[0081] The compounds described herein may also include isotope-labeled compounds of this disclosure. An “isotope-labeled” or “radio-labeled” compound is a compound of this disclosure in which one or more atoms are replaced or substituted by atoms having an atomic mass or mass number different from that normally found in nature (i.e., naturally occurring atomic mass or mass number). Suitable radionuclides that may be incorporated into the compounds of this disclosure include: 2 H (also written as D as Deuterium), 3 H (also written as T for tritium), 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 18 F, 35 S, 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I, 124 I, 125 I and 131 I is included but not limited to. For example, one or more hydrogen atoms in the compounds of this disclosure may be replaced by deuterium atoms (e.g., formula (I), (II), or (III) or C in the compounds of Table 3). 1-6 One or more hydrogen atoms of the alkyl group may be optionally substituted with a deutherium atom such that -CD3 is substituted for -CH3. As used herein, the term “compound” means all stereoisomers, geometric isomers, tautomers, and isotopes of the given structure unless the name indicates a specific stereoisomer. Compounds herein identified by name or structure as a particular tautomer are intended to include other tautomers unless otherwise specified.

[0082] All compounds and their pharmaceutically acceptable salts can be identified together with other substances such as water and solvents (e.g., hydrates and solvates) or isolated.

[0083] In some embodiments, the compounds and salts thereof described herein are substantially isolated. "Substantially isolated" means that the compound is at least partially, or substantially, separated from the environment in which it was formed or detected. Partial isolation may include, for example, a composition in which the compounds described herein are concentrated. Substantial isolation may include a composition containing at least about 50% by weight, at least about 60% by weight, at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 95% by weight, at least about 97% by weight, or at least about 99% by weight of the compounds or salts thereof described herein. Methods for isolating compounds and salts thereof are commonplace in the art.

[0084] The term "pharmaceutically acceptable" is used herein to mean a compound, substance, composition, and / or dosage form that is suitable for use in contact with human and animal tissues within the bounds of appropriate medical judgment, and that provides a reasonable benefit-to-risk ratio without causing excessive toxicity, irritation, allergic reactions, or other problems or complications.

[0085] As used herein, the terms “ambient temperature” and “room temperature” or “rt” are understood in the art and generally refer to a temperature close to the temperature of the room in which the reaction takes place, for example, a temperature between approximately 20°C and approximately 30°C, e.g., the reaction temperature.

[0086] The present invention also includes pharmaceutically acceptable salts of the compounds described herein. As used herein, “pharmaceutically acceptable salt” is a derivative of the disclosed compound, where the parent compound is modified by converting an existing acidic or base moiety to its salt form. Examples of pharmaceutically acceptable salts include, but are not limited to, inorganic or organic salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. The pharmaceutically acceptable salts of the present invention include, for example, conventional non-toxic salts of parent compounds formed from non-toxic inorganic or organic acids. The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acidic or base form of these compounds with a stoichiometric amount of a suitable base or acid in water, an organic solvent, or a mixture thereof, generally in non-aqueous media such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol), or acetonitrile (ACN). A list of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

[0087] As used herein, the terms “subject,” “individual,” or “patient” are interchangeable and refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, most preferably humans. In some embodiments, the “subject,” “individual,” or “patient” is in need of the treatment described above.

[0088] In some embodiments, the inhibitor is administered in a therapeutically effective dose. As used herein, the term “therapeutably effective dose” refers to the amount of an active compound or drug that elicits a desired biological or pharmacokinetic response in a tissue, system, animal, individual, or human, as determined by the researcher, veterinarian, physician, or other clinician.

[0089] As used herein, the term “to treat” or “treatment” means one or more of the following: (1) inhibiting a disease, for example, inhibiting a disease, condition, or disorder in an individual who is experiencing or exhibiting the pathology or overall symptoms of a disease, condition, or disorder (i.e., halting the further progression of the pathology and / or overall symptoms); or (2) improving a disease, for example, improving a disease, condition, or disorder in an individual who is experiencing or exhibiting the pathology or overall symptoms of a disease, condition, or disorder (i.e., improving the pathology and / or overall symptoms), such as reducing the severity of the disease.

[0090] In some embodiments, JAK1 inhibitors can prevent vitiligo in individuals susceptible to the disease. The term “prevent” refers to preventing the development of the disease in patients who may be predisposed to the disease but have not yet experienced or displayed the pathology or symptoms of the disease.

[0091] Combination therapy The methods described herein may further include administering one or more additional therapeutic agents. One or more additional therapeutic agents may be administered to the patient simultaneously or sequentially. One or more additional therapeutic agents may be administered using a method different from that of compound 1 (e.g., topically).

[0092] In some embodiments, additional therapeutic agents are selected from other JAK inhibitors. These additional JAK inhibitors may include ATI-50002 (JAK1 / 3 selective), PF-06651600 (JAK3 selective), PF06700841 (JAK1 / TYK2 selective), upadacitinib, abrocitinib (JAK1 selective), celduratinib (JAK1 / SYK selective), and duklavacitinib (TYK2 selective).

[0093] In some embodiments, additional therapeutic agents are selected from antioxidants. Antioxidants may be selected from pseudocatalase, vitamin E, vitamin C, ubiquinone, lipoic acid, Polypodium leucotomos, catalase / superoxide dismutase combinations, and Ginkgo biloba. In some embodiments, antioxidants may be further administered in combination with phototherapy. Administration of antioxidants during or before phototherapy is intended to counteract oxidative stress induced by ultraviolet light itself and enhance the effects of phototherapy.

[0094] In some embodiments, additional therapeutic agents are wingless-associated embedded site (Wnt) agonists. Wnt agonists may include SKL2001.

