Treatment for Atopic Dermatitis

Abstract

Described herein are treatments and preventions for atopic dermatitis (AD), antibodies and pharmaceutical compositions for use in the treatment or prevention of AD, and the use of anti-IL-31RA antibodies (e.g., nemolizumab) in the manufacture of medicaments for the treatment or prevention of AD. Also described herein are biomarkers for AD, and methods of altering or improving these biomarkers through treatment with an antibody that binds IL-31RA (e.g., nemolizumab).

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority under 35 U.S.C. §119(e) to U.S. Provisional Application No. 63 / 238,643, filed August 30, 2021, the entire contents of which are incorporated herein by reference.

[0002] Field Described herein are treatments and preventions for atopic dermatitis (AD), antibodies and pharmaceutical compositions for use in the treatment or prevention of AD, and the use of anti-IL-31RA antibodies (e.g., nemolizumab) in the manufacture of medicaments for the treatment or prevention of AD. Also described herein are biomarkers for AD, and methods of altering or improving these biomarkers through treatment with an antibody that binds IL-31RA (e.g., nemolizumab). [Background technology]

[0003] background The following discussion is provided merely to aid the reader in understanding the present disclosure and is not admitted to describe or constitute prior art to the present disclosure.

[0004] Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by pruritus (itch), xerosis (dry skin), and eczematous lesions that include erythema, infiltration / papulation, oozing with crusting, desquamation, and lichenification. The prevalence of AD is highest in young children and gradually decreases with age. The prevalence is high in developed countries.

[0005] Although topical treatments are somewhat effective, they are not always sufficient to control moderate to severe AD in patients, especially those who require the addition of phototherapy or systemic treatment to achieve adequate control of AD. Systemic corticosteroids such as cyclosporine and other immunosuppressive treatments can be effective in controlling the disease in some patients, but given the high response variability and known secondary adverse effects of these drugs, new drugs are needed to better control the disease while reducing the risk of secondary adverse effects.

[0006] There remains a need for treatments for AD and for identifying patients who are likely to respond to or are responding to such treatments. Summary of the Invention

[0007] overview Described herein are treatments and preventions for atopic dermatitis (AD) that achieve certain therapeutic results.Generally, the treatments and preventions include administering anti-IL-31RA antibodies (e.g., nemolizumab) to a subject with AD.Also described herein are biomarkers for AD and methods of using the biomarkers of the present disclosure to determine whether a subject responds to treatment.

[0008] In a first aspect, the disclosure provides a method of treating or preventing atopic dermatitis (AD) in a subject, the method comprising administering an anti-IL-31RA antibody to a subject having AD, wherein the subject has lesional skin in which expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is upregulated compared to non-lesional skin of an individual without AD or non-lesional skin of the subject.

[0009] In a second aspect, the disclosure provides a method of altering or reducing expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in lesional skin of a subject with atopic dermatitis (AD), comprising administering an anti-IL-31RA antibody to a subject with AD, wherein at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is upregulated in lesional skin of the subject compared to non-lesional skin of an individual not having AD or non-lesional skin of the subject, and wherein administration of the anti-IL-31RA antibody reduces expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in the lesional skin.

[0010] In some embodiments of the first and second aspects, CCL20, CCL22, CCL27, and VEGF are upregulated in the lesional skin of the subject compared to non-lesional skin of an individual without AD or non-lesional skin of the subject.

[0011] In some embodiments of the first and second aspects, expression of CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18 is determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), DNA or RNA microarray, Western blotting, ELISA, surface plasmon resonance, or mass spectrometry.

[0012] In some embodiments of the first and second aspects, one, two, three, four, five, or six of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 are upregulated in the lesional skin of the subject compared to non-lesional skin of an individual without AD or non-lesional skin of the subject.

[0013] In some embodiments of the first and second aspects, expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is altered or reduced within 2 weeks, 4 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, or 20 weeks of administration of an anti-IL-31RA antibody, compared to baseline expression levels in the subject's skin lesions prior to administration of an anti-IL-31RA antibody.

[0014] In some embodiments of the first and second aspects, expression of CCL20 is reduced by at least about a third, expression of CCL22 is reduced by at least about a 1.1-fold, expression of CCL27 is reduced by at least about a third, expression of VEGF is reduced by at least about a 1.6-fold, expression of CCL18 is reduced by at least about an 8-fold, and / or expression of IL1RA is increased by at least about a 1.1-fold.

[0015] In a third aspect, the disclosure provides a method of reducing expression of inflammatory biomarkers in the skin of a subject having atopic dermatitis (AD), comprising administering to the subject having AD an anti-IL-31RA antibody, thereby reducing the inflammatory response in the skin.

[0016] In some embodiments of any of the preceding aspects, the inflammatory biomarker is selected from at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18. In some embodiments of any of the preceding aspects, the inflammatory biomarker is selected from at least one of CCL20, CCL22, CCL27, and VEGF. In some embodiments of any of the preceding aspects, expression of one, two, three, or all four of CCL20, CCL22, CCL27, and VEGF is reduced.

[0017] In some embodiments of any of the aforementioned aspects, expression of the inflammatory biomarker is reduced in at least one skin lesion of the subject.

[0018] In some embodiments of any of the foregoing aspects, the subject achieves at least a 66.5% reduction in Eczema Area and Severity Index (EASI) scoring after administration of an anti-IL-31RA antibody. In some embodiments of any of the foregoing aspects, the subject achieves at least a 43.2% reduction in Peak Pruritus Numeric Rating Scale (PP-NRS) scoring after administration of an anti-IL-31RA antibody. In some embodiments of any of the foregoing aspects, the subject experiences improved sleep after administration of an anti-IL-31RA antibody.

[0019] In some embodiments of any of the aforementioned aspects, the subject is an adult. In some embodiments of any of the aforementioned aspects, the subject is an adolescent, optionally between the ages of 12 and 17.

[0020] In some embodiments of any of the aforementioned aspects, the anti-IL-31RA antibody is administered subcutaneously.

[0021] In some embodiments of any of the foregoing aspects, the anti-IL-31RA antibody is administered once per week, once per two weeks, once per three weeks, once per four weeks, once per five weeks, once per six weeks, once per seven weeks, or once per eight weeks.

[0022] In some embodiments of any of the two foregoing aspects, the anti-IL-31RA antibody is administered at a dose of about 0.01 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 1.5 mg / kg, about 1.5 mg / kg to about 2.5 mg / kg, or about 2.5 mg / kg to about 10 mg / kg.

[0023] In some embodiments of any of the foregoing aspects, the anti-IL-31RA antibody is administered at a dose of about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, or about 90 mg.

[0024] In some embodiments of any of the foregoing aspects, the anti-IL-31RA antibody comprises a heavy chain variable region comprising an HCDR1 comprising SEQ ID NO:8, an HCDR2 comprising SEQ ID NO:9, and an HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising an LCDR1 comprising SEQ ID NO:12, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:14. In some embodiments of any of the foregoing aspects, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments of any of the foregoing aspects, the anti-IL-31RA antibody is nemolizumab.

[0025] In some embodiments of any of the aforementioned aspects, the anti-IL-31RA antibody is administered subcutaneously at a loading dose of 60 mg, followed by doses of 30 mg every 4 weeks for at least 12, 14, 16, or 18 weeks.

[0026] In some embodiments of any of the aforementioned aspects, the anti-IL-31RA antibody is administered according to a flat dosing regimen.

[0027] In some embodiments of any of the aforementioned aspects, the anti-IL-31RA antibody is administered according to a loading dose regimen.

[0028] In a fourth aspect, the disclosure provides a method of diagnosing atopic dermatitis (AD), comprising detecting expression levels of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a sample obtained from a subject suspected of having AD, and comparing the expression levels of the biomarkers to a reference level, wherein the reference level is the expression level of each corresponding biomarker in a skin sample from an individual not having AD or in a sample of non-lesional skin from the subject.

[0029] In a fifth aspect, the present disclosure provides a method of determining whether a subject with atopic dermatitis (AD) will respond to treatment with an anti-IL-31RA antibody, comprising detecting expression levels of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a sample obtained from a subject suspected of having AD, and comparing the expression levels of the biomarkers to a reference level, wherein if the expression level of the biomarkers is higher than the reference level, the subject will respond to treatment, and the reference level is the expression level of each corresponding biomarker in a skin sample from an individual not having AD or in a sample of non-lesional skin from the subject.

[0030] In some embodiments of the fourth and fifth aspects, the sample obtained from a subject suspected of having AD is a skin sample, optionally including a lesion.

[0031] In some embodiments of the fourth and fifth aspects, the expression level of the biomarker(s) is determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), DNA or RNA microarray, Western blotting, ELISA, surface plasmon resonance, or mass spectrometry.

[0032] In some embodiments of the fourth and fifth aspects, the expression level of CCL20 is increased at least about 1.6-fold higher than the reference level, the expression level of CCL22 is increased at least about 1.02-fold higher than the reference level, the expression level of CCL27 is increased at least about 2.25-fold higher than the reference level, the expression of VEGF is increased at least about 1.05-fold higher than the reference level, the expression of CCL18 is increased at least about 2.45-fold higher than the reference level, and / or the expression of IL1RA is increased at least about 1.1-fold higher than the reference level.

[0033] In a sixth aspect, the present disclosure provides a method of determining whether a subject with atopic dermatitis (AD) responds to treatment with an anti-IL-31RA antibody, comprising detecting expression levels of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a post-treatment sample obtained from a subject with AD who has been administered at least one dose of an anti-IL-31RA antibody, and comparing the expression levels of the biomarkers to baseline expression levels from a sample obtained from the same subject before treatment was initiated, wherein a decrease in expression levels of CCL20, CCL22, CCL27, VEGF, and / or CCL18 and / or an increase in expression levels of IL1RA indicates that the subject responds to treatment.

[0034] In some embodiments of the sixth aspect, the sample is a skin sample, optionally including a lesion.

[0035] In some embodiments of the sixth aspect, the expression level of the biomarker(s) is determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), DNA or RNA microarray, Western blotting, ELISA, surface plasmon resonance, or mass spectrometry.

[0036] In some embodiments of the sixth aspect, compared to baseline levels, post-treatment expression of CCL20 is reduced by at least about a third, expression of CCL22 is reduced by at least about a 1.1-fold, expression of CCL27 is reduced by at least about a third, expression of VEGF is reduced by at least about a 1.6-fold, expression of CCL18 is reduced by at least about an 8-fold, and / or expression of IL1RA is increased by at least about a 1.1-fold.

[0037] In some embodiments of the sixth aspect, the post-treatment sample is obtained about 4 weeks, about 8 weeks, or about 12 weeks after administration of the anti-IL-31RA antibody.

[0038] In some embodiments of the sixth aspect, the anti-IL-31RA antibody comprises a heavy chain variable region comprising an HCDR1 comprising SEQ ID NO:8, an HCDR2 comprising SEQ ID NO:9, and an HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising an LCDR1 comprising SEQ ID NO:12, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:14. In some embodiments of the sixth aspect, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments of the sixth aspect, the anti-IL-31RA antibody is nemolizumab.

[0039] In a seventh aspect, the disclosure provides a pharmaceutical composition comprising an anti-IL-31RA antibody for use in treating or preventing atopic dermatitis (AD) in a subject, wherein the subject has lesional skin in which expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is upregulated compared to non-lesional skin of an individual without AD or non-lesional skin of the subject.

[0040] In an eighth aspect, the present disclosure provides a pharmaceutical composition comprising an anti-IL-31RA antibody for use in altering or reducing expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in lesional skin of a subject with atopic dermatitis (AD), comprising administering an anti-IL-31RA antibody to a subject with AD, wherein at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is upregulated in lesional skin of the subject compared to non-lesional skin of an individual or non-lesional skin of the subject without AD, and wherein administration of the anti-IL-31RA antibody reduces expression of at least one of CCL20, CCL22, CCL27, VEGF, and CCL18 in the lesional skin and / or increases expression of IL1RA in the lesional skin.

[0041] In a ninth aspect, the present disclosure provides a pharmaceutical composition comprising an anti-IL-31RA antibody for use in reducing expression of inflammatory biomarkers in the skin of a subject with atopic dermatitis (AD).

[0042] In some embodiments of the seventh, eighth, or ninth aspect, the inflammatory biomarker is selected from at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18, preferably CCL20, CCL22, CCL27, and / or VEGF.

[0043] In some embodiments of the seventh, eighth, or ninth aspect, the subject is an adult.

[0044] In some embodiments of the seventh, eighth, or ninth aspect, the subject is an adolescent, optionally between the ages of 12 and 17.

[0045] In some embodiments of the seventh, eighth, or ninth aspects, the pharmaceutical composition is formulated for subcutaneous administration.

[0046] In some embodiments of the seventh, eighth, or ninth aspect, the anti-IL-31RA antibody comprises a heavy chain variable region comprising an HCDR1 comprising SEQ ID NO: 8, an HCDR2 comprising SEQ ID NO: 9, and an HCDR3 comprising SEQ ID NO: 10, and a light chain variable region comprising an LCDR1 comprising SEQ ID NO: 12, an LCDR2 comprising SEQ ID NO: 13, and an LCDR3 comprising SEQ ID NO: 14. In some embodiments of the seventh, eighth, or ninth aspect, the anti-IL-31RA antibody is nemolizumab, or a fragment or variant thereof.

[0047] In a tenth aspect, the present disclosure provides a diagnostic agent for use in diagnosing atopic dermatitis (AD), wherein the diagnostic agent is capable of detecting the expression level of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a sample obtained from a subject suspected of having AD, whereby the expression level of the biomarker(s) can be compared to a reference level, the reference level being the expression level of each corresponding biomarker in a skin sample from an individual not having AD or in a sample of non-lesional skin from the subject.

[0048] In an eleventh aspect, the present disclosure provides a diagnostic agent for use in determining whether a subject having atopic dermatitis (AD) will respond to treatment with an anti-IL-31RA antibody, wherein the diagnostic agent is capable of detecting the expression level of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a sample obtained from a subject suspected of having AD, whereby the expression level of the biomarker(s) can be compared to a reference level, where if the expression level of the biomarker(s) is higher than the reference level, the subject will respond to treatment, and the reference level is the expression level of each corresponding biomarker in a skin sample from an individual not having AD or in a sample of non-lesional skin from the subject.

[0049] In a twelfth aspect, the present disclosure provides a diagnostic agent for use in determining whether a subject with atopic dermatitis (AD) responds to treatment with an anti-IL-31RA antibody, the diagnostic agent being capable of detecting expression levels of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a post-treatment sample obtained from a subject with AD administered at least one dose of an anti-IL-31RA antibody, whereby the expression levels of the biomarker(s) can be compared to baseline expression levels from a sample obtained from the same subject before treatment is initiated, and a decrease in the expression levels of CCL20, CCL22, CCL27, VEGF, and / or CCL18 and / or an increase in the expression level of IL1RA indicates that the subject responds to treatment. The foregoing summary and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the present disclosure as claimed. Other objects, advantages and novel features will become readily apparent to those skilled in the art from the following brief description of the drawings and detailed description of the present disclosure. [Brief description of the drawings]

[0050] [Figure 1] Figure 1 shows the study design of a clinical trial for treating atopic dermatitis (AD) with nemolizumab. BL = baseline, n = number of subjects, q4w = every 4 weeks, TCS = topical corticosteroid, W = week. [Diagram 2] The mean ± standard error (SD) of the time concentration profile of nemolizumab (PK population, N=18) is shown. [Diagram 3]Comparison of popPK between simulated and observed nemolizumab concentrations (PK population, N=18) is shown (h=time, CI=confidence interval, PI=prediction interval). Note: Black solid line is observed median, black dashed lines are observed 5th and 95th percentiles, blue solid line is simulated median median, red solid line is simulated 5th and 95th percentile median. Blue area is 95% PI of simulated median, red area is 95% PI of 5th and 95th percentiles. [Figure 4] Pharmacokinetic / pharmacodynamic profiles of predicted population, predicted individual, and observed Adolescent Eczema Area and Severity Index are shown (PK population, N=18). EASI=Eczema Area and Severity Index. Note: Gray points are observed data points, solid black line is observed median, and dashed black lines are observed 2.5 and 97.5 percentiles. Gray area is simulated 95% prediction interval (PI) of median, and blue area is 95% PI of 2.5 and 97.5 percentiles. [Diagram 5] Predicted population, individual predicted population, and observed Adolescent Peak Pruritus Numeric Rating Scale pharmacokinetic / pharmacodynamic profiles are shown (PK population, N=18). PP-NRS=Peak Pruritus Numeric Rating Scale. Note: Gray points are observed data points, solid black line is observed median, and dashed black lines are observed 2.5 and 97.5 percentiles. Gray area is simulated 95% prediction interval (PI) of median, and blue area is 95% PI of 2.5 and 97.5 percentiles. [Figure 6] Predicted and observed adolescent investigator's global assessment pharmacokinetic / pharmacodynamic profiles are shown (PK population, N=18). IGA=Investigator's Global Assessment. Note: The solid black line is the observed median and the grey area is the 95% prediction interval (PI) of the simulated median. [Figure 7] Nemolizumab time concentration profiles of individual subjects with anti-drug antibody results are shown (PK population, N=18). [Figure 8] Figure 1 shows plots of percent change in Eczema Area and Severity Index (EASI) scores over time (last observation carried forward). Intention-to-treat population. Means ± standard errors are reported. [Figure 9] A plot of the proportion of subjects who achieved investigator's global assessment of success (non-responders) is shown (intent-to-treat population). [Figure 10] Figure 1 shows plots of percent change from baseline in weekly mean peak pruritus numerical rating scale scores over time (last observation carried forward) (intention-to-treat population). Means ± standard errors are reported. [Figure 11] Figure 1 shows plots of percent change from baseline in mean weekly Pruritus Numeric Rating Scale scores over time (last observation carried forward) (intention-to-treat population). Means ± standard errors are reported. [Figure 12] Figure 1 shows plots of percent change from baseline in weekly mean sleep disorder numeric rating scale scores over time (last observation carried forward) (intention-to-treat population). Means ± standard errors are reported. [Figure 13-1] Plots of supervised analysis with mixed model repeated measures (MMRM) are shown. Note: Supervised analysis (MMRM) of 6 significantly regulated protein biomarkers in the stratum corneum with respect to EASI75. p-values ​​correspond to the interaction between sample type and EASI75 responders (Y, blue line) vs. non-responders (N, red line). Least squares estimated means (LS-means) with 95% confidence intervals are shown for each treatment in each skin condition (lesional / non-lesional baseline and week 16). For simplicity, only interaction p-values ​​are described in the figure, but the selection criteria were based on the smallest adjusted p-value (interaction, main effect). [Figure 13-2] This is a continuation of Figure 13-1. [Figure 14]Results of an unsupervised analysis on stratum corneum samples of 15 subjects are shown. Note: Unsupervised analysis on stratum corneum samples (NTF) showing the 15 biomarkers that contribute most to the NTF analysis (red=high expression, blue=low expression, X=missing value). p-value=0.0041 corresponds to the association between subject response (EASI-75) and the NTF component used to rank the subjects. Y=EASI-75 responders and N=EASI-75 non-responders. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0051] Detailed Description Described herein is the treatment and prevention of atopic dermatitis (AD) using anti-IL-31RA antibodies (e.g., nemolizumab) and previously unknown biomarkers and gene signatures associated with AD. Biomarkers of the present disclosure include CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18. The treatment and prevention of the present disclosure achieves therapeutic endpoints (e.g., reduction in Eczema Area and Severity Index (EASI), peak Pruritus Numeric Rating Scale (PP-NRS) scoring, and / or improved sleep) previously unknown or unobtainable with conventional treatments for AD.

[0052] I. Definition It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0053] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Unless otherwise specified, materials and / or methodologies known to those of ordinary skill in the art may be utilized in carrying out the methods described herein, based on the guidance provided herein.

