Biomarkers for predicting therapeutic efficacy of GPCR19 agonists in treating atopic dermatitis and methods of treating subjects with these biomarkers

Abstract

Provided herein are biomarkers useful for predicting the efficacy of a composition comprising a GPCR19 agonist in treating atopic dermatitis, and methods for identifying subjects having threshold levels of the biomarkers and treating atopic dermatitis.

Description

[Technical Field]

[0001] This application claims priority to U.S. Provisional Application No. 63 / 599,511, filed November 15, 2023, which claims the benefit of Korean Patent Application Publication No. 10-2022-0152832, filed November 15, 2022, the entire text of each of which is incorporated herein by reference.

[0002] The present disclosure relates to biomarkers useful for predicting the efficacy of a composition containing a GPCR19 agonist for treating atopic dermatitis (AD) in a subject, and methods for identifying subjects exhibiting threshold levels of these biomarkers and treating atopic dermatitis. [Background technology]

[0003] Atopic dermatitis (AD) is a chronic, recurrent skin disease characterized by intense itching. AD exhibits highly heterogeneous clinical phenotypes due to genetic and environmental factors / interactions. The incidence of AD is rapidly increasing worldwide due to increased exposure to environmental pollutants, with the prevalence reaching 20% ​​of the total population. Patients with AD experience reduced quality of life due to restrictions in their daily lives, and the economic burden associated with treatment is also significant. Therefore, measures to effectively manage AD are urgently needed.

[0004] AD typically begins in infancy or childhood and is often associated with familial dynamics. Symptoms generally begin in early childhood, especially around 2 months of age, with approximately 50% of cases occurring before the age of 2 years. While most symptoms appear before the age of 5 years, early symptoms rarely appear in adulthood. In some patients, symptoms improve or resolve spontaneously as they grow older, with more than half of patients with onset of symptoms in infancy showing improvement by the age of 2 years.

[0005] The appearance and distribution of skin lesions in AD are characteristic, with itching (pruritus), dry skin, and a unique eczema. In infants, eczema occurs primarily on the face and extremities, and as they grow, it tends to appear on the crooks of the arms and behind the knees. In adults, lichenification, a process of thickening of the skin due to repeated scratching, frequently occurs in skin creases. Eczema can appear on the face, chest, and neck, as well as the extremities, which differs from the pattern observed in children and infants.

[0006] AD acts as a precursor to allergic diseases such as allergic asthma and rhinitis. However, due to a lack of accurate understanding of the exact cause and mechanism of AD, no definitive treatment has yet been developed. Currently, AD is treated with steroids, PDE4 inhibitors, JAK inhibitors, and antibody therapeutics. Anti-inflammatory drugs and steroid-based immunosuppressants are primarily used to alleviate inflammation and regulate immune responses. While these therapeutic agents have the advantage of rapidly alleviating symptoms, reducing their dosage or discontinuing their use can rapidly worsen symptoms. Furthermore, long-term use of these therapeutic agents can cause secondary systemic side effects such as adrenal insufficiency, diabetes, digestive disorders, ulcers, hirsutism, alopecia, and pigmentation disorders. Cataracts may occur, particularly in children. Steroid ointments can cause serious side effects such as thinning or atrophy of the skin, redness due to vasodilation, and folliculitis. Nonsteroidal immunomodulators, such as pimecrolimus cream and tacrolimus ointment, were developed as alternatives to steroid ointments and have been shown to have fewer side effects associated with existing steroid ointments, even with long-term use. These nonsteroidal immunomodulators are frequently used on sensitive skin areas, such as the neck, and have quickly gained popularity, accounting for approximately 30% of the total atopic dermatitis market. However, concerns about the potential carcinogenic effects of calcineurin inhibitors have led to a decline in sales. As a result, only low-concentration use is recommended for patients under 16 years of age, and low-concentration formulations are not approved for use in children under 2 years of age.

[0007] Crisaborole, a PDE4 inhibitor approved by the FDA in 2016, showed some improvement compared to placebo for patients with mild to moderate severity, but this was not enough to address the unmet medical need. It has also been reported that many patients avoid prescriptions due to the drug-related burning sensation. JAK inhibitors are among the most extensively studied drugs for the treatment of atopic dermatitis. Currently, ruxolitinib, abrocitinib, upadacitinib, and baricitinib are FDA-approved. However, safety issues have been raised during clinical trials for this group of drugs, including serious infections, deaths, cancers, major cardiovascular events, and blood clots. Some JAK inhibitors carry boxed warnings restricting their use, and some are still awaiting review and approval. Due to these safety concerns, the FDA has expressed reservations about the entire JAK inhibitor class, recommending their use only in select patients. In particular, in September 2021, the FDA decided to require certain manufacturers' JAK inhibitor labels to include information about serious side effects and deaths. This has led many experts to suggest restricting the use of JAK inhibitors to the treatment of atopic dermatitis, a non-life-threatening condition.

[0008] Meanwhile, dupilumab, the first monoclonal antibody therapy, received FDA approval in 2017 for patients with moderate to severe atopic dermatitis. Dupilumab controls the symptoms of severe allergic diseases such as atopy and asthma. However, more than 10% of dupilumab users reported a gradual decrease in efficacy and injection site redness due to increased blood levels of antibodies against the drug. Additionally, 1% to 10% of dupilumab users reported insomnia, oral viral infections, gastritis, toothache, eosinophilia, herpes infections, conjunctivitis, sore throat, and joint pain. Less than 1% reported hypersensitivity reactions, xerophthalmia, and eosinophilic side effects such as granulomatosis, eosinophilic pneumonia, and erythema nodosum. Due to these safety and efficacy issues, its use is not recommended for children and adolescents. Additionally, dupilumab is expensive and is only covered by insurance for the treatment of patients who have not responded to steroids, calcineurin inhibitors, crisaborole, and systemic treatments. As a result, patients' access to dupilumab treatment is very limited.

[0009] Other atopic dermatitis treatments include antihistamines, short-term corticosteroid and topical application in severe cases, ultraviolet light therapy, and interferon therapy. However, in most cases, only temporary improvement is observed, and AD recurs when drug treatment is discontinued. Therefore, for most atopic dermatitis patients, there is a pressing need for the development of more effective treatment and prevention drugs with fewer side effects than existing treatments.

[0010] Therefore, there is a need for improved methods of treating atopic dermatitis, particularly for the development of effective treatments with fewer side effects. Summary of the Invention [Problem to be solved by the invention]

[0011] Accordingly, the present disclosure relates to biomarkers for predicting the therapeutic efficacy of GPCR19 agonists in the treatment of atopic dermatitis, and methods for treating subjects with these biomarkers that substantially overcome one or more of the problems caused by the limitations and shortcomings discussed above.

[0012] Additional features and advantages of the present disclosure will be set forth in the description that follows, and in part will be obvious from the description, or may be learned by practice of the present disclosure. Other advantages of the present disclosure will be realized and attained by the structure particularly pointed out in the description, claims and appended drawings. [Means for solving the problem]

[0013] To achieve these and other advantages, as embodied and broadly described by the present disclosure, there is provided a method for predicting the efficacy of a composition comprising a GPCR19 agonist for treating atopic dermatitis in a subject, the method comprising the steps of providing concentrations of one or more biomarkers in a sample from the subject and comparing each concentration to a threshold level, wherein a concentration of at least one of the one or more biomarkers at a threshold level indicates that the composition may be more effective in treating atopic dermatitis, and wherein the one or more biomarkers are selected from the group consisting of IGHA2, ENTP6, SMOC1, ENPL, and CRK.

[0014] In another aspect of the present disclosure, there is provided a method for predicting the efficacy of a composition comprising a GPCR19 agonist for treating atopic dermatitis in a subject, the method comprising the steps of: obtaining a sample from the subject; quantifying the concentration of each of one or more biomarkers in the sample; and comparing each concentration with a threshold level; wherein if the concentration of at least one of the one or more biomarkers is at a threshold level, the composition is more likely to be effective in treating atopic dermatitis, and the one or more biomarkers are selected from the group consisting of IGHA2, ENTP6, SMOC1, ENPL, and CRK.

[0015] In another aspect of the present disclosure, there is provided a composition comprising TDCA for use in treating atopic dermatitis in a subject, wherein the subject suffers from type A or type A' atopy.

[0016] In yet another aspect of the present disclosure, there is provided a use of a composition comprising TDCA for producing a medicament for treating atopic dermatitis in a subject, wherein the subject is characterized by having type A atopy or type A' atopy.

[0017] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the same disclosure as claimed. [Brief explanation of the drawings]

[0018] The accompanying drawings are included to facilitate an understanding of the disclosure, are integrated into and constitute a part of the disclosure, illustrate various aspects of the disclosure, and together with the description, serve to explain the principles of the disclosure.

[0019] In the drawings:

[0020] [Figure 1] FIG. 1 is a schematic diagram of the enrollment, randomization, and inclusion in the primary analysis of a clinical trial for taurodeoxycholic acid (TDCA), a GPCR19 agonist disclosed herein (in a dosage form referred to herein as NUGEL®).

[0021] [Figure 2](A)-(B) show the results of an analysis of the Eczema Area and Severity Index (EASI) and a validated investigator's global assessment (IGA) in patients with AD whose baseline blood SMOC1 levels were 30 pM or higher. In (A), the primary endpoint was defined as the percent change in EASI from baseline (Change EASI). In (B), the secondary endpoint was defined as the percent change in IGA from baseline (Change IGA). Change EASI and Change IGA were compared between the placebo group, the 0.3% taurodeoxycholic acid (TDCA) gel group, and the 0.5% TDCA gel group. A subgroup of patients with baseline blood SMOC1 levels of 30 pM or higher was analyzed. The number (n) and proportion (%) of patients analyzed in each group were the placebo group (n = 12, 46%), the 0.3% TDCA gel group (n = 12, 50%), and the 0.5% TDCA gel group (n = 8, 40%). Boxes indicate the median and interquartile range (1st and 3rd quartiles), and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. †P values ​​were determined by ANOVA test. ‡P values ​​were determined by rank sum test and mean difference (Mean Diff.) of the results between the placebo group and the 0.5% TDCA gel group.