[0095] In some embodiments, additional therapeutic agents are melanocortin 1 receptor (MC1R) agonists.

[0096] In some embodiments, additional therapeutic agents are inflammatory mediators. These inflammatory mediators may include prostaglandin E2 (PGE2) and bimatoprost (a synthetic analog of prostaglandin F2α).

[0097] In some embodiments, additional therapeutic agents are antimetabolites. These antimetabolites may include 5-fluorouracil.

[0098] In some embodiments, additional therapeutic agents include plasmid HSP70i gene therapy. Plasmid HSP70i gene therapy may include HSP70iQ435A.

[0099] In some embodiments, additional therapeutic agents are selected from topical corticosteroids, immunomodulators, calcineurin inhibitors, and phototherapy. In some embodiments, the additional therapeutic agent is a systemic steroid or immunosuppressant.

[0100] In some embodiments, additional therapeutic agents include steroids, including systemic steroids (e.g., orally administered steroids). Steroid therapy may include oral steroid minipulse therapy (e.g., the use of betamethasone and / or dexamethasone).

[0101] In some embodiments, the topical corticosteroid is selected from betamethasone dipropionate, clobetasol propionate, diflorasone diacetate, halobetazole amcinonide propionate, betamethasone valerate, desoxymethasone, diflorasone diacetate, fluocinolone acetonide, halcinonide, and triamcinolone acetonide.

[0102] In some embodiments, additional therapeutic agents include immunomodulators. Immunomodulators may include PDE4 inhibitors (e.g., apremilast (e.g., oral) or crisabolol (e.g., topical)). Immunomodulators may include anti-CD20 therapies (e.g., ofatumumab). Immunomodulators may include anti-CD19 therapies (e.g., tafacitamab). Immunomodulators may include anti-IL15 therapies (e.g., AMG714 monoclonal antibodies). Immunomodulators may include anti-IL36 therapies (e.g., imsidrimab and spesolimab). Immunomodulators may include anti-TNFα therapies (e.g., etanercept and infliximab). Immunomodulators may include anti-CD122 therapies.

[0103] In some embodiments, the immunomodulator is selected from apremilast, crisablol, afamelanotide, rituximab, ofatumumab, tafacitamab, minocycline, latanoprost, zinc, tofacitinib, AMG714 monoclonal antibody, imsidrimab, spesolimab cyclosporine, etanercept, infliximab, cyclophosphamide, cyclosporine, methotrexate, and sodium oxodihydroacridinyl acetate (ODHAA).

[0104] In some embodiments, the calcineurin inhibitor is selected from tacrolimus (FK-506) and pimecrolimus.

[0105] In some embodiments, phototherapy includes exposure to ultraviolet light (e.g., an excimer lamp or laser).

[0106] In some embodiments, the additional therapeutic agent is a Janus kinase inhibitor. In some embodiments, the Janus kinase inhibitor is ruxolitinib or a pharmaceutically acceptable salt thereof. In some embodiments, ruxolitinib is administered topically. Topical administration of ruxolitinib is described in detail, for example, in U.S. Patent Publication No. 2011 / 0288107, filed March 20, 2011, and in Persaud et al., “Plasma pharmacokinetics and distribution of ruxolitinib into skin following oral and topical administration in minipigs,” Int JPharm, 590:119889, November 30, 2020, PMID:32949620, which is incorporated herein by reference in its entirety.

[0107] In some embodiments, the additional therapeutic agent is an IL-6 antagonist or receptor antagonist. In some embodiments, the IL-6 receptor antagonist is tocilizumab.

[0108] In some embodiments, the methods described herein may further include use in combination with surgical techniques. In some embodiments, the surgical techniques include surgical skin grafting.

[0109] Pharmaceutical preparations and dosage forms When used as a pharmaceutical, JAK1 pathway inhibitors or pharmaceutically acceptable salts thereof can be administered in the form of pharmaceutical compositions. These compositions can be prepared by methods well known in the pharmaceutical field and can be administered by various routes depending on whether topical or systemic treatment is preferred and the area to be treated. Administration can be topical (transdermal, epithelial, mucosal, such as intraocular, intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or gas injection of powders or aerosols using a sprayer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular, or injection or infusion, or intracranial administration such as in the subarachnoid space or ventricles. Parenteral administration may be in the form of a single bolus or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, foams, liquids, and powders. Conventional pharmaceutical carriers, aqueous bases, powder bases, or oily bases, thickeners, etc., may be necessary or desirable.

[0110] The present invention also includes pharmaceutical compositions containing, as an active ingredient, a JAK1 pathway inhibitor as described herein or a pharmaceutically acceptable salt thereof, in combination with one or more pharmaceutically acceptable carriers (excipients). In some embodiments, the compositions are suitable for topical administration. When preparing these compositions, the active ingredient is typically mixed with an excipient, diluted by the excipient, or encapsulated in such carriers, for example, in the form of capsules, pouches, paper or other containers. When the excipient functions as a diluent, it may be a solid, semi-solid or liquid material acting as a solvent, carrier or medium for the active ingredient. Thus, the compositions may be in the form of tablets, pills, powders, lozenges, sachets, caches, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solid or liquid media), for example, ointments, soft and hard gelatin capsules, suppositories, sterile injection solutions, and sterile packaging powders containing up to 10% by weight of the active compound.

[0111] When preparing a formulation, the active compound may be ground to provide an appropriate particle size before being combined with other components. If the active compound is substantially insoluble, it may be ground to a particle size of less than 200 mesh. If the active compound is substantially water-soluble, the particle size can be adjusted by grinding to, for example, about 40 mesh to provide a substantially uniform distribution in the formulation.