[0054] As used herein, the singular terms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Reference to an object in the singular is not intended to mean "one and only one," unless expressly so stated, but rather "one or more."

[0055] As used herein, when used in conjunction with a numerical value, "about" means the numerical value set forth, as well as plus or minus 10% of the numerical value. For example, "about 10" should be understood as both "10" and "from 9 to 11."

[0056] As used herein, "and / or" refers to and includes any and all possible combinations with one or more of the associated listed items, as well as the lack of combinations when interpreted in the alternative ("or").

[0057] As used herein, a phrase in the form "A / B" or in the form "A and / or B" means (A), (B), or (A and B), and a phrase in the form "at least one of A, B, and C" means (A), (B), (C), (A and B), (A and C), (B and C), or (A, B and C).

[0058] As used herein, the phrase "therapeutically effective amount" with respect to an anti-IL31R antibody (e.g., nemolizumab) refers to a dose of an antibody that provides the specific pharmacological effect for which the drug is administered to a subject in need of such treatment. A therapeutically effective amount may be effective to reduce, ameliorate, or eliminate one or more of the symptoms / complications of AD, including but not limited to pruritus (itch), xerosis (dry skin), and eczema lesions, and significant sleep disturbance, and / or improve the quality of life in a subject with AD. It is emphasized that a therapeutically effective amount of an anti-IL31R antibody (e.g., nemolizumab) may not always be effective in treating AD in every individual subject, even if such a dose is considered a therapeutically effective amount by those skilled in the art. Those skilled in the art can adjust what is considered a therapeutically effective amount according to standard practices, as necessary to treat a particular subject. A therapeutically effective amount may vary based on, for example, the age and weight of the subject, and / or the general health of the subject, and / or the severity of the subject's AD.

[0059] The terms "treat," "treatment," or "treating" as used herein with respect to AD refer to reducing, ameliorating, or eliminating one or more of the symptoms / complications of AD, including, but not limited to, pruritus (itch), xerosis (dry skin), and eczematous lesions, and significant sleep disturbances, and / or improving the quality of life in a subject with AD.

[0060] The terms "prevent", "preventing", or "prevention" as used herein with respect to AD refer to eliminating or reducing the risk of developing one or more of the symptoms / complications of AD, including but not limited to pruritus (itch), xerosis (dry skin), and eczematous lesions, and significant sleep disturbance, or preventing the development of a biomarker signature of the present disclosure associated with AD. Prevention can also refer to the prevention of relapse of AD or recurrence after an initial relapse has been treated or cured.

[0061] As used herein, the terms "individual," "subject," and "patient" are used interchangeably and refer to any individual mammalian subject, e.g., a bovine, canine, feline, equine, or human. In specific embodiments, a subject, individual, or patient is a human.

[0062] II. Atopic Dermatitis (AD) Atopic dermatitis (or "AD") is a skin disease that causes pruritus (itch), xerosis (dry skin), and / or eczematous lesions, the characteristics of which include erythema, infiltration / papulation, oozing with crusting, peeling, and lichenification. Complications of AD may include asthma, allergic rhinitis (hay fever), chronic itchy or scaly skin, pain, skin infections, irritant hand dermatitis, allergic contact dermatitis, and / or significant sleep disturbances. The prevalence of AD is highest in young children and gradually decreases with age. The prevalence is high in developed countries.

[0063] The causes of AD are not fully understood. The pathophysiology is complex and likely involves genetic predisposition, epidermal dysfunction, and T cell-driven inflammation. AD has a genetic component and is closely associated with and commonly co-occurs with other atopic diseases (such as asthma and allergic rhinitis). Several pathophysiological mechanisms contribute to the pathogenesis and clinical manifestations of AD. For example, impaired epithelial barrier function, due to deficiency of the structural protein filaggrin, may promote inflammation and T cell infiltration. The immune response in AD is tilted toward T helper 2 cell-mediated pathways, which may then favor the disruption of the epidermal barrier. Other contributing factors to AD pathogenesis include dysbiosis of the skin microbiota (especially Staphylococcus aureus overgrowth), systemic immune responses (including immunoglobulin E (IgE)-mediated sensitization), and neuroinflammation involved in itch.

[0064] There is no specific diagnostic test for AD. The diagnosis of the disorder is based on specific criteria that take into account the patient's medical history and clinical symptoms. Various diagnostic criteria for AD have been proposed and / or validated, and are known to those skilled in the art. The following Table 1 provides a set of exemplary diagnostic criteria for AD used in clinical practice (Williams HC, et al., Br J Dermatol. 131:383-396 (1994); Williams HC, et al., Br J Dermatol. 131:397-405 (1994); Williams HC, et al., Br J Dermatol. 131:406-416 (1994)). Using these criteria, the diagnosis of AD requires the presence of an itchy skin condition (or parent / caregiver report of a child scratching or rubbing), plus three or more minor criteria that vary with the patient's age.

[0065] (Table 1) TIFF2024535715000001.tif111128 a Early onset of the disease is not always diagnosed in children under 4 years of age

[0066] The clinical manifestations of AD change with age. In infants, the scalp, face, neck, trunk, and extensor (external) surfaces of the limbs are commonly affected, although the diaper area is usually spared. Children typically have involvement of the flexor surfaces of the limbs (i.e., folds / flexions at the elbows and behind the knees), neck, wrists, and ankles. In adolescence and adulthood, the flexor surfaces of the limbs, hands, and feet are commonly affected. Regardless of age, the itch associated with AD generally lasts throughout the day and worsens at night, resulting in poor sleep and a significant decrease in quality of life. It can be difficult to distinguish AD from other skin disorders (e.g., seborrheic dermatitis, contact dermatitis, psoriasis, scabies). In these cases, a family history of atopy and the distribution of lesions may help make the diagnosis. Patients presenting with atypical forms may require a punch skin biopsy to rule out other skin conditions that may mimic AD.

[0067] Current treatments for AD are directed at restoring the skin barrier, including hydrating and repairing the skin, limiting itch, and reducing inflammation where necessary. Successful management of AD therefore requires a multifaceted approach including patient and caregiver education, optimal skin care practices, anti-inflammatory treatment with topical corticosteroids (TCS, first choice) and / or topical calcineurin inhibitors (TCIs), and treatment of skin infections. Systemic immunosuppressants may also be considered in severe cases that cannot be controlled with proper skin care and topical therapy. First generation antihistamines are not routinely recommended for the management of AD due to their sedative and impairing side effects, but short-term use of these agents may be helpful for individuals experiencing severe AD flare-ups, especially if these flare-ups are associated with significant sleep disturbances. Other therapies that may be beneficial include ultraviolet (UV) phototherapy, allergen-specific immunotherapy, or wet pack therapy (application of wet dressings to AD lesions after application of emollients and / or topical corticosteroids).

[0068] III. Biomarker detection The present disclosure is the first to report a biomarker signature for AD that can not only identify and positively diagnose AD, but also identify subjects with AD who are likely to respond to treatment with anti-IL-31RA antibodies (e.g., nemolizumab) and track the response of treatment with anti-IL-31RA antibodies (e.g., nemolizumab). The polynucleotide or polypeptide biomarkers of the present disclosure include CCL18, CCL20, CCL22, CCL27, CX3CL1, CXCL6, CXCL8, cystatin, FASL, galectin, IL11, IL21, IL1RA, SPD, and VEGF (more particularly, CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18), and these biomarkers can be detected by various methods known in the art. Non-limiting examples of detection methods are described below. The detection assays in the methods of the present disclosure may involve purified or isolated DNA (genomic or cDNA), RNA, or protein (polypeptide), or the detection step may be performed directly from a biological sample without the need for further DNA, RNA, or protein purification / isolation.

[0069] For example, using immunoassays, the cytokines shown in the following table were determined to be significantly upregulated in the lesional skin of subjects with AD compared to their expression levels in non-lesional skin of the same patients. The semi-quantitative values ​​shown in the table are the ratios expressed in parts per million (ppm) of the identified cytokines to the total protein content of the sample. As can be seen in this table, the expression levels of CCL18, CCL20, CCL22, CCL27, CXCL6, CXCL8, cystatin, IL1RA, IL11, IL21, SPD, and VEGF were upregulated compared to their expression levels in non-lesional skin, while CX3CL1 was downregulated. Furthermore, expression levels of CCL18, CCL20, CCL22, CCL27, CXCL6, CXCL8, cystatin, IL21, and VEGF were dramatically decreased after 16 weeks of treatment with anti-IL-31RA antibodies (e.g., nemolizumab), whereas expression levels of CXCL1, IL1RA, IL11, and SPD were significantly increased after this course of treatment. Similarly, expression of CCL17 was significantly different between lesional and non-lesional skin in EASI75 responders, and was concomitantly decreased after treatment in lesional skin at 16 weeks.

[0070] TIFF2024535715000002.tif190167

[0071] As shown in the table above and in the Examples section below, the expression level of a biomarker may be higher in the lesional skin of a subject with AD than in the non-lesional skin of the subject. Furthermore, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may result in a decrease or increase in the expression level of a given biomarker in lesional skin.

[0072] For purposes of the methods, uses, and compositions disclosed herein, expression of CCL18 in lesional skin of a subject having AD (or suspected of having AD) can be about 300 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 310 ppm, about 320 ppm, about 330 ppm, about 340 ppm, about 350 ppm, about 360 ppm, about 370 ppm, about 380 ppm, about 390 ppm, about 400 ppm or more. Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) may be at least 300 ppm (calculated as a ratio of biomarker protein to total protein in the sample), at least 310 ppm, at least 320 ppm, at least 330 ppm, at least 340 ppm, at least 350 ppm, at least 360 ppm, at least 370 ppm, at least 380 ppm, at least 390 ppm, at least 400 ppm or more. Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) may be about 1.5-fold, about 2-fold, about 2.1-fold, about 2.2-fold, about 2.3-fold, about 2.4-fold, about 2.5-fold, about 2.6-fold, about 2.7-fold, about 2.8-fold, about 2.9-fold, or about 3-fold or more than the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.5-fold, at least 2-fold, at least 2.1-fold, at least 2.2-fold, at least 2.3-fold, at least 2.4-fold, at least 2.5-fold, at least 2.6-fold, at least 2.7-fold, at least 2.8-fold, at least 2.9-fold, or at least 3-fold or more higher than the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be about 150%, about 200%, about 210%, about 220%, about 230%, about 240%, about 250%, about 260%, about 270%, about 280%, about 290%, or about 300% or more of the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be at least 150%, at least 200%, at least 210%, at least 220%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, or at least 300% or more of the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0073] For purposes of the methods, uses, and compositions disclosed herein, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce CCL18 expression in lesional skin of a subject with (or suspected of having) AD by about 6-fold, about 6.5-fold, about 7-fold, about 7.5-fold, about 8-fold, about 8.5-fold, or less. The length of treatment required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0074] For purposes of the methods, uses, and compositions disclosed herein, expression of CCL20 in lesional skin of a subject having AD (or suspected of having AD) can be about 15.5 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 15.6 ppm, about 15.7 ppm, about 15.8 ppm, about 15.9 ppm, about 16 ppm, about 16.1 ppm, about 16.2 ppm, about 16.3 ppm, about 16.4 ppm, about 16.5 ppm, about 16.6 ppm, about 16.7 ppm, about 16.8 ppm, about 16.9 ppm, about 17 ppm or more. Additionally or alternatively, expression of CCL20 in lesional skin of a subject having AD (or suspected of having AD) can be at least 15.5 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), at least 15.6 ppm, at least 15.7 ppm, at least 15.8 ppm, at least 15.9 ppm, at least 16 ppm, at least 16.1 ppm, at least 16.2 ppm, at least 16.3 ppm, at least 16.4 ppm, at least 16.5 ppm, at least 16.6 ppm, at least 16.7 ppm, at least 16.8 ppm, at least 16.9 ppm, at least 17 ppm or more. Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.1-fold, about 1.2-fold, about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.6-fold, about 1.7-fold, about 1.8-fold, about 1.9-fold, about 2-fold or more than the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject without AD). Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be at least 1.1-fold, at least 1.2-fold, at least 1.3-fold, at least 1.4-fold, at least 1.5-fold, at least 1.6-fold, at least 1.7-fold, at least 1.8-fold, at least 1.9-fold, at least 2-fold or more than the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject without AD).Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or greater of the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0075] For purposes of the methods, uses, and compositions disclosed herein, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce expression of CCL20 in lesional skin of a subject with (or suspected of having) AD by about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The duration of treatment required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0076] For purposes of the methods, uses, and compositions disclosed herein, expression of CCL22 in lesional skin of a subject having AD (or suspected of having AD) can be about 1550 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 1560 ppm, about 1570 ppm, about 1580 ppm, about 1590 ppm, about 1600 ppm, about 1610 ppm, about 1620 ppm, about 1630 ppm, about 1640 ppm, about 1650 ppm, about 1660 ppm, about 1670 ppm, about 1680 ppm, about 1690 ppm, about 1700 ppm or more. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be at least 1550 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), at least 1560 ppm, at least 1570 ppm, at least 1580 ppm, at least 1590 ppm, at least 1600 ppm, at least 1610 ppm, at least 1620 ppm, at least 1630 ppm, at least 1640 ppm, at least 1650 ppm, at least 1660 ppm, at least 1670 ppm, at least 1680 ppm, at least 1690 ppm, at least 1700 ppm or more. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) may be about 1.01 fold, about 1.02 fold, about 1.03 fold, about 1.04 fold, about 1.05 fold, about 1.06 fold, about 1.07 fold, about 1.08 fold, about 1.09 fold, about 1.1 fold, about 1.2 fold, about 1.3 fold, about 1.4 fold, about 1.5 fold, about 1.6 fold, about 1.7 fold, about 1.8 fold, about 1.9 fold, about 2 fold or more than the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or more of the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0077] For purposes of the methods, uses, and compositions disclosed herein, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce expression of CCL22 in lesional skin of a subject with (or suspected of having) AD by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0078] For purposes of the methods, uses, and compositions disclosed herein, expression of CCL27 in lesional skin of a subject having AD (or suspected of having AD) can be about 50 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 51 ppm, about 52 ppm, about 53 ppm, about 54 ppm, about 55 ppm, about 56 ppm, about 57 ppm, about 58 ppm, about 59 ppm, about 60 ppm, about 61 ppm, about 62 ppm, about 63 ppm, about 64 ppm, about 65 ppm, about 66 ppm, about 67 ppm, about 68 ppm, about 69 ppm, about 70 ppm, about 71 ppm, about 72 ppm, about 73 ppm, about 74 ppm, about 75 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be at least 50 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 51 ppm, at least 52 ppm, at least 53 ppm, at least 54 ppm, at least 55 ppm, at least 56 ppm, at least 57 ppm, at least 58 ppm, at least 59 ppm, at least 60 ppm, at least 61 ppm, at least 62 ppm, at least 63 ppm, at least 64 ppm, at least 65 ppm, at least 66 ppm, at least 67 ppm, at least 68 ppm, at least 69 ppm, at least 70 ppm, at least 71 ppm, at least 72 ppm, at least 73 ppm, at least 74 ppm, at least 75 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.5-fold, about 2-fold, about 2.1-fold, about 2.2-fold, about 2.3-fold, about 2.4-fold, about 2.5-fold, about 2.6-fold, about 2.7-fold, about 2.8-fold, about 2.9-fold, or about 3-fold or more than the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.5-fold, at least 2-fold, at least 2.1-fold, at least 2.2-fold, at least 2.3-fold, at least 2.4-fold, at least 2.5-fold, at least 2.6-fold, at least 2.7-fold, at least 2.8-fold, at least 2.9-fold, or at least 3-fold or more higher than the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be about 150%, about 175%, about 200%, about 210%, about 220%, about 225%, about 230%, about 240%, about 250%, about 260%, about 270%, about 280%, about 290%, or about 300% or more of the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be at least 150%, at least 175%, at least 200%, at least 210%, at least 220%, at least 225%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, or at least 300% or more of the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0079] For purposes of the methods, uses, and compositions disclosed herein, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce CCL27 expression in lesional skin of a subject with (or suspected of having) AD by about 1.5-fold, about 1.75-fold, about 2-fold, about 2.25-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or more. The duration of treatment required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0080] For purposes of the methods, uses, and compositions disclosed herein, expression of VEGF in lesional skin of a subject having AD (or suspected of having AD) can be about 150 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 151 ppm, about 152 ppm, about 153 ppm, about 154 ppm, about 155 ppm, about 156 ppm, about 157 ppm, about 158 ​​ppm, about 159 ppm, about 160 ppm, about 161 ppm, about 162 ppm, about 163 ppm, about 164 ppm, about 165 ppm, about 166 ppm, about 167 ppm, about 168 ppm, about 169 ppm, about 170 ppm, about 171 ppm, about 172 ppm, about 173 ppm, about 174 ppm, about 175 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject having AD (or suspected of having AD) can be at least 150 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 151 ppm, at least 152 ppm, at least 153 ppm, at least 154 ppm, at least 155 ppm, at least 156 ppm, at least 157 ppm, at least 158 ​​ppm, at least 159 ppm, at least 160 ppm, at least 161 ppm, at least 162 ppm, at least 163 ppm, at least 164 ppm, at least 165 ppm, at least 166 ppm, at least 167 ppm, at least 168 ppm, at least 169 ppm, at least 170 ppm, at least 171 ppm, at least 172 ppm, at least 173 ppm, at least 174 ppm, at least 75 ppm or more.Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be about 1.01 fold, about 1.02 fold, about 1.03 fold, about 1.04 fold, about 1.05 fold, about 1.06 fold, about 1.07 fold, about 1.08 fold, about 1.09 fold, about 1.1 fold, about 1.2 fold, about 1.3 fold, about 1.4 fold, about 1.5 fold, about 1.6 fold, about 1.7 fold, about 1.8 fold, about 1.9 fold, about 2 fold or more than the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level can be about the same between the lesional skin and the reference sample. Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level can be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or greater of the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0081] For purposes of the methods, uses, and compositions disclosed herein, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce VEGF expression in lesional skin of a subject with (or suspected of having) AD by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0082] For purposes of the methods, uses, and compositions disclosed herein, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) is about 207000 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 207500 ppm, about 208000 ppm, about 208500 ppm, about 209000 ppm, about 209500 ppm, about 210000 ppm, about 210500 ppm, about 211000 ppm, about 211500 ppm, about 212000 ppm, about 212500 ppm, about 213000 ppm, about 213500 ppm , about 214000ppm, about 214500ppm, about 215000ppm, about 215500ppm, about 216000ppm, about 216500ppm, Approx. 217000ppm, Approx. 217500ppm, Approx. 218000ppm, Approx. 218500ppm, Approx. 219000ppm, Approx. 219500, Approx. 220 000, about 221000, about 222000, about 223000, about 224000, about 225000, about 226000, about 227000, about 228000, about 229000, about 230000, about 231000, about 232000, about 233000, about 234000, about 235000 ppm or more.Additionally or alternatively, expression of CCL27 in lesional skin of a subject having (or suspected of having) AD is at least 207000 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 207500 ppm, at least 208000 ppm, at least 208500 ppm, at least 209000 ppm, at least 209500 ppm, at least 210000 ppm, at least 210500 ppm, at least 211000 ppm, at least 211500 ppm, at least 212000 ppm, at least 212500 ppm, at least 213000 ppm, at least 213500 ppm, at least 214000 ppm, at least 214500 ppm, at least 21500 ppm, at least 216000 ppm, at least 217500 ppm, at least 217600 ppm, at least 217700 ppm, at least 217800 ppm, at least 217900 ppm, at least 218000 ppm, at least 218100 ppm, at least 218200 ppm, at least 218300 ppm, at least 218400 ppm, at least 218500 ppm, at least 218600 ppm, at least 218700 ppm, at least 218800 ppm, at least 218900 ppm, at least 219000 ppm, at least 220000 ppm, at least 2201000 ppm, at least 2202000 ppm, at least 220300 ppm, at least 220400 ppm, at least 220500 ppm, at least 220600 ppm, at least 2207000 ppm, at least It can be 15000 ppm, at least 215500 ppm, at least 216000 ppm, at least 216500 ppm, at least 217000 ppm, at least 217500 ppm, at least 218000 ppm, at least 218500 ppm, at least 219000 ppm, at least 219500, at least 220000, at least 221000, at least 222000, at least 223000, at least 224000, at least 225000, at least 226000, at least 227000, at least 228000, at least 229000, at least 230000, at least 231000, at least 232000, at least 233000, at least 234000, at least 235000 ppm or more. Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) may be about 1.01-fold, about 1.02-fold, about 1.03-fold, about 1.04-fold, about 1.05-fold, about 1.06-fold, about 1.07-fold, about 1.08-fold, about 1.09-fold, about 1.1-fold, about 1.2-fold, about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.6-fold, about 1.7-fold, about 1.8-fold, about 1.9-fold, about 2-fold or more than the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 115%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or more of the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 115%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0083] For purposes of the methods, uses, and compositions disclosed herein, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may increase expression of IL1RA in lesional skin of a subject with (or suspected of having) AD by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or more. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0084] For purposes of the methods, uses, and compositions disclosed herein, the other described biomarkers (e.g., CXCL6, CXCL8, cystatin, FASL, galectin, IL11, IL21, SPD, and CCL17) may show a similar pattern of overexpression in lesional skin of a subject with AD (or suspected of having AD) compared to expression levels in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), and CX3CL1 may show increased expression in lesional skin of a subject with AD (or suspected of having AD) compared to expression levels in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Similarly, CXCL6, CXCL8, cystatin, FASL, galectin, CCL17, and IL21 may show a similar pattern of decreased expression in lesional skin of subjects with AD (or suspected of having AD) after treatment with an anti-IL-31RA antibody (e.g., nemolizumab), and CX3CL1, IL11, and SPD may show increased expression in lesional skin of subjects with AD (or suspected of having AD) after treatment with an anti-IL-31RA antibody (e.g., nemolizumab).