[0022] [Figure 3](A)-(B) show the results of EASI and IGA analysis in AD patients with baseline CRK blood concentrations of 4518.2 pM or less. In (A), the primary outcome variable was defined as Change in EASI. In (B), the secondary outcome variable was defined as Change in IGA. Change in EASI and Change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. A subgroup of patients with baseline CRK blood concentrations of 4518.2 pM or less was analyzed. The number (n) and proportion (%) of patients analyzed in each group were the placebo group (n = 18, 69%), the 0.3% TDCA gel group (n = 12, 50%), and the 0.5% TDCA gel group (n = 6, 30%). †P values ​​were determined by ANOVA test. ‡P values ​​were determined by rank sum test and mean difference of the results between the placebo group and the 0.5% TDCA gel group.

[0023] [Figure 4] (A)-(B) show the results of EASI and IGA analysis in AD patients with baseline ENTP6 blood concentrations of 650 pM or less. In (A), the primary outcome variable was defined as Change in EASI. In (B), the secondary outcome variable was defined as Change in IGA. Change in EASI and Change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. A subgroup of patients with baseline ENTP6 blood concentrations of 650 pM or less was analyzed. The number (n) and percentage (%) of patients analyzed were the placebo group (n = 12, 46%), the 0.3% TDCA gel group (n = 13, 54%), and the 0.5% TDCA gel group (n = 5, 25%). Boxes indicate the median and interquartile range, and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. †P values ​​were determined by ANOVA test. ‡P values ​​were determined by rank sum test and mean difference of the results between the placebo group and the 0.5% TDCA gel group.

[0024] [Figure 5] (A)-(B) show the results of EASI and IGA analysis in AD patients with baseline IGHA2 blood concentrations ≥ 149,579 pM. In (A), the primary outcome variable was defined as Change in EASI. In (B), the secondary outcome variable was defined as Change in IGA. Change in EASI and Change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. A subgroup of patients with baseline IGHA2 blood concentrations ≥ 149,579 pM was analyzed. The number (n) and percentage (%) of patients analyzed were the placebo group (n = 11, 42%), the 0.3% TDCA gel group (n = 16, 67%), and the 0.5% TDCA gel group (n = 11, 55%). †P values ​​were determined by ANOVA test. ‡P values ​​were determined by rank sum test and mean difference of the results between the placebo group and the 0.5% TDCA gel group.

[0025] [Figure 6](A)-(B) show the results of EASI and IGA analysis in AD patients with baseline ENPL blood concentrations of 148.8 pM or less. In (A), the primary outcome variable was defined as Change in EASI. In (B), the secondary outcome variable was defined as Change in IGA. Change in EASI and Change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. A subgroup of patients with baseline ENPL blood concentrations of 148.8 pM or less was analyzed. The number (n) and percentage (%) of patients analyzed were the placebo group (n = 7, 29%), the 0.3% TDCA gel group (n = 8, 34%), and the 0.5% TDCA gel group (n = 6, 30%). †P values ​​were determined by ANOVA test. ‡P values ​​were determined by rank sum test and mean difference of the results between the placebo group and the 0.5% TDCA gel group.

[0026] [Figure 7] 1 is a Venn diagram showing classification criteria for atopic dermatitis patients classified by blood biomarkers.

[0027] [Figure 8](A)-(B) show the results of EASI and IGA analysis in patients with AD classified as type A atopy. In (A), the primary evaluation variable was defined as change in EASI. In (B), the secondary evaluation variable was defined as change in IGA. Change in EASI and change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. Subgroups with baseline SMOC1 blood concentrations of 30 pM or higher or baseline CRK blood concentrations of 4518.2 pM or lower were analyzed. The number (n) and proportion (%) of patients analyzed were the placebo group (n = 20, 77%), the 0.3% TDCA gel group (n = 21, 88%), and the 0.5% TDCA gel group (n = 10, 50%). †P values ​​were determined by ANOVA test. ‡P values ​​were determined by rank sum test and mean difference of the results between the placebo group and the 0.5% TDCA gel group.

[0028] [Figure 9] (A)-(B) show the results of EASI and IGA analysis in patients with AD classified as type A' atopy. In (A), the primary evaluation variable was defined as change in EASI. In (B), the secondary evaluation variable was defined as change in IGA. Change in EASI and change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. A subgroup with baseline blood ENTP6 concentrations of 650 pM or less was analyzed based on type A atopy. The number (n) and proportion (%) of patients analyzed were the placebo group (n = 9, 35%), the 0.3% TDCA gel group (n = 12, 50%), and the 0.5% TDCA gel group (n = 5, 25%). †P values ​​were determined by ANOVA. ‡P values ​​were determined by rank sum test and mean difference of results between the placebo and 0.5% TDCA gel groups.

[0029] [Figure 10] (A)-(B) show the results of EASI and IGA analysis in AD patients with baseline SMOC1 blood concentrations of 30 pM or higher or ENTP6 blood concentrations of 650 pM or lower. In (A), the primary evaluation variable was defined as Change in EASI. In (B), the secondary evaluation variable was defined as Change in IGA. Change in EASI and Change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. Subgroups of patients with baseline SMOC1 blood concentrations of 30 pM or higher or ENTP6 blood concentrations of 650 pM or lower were analyzed. The number (n) and proportion (%) of patients analyzed were the placebo group (n=18, 69%), the 0.3% TDCA gel group (n=19, 79%), and the 0.5% TDCA gel group (n=9, 45%). †P values ​​were determined by ANOVA test. ‡P values ​​were determined by rank sum test and mean difference of the results between the placebo group and the 0.5% TDCA gel group.

[0030] [Figure 11](A)-(B) show the results of EASI and IGA analysis in AD patients with baseline ENTP6 blood concentrations of 650 pM or less or baseline CRK blood concentrations of 4518.2 pM or less. In (A), the primary outcome variable was defined as Change in EASI. In (B), the secondary outcome variable was defined as Change in IGA. Change in EASI and Change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. Subgroups of patients with baseline ENTP6 blood concentrations of 650 pM or less or baseline CRK blood concentrations of 4518.2 pM or less were analyzed. The number (n) and proportion (%) of patients analyzed were the placebo group (n=21, 81%), the 0.3% TDCA gel group (n=16, 67%), and the 0.5% TDCA gel group (n=8, 40%). †P values ​​were determined by ANOVA test. ‡P values ​​were determined by rank sum test and mean difference of the results between the placebo group and the 0.5% TDCA gel group.

[0031] [Figure 12](A)-(B) show the results of EASI and IGA analyses of AD patients with baseline SMOC1 blood concentrations ≥30 pM, ENTP6 blood concentrations ≤650 pM, or CRK blood concentrations ≤4518.2 pM. In (A), the primary outcome variable was defined as Change in EASI. In (B), the secondary outcome variable was defined as Change in IGA. Change in EASI and Change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers extend to the highest and lowest values ​​within 1.5 times the interquartile range. Subgroups of patients with baseline SMOC1 blood concentrations ≥30 pM, ENTP6 blood concentrations ≤650 pM, or CRK blood concentrations ≤4518.2 pM were analyzed. The number (n) and proportion (%) of patients analyzed were the placebo group (n=23, 88%), the 0.3% TDCA gel group (n=22, 92%), and the 0.5% TDCA gel group (n=10, 50%). †P values ​​were determined by ANOVA test. ‡P values ​​were determined by rank sum test and mean difference of the results between the placebo group and the 0.5% TDCA gel group.

[0032] [Figure 13](A)-(B) show the results of EASI and IGA analysis in AD patients with baseline blood FAS levels. In (A), the primary evaluation variable was defined as change in EASI. In (B), the secondary evaluation variable was defined as change in IGA. Change in EASI and change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers indicate the highest and lowest values ​​within 1.5 times the interquartile range. The numbers (n) were the placebo group (n = 27), the 0.3% TDCA gel group (n = 27), and the 0.5% TDCA gel group (n = 25). †P values ​​were determined by ANOVA. ‡P values ​​were determined by rank-sum tests and mean differences between the placebo group and the 0.5% TDCA gel group.

[0033] [Figure 14] (A)-(B) show the results of EASI and IGA analysis of PP. In (A), the primary evaluation variable was defined as Change in EASI. In (B), the secondary evaluation variable was defined as Change in IGA. Change in EASI and Change in IGA were compared between the placebo group, the 0.3% TDCA gel group, and the 0.5% TDCA gel group. Boxes indicate the median and interquartile range, and whiskers indicate the highest and lowest values ​​within 1.5 times the interquartile range. Numbers (n) and proportions (%) were for the placebo group (n = 24, 89%), the 0.3% TDCA gel group (n = 20, 74%), and the 0.5% TDCA gel group (n = 19, 73%). †P values ​​were determined by ANOVA. ‡P values ​​were determined by rank-sum tests and mean differences between the placebo and 0.5% TDCA gel groups. DETAILED DESCRIPTION OF THE INVENTION

[0034] As of 2022, the FDA has approved 149 biomarker-assisted diagnostic treatments, all of which are for cancerous diseases, with the exception of two: obesity and non-transfusion-dependent thalassemia (NTDT). With the exception of one imaging diagnostic agent, the rest are limited to genetic testing or tissue protein staining. Biomarkers and their associated diagnostic treatments are an important concept in precision medicine, drawing attention for their potential to improve the accuracy, efficacy, and safety of treatments. They are recommended or preferred by health regulatory agencies worldwide. A prime example is Iressa, which received conditional approval in 2003 as a third-line treatment for non-small cell lung cancer. However, its approval was revoked in 2005 due to lack of efficacy. However, in 2015, its efficacy was recognized in conjunction with EGFR mutation biomarker diagnosis, and it was reapproved as a first-line treatment. In addition, when atezolizumab was tested in clinical trials for non-small cell lung cancer and all patients were included, no clear efficacy was confirmed compared to existing treatments. However, increased efficacy was confirmed when it was used in combination with PD-L1 in vitro diagnostics, and it was approved as a companion diagnostic treatment.

[0035] Biomarkers are used for various purposes in medicine, particularly as diagnostic tools. In particular, the diagnosis and treatment of atopic dermatitis, unlike other chronic diseases, entirely rely on clinical scores rather than biochemical markers. Therefore, securing reliable biomarkers can reduce observation errors and significantly improve the management of atopic dermatitis through a precision medicine approach. Furthermore, biomarkers can have a variety of effects on preventive approaches and provide useful strategies for future drug development.