[0112] JAK1 pathway inhibitors can be milled using known milling procedures, such as wet milling, to obtain particle sizes suitable for tablet formation and other formulations. Precisely separated (nanoparticle) preparations of JAK1 selective inhibitors can be prepared by methods well known in the art; see, for example, International Patent Application WO2002 / 000196.

[0113] The composition can be formulated into unit dosage forms containing a set amount of the active ingredient in free or salt form. The term "unit dosage form" refers to a physically distinct unit suitable as a unit dose for human subjects and other mammals, each unit containing a predetermined amount of the active substance calculated to produce the desired therapeutic effect in relation to a suitable pharmaceutically acceptable excipient.

[0114] In the methods and uses of the present invention, similar dosages of the compounds described herein can be used.

[0115] Since active compounds can be effective across a wide range of doses, they are generally administered in pharmaceutically effective amounts. However, it should be understood that the actual amount of compound administered is usually determined by the physician, taking into account relevant circumstances including the symptoms being treated, the chosen route of administration, the compound actually administered, the individual patient's age, weight and response, and the severity of the patient's symptoms.

[0116] To prepare solid compositions such as tablets, the main active ingredient is mixed with pharmaceutically acceptable excipients to form a solid preliminary formulation composition containing a homogeneous mixture of the compounds of the present invention. When these preliminary formulation compositions are referred to as homogeneous, the active ingredient is typically uniformly dispersed throughout the composition, thereby allowing the composition to be easily subdivided into equally effective unit dosage forms such as tablets, pills, and capsules. Furthermore, this solid preliminary formulation can be subdivided into the aforementioned unit dosage forms, for example, containing about 0.1 to about 1000 mg of the active ingredient of the present invention.

[0117] The tablets or pills of the present invention can be provided with dosage forms that offer the advantage of prolonged action by coating or other methods of mixing. For example, the tablets or pills may contain internal and external dose components, the latter in the form of an envelope over the former. The two components can be separated by an enteric coating that resists disintegration in the stomach and allows the internal component to pass intact into the duodenum or delays its release. Various materials can be used for such enteric coatings or coatings, and such materials include many polymer acids, as well as mixtures of polymer acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0118] Liquid forms that may be incorporated for oral or injectable administration of the compounds and compositions of the present invention include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, emulsions flavored with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, and elixirs and similar pharmaceutical solvents.

[0119] Compositions for inhalation or gas infusion include solutions and suspensions in pharmaceutically acceptable aqueous solvents or organic solvents or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, compositions are administered by oral or nasal respiratory pathways for topical or systemic effects. Compositions may be sprayed using an inert gas. The sprayed solution may be inhaled directly from a spraying device, or the spraying device may be attached to a face mask, tent, or intermittent positive airway pressure (PAP) device. Compositions in solution, suspension, or powder form may be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0120] Topical formulations may contain one or more conventional carriers. In some embodiments, ointments may contain water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, etc. Carrier compositions for creams may be based on a combination of water, glycerol, and one or more other components, for example, glycerin monostearate, PEG-glycerin monostearate, and cetyl stearyl alcohol. Gels may be formulated using isopropyl alcohol and water, preferably in combination with other components such as glycerol, hydroxyethylcellulose, etc.

[0121] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of administration (such as prevention or treatment), the patient's condition, and the method of administration. For therapeutic use, a composition may be administered to a patient already suffering from the disease in an amount sufficient to cure or at least partially prevent the symptoms and complications of the disease. The effective dose depends on the patient's judgment, based on the disease condition being treated, as well as factors such as the severity of the disease, the patient's age, weight, and general condition.

[0122] The compositions administered to the patient may be in the form of the pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or aseptically filtered. Aqueous solutions may be packaged for immediate use or lyophilized, and the lyophilized preparations may be combined with a sterile aqueous carrier before administration. The pH of the compound preparations will typically be 3 to 11, more preferably 5 to 9, and most preferably 7 to 8. It will be understood that formulations of pharmaceutical salts can be obtained by using certain aforementioned excipients, carriers, or stabilizers.

[0123] The therapeutic dose of the compounds of the present invention may vary, for example, depending on the specific therapeutic use, the method of administering the compound, the patient's health and symptoms, and the judgment of the prescribing physician. The ratio or concentration of the compounds described herein in a pharmaceutical composition may vary depending on many factors, including the dose, chemical properties (e.g., hydrophobicity), and route of administration. The dose may depend on variables such as the type and progression of the disease or disorder, the overall health status of the particular patient, the relative bioefficacy of the selected compound, the formulation of the excipients, and the route of administration. The effective dose can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0124] The compositions of the present invention may further include one or more additional pharmaceuticals, such as chemotherapeutic agents, steroids, anti-inflammatory compounds, or immunosuppressants, examples of which are provided herein.

[0125] kit The present invention also includes, for example, a pharmaceutical kit useful for treating and / or preventing vitiligo, the kit including one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of the compound described herein. As will be apparent to those skilled in the art, such a kit may further include, as necessary, one or more of various conventional pharmaceutical kit components, such as containers having one or more pharmaceutically acceptable carriers, additional containers, etc. Instructions indicating the amount of the component to be administered, guidelines for administration, and / or guidelines for mixing the components may also be included in the kit, either as an insert or a label. [Examples]

[0126] The present invention is described in more detail by specific examples. The following examples are presented for illustrative purposes only and are not intended in any way to limit the scope of the invention. Those skilled in the art will readily recognize various non-essential parameters that can be changed or modified to produce essentially the same results. The compounds of the examples have been found to be JAK inhibitors by at least one assay described herein.