[0085] In some embodiments, protein or polypeptide expression levels of a biomarker of the disclosure (e.g., CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18) may be detected by Western blotting, enzyme-linked immunosorbent assay (ELISA), dot blotting, immunohistochemistry, immunofluorescence, immunoprecipitation, immunoelectrophoresis, or mass spectrometry.

[0086] Additionally or alternatively, in some embodiments, polynucleotides encoding the biomarkers of the disclosure (e.g., CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18) may be detected by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), or DNA or RNA microarrays. The starting material for detecting polynucleotides encoding the biomarkers of the disclosure may be genomic DNA, cDNA, RNA, or mRNA. Nucleic acid amplification may be linear or exponential. Specific variants or mutations may be detected by the use of amplification methods with the aid of oligonucleotide primers or probes designed to interact or hybridize in a specific manner with specific target sequences, thus amplifying only the target variants. The primers and probes may also comprise a detectable label or multiple detectable labels. The detectable label associated with the probe may directly generate a detectable signal. Additionally, the detectable label associated with the probe may be indirectly detected using a reagent, which comprises a detectable label and binds to the label associated with the probe.

[0087] In some embodiments, detectably labeled primers or probes may be used in hybridization assays, including but not limited to, Northern blots, Southern blots, microarrays, dot or slot blots, and in situ hybridization assays such as fluorescent in situ hybridization (FISH) to detect target nucleic acid sequences in biological samples. Detectably labeled probes can also be used to monitor the amplification of target nucleic acid sequences. In some embodiments, detectably labeled probes present in an amplification reaction are suitable for monitoring the amount of amplification product(s) produced as a function of time. Examples of such probes include 5'-exonuclease assays (TAQMAN® probes described herein (see also U.S. Pat. No. 5,538,848)), various stem-loop molecular beacons (see, e.g., U.S. Pat. Nos. 6,103,476 and 5,925,517, and Tyagi and Kramer, 1996, Nature Biotechnology 14:303-308), stemless or linear beacons (see, e.g., WO 99 / 21881), PNA Molecular Beacons™ (see, e.g., U.S. Pat. Nos. 6,355,421 and 6,593,091), linear PNA beacons (see, e.g., Kubista et al., 2001, SPIE 4264:53-58), non-FRET probes (see, e.g., U.S. Pat. No. 6,150,097), Sunrise® / Amplifluor™ probes (U.S. Pat. No. 6,548,250), stem-loop and duplex Scorpion probes (Solinas et al., 2001, Nucleic Acids Research 29:E96 and U.S. Pat. No. 6,589,743), bulge loop probes (U.S. Pat. No. 6,590,091), pseudoknot probes (U.S. Pat. No. 6,589,250), cyclicones (U.S. Pat. No. 6,383,752), MGB Eclipse™ probes (Epoch Biosciences), hairpin probes (U.S. Pat. No. 6,596,490), peptide nucleic acid (PNA) light-up probes, self-assembled nanoparticle probes, as well as those described in, for example, U.S. Pat. No. 6,485,901, Mhlanga et al., 2001, Methods 25:463-471, Whitcombe et al., 1999, Nature Biotechnology. 17:804-807, Isacsson et al., 2000, Molecular Cell Probes. 14:321-328, Svanvik et al., 2000, Anal Biochem. 281:26-35, Wolffs et al., 2001, Biotechniques 766:769-771, Tsourkas et al., 2002, Nucleic Acids Research. 30:4208-4215, Riccelli et al., 2002, Nucleic Acids Research 30:4088-4093, Zhang et al., 2002 Shanghai.34:329-332, Maxwell et al., 2002, J.Am.Chem.Soc.124:9606-9612, Broude et al., 2002, Trends Biotechnol.20:249-56, Huang et al., 2002, Chem.Res.Toxicol.15:118-126, and Yu et al., 2001, J.Am.Chem.Soc 14:11155-11161. In some embodiments, the detectable label is a fluorescent dye molecule. Suitable fluorescent moieties include, but are not limited to, the following fluorescent dye molecules acting individually or in combination: 4-acetamido-4'-isothiocyanatostilbene-2,2' disulfonic acids; acridine and derivatives: acridine, acridine isothiocyanate; Alexa Fluors: Alexa Fluor® 350, Alexa Fluor® 488, Alexa Fluor® 546, Alexa Fluor® 555, Alexa Fluor® 568, Alexa Fluor® 594, Alexa Fluor® 647 (Molecular Probes); 5-(2-aminoethyl)aminonaphthalene-l-sulfonic acid (EDANS); 4-amino-N-[3-vinylsulfonyl)phenyl]naphthalimide-3,5 disulfonate (Lucifer Yellow VS); N-(4-anilino-l-naphthyl)maleimide; anthranilamide; Black Hole Quencher™ (BHQ™) dyes (biosearch Technologies); BODIPY dyes: BODIPY® R-6G, BOPIPY® 530 / 550, BODIPY® FL; Brilliant Yellow; Coumarins and derivatives: Coumarin, 7-amino-4-methylcoumarin (AMC, Coumarin 120), 7-amino-4-trifluoromethylcoumarin (Coumarin 151); Cy2®, Cy3®, Cy3.5®, Cy5®, C y5.5®;cyanosine;4',6-diminidino-2-phenylindole (DAPI);5',5"-dibromopyrogallol-sulfonphthalein (bromopyrogallol red);7-diethylamino-3-(4'-isothiocyanatophenyl)-4-methylcoumarin;diethylenetriaminepentaacetate;4,4'-diisothiocyanatodihydro-stilbene-2,2'-disulfonic acid;4,4'-diisothiocyanatostilbene-2,2'-Disulfonic acid; 5-[dimethylamino]naphthalene-l-sulfonyl chloride (DNS, dansyl chloride); 4-(4'-dimethylaminophenylazo)benzoic acid (DABCYL); 4-dimethylaminophenylazophenyl-4'-isothiocyanate (DABITC); Eclipse™ (Epoch Biosciences Inc.); Eosin and derivatives: Eosin, Eosin isothiocyanate; Erythrosine and derivatives: Erythrosine B, Erythrosine isothiocyanate; Ethidium; Fluorescein and derivatives: 5-carboxyfluorescein (FAM), 5-(4,6-dichlorotriazin-2-yl)aminofluorescein (DTAF), 2',7'-dimethoxy-4',5'-dichloro-6-carboxyfluorescein (JOE), Fluorescein, Fluorescein isothiocyanate (F ITC), hexachloro-6-carboxyfluorescein (HEX), QFITC ​​(XRITC), tetrachlorofluorescein (TET); fluorescamine; IR144; IR1446; lanthamide fluorophores; malachite green isothiocyanate; 4-methylumbelliferone; orthocresolphthalein; nitrotyrosine; pararosaniline; phenol red; B-phycoerythrin, R-phycoerythrin; allophycocyanin; o-phthaldialdehyde; Oregon Green®; propidium iodide; pyrene and derivatives: pyrene, pyrene butyrate, succinimidyl 1-pyrene butyrate; QSY® 7; QSY® 9; QSY® 21; QSY® 35 (Molecular Probes); Reactive Red 4 (Cibacron® Brilliant Red 3B-A); rhodamine and derivatives: 6-carboxy-X-rhodamine (ROX), 6-carboxyrhodamine (R6G), Lissamine rhodamine B sulfonyl chloride, rhodamine (Rhod), rhodamine B, rhodamine 123, rhodamine green, rhodamine X isothiocyanate, riboflavin, rosolinic acid, sulforhodamine B, sulforhodamine 101, sulfonyl chloride derivatives of sulforhodamine 101 (Texas Red); terbium chelate derivatives; N,N,N',N'-tetramethyl-6-carboxyrhodamine (TAMRA); tetramethylrhodamine; tetramethylrhodamine isothiocyanate (TRITC); and VIC®. Detector probes can also include sulfonate derivatives of fluorescein dyes with S03 in place of the carboxylate group, phosphoramidite forms of fluorescein, and phosphoramidite forms of CY5 (e.g., commercially available from Amersham).

[0088] Primers or probes can be designed to selectively hybridize to any portion of the nucleic acid sequence encoding the polypeptide biomarkers of the disclosure (e.g., CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18). Methods for preparing primers or probes are well developed in the art. Exemplary nucleic acid sequences of human orthologs of these genes are provided below.

[0089] Homo sapiens CC motif chemokine ligand 20 (CCL20), transcript variant 1, mRNA (NCBI Reference Sequence: NM_004591.3) (SEQ ID NO: 15) TIFF2024535715000003.tif77139

[0090] Homo sapiens CC motif chemokine ligand 22 (CCL22), mRNA, (NCBI Reference Sequence: NM_002990.5) (SEQ ID NO: 16) TIFF2024535715000004.tif15139TIFF2024535715000005.tif214141TIFF2024535715000006.tif43141

[0091] Homo sapiens CC motif chemokine ligand 27 (CCL27), mRNA (NCBI Reference Sequence: NM_006664.4) (SEQ ID NO: 17) TIFF2024535715000007.tif37139

[0092] Homo sapiens vascular endothelial growth factor A (VEGFA), transcript variant 1, mRNA (NCBI Reference Sequence: NM_001025366.3) (SEQ ID NO: 18) TIFF2024535715000008.tif89139TIFF2024535715000009.tif214141TIFF2024535715000010.tif37141

[0093] Homo sapiens interleukin 1 receptor antagonist (IL1RN), transcript variant 2, mRNA (NCBI Reference Sequence: NM_173841.3) (SEQ ID NO: 19) TIFF2024535715000011.tif163141

[0094] Homo sapiens CC motif chemokine ligand 18 (CCL18), mRNA (NCBI Reference Sequence: NM_002988.4) (SEQ ID NO: 20) TIFF2024535715000012.tif128141

[0095] In another aspect, the disclosure provides a method of diagnosing atopic dermatitis (AD), comprising detecting expression levels of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a sample obtained from a subject suspected of having AD, and comparing the expression levels of the biomarkers to a reference level, where the reference level is the expression level of each corresponding biomarker in a skin sample from an individual without AD or in a sample of non-lesional skin from a subject suspected of having AD. Additionally or alternatively, the present disclosure provides a diagnostic agent for detecting the expression level of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a subject suspected of having AD, wherein the expression level of the biomarkers can be compared to a reference level corresponding to the expression level of each biomarker in a skin sample from an individual without AD or a sample of non-lesional skin from a subject suspected of having AD. In some embodiments, the sample obtained from the subject suspected of having AD is a skin sample, optionally including a lesion. For purposes of the above aspects, the skin sample can comprise, consist of, or consist essentially of the stratum corneum.

[0096] In another aspect, the disclosure provides a method of detecting inflammatory biomarkers in a sample, the method comprising detecting an expression level of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a sample obtained from a subject suspected of having AD. The method may further comprise comparing the expression level of the biomarker(s) to a reference level, where the reference level is a corresponding expression level of each biomarker in a skin sample from an individual without AD or a sample of non-lesional skin from a subject suspected of having AD. Additionally or alternatively, the present disclosure provides a diagnostic agent for detecting inflammatory biomarkers in a sample obtained from a subject suspected of having AD, the inflammatory biomarkers comprising at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18, and the expression levels of the biomarkers can be compared to reference levels corresponding to the expression levels of each biomarker in a skin sample from an individual not having AD. In some embodiments, the sample obtained from a subject suspected of having AD is a skin sample, optionally including a lesion. For purposes of the above aspects, the skin sample can comprise, consist of, or consist essentially of the stratum corneum.

[0097] For purposes of the above embodiment, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) may be about 300 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), about 310 ppm, about 320 ppm, about 330 ppm, about 340 ppm, about 350 ppm, about 360 ppm, about 370 ppm, about 380 ppm, about 390 ppm, about 400 ppm or more. Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) may be at least 300 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 310 ppm, at least 320 ppm, at least 330 ppm, at least 340 ppm, at least 350 ppm, at least 360 ppm, at least 370 ppm, at least 380 ppm, at least 390 ppm, at least 400 ppm or more. Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.5-fold, about 2-fold, about 2.1-fold, about 2.2-fold, about 2.3-fold, about 2.4-fold, about 2.5-fold, about 2.6-fold, about 2.7-fold, about 2.8-fold, about 2.9-fold, or about 3-fold or more higher than the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.5-fold, at least 2-fold, at least 2.1-fold, at least 2.2-fold, at least 2.3-fold, at least 2.4-fold, at least 2.5-fold, at least 2.6-fold, at least 2.7-fold, at least 2.8-fold, at least 2.9-fold, or at least 3-fold or more higher than the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be about 150%, about 200%, about 210%, about 220%, about 230%, about 240%, about 250%, about 260%, about 270%, about 280%, about 290%, or about 300% or more of the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be at least 150%, at least 200%, at least 210%, at least 220%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, or at least 300% or more of the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0098] For purposes of the above embodiments, expression of CCL20 in lesional skin of a subject having (or suspected of having) AD can be about 15.5 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 15.6 ppm, about 15.7 ppm, about 15.8 ppm, about 15.9 ppm, about 16 ppm, about 16.1 ppm, about 16.2 ppm, about 16.3 ppm, about 16.4 ppm, about 16.5 ppm, about 16.6 ppm, about 16.7 ppm, about 16.8 ppm, about 16.9 ppm, about 17 ppm or more. Additionally or alternatively, expression of CCL20 in lesional skin of a subject having AD (or suspected of having AD) can be at least 15.5 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), at least 15.6 ppm, at least 15.7 ppm, at least 15.8 ppm, at least 15.9 ppm, at least 16 ppm, at least 16.1 ppm, at least 16.2 ppm, at least 16.3 ppm, at least 16.4 ppm, at least 16.5 ppm, at least 16.6 ppm, at least 16.7 ppm, at least 16.8 ppm, at least 16.9 ppm, at least 17 ppm or more. Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.1-fold, about 1.2-fold, about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.6-fold, about 1.7-fold, about 1.8-fold, about 1.9-fold, about 2-fold or more than the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject without AD). Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be at least 1.1-fold, at least 1.2-fold, at least 1.3-fold, at least 1.4-fold, at least 1.5-fold, at least 1.6-fold, at least 1.7-fold, at least 1.8-fold, at least 1.9-fold, at least 2-fold or more than the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject without AD).Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or greater of the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0099] For purposes of the above embodiments, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be about 1550 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 1560 ppm, about 1570 ppm, about 1580 ppm, about 1590 ppm, about 1600 ppm, about 1610 ppm, about 1620 ppm, about 1630 ppm, about 1640 ppm, about 1650 ppm, about 1660 ppm, about 1670 ppm, about 1680 ppm, about 1690 ppm, about 1700 ppm or more. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be at least 1550 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), at least 1560 ppm, at least 1570 ppm, at least 1580 ppm, at least 1590 ppm, at least 1600 ppm, at least 1610 ppm, at least 1620 ppm, at least 1630 ppm, at least 1640 ppm, at least 1650 ppm, at least 1660 ppm, at least 1670 ppm, at least 1680 ppm, at least 1690 ppm, at least 1700 ppm or more. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) may be about 1.01 fold, about 1.02 fold, about 1.03 fold, about 1.04 fold, about 1.05 fold, about 1.06 fold, about 1.07 fold, about 1.08 fold, about 1.09 fold, about 1.1 fold, about 1.2 fold, about 1.3 fold, about 1.4 fold, about 1.5 fold, about 1.6 fold, about 1.7 fold, about 1.8 fold, about 1.9 fold, about 2 fold or more than the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or more of the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0100] For purposes of the above embodiments, expression of CCL27 in lesional skin of a subject having (or suspected of having) AD can be about 50 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 51 ppm, about 52 ppm, about 53 ppm, about 54 ppm, about 55 ppm, about 56 ppm, about 57 ppm, about 58 ppm, about 59 ppm, about 60 ppm, about 61 ppm, about 62 ppm, about 63 ppm, about 64 ppm, about 65 ppm, about 66 ppm, about 67 ppm, about 68 ppm, about 69 ppm, about 70 ppm, about 71 ppm, about 72 ppm, about 73 ppm, about 74 ppm, about 75 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be at least 50 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 51 ppm, at least 52 ppm, at least 53 ppm, at least 54 ppm, at least 55 ppm, at least 56 ppm, at least 57 ppm, at least 58 ppm, at least 59 ppm, at least 60 ppm, at least 61 ppm, at least 62 ppm, at least 63 ppm, at least 64 ppm, at least 65 ppm, at least 66 ppm, at least 67 ppm, at least 68 ppm, at least 69 ppm, at least 70 ppm, at least 71 ppm, at least 72 ppm, at least 73 ppm, at least 74 ppm, at least 75 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.5-fold, about 2-fold, about 2.1-fold, about 2.2-fold, about 2.3-fold, about 2.4-fold, about 2.5-fold, about 2.6-fold, about 2.7-fold, about 2.8-fold, about 2.9-fold, or about 3-fold or more than the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.5-fold, at least 2-fold, at least 2.1-fold, at least 2.2-fold, at least 2.3-fold, at least 2.4-fold, at least 2.5-fold, at least 2.6-fold, at least 2.7-fold, at least 2.8-fold, at least 2.9-fold, or at least 3-fold or more higher than the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be about 150%, about 175%, about 200%, about 210%, about 220%, about 225%, about 230%, about 240%, about 250%, about 260%, about 270%, about 280%, about 290%, or about 300% or more of the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be at least 150%, at least 175%, at least 200%, at least 210%, at least 220%, at least 225%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, or at least 300% or more of the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0101] For purposes of the above embodiments, expression of VEGF in lesional skin of a subject having (or suspected of having) AD can be about 150 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 151 ppm, about 152 ppm, about 153 ppm, about 154 ppm, about 155 ppm, about 156 ppm, about 157 ppm, about 158 ​​ppm, about 159 ppm, about 160 ppm, about 161 ppm, about 162 ppm, about 163 ppm, about 164 ppm, about 165 ppm, about 166 ppm, about 167 ppm, about 168 ppm, about 169 ppm, about 170 ppm, about 171 ppm, about 172 ppm, about 173 ppm, about 174 ppm, about 175 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject having AD (or suspected of having AD) can be at least 150 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 151 ppm, at least 152 ppm, at least 153 ppm, at least 154 ppm, at least 155 ppm, at least 156 ppm, at least 157 ppm, at least 158 ​​ppm, at least 159 ppm, at least 160 ppm, at least 161 ppm, at least 162 ppm, at least 163 ppm, at least 164 ppm, at least 165 ppm, at least 166 ppm, at least 167 ppm, at least 168 ppm, at least 169 ppm, at least 170 ppm, at least 171 ppm, at least 172 ppm, at least 173 ppm, at least 174 ppm, at least 75 ppm or more. Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be about 1.01 fold, about 1.02 fold, about 1.03 fold, about 1.04 fold, about 1.05 fold, about 1.06 fold, about 1.07 fold, about 1.08 fold, about 1.09 fold, about 1.1 fold, about 1.2 fold, about 1.3 fold, about 1.4 fold, about 1.5 fold, about 1.6 fold, about 1.7 fold, about 1.8 fold, about 1.9 fold, about 2 fold or more than the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level can be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level can be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or greater of the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0102] For purposes of the above embodiment, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) is about 207000 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), about 207500 ppm, about 208000 ppm, about 208500 ppm, about 209000 ppm, about 209500 ppm, about 210000 ppm, about 210500 ppm, about 211000 ppm, about 211500 ppm, about 212000 ppm, about 212500 ppm, about 213000 ppm, about 213500 ppm, about 214000 ppm, about 21500 ppm, about 216000 ppm, about 217000 ppm, about 218000 ppm, about 219000 ppm, about 220000 ppm, about 221500 ppm, about 222000 ppm, about 223000 ppm, about 224000 ppm, about 22500 ppm, about 226000 ppm, about 227000 ppm, about 228000 ppm, about 229000 ppm, about 23000 ppm, about 231000 ppm, about 232000 ppm, about 23300 ppm, about 234000 ppm, about 23500 ppm, about 236000 ppm, about 237000 ppm, about 23800 ppm, about 23900 ppm, about 24000 ppm, about 241000 ppm, about 242000 ppm, about 24300 ppm, about 244000 ppm, about 24500 ppm, about 24600 ppm, about 247 ppm, about 214500 ppm, about 215000 ppm, about 215500 ppm, about 216000 ppm, about 216500 ppm, about 217000 ppm, about 217500 ppm, about 218000 ppm, about 218500 ppm, about 219000 ppm, about 219500, about 220000, about 221000, about 222000, about 223000, about 224000, about 225000, about 226000, about 227000, about 228000, about 229000, about 230000, about 231000, about 232000, about 233000, about 234000, about 235000 ppm or more.Additionally or alternatively, expression of CCL27 in lesional skin of a subject having (or suspected of having) AD is at least 207000 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 207500 ppm, at least 208000 ppm, at least 208500 ppm, at least 209000 ppm, at least 209500 ppm, at least 210000 ppm, at least 210500 ppm, at least 211000 ppm, at least 211500 ppm, at least 212000 ppm, at least 212500 ppm, at least 213000 ppm, at least 213500 ppm, at least 214000 ppm, at least 214500 ppm, at least 21500 ppm, at least 216000 ppm, at least 217500 ppm, at least 217600 ppm, at least 217700 ppm, at least 217800 ppm, at least 217900 ppm, at least 218000 ppm, at least 218100 ppm, at least 218200 ppm, at least 218300 ppm, at least 218400 ppm, at least 218500 ppm, at least 218600 ppm, at least 218700 ppm, at least 218800 ppm, at least 218900 ppm, at least 219000 ppm, at least 220000 ppm, at least 2201000 ppm, at least 2202000 ppm, at least 220300 ppm, at least 220400 ppm, at least 220500 ppm, at least 220600 ppm, at least 2207000 ppm, at least It can be 15000 ppm, at least 215500 ppm, at least 216000 ppm, at least 216500 ppm, at least 217000 ppm, at least 217500 ppm, at least 218000 ppm, at least 218500 ppm, at least 219000 ppm, at least 219500, at least 220000, at least 221000, at least 222000, at least 223000, at least 224000, at least 225000, at least 226000, at least 227000, at least 228000, at least 229000, at least 230000, at least 231000, at least 232000, at least 233000, at least 234000, at least 235000 ppm or more. Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) may be about 1.01-fold, about 1.02-fold, about 1.03-fold, about 1.04-fold, about 1.05-fold, about 1.06-fold, about 1.07-fold, about 1.08-fold, about 1.09-fold, about 1.1-fold, about 1.2-fold, about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.6-fold, about 1.7-fold, about 1.8-fold, about 1.9-fold, about 2-fold or more than the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 115%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or more of the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 115%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0103] IV. Therapeutic Antibodies and Interleukin-31 Receptor Subunit Alpha (IL-31RA) Interleukin-31 (IL-31) is a neuroinflammatory cytokine that can activate both structural and immune cells and peripheral nerves. IL-31 has been implicated in a number of chronic inflammatory diseases, including atopic dermatitis. IL-31 is produced by a variety of cells, including type 2 helper (Th2) T cells. IL-31 signals through a receptor complex consisting of interleukin-31 receptor subunit alpha ("IL-31RA," also known as NR10, glm-r, and GPL) and oncostatin M receptor beta (OSMRβ), which are expressed in immune and epithelial cells, as well as a subset of neurons.