[0036] Atopic dermatitis has a clinical phenotype influenced by a variety of heterogeneous factors, including environmental, genetic, metabolic, and disease factors. There is currently no treatment that is simultaneously superior in terms of ease of use, cost, and efficacy. While various research efforts to identify biomarkers have been conducted, no clinically meaningful results have been achieved. According to the International Committee on Dermatitis (IEC), which comprises over 100 atopic dermatitis research experts, atopic dermatitis is a polymorphic, heterogeneous disease characterized by the complex manifestation of at least three or more phenotypes. The IEC officially stated the need to improve patient management and treatment by stratifying phenotypes and using them as predictors of treatment response.

[0037] The present inventors have developed a surprising biomarker that can identify patients who will respond significantly to G protein-coupled receptor 19 (GPCR19) agonists, including taurodeoxycholic acid (TDCA). Specifically, the present inventors have developed a biomarker that can identify patients with atopic dermatitis who will respond effectively to TDCA. More specifically, the present inventors have developed a method for selecting patients with atopic dermatitis who will respond effectively to TDCA based on baseline blood protein levels of SMOC1, ENTP6, and CRK. Such biomarkers can be utilized as companion diagnostic compositions that can improve pretreatment prediction and treatment efficacy for patients with atopic dermatitis who use GPCR19 agonists.

[0038] definition The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. As used in this specification and the appended claims, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise.

[0039] When referring to numerical ranges herein, each number therebetween is expressly contemplated with the same precision. For example, in the range of 6 to 9, 7 and 8 are also contemplated in addition to 6 and 9, and in the range of 6.0 to 7.0, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated.

[0040] When referring to protecting a subject from a disease, "treatment" or "treating" means to suppress, conquer, or eliminate the disease. Disease prevention involves administering a composition described herein to a subject before the onset of the disease. Disease suppression involves administering a composition to a subject after the induction of the disease, but before it becomes clinically manifest. Disease suppression involves administering a composition to a subject after the disease has become clinically manifest.

[0041] GPCR19 agonists The present invention provides a compound that may be effective in treating atopic dermatitis. The compound may be a G-protein receptor 19 (GPCR19) agonist, a phanesoid X receptor agonist, or an anti-inflammatory agent. In one example, the compound is a GPCR19 agonist. The GPCR19 agonist may include taurodeoxycholic acid (TDCA), a derivative thereof, or a pharmaceutically acceptable salt thereof. TDCA may have a chemical structure represented by Formula I: [ka]

[0042] TDCA can also have a chemical structure represented by Formula II: [ka]

[0043] In one embodiment, the TDCA is sodium taurodeoxycholate. Examples of TDCA and compositions thereof are disclosed in U.S. Patent No. 9,855,283, the contents of which are incorporated herein by reference.

[0044] Also provided are pharmaceutical compositions comprising the compound and a pharmaceutically acceptable excipient. The pharmaceutical compositions may contain about 0.3 wt / wt%, 0.4 wt / wt%, 0.5 wt / wt%, 1.0 wt / wt%, 1.5 wt / wt%, 2.0 wt / wt%, 3.0 wt / wt%, 4.0 wt / wt%, or 5.0 wt / wt% of the compound, which in one embodiment is TDCA. In one embodiment, the pharmaceutical composition contains 0.3% or 0.5% TDCA. In one embodiment, the pharmaceutical composition is for topical use. The pharmaceutical composition may be applied to skin affected by atopic dermatitis, which may be a lesion. In one embodiment, the pharmaceutical composition is administered once, twice, three times, four times, or five times daily. In one embodiment, the pharmaceutical composition is administered twice daily. The pharmaceutical composition may be administered for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, hi one embodiment, the pharmaceutical composition is administered for at least about 3 weeks.

[0045] In one embodiment, the pharmaceutical composition includes a solvent. The solvent can include polyethylene glycol (PEG), which can be PEG 400. The solvent can also include ethanol. In one embodiment, the pharmaceutical composition includes 70% PEG 400 and 30% ethanol in distilled water.

[0046] The pharmaceutical composition may also include a gel, cream, ointment, tablet, or injection. In one embodiment, the pharmaceutical composition is a hyaluronic acid gel. The gel may be 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10% hyaluronic acid gel. In one embodiment, the pharmaceutical composition contains 5% hyaluronic acid gel. In one embodiment, 0.5 g of sodium taurodeoxycholate is mixed with 20 ml of distilled water to dissolve. 80 mg of chlorobutanol and 70 ml of distilled water can be mixed with a 25% sodium taurodeoxycholate solution. 1 g of hyaluronic acid can be added and mixed with the solution. Distilled water can be added to bring the resulting solution to 100 ml. In one embodiment, the pharmaceutical composition is NUGEL® (Shaperon Inc., Seoul, South Korea). In another example, 0.5g, 1g, 2g, or 4g of sodium taurodeoxycholate is mixed with 0.5g of carbomer 940, 0.3g of diethanolamine, 0.15g of methylparaben, 50mg of propylparaben, and 55mg of ethanol. Distilled water is added and the resulting solution is brought to 100ml. In another example, 0.5g, 1g, 2g, or 4g of sodium taurodeoxycholate is mixed with 50mg of polysorbate 80, 1g of sodium hyaluronate, and 0.2g of methylparaben. Distilled water is added and the resulting solution is brought to 100ml. In another example, 0.5g, 1g, 2g, or 4g of sodium taurodeoxycholate is mixed with 1.5g of carbomer 940, 1g of glycerin, 1.5g of triethanolamine, 0.15g of methylparaben, 50mg of propylparaben, and 1g of ethanol. Distilled water can be added to make the resultant volume up to 100 ml.

[0047] Markers of GPCR19 agonist efficacy Provided herein are biomarkers used to predict the efficacy of the compositions described herein for treating atopic dermatitis, and uses thereof. Also provided are methods for predicting the efficacy of a composition for treating atopic dermatitis, which may include measuring or quantifying the concentration of each of one or more biomarkers in a sample from a subject. A threshold level of the concentration of at least one of the one or more biomarkers may indicate a high likelihood that the composition will be effective in treating atopic dermatitis in the subject. The increased likelihood of efficacy in the subject may be associated with a human population of atopic dermatitis patients in which at least one or all of the one or more biomarkers are at threshold levels.

[0048] In one embodiment, the sample is a blood sample, and may be serum or plasma. The subject may be of any race, but in one embodiment is of Asian descent, and more particularly of Korean descent. In other embodiments, the subject is of American Indian descent, Alaska Native descent, Black American descent, Black British descent, Black Caribbean descent, African descent, Hispanic or Latino descent, Native Hawaiian or other Pacific Islander descent, Middle Eastern descent, or White European descent.

[0049] The biomarkers may include any one of IGHA2 (Immunoglobulin Heavy Constant Alpha 2; UniProt accession no. P01877; SEQ ID NO: 1); ENTP6 (ectonucleoside triphosphate diphosphohydrolase 6; UniProt accession no. O75354; SEQ ID NO: 2) or its isoform; SMOC1 (SPARC-related modular calcium-binding protein 1; UniProt accession no. Q9H4F8; SEQ ID NO: 3) or its isoform; ENPL (Endoplasmin or Heat shock protein 90 kDa beta member 1; UniProt accession no. P14625; SEQ ID NO: 4); and CRK (adapter molecule crk or proto-oncogene c-Crk; UniProt accession no. P46108; SEQ ID NO: 5) or its isoform. The sequences of these biomarkers are known in the art, as are their isoforms. Each biomarker can include the complete sequence of one of the proteins, a fragment thereof, or a protein having a sequence at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the protein.

[0050] The concentration of each biomarker in a patient sample that can predict the efficacy of a composition for treating atopic dermatitis can be a threshold level, which, as described herein, can mean that the concentration is above or below a set concentration. The threshold level of IGHA2 can be a concentration of about 130,000 pM; 135,000 pM; 140,000 pM; 141,000 pM; 142,000 pM, 143,000 pM; 144,000 pM; 145,000 pM; 146,000 pM; 147,000 pM; 148,000 pM; 149,000 pM; 150,000 pM; 151,000 pM; 152,000 pM; 153,000 pM; 154,000 pM; 155,000 pM; 156,000 pM; 157,000 pM; 158,000 pM; 159,000 pM; or 160,000 pM or greater. In one embodiment, the threshold level for IGHA2 is a concentration of about 149,579 pM or greater. The threshold level for ENTP6 can be a concentration of about 600 pM, 610 pM, 620 pM, 630 pM, 640 pM, 650 pM, 660 pM, 670 pM, 680 pM, 690 pM, or 700 pM or less. In one embodiment, the threshold level for ENTP6 is a concentration of about 650 pM or less. The threshold level for SMOC1 can be a concentration of about 20 pM, 21 pM, 22 pM, 23 pM, 24 pM, 25 pM, 26 pM, 27 pM, 28 pM, 29 pM, 30 pM, 31 pM, 32 pM, 33 pM, 34 pM, 35 pM, 36 pM, 37 pM, 38 pM, 39 pM, or 40 pM or greater. In one embodiment, the threshold level for SMOC1 is a concentration of about 30 pM or greater. The threshold level for ENPL can be a concentration of about 135 pM, 136 pM, 137 pM, 138 pM, 139 pM, 140 pM, 141 pM, 142 pM, 143 pM, 144 pM, 145 pM, 146 pM, 147 pM, 148 pM, 149 pM, 150 pM, 151 pM, 152 pM, 153 pM, 154 pM, 155 pM, 156 pM, 157 pM, 158 pM, 159 pM, or 160 pM or less. In one embodiment, the threshold level for ENPL is a concentration of about 148.8 pM or less.The threshold level of CRK can be a concentration of about 4300 pM, 4310 pM, 4320 pM, 4330 pM, 4340 pM, 4350 pM, 4400 pM, 4450 pM, 4500 pM, 4550 pM, 4600 pM, 4650 pM, or 4700 pM or less. In one example, the threshold level of CRK is a concentration of about 4518.2 pM or less. "About" when used with respect to a threshold concentration can mean a value within 0.5%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of the specified concentration.