[0127] Example A: In vitro JAK kinase assay JAK1 pathway inhibitors that may be used to treat cytokine-related diseases or disorders will be tested for their inhibitory activity against JAK targets according to the following in vitro assay described in Park et al., Analytical Biochemistry 1999, 269, 94-104. The catalytic domains of human JAK1 (amino acids 837-1142), JAK2 (amino acids 828-1132), and JAK3 (amino acids 781-1124), which have an N-terminal His tag, were expressed in insect cells using baculovirus and purified. The catalytic activity of JAK1, JAK2, and JAK3 was assayed by measuring the phosphorylation of biotinylated peptides. Phosphorylated peptides were detected by homogeneous time-resolved fluorescence (HTRF). In a 40 μL reaction mixture containing enzymes, ATP, and 500 nM peptides in 50 mM Tris (pH 7.8) buffer containing 100 mM NaCl, 5 mM DTT, and 0.1 mg / mL (0.01%) BSA, the IC50 of each compound was measured for each kinase. 50 The following was measured: 1mM IC 50 For the measurement, the ATP concentration during the reaction was 1 mM. The reaction was carried out at room temperature for 1 hour, and then stopped with 45 mM EDTA, 300 nM SA-APC, and 6 nM Eu-Py20 in 20 μL of assay buffer (Perkin Elmer, Boston, MA). Binding to μ-Europium-labeled antibody was performed for 40 minutes, and the HTRF signal was measured on a Fusion plate reader (Perkin Elmer, Boston, MA). When the compounds listed in Table 3 were tested with this assay, the IC20 levels observed in Table 3 were observed. 50 It was shown that it has the value of [value].

[0128] Example B: In vitro JAK kinase assay using a melanocyte proliferation assay with 20% CD8+ T cell conditioned medium. This assay investigates an in vitro system using CD8+ T cells and melanocytes. It tests whether compound 1 modulates the inflammatory response and how such modulation affects melanocyte biology.

[0129] Methods: CD8+ T cell conditioned medium Steps: Normal human CD8+ T cells were purchased from peripheral blood (STEMCELL Technologies). These were cultured in an Incucyte Live-Cell Analysis System (Essen Bioscience) under activated and deactivated conditions at 37°C and 5% CO2 for 48 hours. Activated T cells were first cultured in flat-bottomed 96-well plates (Costar) coated with 10 μg / ml anti-CD3 (BD ​​Biosciences) at room temperature for 2 hours, then washed once with PBS before being added to the plates. They were grown in melanocyte growth medium (NHM-GM, MatTek) containing 1 μg / ml anti-CD28 (BDBiosciences). Deactivated T cells were treated in the same manner, except they were cultured in a separate plate without anti-CD3 and anti-CD28. Each T cell plate was treated with compound 1 (1000, 100, or 10 nM) or the medium alone at seeding. Each treatment was performed in triplicates. After 48 hours, all samples (T cells and culture medium) were transferred to a new V-bottom plate and centrifuged at 1300 RPM for 10 minutes. The supernatant was carefully collected from the T cell pellet, taking care not to disturb the pellet. The supernatant (CD8+ T cell conditioned medium) was used fresh for processing melanocytes. The remaining volume was stored at -80°C and thawed for protein analysis using Luminex technology (custom Procarta 45-plex, Thermo Fisher Scientific). The processed CD8+ T cell pellet was lysed using Quantigene sample processing kit reagents according to the kit instructions. The lysates were stored at -80°C and thawed for RNA analysis using Luminex technology with two custom Quantigene 80-Plex - a total of 150 target genes (Thermo Fisher Scientific).

[0130] Melanocyte proliferation step: Normal human melanocytes isolated from the foreskin epidermis of newborns of Black donors were purchased (MatTek). These were thawed according to the manufacturer's instructions and seeded in NHM-GM medium in flat-bottom 96-well plates (Costar) at 37°C and 5% CO2 for 18 hours. They were approximately 10% confluent when treated with 20% CD8+ T cell conditioned medium (25 μl of conditioned medium was added to 100 μl of NHM-GM already in the wells). These were cultured in an Incucyte Live-Cell Analysis System (Essen Bioscience) at 37°C and 5% CO2 for 48 hours, with 5 images per well taken every 3 hours. After 48 hours, all supernatant was transferred from the melanocyte plate to a new V-bottom plate and centrifuged at 1300 RPM for 10 minutes. The supernatant was collected, stored at -80°C, and thawed for protein analysis using Luminex technology (custom Procarta 45-plex, Thermo Fisher Scientific). The treated melanocytes were lysed using the Quantigene sample processing kit reagents according to the kit instructions. The lysates were stored at -80°C and thawed for RNA analysis using Luminex technology with two custom Quantigene 80-Plex reagents containing a total of 150 target genes (Thermo Fisher Scientific). Incucyte images were analyzed using Incucyte software to determine melanocyte confluence for each processing method.

[0131] Results: Figure 1 is a graph showing melanocyte proliferation, confluence % versus time (hours). Compound 1 does not directly alter melanocyte proliferation after transfer to 20% inactivated T cell medium. Addition of 20% medium from CD3 / CD28 activated human CD8+ T cells to NHM-GM in vitro cultures significantly inhibited melanocyte proliferation compared to inactivated conditions. Compound 1 (1000 nM) co-cultured with CD3 / CD28 activated human CD8+ T cells partially reversed the suppressed melanocyte proliferation response. This improvement in melanocyte proliferation was statistically significant. Addition of compound 1 to inflammatory medium resulted in a significant decrease in spontaneous proliferation compared to the innate melanocyte proliferation rate. This is a time-dependent decrease in the spontaneous proliferation response.