[0104] When IL-31RA functions as an IL-31 receptor, it forms a heterodimer with oncostatin M receptor (OSMR). There are several known splice variants of human-derived IL-31RA (WO00 / 075314): NR10.1 is composed of 662 amino acids and contains a transmembrane domain. NR10.2 is a soluble receptor-like protein composed of 252 amino acids without a transmembrane domain. Further known IL-31RA splice variants that function as transmembrane receptor proteins include NR10.3 and IL-31RAv3. Preferred IL-31RA variants include NR10.3 (also referred to as ILRAv4 (Nat Immunol 5, 752-60, 2004) and IL-31RAv3. NR10.3 (IL31RAv4) consists of 662 amino acids (WO00 / 075314, Nat Immunol 5, 752-60, 2004), and IL31RAv3 consists of 732 amino acids (GenBank accession number: NM-139017).

[0105] The amino acid sequence of IL31RAv4 is as follows: (SEQ ID NO:1)

[0106] The amino acid sequence of IL31RAv3 is as follows: (SEQ ID NO:2)

[0107] Mouse-derived IL-31RA contains the following amino acid sequence: (SEQ ID NO:3)

[0108] Cynomolgus monkey-derived IL-31RA contains the following amino acid sequence: (SEQ ID NO:4)

[0109] For purposes of this disclosure, an anti-IL-31RA antibody (ie, a therapeutic antibody) such as nemolizumab must bind at least human IL-31RA or a splice variant thereof.

[0110] As used herein, the term "antibody" refers collectively to immunoglobulins or immunoglobulin-like molecules, including IgA, IgD, IgE, IgG, and IgM, combinations thereof, or fragments thereof. Antibody fragments may include, for example, Fab fragments and single-chain variable fragments (scFv). Antibodies generally comprise a heavy (H) chain and a light (L) chain interconnected by disulfide bonds. There are two types of light chains: lambda (λ) and kappa (κ). There are five main heavy chain classes (or isotypes) that determine the functional activity of antibody molecules: IgM, IgD, IgG, IgA, and IgE. Each heavy and light chain contains a constant region and a variable region (also known as a "domain"). In combination, the heavy and light chain variable regions, also referred to as "Fab regions", specifically bind to a given antigen. The light and heavy chain variable regions contain a "framework" region interrupted by three hypervariable regions, also referred to as "complementarity determining regions" or "CDRs". The extent of the framework regions and CDRs are defined (see Kabat et al., Sequences of Proteins of Immunological Interest, USDepartment of Health and Human Services, 1991). The Kabat database is currently maintained online. The sequences of the framework regions of different light or heavy chains are relatively conserved within a species, and the framework regions act to form a scaffold for orienting the CDRs through non-covalent interactions between the chains.

[0111] CDRs are primarily responsible for binding to an epitope on an antigen. The CDRs of each chain are typically referred to as CDR1, CDR2, and CDR3, are numbered sequentially starting from the N-terminus, and are typically identified by the chain in which the particular CDR is located. Thus, HCDR3 is located in the variable domain of the heavy chain of the antibody in which it is found, while LCDR1 is the CDR1 from the variable domain of the light chain of the antibody in which it is found. Antibodies that bind to IL-31RA have a specific VDR1. H Area and V LEach CDR has a specific CDR sequence and therefore a specific CDR sequence. Antibodies with different specificities generally have different CDRs. It is the CDRs that differ from antibody to antibody, but only a limited number of amino acid positions in the CDR are directly involved in antigen binding. These positions in the CDR are called specificity determining residues (SDRs).

[0112] The Fc fragment region (Fc) of an antibody plays a role in regulating immune cell activity. The Fc region functions to ensure that each antibody generates an appropriate immune response to a given antigen by binding to a specific class of proteins found on specific cells, such as B lymphocytes, follicular dendritic cells, natural killer cells, macrophages, and neutrophils, called "Fc receptors." The class of heavy chains in antibodies determines their class effect, since the constant domains of the heavy chains constitute the Fc region of an antibody. Heavy chains in antibodies include alpha, gamma, delta, epsilon, and mu, which correlate with the antibody's isotypes IgA, IgG, IgD, IgE, and IgM, respectively. Thus, different isotypes of antibodies have different class effects, since their different Fc regions bind and activate different types of receptors.

[0113] IgG, the most abundant antibody isotype in human serum, has four subclasses. The four highly conserved subclasses are IgG1, IgG2, IgG3, and IgG4. The amino acid sequences of the constant regions of these peptides are known in the art, see, for example, Rutishauser, U. et al. (1968) "Amino acid sequence of the Fc region of a human gamma G-immunoglobulin" PNAS61(4):1414-1421, Shinoda et al. (1981) "Complete amino acid sequence of the Fc region of a human delta chain" PNAS78(2):785-789, and Robinson et al. (1980) "Complete amino acid sequence of a mouse immunoglobulin alpha chain (MOPC511)" PNAS77(8):4909-4913.

[0114] All therapeutic antibodies for purposes of the methods and pharmaceutical uses of the present disclosure are antibodies or fragments thereof that bind to IL-31RA, but the specific anti-IL-31RA antibody is not limited. Nemolizumab is a preferred anti-IL-31RA antibody, but other anti-IL-31RA antibodies can be used as well. Therapeutic antibodies suitable for use in the methods and pharmaceutical uses of the present disclosure can be human, humanized, or chimeric, and can be IgA, IgG (i.e., IgG1, IgG2, IgG3, and IgG4), IgD, IgE, or IgM.

[0115] Nemolizumab is a humanized monoclonal antibody that binds to IL-31RA. Nemolizumab is annotated as follows: Immunoglobulin G2-kappa, anti-[Homo sapiens IL31RA (Interleukin 31 receptor subunit alpha)], humanized monoclonal antibody; gamma 2 heavy chain (1-445) [humanized VH(Homo sapiens IGHV1-2*02 (83.70%)-(IGHD)-IGHJ5*01)[8.8.14](1-121)-Homo sapiens IGHG2*01 (CH1 C10>S(135), R12>K(137), E16>G(141), S17>G(142)(122-219), hinge C4>S(223)(220-231), CH2 H30>Q(268)(232-340), CH3 R11>Q(355), Q98>E(419)(341-445))(122-445)], (224-214')-disulfide containing kappa light chain, (1'-214') [humanized V-kappa (Homo sapiens IGKV1-39*01(82.10%)-IGKJ4*01)[6.3.9](1'-107')-Homo sapiens IGKC*01(108'-214')], dimeric (227-227":230-230")-bisdisulfide. Nemolizumab has disulfide bridges at the following positions: Intra-H(C23-C104) 22-96 148-204 261-321 367-425 22''-96'' 148''-204'' 261''-321'' 367''-425'', Intra-L(C23-C104) 23'-88' 134'-194' 23'''-88''' 134'''-194'''', Inter-HL(h5-CL126) 224-214' 224''-214'''', Inter-HH(h8,h11) 227-227'' 230-230''. Nemolizumab has N-glycosylation sites at the following positions: H CH2 N84.4: 297,297''. Nemolizumab lacks the C-terminal glycine and lysine of the H chain (CHS G1>del, K2>del).

[0116] Nemolizumab contains the following heavy chain amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYIMNWVRQAPGQGLEWMGLINPYNGGTDYNPQFQDRVTITADKSTSTAYMELSSLRSEDTAVYYCARDGYDDGPYTLETWGQ GTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVE CPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSP (SEQ ID NO: 5).

[0117] Nemolizumab contains the following light chain amino acid sequence: DIQMTQSPSSLSASVGDRVTITCQASEDIYSFVAWYQQKPGKAPKLLIYNAQTEAQGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHHYDSPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 6).

[0118] The heavy chain variable region of nemolizumab comprises the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYIMNWVRQAPGQGLEWMGLINPYNGGTDYNPQFQDRVTITADKSTSTAYMELSSLRSEDTAVYYCARDGYDDGPYTLETWGQGTLVTVSS (SEQ ID NO: 7).

[0119] HCDR1 of nemolizumab comprises the amino acid sequence GYIMN (SEQ ID NO: 8), HCDR2 comprises the amino acid sequence LINPYNGGTDYNPQFQD (SEQ ID NO: 9), and HCDR3 comprises the amino acid sequence DGYDDGPYTLET (SEQ ID NO: 10).

[0120] The light chain variable region of nemolizumab comprises the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCQASEDIYSFVAWYQQKPGKAPKLLIYNAQTEAQGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHHYDSPLTFGGGTKVEIKR (SEQ ID NO: 11).

[0121] LCDR1 of nemolizumab comprises the amino acid sequence QASEDIYSFVA (SEQ ID NO: 12), LCDR2 comprises the amino acid sequence NAQTEAQ (SEQ ID NO: 13), and LCDR3 comprises the amino acid sequence QHHYDSPLT (SEQ ID NO: 14).

[0122] For purposes of this disclosure, "variant antibodies" or "variants" of nemolizumab include (i) antibodies having a heavy chain that comprises at least 55%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% amino acid sequence identity to the heavy chain sequence of nemolizumab; (ii) antibodies having a light chain that comprises at least 55%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% amino acid sequence identity to the light chain sequence of nemolizumab; i) an antibody having a variable region that contains at least 55%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% amino acid sequence identity to the variable region sequence of nemolizumab, (iv) an antibody having a CDR that contains at least 55%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% amino acid sequence identity to the CDR sequence of nemolizumab, and (v) a combination thereof, but is not limited thereto. For example, suitable variants include immunoglobulins or immunoglobulin-like molecules that have heavy and light chain amino acid sequences identical or substantially similar to those of nemolizumab. Other suitable therapeutic antibodies may bind to the same isoform of IL-31RA as nemolizumab (e.g., IL31-RAv3), optionally to the same epitope of IL-31RA, block or neutralize IL-31RA, or a combination thereof. Additional exemplary therapeutic antibodies are described, for example, in WO2010 / 064697.

[0123] The variants of nemolizumab and suitable therapeutic antibodies may be monoclonal or polyclonal antibodies. Such monoclonal antibodies having IL31-RA binding and / or neutralizing activity can be obtained, for example, by the following procedure: anti-IL31-RA monoclonal antibodies are prepared by using IL31-RA or a fragment thereof derived from a mammal such as a human or mouse as an antigen by known methods, and then antibodies having IL31-RA binding and / or neutralizing activity are selected from the anti-IL31-RA monoclonal antibodies thus obtained. Specifically, a desired antigen or a cell expressing a desired antigen is used as a sensitizing antigen for immunization by a conventional immunization method. Anti-IL31-RA monoclonal antibodies can be prepared by fusing the obtained immune cells with a known parent cell using a conventional cell fusion method, and screening for monoclonal antibody-producing cells (hybridomas) by conventional screening methods. Animals to be immunized include, for example, mammals such as mice, rats, rabbits, sheep, monkeys, goats, donkeys, cows, horses, and pigs. Antigens can be prepared using known IL31-RA gene sequences according to known methods, for example, by baculovirus-based methods (e.g., WO98 / 46777). Variants of nemolizumab and suitable therapeutic antibodies include intrabodies, peptibodies, nanobodies, single domain antibodies, multispecific antibodies (e.g., bispecific antibodies, diabodies, triabodies, tetrabodies, tandem di-scFv, tandem tri-scFv), darpins, heavy chain monomers, heavy chain dimers, or single domain antibodies (i.e., V, VL ... H H fragment or "camelid-like" antibodies).

[0124] Hybridomas can be prepared, for example, according to the method of Milstein et al. (Kohler, G. And Milstein, C., Methods Enzymol. (1981) 73:3-46). When the antigen is poorly immunogenic, the antigen may be bound to a macromolecule having immunogenicity such as albumin before immunization. The antigen used to prepare a monoclonal antibody having binding and / or neutralizing activity against human IL31-RA is not particularly limited, so long as it allows the preparation of an antibody having binding and / or neutralizing activity against human IL31-RA. For example, it is known that there are many mutants in human IL31-RA, and any mutant can be used as an immunogen, so long as it allows the preparation of an antibody having binding and / or neutralizing activity against human IL31-RA. Alternatively, under the same conditions, a peptide fragment of IL31-RA or a protein in which an artificial mutation has been introduced into the natural IL31-RA sequence may be used as an immunogen. Human IL31-RA.3 is one of the preferred immunogens for preparing antibodies having IL31-RA binding and / or neutralizing activity in the present disclosure.

[0125] The IL31-RA binding activity of therapeutic antibodies can also be determined by methods known to those skilled in the art. Methods for determining the antigen-binding activity of antibodies include, for example, ELISA (enzyme-linked immunosorbent assay), EIA (enzyme immunoassay), RIA (radioimmunoassay), and fluorescent antibody technique. For example, when using an enzyme immunoassay, an antibody-containing sample, such as purified antibody and culture supernatant of antibody-producing cells, is added to an antigen-coated plate. A secondary antibody labeled with an enzyme such as alkaline phosphatase is added, and the plate is incubated. After washing, an enzyme substrate such as p-nitrophenyl phosphate is added, and the absorbance is measured to evaluate the antigen-binding activity. The binding and / or neutralizing activity of therapeutic antibodies against IL31-RA can be measured, for example, by observing the effect of suppressing the proliferation of IL-31-dependent cell lines. For example, the activity of purified mouse IL-31 antibodies can be assayed by evaluating the IL-31-dependent growth of Ba / F3 cells transfected with mouse IL-31 receptor alpha and mouse OSMR genes.

[0126] Any of the anti-IL31RA antibodies (i.e., "therapeutic antibodies") disclosed herein, including nemolizumab and fragments or variants thereof, can be used to treat and / or prevent AD and achieve the therapeutic endpoints of the disclosure. Optimal dosages and routes of administration may vary.

[0127] V. Pharmaceutical Compositions Provided herein is a pharmaceutical composition for use in the treatment or prevention of atopic dermatitis (AD), including skin lesions, or nodules, or pruritus caused by AD. The pharmaceutical composition comprises an anti-IL31RA antibody (i.e., a "therapeutic antibody"), such as nemolizumab, or a fragment or variant thereof, as an active ingredient. For example, in one aspect, the present disclosure provides a pharmaceutical composition for use in the treatment or prevention of AD in a subject, comprising an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) as an active ingredient, wherein the subject expresses at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in his / her skin at an expression level higher than a reference level. The reference levels may correspond to the expression levels of each biomarker in a skin sample from an individual without AD, or alternatively, in a skin sample from a subject without lesions or pruritus (i.e., non-lesional skin). In some embodiments, the sample obtained from a subject suspected of having AD is a skin sample, optionally including a lesion. For purposes of the above aspects, the skin sample may comprise, consist of, or consist essentially of the stratum corneum.