[0051] If the concentration of a biomarker in a sample is at a threshold level, the sample or subject may be considered "positive" for that biomarker, which may indicate a higher likelihood that the composition will be effective in treating atopic dermatitis in the subject. In one embodiment, the one or more biomarkers include one or more of SMOC1, ENPT6, and CRK. In yet another embodiment, a threshold level of SMOC1 of about ≧30 pM, a threshold level of ENTP6 of about ≦650 pM, and a threshold level of CRK of about ≦4518.2 pM indicates a higher likelihood that the composition will be effective in treating atopic dermatitis.

[0052] The subject may be SMOC1-positive or CRK-positive and may be classified as having type A atopy. In one embodiment, the subject may be classified as having type A atopy and may be more likely to respond to a composition for treating atopic dermatitis. In one embodiment, the subject is classified as having type A atopy and is also ENTP6-positive and is classified as having type A' atopy. In one embodiment, a subject with type A' atopy is SMOC1-positive and ENTP6-positive. In another embodiment, a subject with type A' atopy is CRK-positive and ENTP6-positive. In one embodiment, the subject may be classified as having type A' atopy and may be more likely to respond to a composition for treating atopic dermatitis.

[0053] Methods for detecting and quantifying the concentration of proteins in a sample are known in the art. The concentration of one or more biomarkers can be quantified using mass spectrometry, antibodies, enzyme-linked immunosorbent assays, capillary immunoassay (WES), Western blotting, aptamers, peptides, peptidomimetics, oligonucleotides, or probes. In one embodiment, the concentration of one or more biomarkers is measured using mass spectrometry. The mass spectrometry (MS) can be multiple reaction monitoring MS (MRM-MS). The threshold level of a biomarker can be determined by measuring the maximum area under the curve and comparing the clinical reactivity with a receiver operating characteristic curve.

[0054] Additionally provided herein are kits containing reagents and / or laboratory equipment suitable for quantifying the concentration of one or more biomarkers. The kits may include one or more positive and negative controls. The kits may also include reagents and / or laboratory equipment for collecting a subject sample. The kits may also include reagents and / or laboratory equipment for pre-treating a subject sample. The kits may additionally include instructions for performing the quantification and / or sample collection methods.

[0055] The efficacy of a composition in treating atopic dermatitis can be measured by any method known in the art. In one embodiment, the method is clinical diagnosis. In another embodiment, efficacy is measured by calculating the Eczema Area and Severity Index (EASI) score, where a decrease in the EASI score measured at a later time point compared to an earlier time point indicates improvement. Calculating the EASI score can include assessing the severity of erythema, edema / induration, excoriation, and lichenification on the subject's head / neck, trunk, and lower extremities on a four-point scale (0 = none, 1 = mild, 2 = moderate, 3 = severe). Clinical findings can be scored as a lesion area score (0 = none, 1 = <10%, 2 = 10%-29%, 3 = 30%-49%, 4 = 50%-69%, 5 = 70%-89%, 6 = 90%-100%) of the body area. The scores for each body part can be multiplied by 0.1 for head / neck, 0.2 for upper limbs, 0.3 for trunk, and 0.4 for lower limbs and then added together to calculate the EASI score.

[0056] In yet another embodiment, efficacy is measured by calculating a validated Investigator Global Assessment (IGA) score, where a decrease in the IGA score measured at a later time point compared to an earlier time point indicates improvement. The IGA score can be based on a 5-point scale ranging from 0 to 4 (0 = clear, no inflammatory signs of atopic dermatitis; 1 = almost clear, only noticeable redness and noticeable induration of papules; 2 = mild, slight redness and slight induration of papules, no weeping or crusting; 3 = moderate, moderate redness and moderate induration of papules, possible weeping and crusting; 4 = severe, severe redness and severe induration of papules, weeping and crusting).

[0057] Treatment for atopic dermatitis Provided herein are methods for treating atopic dermatitis in a subject in need thereof. The methods can include administering to the subject a composition comprising a GPCR19 agonist. The composition can include a GPCR19 agonist, particularly TDCA, a derivative thereof, or a pharmaceutically acceptable salt thereof. The composition can be a pharmaceutical composition. Also provided are uses of the composition in treating atopic dermatitis and in the manufacture of a medicament for treating atopic dermatitis. In one embodiment, the composition is administered topically or is intended for topical administration.

[0058] The subject may be identified as being more likely to be effectively treated by the composition, as determined by a method for predicting the efficacy of a composition in treating atopic dermatitis, as described herein. In one embodiment, the method includes administering the composition to a subject identified as being more likely to be effectively treated by a prediction method described herein. The subject may be positive for at least one of IGHA2, ENTP6, SMOC1, ENPL, and CRK, as described above. The subject may have type A atopy and be positive for SMOC1 or CRK. The subject may have type A' atopy and be positive for ENTP6 and positive for at least one of SMOC1 and CRK.

[0059] In other embodiments, the method includes providing a concentration of each of the one or more biomarkers in a sample from the subject, comparing each concentration to a threshold level, and administering a composition to the subject if at least one of the one or more biomarkers is at its respective threshold level. In one embodiment, the method includes obtaining or providing a sample from the subject, quantifying the concentration of each of the one or more biomarkers, and providing the concentration of each of the one or more biomarkers.

[0060] The method can include administering a composition to a subject, wherein the subject is positive for at least one of IGHA2, ENTP6, SMOC1, ENPL, and CRK. In one embodiment, the subject is positive for at least one of ENTP6, SMOC1, and CRK. In one embodiment, the method can include administering a composition to a subject, wherein the subject has type A atopy. In another embodiment, the method can include administering a composition to a subject, wherein the subject has type A' atopy. The method can include determining whether the subject is positive for one or more biomarkers prior to administering the composition, or identifying whether the subject is positive for one or more biomarkers prior to administering the composition. [Example]

[0061] The present invention has various aspects which are illustrated in the following non-limiting examples.

[0062] Example 1 Biomarkers that can be used to identify patients with atopic dermatitis who respond to GPCR19 agonists Atopic dermatitis (AD) is the most common chronic inflammatory skin disease, affecting up to 25% of children and 8% of adults worldwide. Current traditional AD treatment strategies, including corticosteroids, calcineurin inhibitors, PDE4 inhibitors, JAK inhibitors, ultraviolet light therapy, and antibodies targeting the IL-4 / IL-3 pathway, focus on epidermal barrier function, abnormal Th2 / 17 / 22 responses, and late-stage Th1-mediated pathogenesis. Unfortunately, none of these approaches has been able to meet the unmet therapeutic needs or be recognized as a single treatment applicable to all patients. In addition to serious side effects, approximately 20% of AD patients comprise a subset known as treatment-resistant AD, for which existing treatments and management strategies have been unsuccessful. Novel AD treatments based on precision medicine, "the right treatment for the right patient," could address the current unmet needs of AD patients. This example demonstrates that GPCR19, which associates with P2X7R, and the TDCA pharmaceutical composition NUGEL® described herein inhibited P2X7R activation. Noncistronically, NUGEL® inhibited the adenylyl cyclase-PKA pathway and BzATP-mediated Ca activation. 2+NUGEL® inhibited NF-kB activation through the recruitment of NUGEL®. NUGEL® inhibited the expression of P2X7R and NLRP3 inflammasome (N3I) components in keratinocytes. Treatment of keratinocytes with NUGEL® inhibited NLRP3 oligomerization and the production of mature IL-1β and IL-18. Topical treatment with NUGEL® improved features of atopic inflammation in various AD mouse models. Furthermore, a two-phase, double-blind, randomized, placebo-controlled clinical trial evaluated the efficacy of NUGEL® for treating AD in adults with mild to moderate AD, using biomarkers as a patient selection strategy. Blood samples were analyzed during the screening phase of patient recruitment, and only biomarker-positive patients were evaluated. Patients were randomly assigned to receive placebo, NUGEL® 0.3%, or NUGEL® 0.5% twice daily for 8 weeks. In this study, patients were classified as type A or type A' based on the biomarkers used. Among patients with type A and type A' atopic dermatitis, those treated with NUGEL® 0.5% experienced a 53% (p=0.002) and 83% (p=0.008) improvement in the Eczema Area and Severity Index (EASI), respectively, compared to the placebo group. The validated Investigator's Global Assessment (IGA) also achieved a 33% (p=0.007) and 53% (p=0.005) improvement compared to baseline in patients with type A and type A' atopic dermatitis, respectively. No adverse events were reported in this study. These surprising results suggest that NUGEL®, a GPCR19 agonist with a favorable safety profile, can be used in conjunction with molecular markers to identify patients who may benefit from the drug, thereby overcoming the current unmet need for treatment in patients with AD.

[0063] method research design A double-blind, placebo-controlled, parallel-group, multicenter clinical trial was conducted to evaluate the efficacy of the treatment in recruited clinical trial patients (ClinicalTrials.gov no.: NCT04530643). NUGEL® (Shaperon, Korea), a composition containing taurodeoxycholic acid (TDCA) as the active ingredient, was used as the treatment in this study. Enrollment began in August 2020 and ended in July 2021.

[0064] This clinical trial was randomized. Subjects who met the inclusion criteria but not the exclusion criteria were ultimately assigned to a 0.3% TDCA gel treatment group, a 0.5% TDCA gel treatment group, or a placebo group in a 1:1:1 ratio. This randomization method was used to minimize the influence and bias of baseline variables on the study results. This study was designed as a double-blind study, so both the investigators and the subjects were blinded. The type of drug administered to both the experimental and placebo subjects was unknown prior to the completion of the study. Although administering a placebo to the placebo group has the disadvantage of unnecessary administration, a double-blind design was adopted considering the possibility that subjective judgment by the subjects or investigators could affect clinical evaluation. Patients were considered to have mild to moderate atopic dermatitis. The dosage was administered via repeated transdermal administration. Patients applied an appropriate amount of gel to their lesions twice daily, in the morning and evening.

[0065] participants This clinical trial was conducted on male and female patients suffering from atopic dermatitis. Patients who agreed to participate in the clinical trial and met the inclusion criteria but not the exclusion criteria were enrolled. The researchers carefully considered the appropriateness of enrolling subjects, taking into full consideration the purpose of the study, the subjects' rights, and their health status. Subjects who were unable to voluntarily sign the consent form were not selected for this study.