[0132] Figure 2 is a graph showing the effect of compound 1 on JAK1-dependent cytokines, using T cell compartments, protein T cell medium (pg / ml) versus specific JAK1-dependent cytokines. Consistent with previously reported data, compound 1, co-cultured with CD3 / CD28-activated human CD8+ T cells, inhibits JAK-STAT-dependent inflammatory cytokines.

[0133] Figures 3A-C are graphs showing the effects of compound 1 on growth factors and chemokines using T cell compartments, protein T cell medium (pg / ml) versus IL-8 (A), HGF (B), and LIF (C). Compound 1, co-cultured with CD3 / CD28 activated human CD8+ T cells, reduced several additional mediators that did not require the use of JAK1 signaling.

[0134] Figure 4 is a graph showing the effect of compound 1 on JAK1-dependent cytokines, using melanocytes, protein melanocyte medium (pg / ml) versus specific JAK1-dependent cytokines. Compound 1 reduces JAK-STAT-dependent inflammatory cytokines. Some of these cytokines are upregulated by melanocytes under stress conditions (IFNγ, CXCL10, IL-6). Figures 5A-C are graphs showing the effects of compound 1 on growth factors and chemokines using melanocytes, protein melanocyte medium (pg / ml) versus IL-8 (A), HGF (B), and LIF (C). IL-8 is upregulated by melanocytes under stress conditions and increases under compound 1 conditions.

[0135] Discussion: Hepatocyte growth factor (HGF) / mesenchymal epithelial transition factor (c-MET) signaling protects melanocytes from apoptosis and stimulates their proliferation and motility. After transferring activated T cell-conditioned medium, HGF concentration in melanocyte culture supernatant increased. Compound 1 significantly reduced HGF levels (by approximately 67%), but increased melanocyte proliferation. Based on the data (melanocytes and T cells), it is expected that the reduction in HGF expression by compound 1 via the signaling mechanism is detrimental to melanocyte proliferation.

[0136] Interleukin-8 (IL-8 / CXCL8) has been reported to enhance the melanocyte proliferative response to multiple growth factors through its binding to its congener receptors (CXCR1 and CXCR2). Both IL-8 production and CXCR1 / 2 signaling are independent of JAK1. Treatment of CD8+ T cells with compound 1 resulted in a slight decrease in IL-8 concentration (↓22%), but a significant increase (↑450%) in melanocyte cultures. Based on the data (melanocytes and T cells), an unexpected upregulation of IL-8 levels in melanocyte cultures was observed, suggesting that this enhances the proliferative response.

[0137] Leukemia suppressor (LIF) increases the proliferation and differentiation of primary melanocytes. Since LIF receptor signaling is JAK1-dependent, compound 1 is expected to have a detrimental effect on melanocyte proliferation. Unexpectedly, melanocyte proliferation was increased. Based on the data (T cells), on-target inhibition of LIF is not consistent with the enhancement of the melanocyte proliferation response.

[0138] Compound 1 can downregulate inflammatory cytokines in the CD8+ activated T cell microenvironment. It normalizes melanocyte proliferation and also stops the production of inflammatory mediators from melanocytes.

[0139] Example C: Phase 2 test of compound 1 Research design Figure 6 shows a summary of a phase 2 randomized, double-blind, placebo-controlled dose-range trial regarding the efficacy and safety of compound 1. The trial will be conducted using compound 1 to treat participants with non-segmental vitiligo. This trial may include a 28-week double-blind extension period for participants with vitiligo. Participants may include individuals with non-segmental vitiligo with a depigmentation area of ​​systemic BSA ≥ 8%, T-VASI ≥ 8%, facial BSA ≥ 0.5%, and F-VASI ≥ 0.5. Participants will be stratified based on systemic BSA involvement (8–20% and >20%). In some embodiments, participants may include individuals with non-segmental vitiligo with a depigmentation area of ​​systemic BSA ≥ 5%, including T-VASI ≥ 5%, facial BSA ≥ 0.5%, and F-VASI ≥ 0.5. Participants will be stratified based on systemic BSA involvement (8–20% and >20%). This study includes men and women aged 18 to 75.

[0140] In a 24-week placebo-controlled, double-blind period, approximately 160 participants are randomly assigned in a 1:1:1:1 ratio to one of three treatment groups (dose A, dose B, or dose C) or to the placebo group. A 28-week double-blind extension includes all participants who successfully complete the placebo-controlled period. In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered at approximately dose B per day, based on free bases. In some embodiments, the JAK1 pathway inhibitor or a pharmaceutically acceptable salt thereof is administered at approximately dose C per day, based on free bases. The primary endpoint is the percentage change from baseline in the Total Vitiligo Area Scoring Index (T-VASI) at week 24, and a key secondary endpoint is the percentage of participants achieving T-VASI 50 at week 24. In some embodiments, the results of this trial will help inform dose selection to be evaluated in a Phase 3 trial.