[0128] The present disclosure also provides a pharmaceutical composition for use in treating or preventing AD in a subject, comprising as an active ingredient an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof), wherein the subject expresses at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in his / her skin at an expression level higher than a reference level, and the subject is diagnosed as having AD by detecting the expression level of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18 in a sample obtained from a subject suspected of having AD, and optionally comparing the expression level of the biomarker to a reference level. The reference level may correspond to the expression level of each biomarker in a skin sample from an individual without AD, or alternatively in a skin sample from a subject without lesions or pruritus (i.e., non-lesional skin). In some embodiments, the sample obtained from a subject suspected of having AD is a skin sample, optionally including a lesion. For purposes of the above aspects, the skin sample may comprise, consist of, or consist essentially of the stratum corneum.

[0129] For purposes of the above pharmaceutical compositions, expression of CCL18 in lesional skin of a subject having (or suspected of having) AD can be about 300 ppm (calculated as a ratio of biomarker protein to total protein in the sample), about 310 ppm, about 320 ppm, about 330 ppm, about 340 ppm, about 350 ppm, about 360 ppm, about 370 ppm, about 380 ppm, about 390 ppm, about 400 ppm or more. Additionally or alternatively, expression of CCL18 in lesional skin of a subject having (or suspected of having) AD can be at least 300 ppm (calculated as a ratio of biomarker protein to total protein in the sample), at least 310 ppm, at least 320 ppm, at least 330 ppm, at least 340 ppm, at least 350 ppm, at least 360 ppm, at least 370 ppm, at least 380 ppm, at least 390 ppm, at least 400 ppm or more. Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.5-fold, about 2-fold, about 2.1-fold, about 2.2-fold, about 2.3-fold, about 2.4-fold, about 2.5-fold, about 2.6-fold, about 2.7-fold, about 2.8-fold, about 2.9-fold, or about 3-fold or more higher than the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.5-fold, at least 2-fold, at least 2.1-fold, at least 2.2-fold, at least 2.3-fold, at least 2.4-fold, at least 2.5-fold, at least 2.6-fold, at least 2.7-fold, at least 2.8-fold, at least 2.9-fold, or at least 3-fold or more higher than the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be about 150%, about 200%, about 210%, about 220%, about 230%, about 240%, about 250%, about 260%, about 270%, about 280%, about 290%, or about 300% or more of the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be at least 150%, at least 200%, at least 210%, at least 220%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, or at least 300% or more of the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0130] For purposes of the above embodiments, expression of CCL20 in lesional skin of a subject having (or suspected of having) AD can be about 15.5 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 15.6 ppm, about 15.7 ppm, about 15.8 ppm, about 15.9 ppm, about 16 ppm, about 16.1 ppm, about 16.2 ppm, about 16.3 ppm, about 16.4 ppm, about 16.5 ppm, about 16.6 ppm, about 16.7 ppm, about 16.8 ppm, about 16.9 ppm, about 17 ppm or more. Additionally or alternatively, expression of CCL20 in lesional skin of a subject having AD (or suspected of having AD) can be at least 15.5 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), at least 15.6 ppm, at least 15.7 ppm, at least 15.8 ppm, at least 15.9 ppm, at least 16 ppm, at least 16.1 ppm, at least 16.2 ppm, at least 16.3 ppm, at least 16.4 ppm, at least 16.5 ppm, at least 16.6 ppm, at least 16.7 ppm, at least 16.8 ppm, at least 16.9 ppm, at least 17 ppm or more. Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.1-fold, about 1.2-fold, about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.6-fold, about 1.7-fold, about 1.8-fold, about 1.9-fold, about 2-fold or more than the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject without AD). Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be at least 1.1-fold, at least 1.2-fold, at least 1.3-fold, at least 1.4-fold, at least 1.5-fold, at least 1.6-fold, at least 1.7-fold, at least 1.8-fold, at least 1.9-fold, at least 2-fold or more than the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject without AD).Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or greater of the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0131] For purposes of the above pharmaceutical compositions, expression of CCL22 in lesional skin of a subject having (or suspected of having) AD can be about 1550 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 1560 ppm, about 1570 ppm, about 1580 ppm, about 1590 ppm, about 1600 ppm, about 1610 ppm, about 1620 ppm, about 1630 ppm, about 1640 ppm, about 1650 ppm, about 1660 ppm, about 1670 ppm, about 1680 ppm, about 1690 ppm, about 1700 ppm or more. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be at least 1550 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), at least 1560 ppm, at least 1570 ppm, at least 1580 ppm, at least 1590 ppm, at least 1600 ppm, at least 1610 ppm, at least 1620 ppm, at least 1630 ppm, at least 1640 ppm, at least 1650 ppm, at least 1660 ppm, at least 1670 ppm, at least 1680 ppm, at least 1690 ppm, at least 1700 ppm or more. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) may be about 1.01 fold, about 1.02 fold, about 1.03 fold, about 1.04 fold, about 1.05 fold, about 1.06 fold, about 1.07 fold, about 1.08 fold, about 1.09 fold, about 1.1 fold, about 1.2 fold, about 1.3 fold, about 1.4 fold, about 1.5 fold, about 1.6 fold, about 1.7 fold, about 1.8 fold, about 1.9 fold, about 2 fold or more than the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or more of the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0132] For purposes of the above pharmaceutical compositions, expression of CCL27 in lesional skin of a subject having (or suspected of having) AD can be about 50 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 51 ppm, about 52 ppm, about 53 ppm, about 54 ppm, about 55 ppm, about 56 ppm, about 57 ppm, about 58 ppm, about 59 ppm, about 60 ppm, about 61 ppm, about 62 ppm, about 63 ppm, about 64 ppm, about 65 ppm, about 66 ppm, about 67 ppm, about 68 ppm, about 69 ppm, about 70 ppm, about 71 ppm, about 72 ppm, about 73 ppm, about 74 ppm, about 75 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be at least 50 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 51 ppm, at least 52 ppm, at least 53 ppm, at least 54 ppm, at least 55 ppm, at least 56 ppm, at least 57 ppm, at least 58 ppm, at least 59 ppm, at least 60 ppm, at least 61 ppm, at least 62 ppm, at least 63 ppm, at least 64 ppm, at least 65 ppm, at least 66 ppm, at least 67 ppm, at least 68 ppm, at least 69 ppm, at least 70 ppm, at least 71 ppm, at least 72 ppm, at least 73 ppm, at least 74 ppm, at least 75 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.5-fold, about 2-fold, about 2.1-fold, about 2.2-fold, about 2.3-fold, about 2.4-fold, about 2.5-fold, about 2.6-fold, about 2.7-fold, about 2.8-fold, about 2.9-fold, or about 3-fold or more than the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.5-fold, at least 2-fold, at least 2.1-fold, at least 2.2-fold, at least 2.3-fold, at least 2.4-fold, at least 2.5-fold, at least 2.6-fold, at least 2.7-fold, at least 2.8-fold, at least 2.9-fold, or at least 3-fold or more higher than the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be about 150%, about 175%, about 200%, about 210%, about 220%, about 225%, about 230%, about 240%, about 250%, about 260%, about 270%, about 280%, about 290%, or about 300% or more of the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be at least 150%, at least 175%, at least 200%, at least 210%, at least 220%, at least 225%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, or at least 300% or more of the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0133] For purposes of the above pharmaceutical compositions, expression of VEGF in lesional skin of a subject having AD (or suspected of having AD) can be about 150 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 151 ppm, about 152 ppm, about 153 ppm, about 154 ppm, about 155 ppm, about 156 ppm, about 157 ppm, about 158 ​​ppm, about 159 ppm, about 160 ppm, about 161 ppm, about 162 ppm, about 163 ppm, about 164 ppm, about 165 ppm, about 166 ppm, about 167 ppm, about 168 ppm, about 169 ppm, about 170 ppm, about 171 ppm, about 172 ppm, about 173 ppm, about 174 ppm, about 175 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject having AD (or suspected of having AD) can be at least 150 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 151 ppm, at least 152 ppm, at least 153 ppm, at least 154 ppm, at least 155 ppm, at least 156 ppm, at least 157 ppm, at least 158 ​​ppm, at least 159 ppm, at least 160 ppm, at least 161 ppm, at least 162 ppm, at least 163 ppm, at least 164 ppm, at least 165 ppm, at least 166 ppm, at least 167 ppm, at least 168 ppm, at least 169 ppm, at least 170 ppm, at least 171 ppm, at least 172 ppm, at least 173 ppm, at least 174 ppm, at least 75 ppm or more. Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be about 1.01 fold, about 1.02 fold, about 1.03 fold, about 1.04 fold, about 1.05 fold, about 1.06 fold, about 1.07 fold, about 1.08 fold, about 1.09 fold, about 1.1 fold, about 1.2 fold, about 1.3 fold, about 1.4 fold, about 1.5 fold, about 1.6 fold, about 1.7 fold, about 1.8 fold, about 1.9 fold, about 2 fold or more than the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level can be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level can be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or greater of the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0134] For purposes of the above pharmaceutical compositions, the expression of IL1RA in lesional skin of a subject having (or suspected of having) AD is about 207000 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 207500 ppm, about 208000 ppm, about 208500 ppm, about 209000 ppm, about 209500 ppm, about 210000 ppm, about 210500 ppm, about 211000 ppm, about 211500 ppm, about 212000 ppm, about 212500 ppm, about 213000 ppm, about 213500 ppm, about 2140 00ppm, approx. 214500ppm, approx. 215000ppm, approx. 215500ppm, approx. 216000ppm, approx. 216500ppm, approx. 217 000ppm, about 217500ppm, about 218000ppm, about 218500ppm, about 219000ppm, about 219500, about 220000 , about 221000, about 222000, about 223000, about 224000, about 225000, about 226000, about 227000, about 228000, about 229000, about 230000, about 231000, about 232000, about 233000, about 234000, about 235000 ppm or more.Additionally or alternatively, expression of CCL27 in lesional skin of a subject having (or suspected of having) AD is at least 207000 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 207500 ppm, at least 208000 ppm, at least 208500 ppm, at least 209000 ppm, at least 209500 ppm, at least 210000 ppm, at least 210500 ppm, at least 211000 ppm, at least 211500 ppm, at least 212000 ppm, at least 212500 ppm, at least 213000 ppm, at least 213500 ppm, at least 214000 ppm, at least 214500 ppm, at least 21500 ppm, at least 216000 ppm, at least 217500 ppm, at least 217600 ppm, at least 217700 ppm, at least 217800 ppm, at least 217900 ppm, at least 218000 ppm, at least 218100 ppm, at least 218200 ppm, at least 218300 ppm, at least 218400 ppm, at least 218500 ppm, at least 218600 ppm, at least 218700 ppm, at least 218800 ppm, at least 218900 ppm, at least 219000 ppm, at least 220000 ppm, at least 2201000 ppm, at least 2202000 ppm, at least 220300 ppm, at least 220400 ppm, at least 220500 ppm, at least 220600 ppm, at least 2207000 ppm, at least It can be 15000 ppm, at least 215500 ppm, at least 216000 ppm, at least 216500 ppm, at least 217000 ppm, at least 217500 ppm, at least 218000 ppm, at least 218500 ppm, at least 219000 ppm, at least 219500, at least 220000, at least 221000, at least 222000, at least 223000, at least 224000, at least 225000, at least 226000, at least 227000, at least 228000, at least 229000, at least 230000, at least 231000, at least 232000, at least 233000, at least 234000, at least 235000 ppm or more. Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) may be about 1.01-fold, about 1.02-fold, about 1.03-fold, about 1.04-fold, about 1.05-fold, about 1.06-fold, about 1.07-fold, about 1.08-fold, about 1.09-fold, about 1.1-fold, about 1.2-fold, about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.6-fold, about 1.7-fold, about 1.8-fold, about 1.9-fold, about 2-fold or more than the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 115%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or more of the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 115%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0135] Use or administration of a pharmaceutical composition of the present disclosure may also result in a decrease in expression of CCL20, CCL22, CCL27, VEGF, and / or CCL18, and / or an increase in expression of IL1Ra in the skin (particularly lesional skin) of a subject receiving the pharmaceutical composition.

[0136] For purposes of the pharmaceutical compositions of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce CCL18 expression in lesional skin of a subject with (or suspected of having) AD by about 6-fold, about 6.5-fold, about 7-fold, about 7.5-fold, about 8-fold, about 8.5-fold, or less. The length of treatment required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0137] For purposes of the pharmaceutical compositions of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce expression of CCL20 in lesional skin of a subject with (or suspected of having) AD by about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0138] For purposes of the pharmaceutical compositions of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce the expression of CCL22 in the lesional skin of a subject with (or suspected of having) AD by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0139] For purposes of the pharmaceutical compositions of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce the expression of CCL27 in the lesional skin of a subject with (or suspected of having) AD by about 1.5-fold, about 1.75-fold, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0140] For purposes of the pharmaceutical compositions of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce VEGF expression in lesional skin of a subject with (or suspected of having) AD by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0141] For purposes of the pharmaceutical compositions of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may increase expression of IL1RA in the lesional skin of a subject with (or suspected of having) AD by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or more. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0142] For purposes of the pharmaceutical compositions of the present disclosure, the other described biomarkers (e.g., CXCL6, CXCL8, cystatin, FASL, galectin, IL11, IL21, SPD, and CCL17) may show a similar pattern of overexpression in lesional skin of a subject with AD (or suspected of having AD) compared to expression levels in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), and CX3CL1 may show increased expression in lesional skin of a subject with AD (or suspected of having AD) compared to expression levels in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Similarly, CXCL6, CXCL8, cystatin, FASL, galectin, CCL17, and IL21 may show a similar pattern of decreased expression in lesional skin of subjects with AD (or suspected of having AD) after treatment with an anti-IL-31RA antibody (e.g., nemolizumab), and CX3CL1, IL11, and SPD may show increased expression in lesional skin of subjects with AD (or suspected of having AD) after treatment with an anti-IL-31RA antibody (e.g., nemolizumab).

[0143] The phrase "comprising nemolizumab or a fragment or variant thereof as an active ingredient" means that nemolizumab or a fragment or variant thereof is included as at least one of the active ingredients, and does not limit the proportion of the antibody. In addition, the therapeutic agent for AD in the present disclosure may include other ingredients that enhance the treatment or prevention of AD in combination with nemolizumab or a fragment or variant thereof. For example, the composition may include one or more of a topical corticosteroid cream or injection, an ointment containing menthol or phenol for cooling and soothing itchy skin, a capsaicin cream, an oral corticosteroid, a selective serotonin reuptake inhibitor (SSRI), and an oral antihistamine.

[0144] Pharmaceutical compositions of the anti-IL31RA antibodies (e.g., nemolizumab or fragments or variants thereof) of the present disclosure can be prepared as formulations according to standard methods (see, e.g., Remington's Pharmaceutical Science, Mark Publishing Company, Easton, USA). Pharmaceutical compositions generally include carriers and / or excipients in addition to the antibody. For example, in some embodiments, the pharmaceutical composition comprises one or more surfactants (e.g., PEG and Tween), excipients, antioxidants (e.g., ascorbic acid), colorants, flavoring agents, preservatives, stabilizers, buffers (e.g., phosphoric acid, citric acid, and other organic acids), chelating agents (e.g., EDTA), suspending agents, isotonizing agents, binders, disintegrants, lubricants, flow enhancers, Corrigent, light anhydrous silicic acid, lactose, crystalline cellulose, mannitol, starch, carmellose calcium, carmellose sodium, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinyl acetal diethylaminoacetate, polyvinylpyrrolidone, gelatin, medium chain triglycerides, polyoxyethylene hydrogenated castor oil 60, sucrose, carboxymethyl cellulose, corn starch, and inorganic salts. In some embodiments, the pharmaceutical composition comprises one or more other low molecular weight polypeptides, proteins such as serum albumin, gelatin, and immunoglobulins, and amino acids such as glycine, glutamine, asparagine, arginine, and lysine.

[0145] The anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) may be prepared as an injectable aqueous solution, and the anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) may be dissolved in an isotonic solution containing, for example, saline, dextrose, or other excipients or isotonic agents (i.e., isotonicity agents). Isotonicity agents may include, for example, D-sorbitol, D-mannose, D-mannitol, and sodium chloride. In addition, suitable solubilizers, such as alcohols (e.g., ethanol), polyalcohols (e.g., propylene glycol and PEG), and non-ionic detergents (polysorbate 80 and HCO-50) may be used simultaneously.

[0146] In some embodiments, an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) can be encapsulated in microcapsules (microcapsules made of hydroxymethylcellulose, gelatin, polymethylmethacrylate, etc.) and made into components of colloid drug delivery systems (liposomes, albumin microparticles, microemulsions, nanoparticles, and nanocapsules) (see, e.g., "Remington's Pharmaceutical Science 16th edition" &, Oslo Ed. (1980)). Furthermore, methods for making sustained release drugs are known and can be applied to nemolizumab or a fragment or variant thereof (Langer et al., J. Biomed. Mater. Res. (1981) 15, 167-277; Langer, Chem. Tech. (1982) 12, 98-105; U.S. Pat. No. 3,773,919; European Patent Application (EP) No. 58,481; Sidman et al., Biopolymers (1983) 22, 547-56; EP 133,988).

[0147] The pharmaceutical composition of the present disclosure may be administered orally or parenterally, but is preferably administered parenterally. Specifically, the pharmaceutical composition is administered to the patient by injection or transdermal administration. Injection includes, for example, intravenous injection, intramuscular injection, and subcutaneous injection for systemic or local administration. The pharmaceutical composition may be given to the site where inflammation and / or itch is suppressed, or the area surrounding the site by local injection or intramuscular or subcutaneous injection. In some embodiments, the pharmaceutical composition is administered at the site of one or more skin peels, lesions, or nodules, or proximal to the site of one or more skin peels, lesions, or nodules.

[0148] The administration method can be appropriately selected depending on the age, body weight, and condition of the patient. The single administration dose can be selected, for example, within the range of 0.0001 to 100 mg of antibody (e.g., nemolizumab or a fragment or variant thereof) per kg of body weight. Alternatively, for example, when the antibody is administered to a human patient, the dose of the antibody can be selected from the range of 0.001 to 1,000 mg / kg of body weight. In some embodiments, the composition may be administered in a concentration of, for example, about 0.01 to 50 mg / kg, about 0.01 mg / kg to about 0.1 mg / kg, about 0.05 mg / kg to 0.15 mg / kg, about 0.1 mg / kg to about 0.6 mg / kg, about 0.1 mg / kg to about 1 mg / kg, about 0.25 mg / kg to about 0.75 mg / kg, about 0.4 mg / kg to about 0.8 mg / kg, about 0.4 mg / kg to about 1.8 mg / kg, about 0.5 to about 2.5 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.5 mg / kg, about 1 mg / kg to about 3.5 mg / kg, The composition is formulated to administer a dose containing nemolizumab or a fragment or variant thereof of about 1 mg / kg to about 5 mg / kg, about 2 mg / kg to about 4 mg / kg, about 2.5 mg / kg to about 10 mg / kg, about 5 mg / kg to about 10 mg / kg, about 10 mg / kg to about 20 mg / kg, about 10 mg / kg to about 40 mg / kg, about 20 mg / kg to about 50 mg / kg, about 25 mg / kg to about 75 mg / kg, about 50 mg / kg to about 100 mg / kg, or about 100 mg / kg to about 500 mg / kg, or about 100 mg / kg to about 1000 mg / kg of body weight. In preferred embodiments, the dosage ranges from about 0.01 mg / kg to about 0.1 mg / kg, from about 0.1 mg / kg to about 0.5 mg / kg, from about 0.5 mg / kg to about 1.5 mg / kg, from about 1.5 mg / kg to about 2.5 mg / kg, or from about 2.5 mg / kg to about 10 mg / kg.In some embodiments, the dose is about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about g / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3 mg / kg, about 3.5 mg / kg, about 4 mg / kg, about 4.5 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 500 mg / kg or about 1,000 mg / kg. In certain embodiments, the effective amount of nemolizumab or a fragment or variant thereof is about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 1.5 mg / kg, about 2 mg / kg, or about 2.5 mg / kg. In a preferred embodiment, the dose is about 0.5 mg / kg.