[0066] Participants who met the following inclusion criteria were included in the study: minimum age 19 years or older, clinically diagnosed with atopic dermatitis according to the Hanifin and Rajka criteria, IGA score of 2 or 3 at the baseline visit (day 1), and a minimum of 5% and a maximum of 40% BSA covered by AD at the baseline visit.

[0067] Exclusion criteria were as follows: those who had received steroids, oral antibiotics, photochemotherapy, or immunosuppressants before the baseline visit, and those whose AST / ALT or creatinine values ​​exceeded twice the upper limit of the normal range in the screening test.

[0068] During the clinical trial period, the use of prescription moisturizers and moisturizers containing additives such as ceramide, hyaluronic acid, urea, and filaggrin was prohibited. Also prohibited during the clinical trial period were immunomodulators, corticosteroids, antihistamines, photochemotherapy, existing atopic dermatitis treatments, and other drugs thought to affect immune function.

[0069] Concomitant medications taken by subjects prior to participating in this clinical trial that were not expected to affect the interpretation of the results of this clinical trial were permitted at the discretion of the clinician. Medications temporarily used to treat other diseases or abnormal reactions were administered in consultation with the attending physician.

[0070] Study protocol and measurements From days -21 to 0, patients were screened to assess inclusion and exclusion eligibility and enrolled into one of three treatment groups on day 1. During the treatment period, patients received 0.3% TDCA gel, 0.5% TDCA gel, or placebo. Study drug safety was monitored through laboratory testing at the discretion of the principal investigator. Routine laboratory testing was performed at each study site's central laboratory.

[0071] Evaluation variables The primary outcome was the percentage change from baseline in the EASI clinical score. A negative change from baseline indicated improvement. Researchers measured patients' EASI clinical scores at baseline, 2-week visit, and 4-week visit. The same examiner assessed the severity of erythema, edema / induration, excoriation, and lichenification on the subjects' head / neck, trunk, and lower extremities on a 4-point scale (0 = none, 1 = mild, 2 = moderate, 3 = severe). Clinical findings were scored as a lesion area score (0 = none, 1 = <10%, 2 = 10%-29%, 3 = 30%-49%, 4 = 50%-69%, 5 = 70%-89%, 6 = 90%-100%) of the body area. The scores for each body part were then multiplied by 0.1 for head / neck, 0.2 for upper limbs, 0.3 for trunk, and 0.4 for lower limbs, and then summed to calculate a total EASI score.

[0072] The secondary outcome was the percentage change from baseline in the IGA clinical score. A negative change from baseline indicates improvement. Researchers measured patients' IGA clinical scores at the start date (baseline) and during the 2-week and 4-week visits. The IGA is a rating scale used in clinical trials to measure the severity of atopy and clinical response to treatment, and is based on a 5-point scale ranging from 0 to 4 (0 = clear, no inflammatory signs of atopic dermatitis; 1 = almost clear, only noticeable redness and noticeable induration of papules; 2 = mild, slight redness and slight induration of papules, no weeping or crusting; 3 = moderate, moderate redness and moderate induration of papules, possible weeping and crusting; 4 = severe, severe redness and severe induration of papules, weeping and crusting).

[0073] Proteome analysis To identify potential biomarkers for predicting therapeutic drug response, we quantitatively analyzed 802 proteins using MRM-MS in patient plasma samples collected before drug administration.

[0074] Based on existing research and expert opinions, a total of 802 target proteins detectable in plasma were selected. These proteins consist of disease biomarkers approved by the FDA (Food and Drug Administration) / LDT (Laboratory Developed Test) and a group of biomarker candidates confirmed by our research team. To identify biomarker candidates that could be analyzed by MRM-MS using a 6495 QQQ mass spectrometer, we analyzed each pooled sample of each biomarker candidate protein alongside an internal standard peptide (Stable isotope-23 labeled synthetic (SIS)-peptide, SIS peptide) to confirm the detected markers.

[0075] Targets with intensities below 100 when analyzed with SIS alone, targets that did not co-elute with SIS in plasma, and targets with intensities below 100 for both endogenous peptides and SIS were excluded. A final 502 proteins / peptides were selected as candidate target biomarkers for MRM-MS analysis of individual samples. The amount of SIS for analysis was determined by analyzing pooled plasma endogenous peptides and SIS and selecting the amount whose peak area ratio (= amount of endogenous peptide / amount of SIS) was closest to 1 while satisfying the range of 0.1–10. This amount was used for analysis of individual plasma samples. To obtain the final results, plasma samples were first pretreated with trypsin. Then, plasma samples from 26 patients in the placebo group, 24 patients in the 0.3% TDCA gel-treated group, and 20 patients in the 0.5% TDCA gel-treated group were analyzed. To prevent experimental bias, random pretreatment batches were set up using block randomization. Each batch contained 23 or 24 randomly selected samples. For targeted proteome analysis, plasma samples were thawed on ice and centrifuged at 4°C and 10,000 × g for 10 min. The supernatant was then centrifuged through a 0.22 μm filter (12,000 g, room temperature). The centrifuged sample was transferred to a new tube. Sample proteins were then analyzed using Pierce TMQuantification was performed by bicinchoninic acid assay (BCA assay) using a BCA protein assay kit (Thermo Scientific).

[0076] For each crude plasma sample, 200 micrograms of protein were digested with RapiGest detergent and trypsin. More specifically, 40 μL of 0.2% RapiGest, 20 mM dithiothreitol (DTT), and 100 mM ABC buffer (pH 8.0) were added to 200 μg of plasma protein, and the volume was adjusted to 40 μL with HPLC-grade water. After incubation at 60°C for 1 hour, 20 μL of 100 mM iodoacetamide (IAA) was added, and the sample was further incubated at room temperature for 30 minutes in the dark. Trypsin dissolved in 50 mM ABC buffer (pH 8.0) was added to the sample, and the sample was incubated at 37°C for 4 hours. To terminate the enzymatic reaction, 10% formic acid solution was added to the sample to a final concentration of 1%. The sample was then incubated at 37°C for 30 minutes to hydrolyze the RapiGest detergent. After centrifugation at 15,000 rpm and 4°C for 1 hour to precipitate the cleaved RapiGest detergent, the supernatant was transferred to a new tube. Finally, crude stable isotope-labeled internal standard (SIS) peptides were added to the plasma peptide samples.

[0077] For MRM-MS analysis, an Agilent 6495 triple quadrupole mass spectrometer coupled with an Agilent 1260 Infinity HPLC system was used. HPLC solvents A and B consisted of 0.1% formic acid (v / v) in water and 0.1% formic acid (v / v) in acetonitrile, respectively. A total of 40 μL of digested sample was injected onto a guard column (2.1 × 15.0 mm, 1.8 μm, 80 Å, Agilent). Online desalting was performed at 40 °C and 3% solvent B for 10 min at a rate of 5 μL / min, with the effluent sent to waste. The desalted sample was then transferred to an analytical column (0.5 × 35.0 mm, 3.5 μm, 80 Å, Agilent) at a flow rate of 40 μL / min in 3% solvent B for 5 min. The analytical column was heated and maintained at 40°C in an oven. The total run time per MRM-MS analysis was 140 min. Approximately 10 μg of digested peptides was injected per MRM-MS run. Peptides were separated on the column and eluted with a linear gradient of 3% to 40% acetonitrile with 0.1% formic acid (FA) at a flow rate of 40 μL / min for 125 min. Separated peptides were monitored using the mass spectrometer in the MRM mode scheduled for each protein conversion. Peak integration was performed using the Skyline tool, an MRM-MS data preprocessing program. The peak area ratio value for each target was derived during the peak integration process. The final quantification value was determined by multiplying the peak area ratio value of each target by the amount of internal standard injected. To identify targets suitable for stable quantification, 502 targets were first screened in 70 individual samples prior to administration. Targets with unstable peak shapes (slanted peaks) or intensities less than 100% relative to the internal standard and endogenous peptides were excluded. As a result, 33 proteins were excluded, and 469 proteins were finally selected for statistical analysis. The dynamic range of the quantitative values ​​(fmol) of the 469 target proteins was approximately 7 orders of magnitude. Approximately 90% of the protein targets had a dynamic range of 5 orders of magnitude, while the top 5% of protein targets, including the abundant protein human albumin, had a dynamic range of 2.5 orders of magnitude. The bottom 5% of low-abundance proteins had a dynamic range of approximately 1 order of magnitude.Considering the dynamic range of the 6495 QQQ instrument used for the analysis, which spans six orders of magnitude, the optimized MRM-MS analytical method was able to stably quantify 469 target proteins in blood samples. A total of 70 plasma samples were collected at five centers prior to administration. To determine whether technical variability between hospitals might affect the quantitative values ​​of the protein targets, we examined the distribution of protein quantification values ​​by institution. The distribution of protein quantification values ​​showed no statistically significant differences between institutions, and PCA confirmed that samples were uniformly distributed between hospitals. Therefore, we concluded that technical variability between hospitals had a negligible impact on the quantitative values ​​of the protein targets.

[0078] Subsequently, stratified analysis was performed to investigate the correlation between baseline blood plasma protein levels and drug therapeutic efficacy, as indicated by EASI and IGA scores. Potential biomarker candidates and effective cutoff values, which play an important role in determining drug responsiveness, were established. Among groups with positive biomarker levels, the TDCA-treated group showed statistically greater therapeutic efficacy than the placebo-treated group. Receiver operating characteristic (ROC) analysis was used to confirm whether there was a correlation between protein levels and therapeutic response.

[0079] statistical analysis The Full Analysis Set (FAS) consists of subjects who received at least one dose and whose EASI value, the primary endpoint, was measured at baseline within four weeks of receiving the clinical trial drug (n=79). The Per-Protocol (PP) included FAS patients without major protocol deviations described in the approved synopsis (n=63). The Biomarker Analysis Set (BAS) consists of subjects who completed the final schedule and whose plasma baseline biomarkers were measured, excluding subjects who discontinued the clinical trial from the FAS analysis group (n=70). In this analysis set, biomarkers are used as stratification factors along with clinical efficacy analysis.