[0141] In some embodiments, the criteria for exclusion of participants from the study include any one of the following: other depigmentation disorders of the skin (e.g., macular baldness, pityriasis leucoma, leprosy, post-inflammatory hypopigmentation, progressive macular hypomelanosis, anemic nevus, chemical albinism, and tinea versicolor); uncontrolled thyroid function at screening as determined by the principal investigator (Note: If a participant has a history of thyroid disease and is receiving treatment, the participant must have received stable thyroid therapy for at least three months prior to day 1); use of laser or light-based treatment (phototherapy), including tanning beds, within eight weeks prior to day 1; use of dihydroxyacetone (typically found in self-tanning products) within four weeks prior to day 1. Current or past use of hydroquinone bleaching agents such as Benoquin® (monobenzone) monobenzyl ether; a history of melanocyte keratinocyte transplantation (MKTP) or other surgical treatment for vitiligo; spontaneous and significant repigmentation within 6 months prior to screening (e.g., significant repigmentation in an amount determined by the researcher without treatment); pregnant women, women considering pregnancy, or breastfeeding women. History of treatment failure for vitiligo or other inflammatory conditions with systemic or topical JAK or TYK2 inhibitors (e.g., ruxolitinib, tofacitinib, baricitinib, filgotinib, restaurtinib, pacritinib, abrocitinib, prepocitinib); current use of anticoagulants or medications known to cause thrombocytopenia; hepatitis: evidence of HBV or HCV infection or risk of reactivation (participants must not be positive for HBV DNA, HCV RNA, HBsAg, or anti-hepatitis B core antibodies, however, participants with no history of HBV infection and who have received the HBV vaccine are eligible); individuals with positive antibodies against HBV surface antigen (anti-HBs+) as the sole evidence of past exposure may participate in the study; known HIV infection.Known hypersensitivity or severe reaction to compound 1 or its excipients; the investigator's opinion that the participant is unable or unlikely to be able to follow the dose schedule and study evaluations; and, at the discretion of the principal investigator, the administration of the investigational drug or attendance at required trial visits; any condition that poses a significant risk to the participant or interferes with full participation in the trial, including interference with the interpretation of study data.

[0142] In some embodiments, criteria for participant exclusion from the study include participants with a history or co-existence of other conditions, including any one of the following: a history of thrombocytopenia, coagulation disorders, platelet dysfunction, or thrombotic events; a clinically significant condition other than vitiligo as determined by the researcher that is not adequately controlled with appropriate treatment or that may interfere with the course, severity, or evaluation of this study; any other active skin disease or condition that may interfere with the course, severity, or evaluation of this study; a participant will be an unsuitable candidate for the study if, based on the researcher's clinical assessment, they have a bacterial, fungal, or viral infection; a current herpes zoster infection, a history of disseminated herpes simplex, or a history of herpes zoster; a history of malignancies, including melanoma, lymphoma, and leukemia, within 5 years prior to day 1, excluding non-metastatic cutaneous squamous cell carcinoma, basal cell carcinoma, or focal cervical epithelial carcinoma that has been successfully treated; and albinism.

[0143] In some embodiments, the criteria for exclusion of participants from the study include receiving the drug or investigational drug within the following intervals prior to day 1 (first dose of the investigational drug): <12 weeks or 5 half-lives (if known); longer for topical or systemic JAK or TYK2 inhibitors; less than 12 weeks or 5 half-lives (if known) for research or experimental treatments; less than 12 weeks or 5 half-lives (if known) for systemic immunosuppressive or immunomodulatory biological agents (e.g., egadalimumab, etanercept, infliximab, golimumab, certolizumab, ustekinumab, secukinumab, brodalumab, ikikizumab, risankizumab, guselkumab, bimekizumab, iscarimab, vermekimab, rituximab, anakinra). Live vaccine administration is scheduled less than 6 weeks prior to the trial, or within 6 weeks of the last dose of the investigational drug during the trial period or after the last dose of the investigational drug. For systemic immunosuppressants or immunomodulatory small molecule drugs (e.g., oral or injectable corticosteroids, methotrexate, cyclosporine, dapsone, azathioprine), the interval is less than 4 weeks. For oral or topical PDE-4 inhibitors (e.g., apremilast, crisabolol), the interval is less than 3 weeks. For topical application of medication to vitiligo lesions, the interval is less than 2 weeks. For over-the-counter (OTC) therapies used to treat vitiligo, the interval is less than 2 weeks. For potent and moderate systemic CYP3A4 inhibitors and potent systemic CYP3A4 inducers, the interval is less than 2 weeks or 5 half-lives (if known), whichever is longer. Examples of medications that may cause adverse reactions include, but are not limited to, erythromycin, rifampicin / rifampin, ciprofloxacin, some azole antifungals (such as ketoconazole and fluconazole), nefazodone, St. John's wort, diltiazem, mibeflazil, verapamil, grapefruit / grapefruit juice, and Seville orange; and in the case of antiplatelet drugs, a duration of less than one week.

[0144] In some embodiments, criteria for exclusion of participants from the study include the use of systemic immunosuppressants or immunomodulatory small molecule drugs. Note: Use of corticosteroid inhalers and intranasal sprays is permitted; use of oral corticosteroids for non-skin conditions (e.g., asthma exacerbations, bronchitis) is permitted for less than 7 days if deemed acceptable by the principal investigator and sponsor; use of topical corticosteroids for skin conditions other than vitiligo (e.g., atopic dermatitis and psoriasis) is permitted in areas not treated for vitiligo (the total involvement of BSA in other skin conditions other than the area treated for vitiligo should not exceed 10%).

[0145] In some embodiments, criteria for exclusion from the study include the use of systemic immunosuppressive or immunomodulatory small molecule drugs, but it should be noted that, at the discretion of the researchers, low doses of acetylsalicylic acid (≤100 mg QD) may be permitted for cardiovascular prophylaxis.