[0149] The present disclosure provides a pharmaceutical composition for use in the treatment or prevention of atopic dermatitis (AD) in a subject, the pharmaceutical composition comprising an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) as an active ingredient, wherein the subject differentially expresses at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 compared to a reference expression level of at least one gene.

[0150] Any of the pharmaceutical compositions disclosed herein comprising nemolizumab and fragments or variants thereof can be used to treat and / or prevent AD and achieve the therapeutic endpoints of the disclosure. Optimal dosages and routes of administration may vary.

[0151] VI. Treatment / Prevention of Atopic Dermatitis The present disclosure provides a method of treating or preventing pruritus in a subject with atopic dermatitis (AD), the method comprising, consisting of, or consisting essentially of administering to the subject an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof (i.e., a "therapeutic antibody"). The disclosed methods may be carried out to achieve certain therapeutic endpoints, which are described in more detail below. Also disclosed herein is the use of an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof (i.e., a "therapeutic antibody"), to a subject to treat or prevent AD and / or achieve the therapeutic endpoints of the disclosure. Also disclosed herein is the use of an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof (i.e., a "therapeutic antibody"), to a subject to treat or prevent AD and / or achieve the therapeutic endpoints of the disclosure. In addition, certain subgroups of subjects with AD may be particularly suitable for treatment with the methods and uses of the present disclosure. For example, patients exhibiting increased expression of the biomarkers of the present disclosure, such as CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18, have been found to respond particularly well to treatment with anti-IL-31RA antibodies (e.g., nemolizumab). Similarly, the present disclosure is the first to show that adolescents with moderate to severe AD respond to treatment with anti-IL-31RA antibodies (e.g., nemolizumab). Thus, in some embodiments of the methods and uses of the present disclosure, the subject being treated may exhibit increased expression of CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18 in their skin, and in some embodiments, the subject may be an adolescent, and in some embodiments, the subject may be an adolescent exhibiting increased expression of CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18 in their skin.

[0152] The present disclosure is the first to report a biomarker signature for AD that can not only identify and positively diagnose AD, but also identify subjects with AD who will likely respond to treatment with an anti-IL-31RA antibody (e.g., nemolizumab) and track the response of treatment with an anti-IL-31RA antibody (e.g., nemolizumab).

[0153] In one aspect, the disclosure provides a method of treating or preventing atopic dermatitis (AD) in a subject, comprising administering an anti-IL-31RA antibody to a subject with AD, wherein the subject has skin lesions in which expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is upregulated compared to non-lesional skin of an individual without AD. In some embodiments, "preventing" AD can refer simply to reducing the risk of flare-ups or to reducing the risk of developing skin lesions.

[0154] In another aspect, the disclosure provides a method of reducing expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in lesional skin of a subject with atopic dermatitis (AD), comprising administering an anti-IL-31RA antibody to a subject with AD, wherein at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is upregulated in lesional skin of the subject compared to non-lesional skin of the subject or an individual without AD, and wherein administration of the anti-IL-31RA antibody reduces expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in the lesional skin.

[0155] For purposes of the uses and methods of the present disclosure, one, two, three, four, five, or six of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 may be upregulated in at least one lesional skin of a subject compared to non-lesional skin of a subject or individual without AD. In some embodiments, CCL20, CCL22, CCL27, and VEGF are upregulated in the lesional skin of a subject compared to non-lesional skin of a subject or individual without AD.

[0156] Additionally or alternatively, in some embodiments, expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is reduced within 2 weeks, 4 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, or 20 weeks of administration of an anti-IL-31RA antibody, compared to baseline expression levels in the subject's skin lesion prior to administration of an anti-IL-31RA antibody. Additionally or alternatively, in some embodiments, expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is reduced or increased, as described herein.

[0157] For example, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be about 300 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), about 310 ppm, about 320 ppm, about 330 ppm, about 340 ppm, about 350 ppm, about 360 ppm, about 370 ppm, about 380 ppm, about 390 ppm, about 400 ppm or more. Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be at least 300 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 310 ppm, at least 320 ppm, at least 330 ppm, at least 340 ppm, at least 350 ppm, at least 360 ppm, at least 370 ppm, at least 380 ppm, at least 390 ppm, at least 400 ppm or more. Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.5-fold, about 2-fold, about 2.1-fold, about 2.2-fold, about 2.3-fold, about 2.4-fold, about 2.5-fold, about 2.6-fold, about 2.7-fold, about 2.8-fold, about 2.9-fold, or about 3-fold or more higher than the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.5-fold, at least 2-fold, at least 2.1-fold, at least 2.2-fold, at least 2.3-fold, at least 2.4-fold, at least 2.5-fold, at least 2.6-fold, at least 2.7-fold, at least 2.8-fold, at least 2.9-fold, or at least 3-fold or more higher than the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be about 150%, about 200%, about 210%, about 220%, about 230%, about 240%, about 250%, about 260%, about 270%, about 280%, about 290%, or about 300% or more of the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL18 in lesional skin of a subject with AD (or suspected of having AD) can be at least 150%, at least 200%, at least 210%, at least 220%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, or at least 300% or more of the expression level of CCL18 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0158] Expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be about 15.5 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 15.6 ppm, about 15.7 ppm, about 15.8 ppm, about 15.9 ppm, about 16 ppm, about 16.1 ppm, about 16.2 ppm, about 16.3 ppm, about 16.4 ppm, about 16.5 ppm, about 16.6 ppm, about 16.7 ppm, about 16.8 ppm, about 16.9 ppm, about 17 ppm or more. Additionally or alternatively, expression of CCL20 in lesional skin of a subject having AD (or suspected of having AD) can be at least 15.5 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), at least 15.6 ppm, at least 15.7 ppm, at least 15.8 ppm, at least 15.9 ppm, at least 16 ppm, at least 16.1 ppm, at least 16.2 ppm, at least 16.3 ppm, at least 16.4 ppm, at least 16.5 ppm, at least 16.6 ppm, at least 16.7 ppm, at least 16.8 ppm, at least 16.9 ppm, at least 17 ppm or more. Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.1-fold, about 1.2-fold, about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.6-fold, about 1.7-fold, about 1.8-fold, about 1.9-fold, about 2-fold or more than the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject without AD). Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be at least 1.1-fold, at least 1.2-fold, at least 1.3-fold, at least 1.4-fold, at least 1.5-fold, at least 1.6-fold, at least 1.7-fold, at least 1.8-fold, at least 1.9-fold, at least 2-fold or more than the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject without AD).Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or greater of the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL20 in lesional skin of a subject with AD (or suspected of having AD) can be at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of CCL20 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0159] Expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be about 1550 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 1560 ppm, about 1570 ppm, about 1580 ppm, about 1590 ppm, about 1600 ppm, about 1610 ppm, about 1620 ppm, about 1630 ppm, about 1640 ppm, about 1650 ppm, about 1660 ppm, about 1670 ppm, about 1680 ppm, about 1690 ppm, about 1700 ppm or more. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be at least 1550 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), at least 1560 ppm, at least 1570 ppm, at least 1580 ppm, at least 1590 ppm, at least 1600 ppm, at least 1610 ppm, at least 1620 ppm, at least 1630 ppm, at least 1640 ppm, at least 1650 ppm, at least 1660 ppm, at least 1670 ppm, at least 1680 ppm, at least 1690 ppm, at least 1700 ppm or more. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) may be about 1.01 fold, about 1.02 fold, about 1.03 fold, about 1.04 fold, about 1.05 fold, about 1.06 fold, about 1.07 fold, about 1.08 fold, about 1.09 fold, about 1.1 fold, about 1.2 fold, about 1.3 fold, about 1.4 fold, about 1.5 fold, about 1.6 fold, about 1.7 fold, about 1.8 fold, about 1.9 fold, about 2 fold or more than the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or more of the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL22 in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of CCL22 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0160] Expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be about 50 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 51 ppm, about 52 ppm, about 53 ppm, about 54 ppm, about 55 ppm, about 56 ppm, about 57 ppm, about 58 ppm, about 59 ppm, about 60 ppm, about 61 ppm, about 62 ppm, about 63 ppm, about 64 ppm, about 65 ppm, about 66 ppm, about 67 ppm, about 68 ppm, about 69 ppm, about 70 ppm, about 71 ppm, about 72 ppm, about 73 ppm, about 74 ppm, about 75 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be at least 50 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 51 ppm, at least 52 ppm, at least 53 ppm, at least 54 ppm, at least 55 ppm, at least 56 ppm, at least 57 ppm, at least 58 ppm, at least 59 ppm, at least 60 ppm, at least 61 ppm, at least 62 ppm, at least 63 ppm, at least 64 ppm, at least 65 ppm, at least 66 ppm, at least 67 ppm, at least 68 ppm, at least 69 ppm, at least 70 ppm, at least 71 ppm, at least 72 ppm, at least 73 ppm, at least 74 ppm, at least 75 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be about 1.5-fold, about 2-fold, about 2.1-fold, about 2.2-fold, about 2.3-fold, about 2.4-fold, about 2.5-fold, about 2.6-fold, about 2.7-fold, about 2.8-fold, about 2.9-fold, or about 3-fold or more than the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.5-fold, at least 2-fold, at least 2.1-fold, at least 2.2-fold, at least 2.3-fold, at least 2.4-fold, at least 2.5-fold, at least 2.6-fold, at least 2.7-fold, at least 2.8-fold, at least 2.9-fold, or at least 3-fold or more higher than the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be about 150%, about 175%, about 200%, about 210%, about 220%, about 225%, about 230%, about 240%, about 250%, about 260%, about 270%, about 280%, about 290%, or about 300% or more of the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of CCL27 in lesional skin of a subject with AD (or suspected of having AD) can be at least 150%, at least 175%, at least 200%, at least 210%, at least 220%, at least 225%, at least 230%, at least 240%, at least 250%, at least 260%, at least 270%, at least 280%, at least 290%, or at least 300% or more of the expression level of CCL27 in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0161] Expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be about 150 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 151 ppm, about 152 ppm, about 153 ppm, about 154 ppm, about 155 ppm, about 156 ppm, about 157 ppm, about 158 ​​ppm, about 159 ppm, about 160 ppm, about 161 ppm, about 162 ppm, about 163 ppm, about 164 ppm, about 165 ppm, about 166 ppm, about 167 ppm, about 168 ppm, about 169 ppm, about 170 ppm, about 171 ppm, about 172 ppm, about 173 ppm, about 174 ppm, about 175 ppm or more. Additionally or alternatively, expression of CCL27 in lesional skin of a subject having AD (or suspected of having AD) can be at least 150 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 151 ppm, at least 152 ppm, at least 153 ppm, at least 154 ppm, at least 155 ppm, at least 156 ppm, at least 157 ppm, at least 158 ​​ppm, at least 159 ppm, at least 160 ppm, at least 161 ppm, at least 162 ppm, at least 163 ppm, at least 164 ppm, at least 165 ppm, at least 166 ppm, at least 167 ppm, at least 168 ppm, at least 169 ppm, at least 170 ppm, at least 171 ppm, at least 172 ppm, at least 173 ppm, at least 174 ppm, at least 75 ppm or more. Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be about 1.01 fold, about 1.02 fold, about 1.03 fold, about 1.04 fold, about 1.05 fold, about 1.06 fold, about 1.07 fold, about 1.08 fold, about 1.09 fold, about 1.1 fold, about 1.2 fold, about 1.3 fold, about 1.4 fold, about 1.5 fold, about 1.6 fold, about 1.7 fold, about 1.8 fold, about 1.9 fold, about 2 fold or more than the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level can be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level can be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or greater of the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of VEGF in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of VEGF in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0162] Expression of IL1RA in lesional skin of subjects with AD (or suspected of having AD) is about 207000 ppm (when calculated as the ratio of biomarker protein to total protein in the sample), about 207500 ppm, about 208000 ppm, about 208500 ppm, about 209000 ppm, about 209500 ppm, about 210000 ppm, about 210500 ppm, about 211000 ppm, about 211500 ppm, about 212000 ppm, about 212500 ppm, about 213000 ppm, about 213500 ppm, about 214000 ppm, about 21500 ppm, about 216000 ppm, about 217000 ppm, about 218000 ppm, about 219000 ppm, about 220000 ppm, about 221000 ppm, about 222500 ppm, about 223000 ppm, about 223500 ppm, about 224000 ppm, about 22500 ppm, about 226000 ppm, about 227000 ppm, about 228000 ppm, about 229000 ppm, about 23000 ppm, about 231000 ppm, about 232000 ppm, about 233000 ppm, about 234000 ppm, about 23500 ppm, about 236000 ppm, about 237000 ppm, about 238000 ppm, about 239000 ppm, about 24000 ppm, about 241000 ppm, about 242000 ppm, about 24300 ppm, about 244000 ppm, about 24500 ppm, about 246000 ppm 4500 ppm, about 215000 ppm, about 215500 ppm, about 216000 ppm, about 216500 ppm, about 217000 ppm, about 217500 ppm, about 218000 ppm, about 218500 ppm, about 219000 ppm, about 219500, about 220000, about 221000, about 222000, about 223000, about 224000, about 225000, about 226000, about 227000, about 228000, about 229000, about 230000, about 231000, about 232000, about 233000, about 234000, about 235000 ppm or more.Additionally or alternatively, expression of CCL27 in lesional skin of a subject having (or suspected of having) AD is at least 207000 ppm (when calculated as a ratio of biomarker protein to total protein in the sample), at least 207500 ppm, at least 208000 ppm, at least 208500 ppm, at least 209000 ppm, at least 209500 ppm, at least 210000 ppm, at least 210500 ppm, at least 211000 ppm, at least 211500 ppm, at least 212000 ppm, at least 212500 ppm, at least 213000 ppm, at least 213500 ppm, at least 214000 ppm, at least 214500 ppm, at least 21500 ppm, at least 216000 ppm, at least 217500 ppm, at least 217600 ppm, at least 217700 ppm, at least 217800 ppm, at least 217900 ppm, at least 218000 ppm, at least 218100 ppm, at least 218200 ppm, at least 218300 ppm, at least 218400 ppm, at least 218500 ppm, at least 218600 ppm, at least 218700 ppm, at least 218800 ppm, at least 218900 ppm, at least 219000 ppm, at least 220000 ppm, at least 2201000 ppm, at least 2202000 ppm, at least 220300 ppm, at least 220400 ppm, at least 220500 ppm, at least 220600 ppm, at least 2207000 ppm, at least It can be 15000 ppm, at least 215500 ppm, at least 216000 ppm, at least 216500 ppm, at least 217000 ppm, at least 217500 ppm, at least 218000 ppm, at least 218500 ppm, at least 219000 ppm, at least 219500, at least 220000, at least 221000, at least 222000, at least 223000, at least 224000, at least 225000, at least 226000, at least 227000, at least 228000, at least 229000, at least 230000, at least 231000, at least 232000, at least 233000, at least 234000, at least 235000 ppm or more. Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) may be about 1.01-fold, about 1.02-fold, about 1.03-fold, about 1.04-fold, about 1.05-fold, about 1.06-fold, about 1.07-fold, about 1.08-fold, about 1.09-fold, about 1.1-fold, about 1.2-fold, about 1.3-fold, about 1.4-fold, about 1.5-fold, about 1.6-fold, about 1.7-fold, about 1.8-fold, about 1.9-fold, about 2-fold or more than the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be about the same between the lesional skin and the reference sample.Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) may be at least 1.01 fold, at least 1.02 fold, at least 1.03 fold, at least 1.04 fold, at least 1.05 fold, at least 1.06 fold, at least 1.07 fold, at least 1.08 fold, at least 1.09 fold, at least 1.1 fold, at least 1.2 fold, at least 1.3 fold, at least 1.4 fold, at least 1.5 fold, at least 1.6 fold, at least 1.7 fold, at least 1.8 fold, at least 1.9 fold, at least 2 fold or more than the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), or the expression level may be approximately the same between the lesional skin and the reference sample. Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) can be about 101%, about 102%, about 103%, about 104%, about 105%, about 106%, about 107%, about 108%, about 109%, about 110%, about 115%, about 120%, about 125%, about 130%, about 135%, about 140%, about 145%, about 150%, about 155%, about 160%, about 165%, about 170%, or about 175% or more of the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Additionally or alternatively, expression of IL1RA in lesional skin of a subject with AD (or suspected of having AD) can be at least 101%, at least 102%, at least 103%, at least 104%, at least 105%, at least 106%, at least 107%, at least 108%, at least 109%, at least 110%, at least 115%, at least 120%, at least 125%, at least 130%, at least 135%, at least 140%, at least 145%, at least 150%, at least 155%, at least 160%, at least 165%, at least 170%, or at least 175% or more of the expression level of IL1RA in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD).

[0163] In another aspect, the disclosure provides a method of reducing expression of an inflammatory biomarker in the skin of a subject with atopic dermatitis (AD), comprising administering an anti-IL-31RA antibody to a subject with AD, thereby reducing expression of the inflammatory biomarker in the skin. In some embodiments, the inflammatory biomarker is selected from at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18. In some embodiments, the inflammatory biomarker is selected from at least one of CCL20, CCL22, CCL27, and VEGF. In some embodiments, expression of one, two, three, or all four of CCL20, CCL22, CCL27, and VEGF is reduced. In some embodiments, expression of the inflammatory biomarker is reduced in at least one skin lesion of the subject. Reduction of a biomarker may be measured by taking a baseline measurement of expression of the biomarker(s) at time 0 (i.e., prior to administration of an anti-IL-31RA antibody (e.g., nemolizumab) and comparing this initial measurement to a later time point after initiation of treatment, such as 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14 weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21 weeks, 22 weeks, 23 weeks, or 24 weeks or more after initiation of treatment. In some embodiments, treatment with an anti-IL31RA antibody may include repeated administration of the antibody once every 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, or 6 weeks, and the dosing regimen may be "flat" or may include a loading dose, as described in more detail below.

[0164] For purposes of the disclosed methods and uses, administering to a subject an anti-IL-31RA antibody (i.e., a "therapeutic antibody"), such as nemolizumab or a fragment or variant thereof, may be effective in preventing, reducing, ameliorating, or eliminating one or more of the symptoms / complications caused by AD, including but not limited to pruritus (itch), dryness (dry skin), eczematous lesions, and significant sleep disturbances, and improving quality of life in a subject with AD. It is understood that not all subjects with AD will respond to treatment, but subjects with skin lesions that have increased expression of CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18 (compared to non-lesional samples of the subject's skin or non-lesional samples of the skin of individuals without AD) are more likely to respond favorably to treatment with an anti-IL31RA antibody, such as nemolizumab.

[0165] For purposes of the disclosed methods and uses, a subject may achieve at least a 66.5% reduction in Eczema Area and Severity Index (EASI) scoring after administration of an anti-IL-31RA antibody. In some embodiments, the reduction in EASI scoring may be at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% or more. Additionally or alternatively, in some embodiments, a subject may achieve at least a 53.3% reduction in AD-involved body surface area (EASI) after administration of an anti-IL-31RA antibody. In some embodiments, the reduction in body surface area EASI scoring can be at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% or more.

[0166] For purposes of the disclosed methods and uses, a subject may achieve at least a 31.2% reduction in Scoring Atopic Dermatitis (SCORAD) scoring following administration of an anti-IL-31RA antibody. In some embodiments, the reduction in SCORAD scoring may be at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% or more.