[0080] The mean and standard error of the mean (SEM) of efficacy variables were determined for each group. Differences in efficacy between groups were assessed using ANOVA. Post-hoc analysis was also performed to determine whether there were significant differences between groups using the rank sum test. Statistical analysis of this clinical trial was performed using SAS version 9.4.

[0081] In the safety analysis, abnormal reactions were classified and analyzed by treatment group, affected organ, and preferred term, including abnormal reactions, abnormal drug reactions, serious abnormal reactions, and abnormal reactions that led to participant withdrawal. Differences in the incidence of abnormal reactions between treatment groups were summarized as frequencies and percentages. Technical statistics for laboratory results before treatment (screening) and after completion of study drug or placebo administration were presented for each treatment group. In addition, for each laboratory result, the frequency and proportion of subjects who had normal / clinically insignificant abnormalities before treatment (screening) but changed to clinically significant abnormalities after completion of study drug administration were presented by treatment group. Technical statistics (number of subjects, mean, standard deviation, minimum, median, maximum) for vital signs (blood pressure, heart rate, body temperature) were calculated and presented for each treatment group. Physical examination results for each treatment group were summarized as frequencies and percentages to identify changes due to study drug administration. Electrocardiogram (ECG) results were compared before treatment (screening) and after study drug administration, and observed changes were summarized by treatment group. Percentages for each treatment group were presented. All reported abnormal reactions were reported using MedDRA Version 24.0.

[0082] result Research Subjects Of the 94 eligible patients, 80 were randomly assigned: 27 to the 0.3% TDCA gel group, 26 to the 0.5% TDCA gel group, and 27 to the placebo group (Figure 1). The median age of patients was 28 years in the 0.3% TDCA gel group, 26 years in the 0.5% TDCA gel group, and 26 years in the placebo group. 37% of the 0.3% TDCA gel group, 56% of the 0.5% TDCA gel group, and 52% of the placebo group were male (Table 1). [Table 1]

[0083] The baseline EASI scores (mean ± standard deviation) were 7.41 ± 3.77 in the 0.3% TDCA gel group, 8.96 ± 4.47 in the 0.5% TDCA gel group, and 8.50 ± 4.80 in the placebo group. The baseline IGA scores (mean ± standard deviation) were 2.30 ± 0.47 in the 0.3% TDCA gel group, 2.56 ± 0.51 in the 0.5% TDCA gel group, and 2.37 ± 0.49 in the placebo group (Table 2). [Table 2]

[0084] Biomarker discovery To explore specific blood biomarkers associated with predicting NUGEL® drug response in patients with atopic dermatitis, we used NUGEL®, a clinical drug formulation containing TDCA, a GPCR19 agonist. In a phase 2 clinical trial, we analyzed the correlation between baseline blood protein levels and treatment response after four weeks of drug treatment in patients prescribed high-dose treatment (0.5% TDCA gel).

[0085] Specifically, we performed ROC curve analysis to evaluate the pre-treatment serum quantitative values ​​of each protein and the clinical improvement of EASI in patients prescribed high-dose treatment, and to confirm the improvement of atopic symptoms. We then selected 42 proteins with a sensitivity / specificity of AUC (area under the ROC curve) > 0.7, and predicted the criterion value (cutoff) for stratification analysis (Table 3). [Table 3]

[0086] Biomarker stratification analysis In the FAS population, the mean ± SEM change in EASI (primary outcome) was -12.2 ± 8.8%, -11.9 ± 11.1%, and -2.91 ± 9.0% for the 0.3% TDCA gel group, the 0.5% TDCA gel group, and the placebo group, respectively. The difference between the placebo group and the 0.5% TDCA gel group was not significant by rank sum test (p = 0.531). The mean ± SEM change in IGA (secondary outcome) was also -1.9 ± 4.7% for the placebo group and -9.3 ± 5.0% for the 0.5% TDCA gel group. The difference between the placebo group and the 0.5% TDCA gel group was not significant by rank sum test (p = 0.299) (Figure 13).

[0087] In the PP population, the mean ± SEM changes in EASI were -18.0 ± 8.9%, -9.4 ± 14.1%, and -3.0 ± 10.2% for the 0.3% TDCA gel group, the 0.5% TDCA gel group, and the placebo group, respectively. The difference between the placebo group and the 0.5% TDCA gel group was not significant by rank sum test (p = 0.669). The mean ± SEM changes in IGA, the secondary outcome, were also -2.1 ± 5.3% for the placebo group and -7.9 ± 6.0% for the 0.5% TDCA gel-treated group. The difference between the placebo group and the 0.5% TDCA gel group was not significant by rank sum test (p = 0.506) (Figure 14).

[0088] Surprisingly, five protein biomarkers (IGHA2, ENTP6, SMOC1, ENPL, and CRK) showed significant ROC responses with clinical improvement in EASI and / or IGA (Table 4). Based on the concentration cutoff values ​​of each biomarker, we filtered out patients with SMOC1 positivity (defined as ≥ 30 pM), CRK positivity (defined as ≤ 4518.2 pM), ENTP6 positivity (defined as ≤ 650 pM), IGHA2 positivity (defined as ≥ 149,579 pM), and ENPL positivity (defined as ≤ 148.8 pM). [Table 4]

[0089] The predictive ability of each biomarker for therapeutic efficacy was evaluated. A stratified analysis was performed for patients with SMOC1 threshold levels. As shown in Figure 2, the therapeutic efficacy of 0.5% NUGEL® was statistically greater than that of placebo. The validity of biomarker-based therapeutic efficacy prediction was confirmed. Specifically, the number of SMOC1-positive patients per group (n) and their proportion (%) of the total analyzed patients were as follows: placebo group (n = 12, 46%), 0.3% TDCA gel treatment group (n = 12, 50%), and 0.5% TDCA gel treatment group (n = 8, 40%). The primary efficacy evaluation variable (EASI change rate; mean ± SEM) was 2.1 ± 10.7% in the placebo group and -51.3 ± 10.9% in the 0.5% TDCA gel treatment group, demonstrating a significant therapeutic synergy effect of 53.4% ​​(p = 0.005), confirming the difference in therapeutic efficacy. The IGA change rate (mean ± SEM) was 1.4 ± 8.1% in the placebo group and -33.3 ± 8.9% in the 0.5% TDCA gel-treated group, demonstrating a significant therapeutic synergy effect with a difference in treatment effect of 34.7% (p = 0.014).

[0090] In Figure 3, a stratified analysis was performed on patients with positive baseline blood CRK levels to compare the therapeutic effects of placebo and 0.5% TDCA gel. The validity of biomarker-based treatment efficacy prediction was confirmed by analyzing the treatment effects of placebo and 0.5% TDCA gel. Specifically, the number of CRK-positive patients per group (n) and their proportion (%) of the total analyzed patients were as follows: placebo group (n = 18, 69%), 0.3% TDCA gel treatment group (n = 12, 50%), and 0.5% TDCA gel treatment group (n = 6, 30%). The primary efficacy variable (EASI change rate; mean ± SEM) was 6.4 ± 11.0% in the placebo group and -52.6 ± 9.9% in the 0.5% TDCA gel treatment group, demonstrating a significant therapeutic synergy effect with a difference of 59.1% (p = 0.005). The IGA change rate (mean ± SEM) was 4.6 ± 6.8% in the placebo group and -38.9 ± 10.3% in the 0.5% TDCA gel-treated group, demonstrating a significant therapeutic synergy effect with a difference in treatment effect of 43.5% (p = 0.005).

[0091] In Figure 4, a stratified analysis was performed on patients with positive baseline blood levels of ENTP6. The effectiveness of the placebo-treated group and the 0.5% TDCA gel treatment group were compared to confirm the validity of biomarker-based treatment efficacy prediction. Specifically, the number of ENTP6-positive patients per group (n) and their proportion (%) of the total analyzed patients were as follows: placebo group (n = 12, 46%), 0.3% TDCA gel treatment group (n = 13, 54%), and 0.5% TDCA gel treatment group (n = 5, 25%). The primary efficacy variable (EASI change rate; mean ± SEM) was 8.5 ± 15.1% in the placebo group and -61.3 ± 11.2% in the 0.5% TDCA gel treatment group, demonstrating a significant therapeutic synergy of 69.8% (p = 0.007), confirming the difference in treatment efficacy. The IGA change rate (mean ± SEM) was 9.7 ± 7.5% in the placebo group and -40.0 ± 6.7% in the 0.5% TDCA gel-treated group, demonstrating a significant therapeutic synergy effect with a difference in treatment effect of 49.7% (p = 0.002).

[0092] In Figure 5, a stratified analysis was performed on patients with positive baseline blood levels of IGHA2, comparing the treatment effects of placebo and 0.5% TDCA gel to confirm the efficacy of biomarker-based treatment efficacy prediction. Specifically, the number of IGHA2-positive patients per group (n) and their proportion (%) of the total analyzed patients were as follows: placebo group (n = 11, 42%), 0.3% TDCA gel treatment group (n = 16, 67%), and 0.5% TDCA gel treatment group (n = 11, 55%). The primary efficacy outcome variable (EASI percentage change; mean ± SEM) was -5.0 ± 3.0% in the placebo group and -45.4 ± 9.6% in the 0.5% TDCA gel treatment group, demonstrating a significant therapeutic synergy effect with a difference of 40.4% (p = 0.036). The IGA change rate (mean ± SEM) was 6.1 ± 7.2% in the placebo group and -24.2 ± 9.1% in the 0.5% TDCA gel-treated group, demonstrating a significant therapeutic synergy effect with a difference in treatment effect of 30.3% (p = 0.017).

[0093] In Figure 6, a stratified analysis was performed on patients with positive baseline blood levels of ENPL to compare the therapeutic effects of placebo and 0.5% TDCA gel. The validity of biomarker-based treatment efficacy prediction was confirmed by analyzing the treatment effects of placebo and 0.5% TDCA gel. Specifically, the number of ENPL-positive patients per group (n) and their proportion (%) of the total analyzed patients were as follows: placebo group (n = 7, 29%), 0.3% TDCA gel treatment group (n = 8, 34%), and 0.5% TDCA gel treatment group (n = 6, 30%). The primary efficacy variable (EASI change rate; mean ± SEM) was 39.6 ± 20.3% in the placebo group and -53.3 ± 11.6% in the 0.5% TDCA gel treatment group, demonstrating a significant therapeutic synergy of 92.9% (p = 0.005), confirming the difference in treatment efficacy. The IGA change rate (mean ± SEM) was 16.7 ± 12.6% in the placebo group and -33.3 ± 12.2% in the 0.5% TDCA gel-treated group, demonstrating a significant therapeutic synergy effect with a difference in treatment effect of 50.0% (p = 0.035).