[0146] In some embodiments, the criteria for exclusion of participants from the study include evidence of an active, latent, or poorly treated infection with Mycobacterium tuberculosis (i.e., TB) as defined by at least one of the following: a positive QuantiFERON®-TB GoldIn-Tube test performed within 12 weeks up to day 1. Tests (QFT-GIT) or positive Mantoux / PPD tuberculin skin tests are excluded, and a negative test is required to be eligible, unless other criteria described here apply (Participants with a history of Calmette-Guérin vaccination are recommended to undergo QFT-GIT testing as vaccination may cause a positive Mantoux / PPD tuberculin skin test (Note: QFT-GIT or Mantoux / PPD tuberculin skin test is not required if the participant has previously received a documented appropriate course of treatment for latent or active tuberculosis infection); History of untreated or inadequately treated latent or active tuberculosis infection; Participant has latent tuberculosis infection (initial polydrug complications) If you have previously received an appropriate course of treatment for either 9 months of isoniazid administration or an acceptable alternative regimen in a region with a tuberculosis resistance rate of <5%, or active tuberculosis infection (an acceptable multidrug regimen), neither the QFT-GIT nor the Mantoux / PPD tuberculin skin test is required, but a chest X-ray or other appropriate diagnostic imaging performed within 3 months up to day 1 is required (to be considered for the study, participants must have negative X-rays(or more) for active tuberculosis infection as determined by a qualified radiologist, have a record of appropriate treatment for tuberculosis, and have obtained negative chest X-ray(or more) results by day 1); and participants currently receiving treatment for active tuberculosis infection.

[0147] In some embodiments, the criteria for excluding participants from the study include participants having the test values ​​at the time of screening as defined in Table 4. [Table 4]

[0148] Example D: In vitro JAK kinase assay using a melanocyte proliferation assay with 20% CD8+ T cell-conditioned medium. This assay investigates an in vitro system using CD8+ T cells and melanocytes. It tests whether ruxolitinib modulates the inflammatory response and how such modulation affects melanocyte biology.

[0149] Methods: CD8+ T cell conditioned medium Steps: Normal human CD8+ T cells were purchased from peripheral blood (STEMCELL Technologies). These were cultured in an Incucyte Live-Cell Analysis System (Essen Bioscience) under activated and deactivated conditions at 37°C and 5% CO2 for 48 hours. Activated T cells were first cultured in flat-bottomed 96-well plates (Costar) coated with 10 μg / ml anti-CD3 (BD ​​Biosciences) at room temperature for 2 hours, then washed once with PBS before being added to the plates. They were grown in melanocyte growth medium (NHM-GM, MatTek) containing 1 μg / ml anti-CD28 (BD Biosciences). Deactivated T cells were treated in the same manner, except they were cultured in a separate plate without anti-CD3 and anti-CD28. Each T cell plate was treated with ruxolitinib (1000, 100, or 10 nM) or the medium alone at seeding. Each treatment was performed in triplicates. After 48 hours, all samples (T cells and culture medium) were transferred to new V-bottom plates and centrifuged at 1300 RPM for 10 minutes. The supernatant was carefully collected from the T cell pellet, taking care not to disturb the pellet. The supernatant (CD8+ T cell conditioned medium) was used fresh for processing melanocytes. The remaining volume was stored at -80°C and thawed for protein analysis using Luminex technology (custom Procarta 45-plex, Thermo Fisher Scientific). The processed CD8+ T cell pellet was lysed using Quantigene sample processing kit reagents according to the kit instructions. The lysates were stored at -80°C and thawed for RNA analysis using Luminex technology with two custom Quantigene 80-Plex - a total of 150 target genes (Thermo Fisher Scientific).

[0150] Melanocyte proliferation step: Normal human melanocytes isolated from the foreskin epidermis of newborns of Black donors were purchased (MatTek). These were thawed according to the manufacturer's instructions and seeded in NHM-GM medium in flat-bottom 96-well plates (Costar) at 37°C and 5% CO2 for 18 hours. They were approximately 10% confluent when treated with 20% CD8+ T cell conditioned medium (25 μl of conditioned medium was added to 100 μl of NHM-GM already in the wells). These were cultured in an Incucyte Live-Cell Analysis System (Essen Bioscience) at 37°C and 5% CO2 for 48 hours, with 5 images per well taken every 3 hours. After 48 hours, all supernatant was transferred from the melanocyte plate to a new V-bottom plate and centrifuged at 1300 RPM for 10 minutes. The supernatant was collected, stored at -80°C, and thawed for protein analysis using Luminex technology (custom Procarta 45-plex, Thermo Fisher Scientific). The treated melanocytes were lysed using the Quantigene sample processing kit reagents according to the kit instructions. The lysates were stored at -80°C and thawed for RNA analysis using Luminex technology with two custom Quantigene 80-Plex reagents containing a total of 150 target genes (Thermo Fisher Scientific). Incucyte images were analyzed using Incucyte software to determine melanocyte confluence for each processing method.

[0151] Results: Figure 7 is a graph showing melanocyte proliferation, confluence % versus time (hours). Addition of 20% medium from CD3 / CD28-activated human CD8+ T cells to NHM-GM in vitro cultures significantly inhibited melanocyte proliferation compared to inactivated conditions. Ruxolitinib (1000 nM) co-cultured with CD3 / CD28-activated human CD8+ T cells appeared to completely reverse the suppressed melanocyte proliferation response. This improvement in melanocyte proliferation was statistically significant. Ruxolitinib added to inflammatory media resulted in a significant decrease in spontaneous proliferation compared to the innate melanocyte proliferation rate. This is a time-dependent decrease in the spontaneous proliferation response.

[0152] Figure 8 is a graph showing the effect of ruxolitinib on JAK1-dependent cytokines, using T cell compartments, protein T cell medium (pg / ml) versus specific JAK1-dependent cytokines. Consistent with previously reported data, ruxolitinib co-cultured with CD3 / CD28-activated human CD8+ T cells inhibits JAK-STAT-dependent inflammatory cytokines, with the exception of the upregulated inflammatory cytokine IL-2 (it appears to have no effect on inflammatory cytokines IL-4, IL-5 (note that IL-5 is JAK2 / 2), and IL-6).