[0167] For purposes of the disclosed methods and uses, a subject may achieve at least a 43.2% reduction in peak pruritus numerical rating scale (PP-NRS) score after administration of an anti-IL-31RA antibody. In some embodiments, the reduction in PP-NRS score may be at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% or more. In some embodiments, a subject may achieve at least a 40.9% reduction in mean pruritus numerical rating scale (AP NRS) score after administration of an anti-IL-31RA antibody. In some embodiments, the reduction in AP NRS scoring can be at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% or more.

[0168] Additionally or alternatively, in some embodiments, the subject experiences improved sleep after administration of an anti-IL-31RA antibody. In some embodiments, the subject achieves at least a 53.5% reduction in a sleep disorder numerical rating scale after administration of an anti-IL-31RA antibody. In some embodiments, the reduction in a sleep disorder numerical rating scale can be at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% or more.

[0169] Additionally or alternatively, in some embodiments, the subject experiences an increase in the number of medication-free days following administration of an anti-IL-31RA antibody. For example, the subject may not require any other medication for 1, 2, 3, 4, 5, 6, or 7 days, or for 1, 2, 3, 4, or more weeks. Additionally or alternatively, in some embodiments, the subject achieves an improvement in Dermatology Life Eligibility Index (DLQI) or Pediatric Dermatology Life Quality Index (cDLQI) scoring.

[0170] In some embodiments of the methods disclosed herein, the subject is an adult (i.e., 18 years of age or older). In some embodiments, the subject is an adolescent, e.g., 12-17 years of age. In some embodiments, the subject is under the age of 12, such as 7-11 years of age or 2-6 years of age. In some embodiments, the subject has been diagnosed with AD. In some embodiments, the subject is suspected of having AD or is at risk of developing AD.

[0171] In some embodiments, the anti-IL-31RA antibody is administered subcutaneously. In some embodiments, the anti-IL-31RA antibody is administered intravenously, intramuscularly, intraperitoneally, or otherwise via injection. In some embodiments, the anti-IL-31RA antibody may be administered once per week, once per two weeks, once per three weeks, once per four weeks, once per five weeks, once per six weeks, once per seven weeks, or once per eight weeks. In some embodiments, the anti-IL-31RA antibody is administered once per four weeks.

[0172] In some embodiments, the anti-IL-31RA antibody is administered at a dose of about 0.01 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 1.5 mg / kg, about 1.5 mg / kg to about 2.5 mg / kg, or about 2.5 mg / kg to about 10 mg / kg. In some embodiments, the anti-IL-31RA antibody is administered at a dose of about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, or about 90 mg. In some embodiments, the anti-IL-31RA antibody is administered at 30 mg to 60 mg. In some embodiments, the anti-IL-31RA antibody is administered at 30 mg. In some embodiments, the anti-IL-31RA antibody is administered at 60 mg.

[0173] In some embodiments, the anti-IL-31RA antibody comprises a heavy chain variable region comprising HCDR1 comprising SEQ ID NO:8, HCDR2 comprising SEQ ID NO:9, and HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising LCDR1 comprising SEQ ID NO:12, LCDR2 comprising SEQ ID NO:13, and LCDR3 comprising SEQ ID NO:14. In some embodiments, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments, the anti-IL-31RA antibody is nemolizumab.

[0174] In some embodiments, the anti-IL-31RA antibody (e.g., nemolizumab) is administered according to a loading dose regimen. In some embodiments, the anti-IL-31RA antibody (e.g., nemolizumab) is administered subcutaneously at a loading dose of 60 mg, followed by doses of 30 mg every 4 weeks for at least 8, 12, 14, 16, 18, 20, 22, or 24 weeks.

[0175] In some embodiments, the anti-IL-31RA antibody (e.g., nemolizumab) is administered according to a flat dosing regimen. For example, in some embodiments, the anti-IL-31RA antibody (e.g., nemolizumab) is administered subcutaneously at a dose of 30 mg or 60 mg every 4 weeks for at least 8, 12, 14, 16, 18, 20, 22, or 24 weeks.

[0176] In another aspect, the present disclosure provides a method for determining whether a subject with atopic dermatitis (AD) responds to treatment with an anti-IL-31RA antibody, comprising: detecting the expression level of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a sample obtained from a subject suspected of having AD; and comparing the expression level of the biomarker to a reference level, where if the expression level of the biomarker is higher than the reference level, the subject responds to treatment; and the reference level is the expression level of each corresponding biomarker in a skin sample from an individual without AD or a sample of non-lesional skin from the subject. In some embodiments, the sample obtained from a subject suspected of having AD is a skin sample, optionally including a lesion. For purposes of the above aspects, the skin sample may comprise, consist of, or essentially consist of the stratum corneum.

[0177] In yet another aspect, the disclosure provides a method of determining whether a subject with atopic dermatitis (AD) responds to treatment with an anti-IL-31RA antibody, comprising detecting expression levels of at least one, at least two, at least three, at least four, at least five, or all six biomarker(s) selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a post-treatment sample obtained from a subject with AD administered at least one dose of an anti-IL-31RA antibody, and comparing the expression levels of the biomarkers to baseline expression levels from a sample obtained from the same subject before treatment was initiated, wherein a decrease in the expression levels of CCL20, CCL22, CCL27, VEGF, and / or CCL18 indicates that the subject responds to treatment, and / or an increase in the expression level of IL1RA indicates that the subject responds to treatment. In some embodiments, the sample is a skin sample, and optionally includes a lesion. In some embodiments, the post-treatment sample is obtained about 4 weeks, about 8 weeks, about 12 weeks, about 14 weeks, about 16 weeks, about 18 weeks, or about 20 weeks after administration of the anti-IL-31RA antibody. In some embodiments, the anti-IL-31RA antibody comprises a heavy chain variable region comprising an HCDR1 comprising SEQ ID NO:8, an HCDR2 comprising SEQ ID NO:9, and an HCDR3 comprising SEQ ID NO:10, and a light chain variable region comprising an LCDR1 comprising SEQ ID NO:12, an LCDR2 comprising SEQ ID NO:13, and an LCDR3 comprising SEQ ID NO:14. In some embodiments, the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof. In some embodiments, the anti-IL-31RA antibody is nemolizumab. For purposes of the above aspects, the skin sample may comprise, consist of, or consist essentially of the stratum corneum.

[0178] For therapeutic and prophylactic purposes of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce CCL18 expression in lesional skin of a subject with (or suspected of having) AD by about 6-fold, about 6.5-fold, about 7-fold, about 7.5-fold, about 8-fold, about 8.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0179] For therapeutic and prophylactic purposes of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce CCL20 expression in lesional skin of a subject with (or suspected of having) AD by about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0180] For therapeutic and prophylactic purposes of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce expression of CCL22 in lesional skin of a subject with (or suspected of having) AD by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0181] For therapeutic and prophylactic purposes of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce CCL27 expression in lesional skin of a subject with (or suspected of having) AD by about 1.5-fold, about 1.75-fold, about 2-fold, about 2.25-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0182] For therapeutic and prophylactic purposes of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may reduce VEGF expression in lesional skin of a subject with (or suspected of having) AD by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or less. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0183] For therapeutic and prophylactic purposes of the present disclosure, treatment with an anti-IL-31RA antibody (e.g., nemolizumab) may increase expression of IL1RA in lesional skin of a subject with (or suspected of having) AD by about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2-fold, about 2.25-fold, about 2.5-fold, about 2.75-fold, about 3-fold, about 3.25-fold, about 3.5-fold, or more. The treatment period required to achieve such a reduction may be 4 weeks, 6 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, 20 weeks, or more. Suitable dosing and administration regimens are described in more detail below.

[0184] For therapeutic and preventative purposes of the present disclosure, the other described biomarkers (e.g., CXCL6, CXCL8, cystatin, FASL, galectin, IL11, IL21, SPD, and CCL17) may show similar patterns of overexpression in lesional skin of subjects with AD (or suspected of having AD) compared to expression levels in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD), and CX3CL1 may show increased expression in lesional skin of subjects with AD (or suspected of having AD) compared to expression levels in a reference sample (e.g., non-lesional skin of the subject or skin from a subject not having AD). Similarly, CXCL6, CXCL8, cystatin, FASL, galectin, CCL17, and IL21 may show a similar pattern of decreased expression in lesional skin of subjects with AD (or suspected of having AD) after treatment with an anti-IL-31RA antibody (e.g., nemolizumab), and CX3CL1, IL11, and SPD may show increased expression in lesional skin of subjects with AD (or suspected of having AD) after treatment with an anti-IL-31RA antibody (e.g., nemolizumab).

[0185] In any of the foregoing aspects or embodiments, the expression levels of these genes (e.g., CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18) in the skin lesions may be detected using any nucleic acid or polypeptide detection assay known in the art. In some embodiments, expression of one or more of CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18 is determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), DNA or RNA microarrays, Western blotting, ELISA, surface plasmon resonance, or mass spectrometry.

[0186] VII. Dosage and Administration Regimens for the Methods and Uses of the Disclosure An effective amount of an anti-IL-31RA antibody, such as nemolizumab or a fragment or variant thereof, is an amount sufficient to produce a beneficial or desired result, such as alleviating at least one or more symptoms of AD. As used herein, an effective amount may also include an amount sufficient to delay or prevent the onset, alter the course of AD symptoms, or reverse AD symptoms. Thus, an exact "effective amount" cannot be specified. However, the appropriate "effective amount" in any given case can be determined by one of ordinary skill in the art using only routine experimentation.

[0187] An effective amount can be administered in one or more administrations, applications, or dosages. Such delivery depends on several variables, including the period for which the individual dosage units are used, the bioavailability of the therapeutic agent, the route of administration, and the like. However, it is understood that the specific dosage level of the therapeutic agent of the present disclosure for any particular subject will depend on a variety of factors, including the activity of the particular compound used, the age, weight, general health, sex, and diet of the subject, the time of administration, the rate of excretion, the drug combination, and the severity and form of administration of the particular disorder being treated. Treatment and prophylactic dosages can generally be titrated to optimize safety and efficacy. Dosages are determined by the physician and can be adjusted, if necessary, to the observed effects of the treatment. Typically, dosage-effect relationships from in vitro and / or in vivo testing can initially provide useful guidance regarding appropriate doses for patient administration. In general, it is desired to administer an amount of the compound effective to achieve serum levels commensurate with the concentrations found to be effective in vitro. Determination of these parameters is within the skill of the art. These considerations, as well as effective formulation and administration procedures, are well known in the art and are described in standard textbooks.

[0188] Dosage regimens for treating or preventing AD can include flat dosing (i.e., administering the same dose repeatedly at predetermined intervals) or loading doses (i.e., administering an initial dose that is higher or different than subsequent successive doses). For purposes of either type of dosing regimen, an effective dose can be administered topically, parenterally, subcutaneously, intradermally, or intramuscularly. In a preferred embodiment, administration includes subcutaneous injection.

[0189] In some embodiments, the loading dose and the subsequent consecutive doses may be administered via the same route (e.g., subcutaneously), while in some embodiments, the loading dose and the subsequent consecutive doses may be administered via different routes (e.g., parenteral and subcutaneous, respectively). In some embodiments, the loading dose may be about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, or more. In some embodiments, the loading dose can be 5 mg, 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, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg or more.In some embodiments, the loading dose is about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about g / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3 mg / kg, about 3.5 mg / kg, about 4 mg / kg, about 4.5 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 500 mg / kg or about 1,000 mg / kg.In some embodiments, the loading dose is 0.01 mg / kg, 0.02 mg / kg, 0.03 mg / kg, 0.04 mg / kg, 0.05 mg / kg, 0.06 mg / kg, 0.07 mg / kg, 0.08 mg / kg, 0.09 mg / kg, 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 1 mg / kg, 1.1 mg / kg, 1.2 mg / kg, 1.3 mg / kg, 1.4 mg / kg, 1.5 mg / kg, 1.6 mg / kg, 1. The loading dose may be 7 mg / kg, 1.8 mg / kg, 1.9 mg / kg, 2 mg / kg, 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg, 2.6 mg / kg, 2.7 mg / kg, 2.8 mg / kg, 2.9 mg / kg, 3 mg / kg, 3.5 mg / kg, 4 mg / kg, 4.5 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 15 mg / kg, 25 mg / kg, 50 mg / kg, 75 mg / kg, 100 mg / kg, 500 mg / kg, or 1,000 mg / kg. In some embodiments, the loading dose is administered as a single injection. In some embodiments, the loading dose is administered as multiple injections that may be administered simultaneously or spaced apart by defined intervals.

[0190] The subsequent successive doses of the loading dose regimen are generally lower than the loading dose. For example, in some embodiments, the dosing regimen may include a loading dose of 60 mg and a successive dose of 30 mg, which may be administered at a defined interval, for example, once every 4 weeks. In some embodiments, the successive doses of the dosing regimen may be about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg or more. In some embodiments, the sequential dose can be 5 mg, 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, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg or more.In some embodiments, the continuous dose is about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about g / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3 mg / kg, about 3.5 mg / kg, about 4 mg / kg, about 4.5 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 500 mg / kg or about 1,000 mg / kg.In some embodiments, the sequential doses are 0.01 mg / kg, 0.02 mg / kg, 0.03 mg / kg, 0.04 mg / kg, 0.05 mg / kg, 0.06 mg / kg, 0.07 mg / kg, 0.08 mg / kg, 0.09 mg / kg, 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 1 mg / kg, 1.1 mg / kg, 1.2 mg / kg, 1.3 mg / kg, 1.4 mg / kg, 1.5 mg / kg, 1.6 mg / kg, 1.7 mg / kg, 1.8 mg / kg, 1.9 mg / kg, 2.0 mg / kg, 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg, 2.6 mg / kg, 2.7 mg / kg, 2.8 mg / kg, 2.9 ...9 mg / kg, 2.1 mg / kg, 2.2 mg / kg, 2.3 mg / kg, 2.4 mg / kg, 2.5 mg / kg, 2.6 mg / kg, 2.7 mg / kg, 2.8 mg / kg, 2.9 mg / kg, 2.8 mg / kg The dose may be 7mg / kg, 1.8mg / kg, 1.9mg / kg, 2mg / kg, 2.1mg / kg, 2.2mg / kg, 2.3mg / kg, 2.4mg / kg, 2.5mg / kg, 2.6mg / kg, 2.7mg / kg, 2.8mg / kg, 2.9mg / kg, 3mg / kg, 3.5mg / kg, 4mg / kg, 4.5mg / kg, 5mg / kg, 6mg / kg, 7mg / kg, 8mg / kg, 9mg / kg, 10mg / kg, 15mg / kg, 25mg / kg, 50mg / kg, 75mg / kg, 100mg / kg, 500mg / kg or 1,000mg / kg.

[0191] For the purpose of the loading dose regimen, the first consecutive dose may be administered 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or 10 weeks after the initial loading dose. In some embodiments, the first consecutive dose is administered 4 weeks after the initial loading dose. In some embodiments, the subsequent consecutive doses are administered once every 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or 10 weeks. In some embodiments, the successive doses are spaced four weeks apart (i.e., nemolizumab, or a fragment or variant thereof, is administered once every four weeks).

[0192] In some embodiments, the dose of an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) administered to a subject can be in the range of 0.001 to 1,000 mg / kg of the subject's body weight. In some embodiments, the dose can be about 0.01 to 50 mg / kg, about 0.01 mg / kg to about 0.1 mg / kg, about 0.05 mg / kg to 0.15 mg / kg, about 0.1 mg / kg to about 0.6 mg / kg, about 0.1 mg / kg to about 1 mg / kg, about 0.25 mg / kg to about 0.75 mg / kg, about 0.4 mg / kg to about 0.8 mg / kg, about 0.4 mg / kg to about 1.8 mg / kg, about 0.5 to about 2.5 mg / kg, about 0.8 mg / kg to about 2.2 mg / kg, about 1 mg / kg to about 2.5 mg / kg, about 1 mg / kg to about 3.5 mg / kg, about The range is from 1 mg / kg to about 5 mg / kg, from about 2 mg / kg to about 4 mg / kg, from about 2.5 mg / kg to about 10 mg / kg, from about 5 mg / kg to about 10 mg / kg, from about 10 mg / kg to about 20 mg / kg, from about 10 mg / kg to about 40 mg / kg, from about 20 mg / kg to about 50 mg / kg, from about 25 mg / kg to about 75 mg / kg, from about 50 mg / kg to about 100 mg / kg, or from about 100 mg / kg to about 500 mg / kg, or from about 100 mg / kg to about 1000 mg / kg body weight of an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof). In preferred embodiments, the dosage ranges from about 0.01 mg / kg to about 0.1 mg / kg, from about 0.1 mg / kg to about 0.5 mg / kg, from about 0.5 mg / kg to about 1.5 mg / kg, from about 1.5 mg / kg to about 2.5 mg / kg, or from about 2.5 mg / kg to about 10 mg / kg.In some embodiments, the dose is about 0.01 mg / kg, about 0.02 mg / kg, about 0.03 mg / kg, about 0.04 mg / kg, about 0.05 mg / kg, about 0.06 mg / kg, about 0.07 mg / kg, about 0.08 mg / kg, about 0.09 mg / kg, about 0.1 mg / kg, about 0.2 mg / kg, about 0.3 mg / kg, about 0.4 mg / kg, about 0.5 mg / kg, about 0.6 mg / kg, about 0.7 mg / kg, about 0.8 mg / kg, about 0.9 mg / kg, about 1 mg / kg, about 1.1 mg / kg, about 1.2 mg / kg, about 1.3 mg / kg, about 1.4 mg / kg, about 1.5 mg / kg, about 1.6 mg / kg, about 1.7 mg / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2.0 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3.0 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about 3.8 mg / kg, about 3.9 mg / kg, about 3.1 mg / kg, about 3.2 mg / kg, about 3.3 mg / kg, about 3.4 mg / kg, about 3.5 mg / kg, about 3.6 mg / kg, about 3.7 mg / kg, about g / kg, about 1.8 mg / kg, about 1.9 mg / kg, about 2 mg / kg, about 2.1 mg / kg, about 2.2 mg / kg, about 2.3 mg / kg, about 2.4 mg / kg, about 2.5 mg / kg, about 2.6 mg / kg, about 2.7 mg / kg, about 2.8 mg / kg, about 2.9 mg / kg, about 3 mg / kg, about 3.5 mg / kg, about 4 mg / kg, about 4.5 mg / kg, about 5 mg / kg, about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 15 mg / kg, about 25 mg / kg, about 50 mg / kg, about 75 mg / kg, about 100 mg / kg, about 500 mg / kg or about 1,000 mg / kg. In certain embodiments, the dose of an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) is about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 1.5 mg / kg, about 2 mg / kg, or about 2.5 mg / kg. In a preferred embodiment, the dose is about 0.5 mg / kg.

[0193] In some embodiments, the dose of an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) administered to a subject is within the range of 1-100 mg, 25-75 mg, 30-60 mg, 40-80 mg, 20-80 mg, 1-25 mg, 1-50 mg, 10-90 mg, 15-85 mg, or any range therebetween. In some embodiments, the dose can be about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg or more. In some embodiments, the dose can be 5 mg, 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, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg or more.

[0194] In some embodiments of the methods and uses of the present disclosure, a loading dose of about 60 mg of an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) may be administered to a subject with AD, followed by a subsequent consecutive dose of an anti-IL31RA antibody (e.g., a fragment of nemolizumab or a variant thereof) at about 30 mg once every four weeks. In some embodiments of the methods and uses of the present disclosure, a first dose of about 60 mg of an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) may be administered to a subject with AD, followed by a subsequent consecutive dose of an anti-IL31RA antibody (e.g., a fragment of nemolizumab or a variant thereof) at about 60 mg once every four weeks (i.e., the dose is constant or is a "flat" dosing regimen). In some embodiments of the methods and uses of the present disclosure, a first dose of an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) of about 30 mg may be administered to a subject with AD, followed by subsequent consecutive doses of an anti-IL31RA antibody (e.g., a fragment of nemolizumab or a variant thereof) at about 30 mg once every four weeks.