[0094] Additional classification and analysis These results suggest that biomarkers can be used to predict the efficacy of TDCA treatment for AD, including 0.5% TDCA gel. The baseline blood protein levels of all of the above are potentially useful for accompanying diagnostic treatment using TDCA gel. In particular, SMOC1, ENTP6, and CRK are considered the most useful biomarkers because they facilitate patient recruitment without stringent filtering and are statistically significant stratification factors for drug response. Therefore, 0.5% TDCA gel was confirmed to exhibit excellent efficacy in atopic dermatitis patients whose blood baseline levels were positive for SMOC1, CRK, and ENTP6.

[0095] More specifically, patients who were SMOC1 or CRK positive and showed a high response to TDCA gel therapy were classified as "type A atopy." Furthermore, patients who were type A and ENTP6 positive, who responded best to TDCA gel therapy, were classified as "type A' atopy" (Figure 7). Furthermore, the diagnostic value of biomarkers in predicting drug efficacy increased not only when biomarkers were used individually, but also when two or more were combined.

[0096] Specifically, the number of SMOC1- or CRK-positive "type A atopic dermatitis" patients per group (n) and their proportion (%) of the total analyzed patients were as follows: placebo group (n = 20, 77%), 0.3% TDCA gel treatment group (n = 21, 88%), and 0.5% TDCA gel treatment group (n = 11, 55%). The primary efficacy variable (EASI change rate; mean ± SEM) was 2.7 ± 10.4% in the placebo group and -50.0 ± 8.7% in the 0.5% TDCA gel treatment group, demonstrating a significant therapeutic synergy effect with a difference of 52.7% (p = 0.002). The IGA change rate (mean ± SEM) was 2.5 ± 6.4% in the placebo group and -30.0 ± 7.8% in the 0.5% TDCA gel-treated group, demonstrating a significant therapeutic synergy effect with a difference in treatment effect of 32.5% (p = 0.007) (Figure 8).

[0097] Among the patients analyzed who were classified as "type A' atopy," the number of patients per group who were also type A and ENTP6 positive (n) and their ratio (%) to the total number of patients analyzed were as follows: placebo group (n=9, 35%), TDCA gel treatment group (n=12, 50%), and TDCA gel treatment group (n=5, 25%). The primary efficacy evaluation variable (EASI change rate; mean ± SEM) was 22.1 ± 18.0% in the placebo group and -61.3 ± 11.2% in the 0.5% TDCA gel treatment group, demonstrating a significant therapeutic synergy effect with a difference in treatment effect of 83.4% (p=0.008). The IGA change rate (mean ± SEM) was 13.0 ± 9.9% in the placebo group and -40.0 ± 6.7% in the 0.5% TDCA gel-treated group, demonstrating a significant therapeutic synergy effect with a difference in treatment efficacy of 53% (p = 0.005) (Figure 9).

[0098] A stratified analysis was performed on SMOC1- or ENTP-positive patients to compare the therapeutic effects of placebo and 0.5% TDCA gel to confirm the validity of the combination of the two biomarkers for predicting therapeutic efficacy. Specifically, the number of SMOC1- or ENTP-positive patients per group (n) and their proportion (%) of the total analyzed patients were as follows: placebo group (n=18, 69%), 0.3% TDCA gel treatment group (n=19, 79%), and 0.5% TDCA gel treatment group (n=9, 45%). The primary efficacy outcome variable (EASI change rate, mean ± SEM) was -1.2 ± 10.9% in the placebo group and -51.4 ± 9.6% in the 0.5% TDCA gel treatment group, demonstrating a significant therapeutic synergy effect with a difference of 50.2% (p=0.006). The IGA change rate (mean ± SEM) was 1.9 ± 6.3% in the placebo group and -33.3 ± 7.9% in the 0.5% TDCA gel-treated group, demonstrating a significant therapeutic synergy effect with a difference in treatment effect of 35.2% (p = 0.003) (Figure 10).

[0099] The effectiveness of biomarker-based treatment efficacy prediction was confirmed by comparing the therapeutic effect of 0.5% TDCA gel with placebo in patients stratified by ENTP6- or CRK-positive status. Specifically, the number of ENTP6- or CRK-positive patients per group (n) and their proportion (%) of the total analyzed patients were as follows: placebo (n=21, 81%), 0.3% TDCA gel (n=16, 67%), and 0.5% TDCA gel (n=8, 40%). The primary efficacy outcome (EASI percentage change; mean ± SEM) was 0.9 ± 9.9% in the placebo group and -50.9 ± 8.5% in the 0.5% TDCA gel group, demonstrating a significant therapeutic synergy effect with a difference of 51.8% (p=0.003). The IGA change rate (mean ± SEM) was 4.0 ± 5.9% in the placebo group and -37.5 ± 7.6% in the 0.5% TDCA gel-treated group, demonstrating significant therapeutic synergy with a treatment difference of 41.5% (p < 0.001) (Figure 11).

[0100] The efficacy of biomarker-based treatment efficacy prediction was confirmed by comparing the therapeutic effect of 0.5% TDCA gel with placebo in patients stratified by SMOC1-, ENTP6-, or CRK-positive status. Specifically, the number of SMOC1-, ENTP6-, or CRK-positive patients per group (n) and their proportion (%) of the total analyzed patients were as follows: placebo (n=23, 88%), 0.3% TDCA gel (n=22, 92%), and 0.5% TDCA gel (n=10, 50%). The primary efficacy variable (EASI percentage change; mean ± SEM) was -1.8 ± 9.3% in the placebo group and -50.0 ± 8.7% in the 0.5% TDCA gel group, demonstrating a significant therapeutic synergy effect with a difference of 48.1% (p=0.003). The IGA change rate (mean ± SEM) was 2.2 ± 5.6% in the placebo group and -30.0 ± 7.8% in the 0.5% TDCA gel-treated group, demonstrating a significant therapeutic synergy effect with a treatment difference of 32.2% (p = 0.004) (Figure 12).

[0101] Through a biomarker-based clinical phase 2 efficacy analysis, it was confirmed that the clinical efficacy of TDCA gel could be predicted by the baseline levels of biomarker proteins, particularly SMOC1, ENTP6, and CRK.

[0102] safety results In the FAS population, which included all 80 patients, no serious side effects were reported with the administration of 0.3% TDCA gel or 0.5% TDCA gel during the clinical trial, and no abnormal drug reactions, abnormal reactions that led to permanent discontinuation of administration, or dropouts were reported. There were few side effects, and there was no difference compared to the placebo group. Any side effects that occurred were very mild and were temporary symptoms that all resolved during the clinical trial. Furthermore, the causality of all abnormal reactions was assessed as "not related" by the clinician. This confirmed the safety and excellent tolerability of TDCA gel, the therapeutic agent used in this study (Table 5). [Table 5]

[0103] As shown in the table below, when compared with existing approved treatments such as the PDE4 inhibitor crisaborole, the JAK inhibitor ruxolitinib, and topical steroids, TDCA targets "type A atopic dermatitis" patients, who account for 73% of mild to moderate atopic dermatitis patients. 0.5% TDCA gel has therapeutic effects equivalent to or even superior to existing drugs. Furthermore, for "type A' atopic dermatitis" patients, who account for 37% of patients with the highest drug response, 0.5% TDCA gel demonstrates far superior efficacy and safety compared to existing drugs. [Table 6]

[0104] In a small, multi-institutional, phase 2 clinical trial involving 80 hospitalized patients, the differences in primary outcome variables between groups did not reach statistical significance at the 5% threshold. However, biomarker stratification analysis revealed significantly higher clinical outcomes in the 0.5% TDCA gel group compared to the placebo group. This suggests that the use of biomarkers as a companion diagnostic method may further increase the potential for clinical improvement with TDCA gel. Considering cancer treatment areas where companion diagnostics are firmly established, triple-negative breast cancer (TNBC) is the type of breast cancer with the poorest prognosis. Triple-negative breast cancer is breast cancer that is negative for estrogen receptors (ER), progesterone receptors (PR), and epidermal growth factor receptors (HER2). Currently, three receptor biomarkers are used to predict prognosis and drug response to anticancer drugs. Based on the above precedent, it is considered that screening for the three biomarkers SMOC1, CRK, and ENTP6 before atopic treatment and using them to predict treatment prognosis and drug response is sufficiently valid. Therefore, in the pre-treatment screening stage of atopic dermatitis patients, a diagnostic system can be established to classify atopic patients using the baseline values ​​of the three protein biomarkers.

[0105] The present disclosure relates to biomarkers that predict responsiveness to TDCA treatment for atopic dermatitis. The present disclosure relates to a companion diagnostic composition that predicts a patient group likely to respond to TDCA treatment for atopic dermatitis based on baseline levels of SMOC1, CRK, and / or ENTP6 in plasma. The present invention confirms that such biomarkers can be useful as companion diagnostic compositions that can predict atopic dermatitis patients who will respond better to TDCA.

[0106] It will be apparent to those skilled in the art that various modifications and variations of the biomarkers for predicting the therapeutic efficacy of GPCR19 agonists in treating atopic dermatitis and the methods for treating subjects having such biomarkers can be made without departing from the spirit or scope of the present disclosure. Accordingly, the present disclosure is intended to cover such modifications and variations as fall within the scope of the appended claims and their equivalents.