[0153] Figures 9A and 9B are graphs showing the effects of ruxolitinib on growth factors and chemokines using T cell compartments, protein T cell medium (pg / ml) versus IL-8 (A), and LIF (B). Ruxolitinib co-cultured with CD3 / CD28 activated human CD8+ T cells reduced several additional mediators that did not require the use of JAK1 signaling.

[0154] Figure 10 is a graph showing the effect of ruxolitinib on JAK1-dependent cytokines, using melanocytes, protein melanocyte medium (pg / ml) versus specific JAK1-dependent cytokines. Ruxolitinib reduces JAK-STAT-dependent inflammatory cytokines (it appears to have no effect on inflammatory cytokines IL-13 and IL-10). Some of these cytokines are upregulated by melanocytes under stress conditions (IFNγ, CXCL10, IL-6). Figures 11A and 11B are graphs showing the effects of ruxolitinib on growth factors and chemokines using melanocytes, protein melanocyte medium (pg / ml) versus IL-8 (A), and LIF (B). IL-8 is upregulated by melanocytes under stress conditions and decreases under ruxolitinib conditions.

[0155] Discussion: Interleukin-8 (IL-8 / CXCL8) has been reported to enhance the melanocyte proliferative response to multiple growth factors through its binding to its congener receptors (CXCR1 and CXCR2). Both IL-8 production and CXCR1 / 2 signaling are independent of JAK1. Ruxolitinib treatment of CD8+ T cells slightly reduced IL-8 levels.

[0156] Leukemia suppressor (LIF) increases the proliferation and differentiation of primary melanocytes. Since LIF receptor signaling is JAK1-dependent, ruxolitinib was expected to have a detrimental effect on melanocyte proliferation. Unexpectedly, melanocyte proliferation increased. Based on the data (T cells), on-target inhibition of LIF is not consistent with the enhancement of the melanocyte proliferation response.

[0157] Ruxolitinib can downregulate inflammatory cytokines in the CD8+ activated T cell microenvironment. It normalizes melanocyte proliferation and also stops the production of inflammatory mediators from melanocytes.

[0158] In addition to those described herein, various modifications of the present invention will be apparent to those skilled in the art from the foregoing description. Such modifications are also intended to fall within the scope of the appended claims. All references, including all patents, patent applications, and publications cited herein, are incorporated herein by reference in their entirety.

Claims

1. 1. A pharmaceutical composition for treating vitiligo in a subject, comprising a JAK1 pathway inhibitor or a pharma- ceutical acceptable salt thereof.

2. 2. The pharmaceutical composition of claim 1, wherein the JAK1 pathway inhibitor or a pharma- ceutically acceptable salt thereof is selective for JAK1 over JAK2, JAK3, and Tyk2.

3. 2. The pharmaceutical composition of claim 1, wherein the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide or a pharma- ceutically acceptable salt thereof.

4. The pharmaceutical composition of claim 1, wherein the JAK1 pathway inhibitor is a pharma- ceutically acceptable salt of 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide.

5. 5. The pharmaceutical composition of claim 4, wherein the JAK1 pathway inhibitor is 4-[3-(cyanomethyl)-3-(3',5'-dimethyl-1H,1'H-4,4'-bipyrazol-1-yl)azetidin-1-yl]-2,5-difluoro-N-[(1S)-2,2,2-trifluoro-1-methylethyl]benzamide phosphate.

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the vitiligo is non-segmental vitiligo.

7. 7. The pharmaceutical composition of any one of claims 1 to 6, wherein the JAK1 pathway inhibitor, or a pharma- ceutical acceptable salt thereof, is configured to be administered in an amount of about 5 mg to about 95 mg per day on a free base basis.

8. The pharmaceutical composition of any one of claims 1 to 6, wherein the JAK1 pathway inhibitor or a pharma- ceutically acceptable salt thereof is configured to be administered in an amount of about 10 mg to about 80 mg per day on a free base basis.

9. The pharmaceutical composition of any one of claims 1 to 6, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is configured to be administered in an amount of about 15 mg per day on a free base basis.

10. The pharmaceutical composition of any one of claims 1 to 6, wherein the JAK1 pathway inhibitor or a pharma- ceutically acceptable salt thereof is configured to be administered in an amount of about 45 mg per day on a free base basis.

11. The pharmaceutical composition of any one of claims 1 to 6, wherein the JAK1 pathway inhibitor or a pharma- ceutically acceptable salt thereof is configured to be administered in an amount of about 75 mg per day on a free base basis.

12. The pharmaceutical composition of any one of claims 1 to 6, wherein the JAK1 pathway inhibitor or a pharma- ceutically acceptable salt thereof is configured to be administered in an amount of about 30 mg per day on a free base basis.

13. The pharmaceutical composition of any one of claims 1 to 6, wherein the JAK1 pathway inhibitor or a pharma- ceutically acceptable salt thereof is configured to be administered in an amount of about 60 mg per day on a free base basis.

14. The pharmaceutical composition of any one of claims 1 to 13, wherein the JAK1 pathway inhibitor, or a pharma- ceutically acceptable salt thereof, is adapted to be administered in combination with a further therapeutic agent.

15. 15. The pharmaceutical composition of claim 14, wherein the additional therapeutic agent comprises a Janus kinase inhibitor.

16. 16. The pharmaceutical composition of claim 15, wherein the Janus kinase inhibitor comprises ruxolitinib or a pharma- ceutically acceptable salt thereof.

17. The pharmaceutical composition of any one of claims 1 to 16, wherein the JAK1 pathway inhibitor or a pharma- ceutically acceptable salt thereof is configured to be administered together with at least one pharma- ceutically acceptable carrier or excipient.

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