[0107] SEQ ID NO: 1 (IGHA2) ASPTSPKVFPLSLDSTPQDGNVVVACLVQGFFPQEPLSVTWSESGQNVTARNFPPSQDASGDLYTTSSQLTLPATQCPDGKSVTCHVKHYTNSSQDVTVPCRVPPPPPCCHPRLSLHRPALEDLLLGSEANLTCTLTGLRDASGATFTWTPSSGKSAVQGPPERDLCGCY SVSSVLPGCAQPWNHGETFTCTAAHPELKTPLTANITKSGNTFRPEVHLLPPPSEELALNELVTLTCLARGFSPKDVLVRWLQGSQELPREKYLTWASRQEPSQGTTTYAVTSILRVAAEDWKKGETFSCMVGHEALPLAFTQKTIDRMAGKPTHINVSVVMAEADGTCY SEQ ID NO: 2 (ENTP6) MKKGIRYETSRKTSYIFQQPQHGPWQTRMRKISNHGSLRVAKVAYPLGLCVGVFIYVAYIKWHRATATQAFFSITRAAPGARWGQQAHSPLGTAADGHEVFYGIMFDAGSTGTRVHVFQFT RPPRETPTLTHETFKALKPGLSAYADDVEKSAQGIRELLDVAKQDIPFDFWKATPLVLKATAGLRLLPGEKAQKLLQKVKKVFKASPFLVGDDCVSIMNGTDEGVSAWITINFLTGSLKTP GGSSVGMLDLGGGSTQIAFLPRVEGTLQASPPGYLTALRMFNRTYKLYSYSYLGLGLMSARLAILGGVEGQPAKDGKELVSPCLSPSFKGEWEHAEVTYRVSGQKAAASLHELCAARVSEV LQNRVHRTEEVKHVDFYAFSYYYDLAAGVGLIDAEKGGSLVVGDFEIAAKYVCRTLETQPQSSPFSCMDLTYVSLLLQEFGFPRSKVLKLTRKIDNVETSWALGAIFHYIDSLNRQKSPAS SEQ ID NO: 3 (SMOC1) MLPARCARLLTPHLLLVLVQLSPARGHRTTGPRFLISDRDPQCNLHCSRTQPKPICASDGRSYESMCEYQRAKCRDPTLGVVHRGRCKDAGQSKCRLERAQALEQAKK PQEAVFVPECGEDGSFTQVQCHTYTGYCWCVTPDGKPISGSSVQNKTPVCSGSVTDKPLSQGNSGRKDDGSKPTPTMETQPVFDGDEITAPTLWIKHLVIKDSKLNNTN IRNSEKVYSCDQERQSALEEAQQNPREGIVIPECAPGGLYKPVQCHQSTGYCWCVLVDTGRPLPGTSTRYVMPSCESDARAKTTEADDPFKDRELPGCPEGKKMEFIT SLLDALTTDMVQAINSAAPTGGGRFSEPDPSHTLEERVVHWYFSQLDSNSSNDINKREMKPFKRYVKKKAKPKKCARRFTDYCDLNKDKVISLPELKGCLGVSKEGRLV SEQ ID NO: 4 (ENPL) MRALWVLGLCCVLLTFGSVRADDEVDVDGTVEEDLGKSREGSRTDDEVVQREEEAIQLDGLNASQIRELREKSEKFAFQAEVNRMMMKLIINSLYKNKEIFLRELISNASDALDKIRLISLTDENALSGNEELTVKIKCDKEKNLLHVTDTGVGMTREELVKNLGTIAKSGTSEFLNKMTEAQEDGQSTSELIGQFGVGFYSAFLVADKVIVTSKHNNDTQHIWESDSNEFSVIADPRGNTLGRGTTITLVLKEEASDYLELDTIKNLVKKYSQFINFPIYVWSSKTETVEEPMEEEAAKEEKEESDDEAAVEEEEEEKKPKTKKVEKTVWDWELMNDIKPIWQRPSKEVEEDEYKAFYKSFSKSEDDPAYIHFTAEGEVTFSILIFVPTSAPRGLFDEY GSKKSDYIKLYVRRVFITDDFHDMMPKYLNFVKGVVDSDDLPLNVSRETLQQHKLLKVIRKKLVRKTLDMIKIADDKYNDTFWKEFGTNIKLGVIEDHSNRTRLAKLLRFQSSHHPTDITSLDQYVERMKEKQDKIYFMAGSSRKEAESSPFVERLLKKGYEVIYLTEPVDEYCIQALPEFDGKRFQNVAKEGVKFDESEKTKESREAVEKEFEPLLNWMKDKALKDKIEKAVVSQRLTESPCALVASQYGWSGNMERIMKAQAYQTGKDISTNYYASQKKTFEINPRHPLIRDMLRRIKEDEDDKTVLDLAVVLFETATLRSGYLLPDTKAYGDRIERMLRLSLNIDPDAKVEEEEEPEETAEDTTEDTEQDEDEEMDVGTDEEEETAKESTAEKDEL sequence number 5(CRK) MAGNFDSEERSSWYWGRLSRQEAVALLQGQRHGVFLVRDSSTSPGDYVLSVSENSRVSHYIINSSGPRPPVPPSPAQPPPGVSPSRLRIGDQEFDSLPALLEFYKIHYLDTTTLIEPVSRSRQGSGVILRQEEAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQWWNAEDSEGKRGMIPVPYVEKYRPASASVSALIGGNQEGSHPQPLGGPEPGPYAQPSVNTPLPNLQNGPIYARVIQKRVPNAYDKTALALEVGELVKVTKINVSGQWEGECNGKRGHFPFTHVRLLDQQNPDEDFS

Claims

1. A method for predicting the efficacy of a composition comprising a GPCR19 agonist for treating atopic dermatitis in a subject, the method comprising the steps of providing concentrations of one or more biomarkers in a sample from the subject and comparing each concentration to a threshold level, wherein a concentration of at least one of the one or more biomarkers being at the threshold level indicates that the composition may be more effective in treating atopic dermatitis, and the one or more biomarkers are selected from the group consisting of IGHA2, ENTP6, SMOC1, ENPL, and CRK.

2. 1. A method for predicting the efficacy of a composition comprising a GPCR19 agonist for treating atopic dermatitis in a subject, comprising: obtaining a sample from a subject; quantitating the concentration of each of one or more biomarkers in the sample; and comparing each concentration to a threshold level. wherein, when the concentration of at least one of the one or more biomarkers is at a threshold level, the composition is more likely to be effective in treating atopic dermatitis; and the one or more biomarkers are selected from the group consisting of IGHA2, ENTP6, SMOC1, ENPL, and CRK.

3. The method of claim 1 or 2, wherein the one or more biomarkers include one or more of SMOC1, ENTP6, and CRK.

4. The method of claim 3, wherein the concentration of SMOC1 or CRK is at a threshold value, which indicates that the composition is more likely to be effective in treating atopic dermatitis.

5. The method of claim 3, wherein the one or more biomarkers include ENTP6 and at least one of SMOC1 and CRK.

6. The method of claim 4, wherein the concentration of ENTP6 is at a threshold value, which additionally indicates that the composition is more likely to be effective in treating atopic dermatitis.

7. The method according to any one of claims 1 to 6, wherein the threshold level of IGHA2 is at a concentration of about 149,579 pM or more, the threshold level of ENTP6 is at a concentration of about 650 pM or less, the threshold level of SMOC1 is at a concentration of about 30 pM or more, the threshold level of ENPL is at a concentration of about 148.8 pM or less, and the threshold level of CRK is at a concentration of about 4518.2 pM or less.

8. The method according to any one of claims 1 to 7, characterized in that the sample is blood, serum or plasma.

9. 9. The method according to any one of claims 1 to 8, characterized in that the concentration of the biomarker is measured via mass spectrometry, enzyme-linked immunosorbent assay (ELISA), capillary Western immunoassay (WES), bead-based flow cytometry or Western blotting.

10. 10. The method of claim 9, wherein the mass spectrometry is multiple reaction monitoring MS (MRM-MS).

11. 11. The method according to any one of claims 1 to 10, characterized in that the efficacy of the composition is determined by calculating an Eczema Area and Severity Index (EASI) score or a validated Investigator Global Assessment (IGA) score.

12. The method according to any one of claims 1 to 11, wherein the GPCR19 agonist comprises taurodeoxycholic acid (TDCA), a derivative thereof, or a pharmaceutically acceptable salt thereof.

13. 13. The method of claim 12, wherein the TDCA comprises sodium taurodeoxycholate.

14. 14. The method according to claim 12 or 13, characterized in that the composition is a pharmaceutical composition (drug dosage form) selected from the group consisting of oral forms, topical forms, and injectables.

15. 15. The method of claim 14, wherein the composition is a topical form containing TDCA.

16. 16. The method of claim 15, wherein the composition is a gel comprising TDCA.

17. 17. The method of claim 16, wherein the composition is NUGEL®.

18. A method for treating atopic dermatitis in a subject in need thereof, comprising administering to the subject a composition comprising a GPCR19 agonist, wherein the subject has been identified as being more likely to be effectively treated by the composition using the method of any one of claims 1 to 17.

19. 20. The method of claim 18, wherein the composition comprises TDCA.

20. 20. The method of claim 19, wherein the composition is a TDCA gel.

21. 21. The method of claim 20, wherein the composition is NUGEL®.

22. The method according to any one of claims 18 to 21, wherein the concentration of one or more of IGHA2, ENTP6, SMOC1, ENPL and CRK in the subject's blood sample is at a threshold level.

23. 23. The method of claim 22, wherein the concentration of one or more of ENTP6, SMOC1, and CRK in the blood sample is at a threshold level.

24. 24. The method of claim 23, wherein the concentration of SMOC1 or CRK in the blood sample is at a threshold level.

25. 24. The method of claim 23, wherein the concentration of ENTP6 and the concentration of at least one of SMOC1 and CRK in the blood sample are at threshold levels.

26. 25. The method of claim 24, wherein the concentration of ENTP6 in the blood sample is at a threshold level.

27. The method of any one of claims 18 to 24, wherein the threshold level of IGHA2 is at a concentration of about 149,579 pM or more, the threshold level of ENTP6 is at a concentration of about 650 pM or less, the threshold level of SMOC1 is at a concentration of about 30 pM or more, the threshold level of ENPL is at a concentration of about 148.8 pM or less, and the threshold level of CRK is at a concentration of about 4518.2 pM or less.

28. Use of a composition comprising TDCA in the manufacture of a medicament for treating atopic dermatitis in a subject with type A atopy or type A' atopy.

29. A composition comprising TDCA for use in treating atopic dermatitis in a subject with type A atopy or type A' atopy.

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