[0195] In some embodiments of the methods and uses of the present disclosure, the anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) is administered by a topical or parenteral route. In some embodiments, the anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) is administered subcutaneously. In some embodiments, a dose is administered subcutaneously at or proximal to the site of one or more nodules, lesions, or denudations.

[0196] In some embodiments of the methods and uses of the present disclosure, the anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) is administered daily, every other day, twice a week, three times a week, four times a week, five times a week, six times a week, once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, once every eight weeks, once every nine weeks, once every ten weeks, once every eleven weeks, once every twelve weeks, twice a year, once a year, and / or as needed based on the appearance of AD symptoms. In a preferred embodiment, the anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) is administered every four weeks or every eight weeks.

[0197] In some embodiments of the disclosed methods and uses, the duration of treatment or prevention is about 1 day, about 1 week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 6 weeks, about 7 weeks, about 8 weeks, about 9 weeks, about 10 weeks, about 11 weeks, about 12 weeks, about 13 weeks, about 14 weeks, about 15 weeks, about 16 weeks, about 17 weeks, about 18 weeks, about 19 weeks, about 20 weeks, about 24 weeks, about 30 weeks, about 36 weeks, about 40 weeks, about 48 weeks, about 50 weeks, about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, or as needed based on the appearance of AD symptoms. In preferred embodiments, the duration of treatment or prevention is about 12 weeks to about 24 weeks, about 12 weeks to about 36 weeks, about 12 weeks to about 48 weeks, or about 24 weeks to about 36 weeks.

[0198] The present disclosure provides the use of an anti-IL31RA antibody (e.g., nemolizumab or a fragment or variant thereof) in the manufacture of a medicament for treating or preventing AD, for normalizing expression of one or more of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a subject with AD, and / or for reducing an inflammatory response in the skin. All of the doses, administration regimens, administration routes, biomarkers, and treatment endpoints of the present disclosure are also applicable to these uses.

[0199] The following examples are provided to illustrate the present disclosure, it being understood that the invention should not be limited to the specific conditions or details described in these examples. EXAMPLES

[0200] Example 1 - Clinical trial to determine the efficacy of nemolizumab in treating AD. Introduction and Methods A Phase 2, single-arm, open-label study was conducted to evaluate the pharmacokinetics (PK), safety, and clinical efficacy of nemolizumab in 18 subjects with moderate to severe AD, as described below. The effect of nemolizumab on biomarkers of AD was also investigated.

[0201] Overall Study Design. This was an open-label, single-arm study to evaluate the PK and safety of nemolizumab in adolescent subjects (ages 12-17) with moderate to severe AD and associated pruritus. Approximately 20 eligible subjects were enrolled and received subcutaneous injections of nemolizumab (30 mg) every 4 weeks over a 16-week treatment period, with LD of 60 mg on day 1. The final dose of study drug was administered at week 12, and subjects completed the treatment period at the week 16 visit. An 8-week follow-up period (week 24 visit) was scheduled for subjects who did not roll over into the nemolizumab long-term extension study. Subjects who discontinued the study early before the week 16 visit had to be followed for 12 weeks after their last dose of study drug. Figure 1 and Table 2 provide an overview of the clinical study.

[0202] (Table 2) Overview of clinical studies TIFF2024535715000013.tif61167LD=loading dose, n=number of subjects, Q4W=every 4 weeks.

[0203] Nemolizumab serum concentrations were assessed at baseline, weeks 1-2, 4, 8, 12, 16, and 24, and at any unscheduled visit for safety reasons. Previous PK data from phase 1 and phase 2a studies were used to develop a population PK model that allows accurate simulation of nemolizumab serum concentrations at different doses and dose regimens. The population PK model was used to simulate systemic exposure using several fixed doses for the phase 2b study. Phase 2b results support the predictability of the popPK model for the 30 mg dose in adults (Table 3).

[0204] (Table 3) Nemolizumab has an actual C trough Mean serum concentrations (μg / mL) compared with predicted levels TIFF2024535715000014.tif65167 a N represents the number of observations. Concentrations are the individual predictions derived from the model. b n represents the number of adult subjects providing PK data for the interim PK analysis of Study 114322.

[0205] the purpose: The primary objective of the study was to evaluate the PK and safety of nemolizumab, when administered in combination with TCS, in adolescent subjects with moderate to severe atopic AD and associated pruritus not adequately controlled with topical treatments.

[0206] The secondary objective of the study was to evaluate the efficacy of nemolizumab and to further characterize the relationship between nemolizumab systemic exposure and clinical efficacy endpoints (PK / PD relationship).

[0207] Inclusion criteria. To participate in the study, subjects must meet the following criteria: 1. Subjects aged 12 to 17 years at the time of screening visit. 2. Chronic AD for at least 2 years prior to the screening visit, confirmed according to the American Academy of Dermatology consensus criteria at the time of the screening visit. 3. EASI score ≥ 16 at both screening and baseline visits. 4. IGA score ≥ 3 at both screening and baseline visits (based on an IGA scale ranging from 0 to 4, with 3 being moderate and 4 being severe). 5. AD involvement of ≥10% of BSA at both screening and baseline visits. 6. Peak (maximum) Pruritus Numeric Rating Scale (NRS) score of at least 4.0 at both Screening and Baseline visits Screening PP NRS score was determined by a single PP NRS assessment (score ranging from 0 to 10) during the 24 hours immediately preceding the screening visit. Baseline PP NRS scores were determined based on the average of daily PP NRS scores (scores ranging from 0 to 10) for the 7 days immediately preceding baseline (rounding was not allowed). A minimum of 4 daily scores for the 7 days immediately preceding baseline were required for this calculation. 7. Documented recent medical history (within 6 months prior to the screening visit) of inadequate response to local drug treatment (TCS with or without TCI). Acceptable documentation includes patient records containing information on prescription of TCS (with or without TCI) and treatment outcome, or documentation of a conversation with the subject's treating physician if different from the Investigator. In cases of insufficient documentation, the subject may have been rescreened after such documentation was obtained. Inadequate response to TCS treatment (with or without TCI) was defined as: 7a. Failure to achieve or maintain remission or low disease activity (equivalent to IGA ≤ 2, mild) despite treatment with a daily regimen of medium- or high-potency TCS (with or without a TCI) applied for at least 4 weeks or for the longest period according to the prescribing information; or 7b.Need for long-term treatment (>4 weeks) with high-potency TCS (with or without a TCI) to achieve or maintain remission or low disease activity (corresponding to an IGA ≤ 2). Note: If the subject was receiving a TCI in addition to a TCS, documentation of failure to achieve or maintain remission or low disease activity (corresponding to an IGA ≤ 2) was also required. 7c. If documentation of an inadequate response to topical medical treatment was not available, subjects with a documented recent history of systemic treatment or phototherapy for AD (within 6 months prior to the visit) were also considered to be inadequate responders to topical treatment. 8. Agreed to apply moisturizer ad libitum throughout the study, daily from the Screening Visit and as needed. Agreed to apply approved TCS from the Screening Visit throughout the study, as determined appropriate by the Investigator. 9. Women of childbearing potential (WOCBP) must agree to abstain throughout the study and for 12 weeks after the last study drug injection, or to use an effective, approved method of contraception throughout the study and for 12 weeks after the last study drug injection. This criterion also applied to prepubertal female subjects who began menstruating during the study.

[0208] Effective and approved contraceptive methods available to the subject and / or his / her partner are defined as follows: Progestogen-only oral hormonal contraception ● Male or female condoms Spermicide cap, diaphragm, or sponge - Male or female condoms in combination with spermicide caps, diaphragms, or sponges Combined (estrogen and progestogen-containing) oral, intravaginal, or transdermal hormonal contraception Injectable or implantable hormonal contraception Intrauterine contraceptive device 10. Subject and guardian are willing and able to comply with all time commitments and procedural requirements of the clinical research protocol, including the recording of a daily diary by the subject using the electronic handheld device provided for this study. 11. Understood and signed the ICF and consent form before any study procedures were performed.

[0209] Exclusion Criteria. To participate in the study, subjects must not meet any of the following criteria: 1.Weight < 30kg. 2.Subjects who meet one or more of the following criteria at screening or baseline: 2a. Have had an asthma exacerbation requiring hospitalization within the past 12 months. 2b. Report of poorly controlled asthma (i.e., symptoms more than 2 days per week, nighttime awakenings more than 1-3 times per week, or any interference with normal activities) in the past 3 months. 2c. Asthma Control Test (ACT) ≦19 (only applicable to subjects with a history of asthma). 2d. Peak expiratory flow (PEF) <80% of predicted. 3. Subject with a current history of chronic obstructive pulmonary disease (COPD) and / or chronic bronchitis. 4. Skin infection within 1 week prior to the screening visit, or infection requiring treatment with oral or parenteral antibiotics, antivirals, antiparasitics, or antifungals within 1 week prior to the screening visit. Subjects may be rescreened once the infection has resolved. 5. Requires rescue therapy for AD during the trial period or is anticipated to require rescue therapy within 2 weeks after the baseline visit. 6. Positive serologic results for hepatitis B surface antigen [HBsAg] or hepatitis B core antibody [HBcAb], hepatitis C antibody, or human immunodeficiency virus [HIV] antibody at the screening visit. Note: Subjects who were HBcAb positive and HBsAg negative could be included in this clinical study if they had positive hepatitis B surface antibody (HBsAb) (considered immune after natural infection). 7. Received any of the following treatments in Table 4 within the specified time frame prior to the Baseline Visit:

[0210] (Table 4) Previous treatments TIFF2024535715000015.tif166167AD=atopic dermatitis, JAK=Janus kinase, TCS=topical corticosteroids NOTE: These treatments were not to be discontinued for reasons related to this clinical study.

[0211] Treatment. Nemolizumab is a humanized monoclonal modified immunoglobulin G (IgG)2 antibody that contains a structure of two H chains (445 amino acid residues) and two L chains (214 amino acid residues) connected by 16 disulfide bonds. The drug product used in this study was a vial containing 153 mg of lyophilized nemolizumab powder, which was stored protected from light at 2-8 °C until reconstituted with 1.3 mL of sterile water, then diluted using nemolizumab placebo to obtain an injection solution containing 100 mg / mL of nemolizumab, which was further diluted to obtain the appropriate dosing solution. Subsequently, each 100 mg / mL vial was diluted with reconstituted placebo to obtain 30 mg / mL or 60 mg / mL (LD). By design, a volume of 1 mL was consistently injected. Reconstitution with sterile water for injection and dilution with nemolizumab placebo were identical to the steps used in the adult Phase 2b study (SPR.114322). The drug administration scheme and detailed description are shown in Tables 5 and 6.

[0212] Table 5. Dosing scheme TIFF2024535715000016.tif15128

[0213] (Table 6) Description of Nemolizumab TIFF2024535715000017.tif63167

[0214] Prescribed use of background therapy was documented on the Case Report Form (CRF). Background therapy was required throughout the study (from screening to follow-up visits) as described below.

[0215] Moisturizers: Subjects were required to apply moisturizer daily and ad libitum as needed to dry skin and AD lesions throughout the study. The subject's current moisturizer or a moisturizer recommended by the investigator may have been used. It should not be used within 8 hours prior to each clinic visit. Moisturizers were not considered topical AD medications.

[0216] Topical Corticosteroid Therapy (TCS): Subjects were required to apply an approved basal TCS regimen to all AD lesions throughout the study as directed by the investigator, beginning within the screening period and no less than 14 days prior to Day 1. Subjects were required to apply medium-potency TCS to areas of the body where use of medium-potency TCS was deemed safe (e.g., trunk and extremities). Low-potency TCS was used in TCS-sensitive areas (e.g., face, neck, intertriginous areas). Subjects were required to apply a thin layer of approved TCS to all AD lesions as frequently as necessary to ensure disease stability and prevent AD flares, but not more frequently than the daily frequency recommended on the product label. It should be noted that "as needed" use of TCS was not permitted.

[0217] Rescue Therapy: Rescue therapy was prescribed to subjects at any time during the study except during the trial period if deemed medically necessary by the investigator (e.g., to control intolerable AD signs / symptoms). Subjects who received rescue therapy during the trial period were not eligible to participate in the study. As a general guideline and according to the individual investigator's judgment, rescue therapy was not prescribed within the first 2 weeks after baseline (i.e., week 2 visit) to allow for a minimum period of study drug exposure in the presence of background therapy. Rescue therapies were only those that directly treated AD (primarily those that were approved or standard of care) and included topical and systemic therapies as outlined. Some rescue therapies included: TCIs, higher potency TCS (class I-II according to US classification), oral corticosteroids, biologics (including their biosimilars), systemic nonsteroidal immunosuppressants / immunomodulators, and phototherapy.

[0218] Efficacy, safety, and pharmacokinetic variables PK, safety, and efficacy assessments were performed throughout the study. Subject-reported assessments of topical AD medication use for pruritus, sleep disturbance, and eczema were collected daily. The assessment schedule is provided in Table 7.

[0219] (Table 7) Evaluation schedule TIFF2024535715000018.tif240141TIFF2024535715000019.tif245156TIFF2024535715000020.tif240147TIFF2024535715000021.tif241134

[0220] Efficacy measures were performed by the investigator (or trained designee) and by the subject (for subject-reported efficacy measures) according to Error! Reference source not found. . Whenever possible, the same assessors performed the assessments throughout the study.

[0221] Eczema Area and Severity Index. The Eczema Area and Severity Index is a validated scale commonly used in clinical research and clinical practice to assess the severity and extent of AD signs. The Eczema Area and Severity Index is a composite score ranging from 0 to 72. The severity of erythema, induration / papulation, peeling, and lichenification was assessed by the investigator or a trained designee on a scale of 0 (absent) to 3 (severe), with half points allowed, for each of four body regions: head / neck, trunk, upper limbs, and lower limbs. In addition, the extent of AD involvement in each of the four body regions was assessed as a percentage by body region for head, trunk, upper limbs, and lower limbs and converted to a score of 0 to 6. EASI scores were calculated on the CRF.

[0222] Investigator's Global Assessment (IGA). The IGA is a 5-point scale ranging from 0 (clear) to 4 (severe) used by the investigator or a trained designee to assess the overall severity of AD and the clinical response to treatment. Treatment success is defined as 0 (clear) or 1 (almost clear), and a 2-grade change from baseline.

[0223] Body surface area (BSA). BSA involvement of AD was assessed by the investigator or trained designee for each body part (maximum possible scores for each region were head and neck [9%], anterior trunk [18%], back [18%], upper extremities [18%], lower extremities [36%], and genitals [1%]) and reported as a combined percentage of all major body sections calculated on the CRF.

[0224] Scoring Atopic Dermatitis (SCORAD). SCORAD is a validated scale commonly used in clinical research and clinical practice to assess the severity and extent of AD signs and symptoms. The SCORAD score ranges from 0 to 103 and has three components: degree of AD (BSA), signs, and symptoms, and subject-reported symptoms of pruritus and sleep deprivation. The investigator or designee assessed the severity of six signs of AD (erythema / darkening, edema / papulation, oozing / scabbing, peeling, lichenification / prurigo, and dryness), each on a scale ranging from 0 (none) to 3 (severe). The investigator or designee also asked the subject to rate symptoms of pruritus and sleep deprivation (average of the past 3 days / nights) on a VAS ranging from 0 to 10. SCORAD scores were calculated on the CRF.

[0225] Pruritus Numeric Rating Scale (NRS). The Pruritus NRS is a scale used by subjects to report the intensity of pruritus (itch) over the past 24 hours. The Pruritus NRS has been validated in other AD clinical studies in adults, and the minimal clinically important difference has been shown to be 3-4.

[0226] The screening PP NRS score, measuring maximum pruritus intensity, was determined by a single PP NRS assessment (score ranging from 0 to 10) during the 24 hours immediately preceding the screening visit. The baseline PP NRS score was determined based on the average of the daily PP NRS scores (score ranging from 0 to 10) during the 7 days immediately preceding baseline (rounding was not allowed). A minimum of four daily scores during the 7 days immediately preceding baseline were required for this calculation. Subjects received instructions on how to use and record their pruritus NRS score on the electronic device and completed the assessment once each evening throughout the entire clinical study (including the trial and follow-up periods). Subjects were asked the following questions in their local language: · Average itch intensity: "On a scale of 0 to 10, with 0 being 'no itch' and 10 being 'the worst itch imaginable,' how would you rate your overall itch over the past 24 hours?" · Regarding maximum itch intensity: "On a scale of 0 to 10, with 0 being 'no itch' and 10 being 'the worst itch imaginable,' how would you rate the worst itch you've experienced in the past 24 hours?"

[0227] Application of topical medication for atopic dermatitis: diary. Subjects were instructed to use the approved basal TCS as prescribed by the investigator. Every evening, subjects were asked to record in an electronic device (i.e. diary) the answer (yes / no) to the following question throughout the entire clinical study (including trial and follow-up periods): "Did you apply the eczema medication given to you by your doctor to your skin today?"

[0228] Sleep Disorder NRS. The Sleep Disorder NRS is a scale used by subjects to report the degree of sleep deprivation associated with AD. Subjects received instructions on how to use and record their Sleep Disorder NRS score on an electronic device and completed the assessment once each morning throughout the clinical study (including trial and follow-up periods). Subjects were asked the following question in their local language: On a scale of 0 to 10, with 0 being "no sleep deprivation associated with AD signs / symptoms" and 10 being "unable to sleep at all due to AD signs / symptoms," how would you rate your sleep last night?"

[0229] Dermatology Life Quality Index / Pediatric Dermatology Life Quality Index. The Dermatology Life Quality Index (DLQI) is a validated 10-item questionnaire for subjects over 16 years of age, covering domains including symptoms / emotions, daily activities, leisure, work / school, personal relationships, and treatments. The Pediatric DLQI (cDLQI) is an equivalent validated 10-item questionnaire designed for pediatric subjects 16 years of age and younger. Subjects rated each question on a scale ranging from 0 (not at all) to 3 (very much). A higher total score indicates a poorer QoL.

[0230] Safety assessments were performed for all subjects at the screening visit (signing of the ICF) and at each visit thereafter. An adverse event (AE) is defined as an undesirable medical occurrence in a clinical study subject receiving a medicinal product, not necessarily having a causal relationship to this treatment. Thus, an AE can be any undesirable and unintended sign (including abnormal laboratory findings), symptom, or disease that is primarily related to the use of a medicinal product, whether or not related to the medicinal product. This includes a worsening of a pre-existing condition or event, an intercurrent illness, a drug interaction, or a significant worsening of the indication under investigation that is not recorded elsewhere in the CRF under a specific efficacy evaluation. Each AE was assigned by the investigator to a category as follows: • Mild: An AE that is easily tolerated by the subject, causes minimal discomfort, and does not interfere with daily activities. Moderate: An AE that is sufficiently unpleasant to interfere with normal daily activities and may require intervention. Severe: An AE that interferes with normal daily activities and usually requires medical treatment or other intervention.

[0231] If there is a change in severity of an AE, it should be recorded as a separate event.

[0232] Adverse events of special interest (AESIs) are events of note for an investigational drug that should be closely monitored and reported immediately. AESIs were either serious or non-serious. Based on the potential risks of nemolizumab and the risks associated with biologics in general (and their biosimilar equivalents) (i.e., class effects), the following AEs are considered AESIs: Injection-related reactions, including anaphylactic reactions, acute allergic reactions requiring medical treatment, and severe injection site reactions (ISR) (i.e., lasting >24 hours) Newly diagnosed asthma or worsening asthma - More specifically, subjects with a history of asthma were referred to a physician for management of asthma if: ACT score ≤ 19; an ACT score ≤ 19 conveyed inadequately controlled asthma. AESIs were reported based on the investigator's clinical judgment versus the supervising physician's report. □PEF <80% of predicted value, AESI reported. Unexpected asthma exacerbations were observed or reported. AESIs were reported based on the investigator's clinical judgment. - Subjects without a history of asthma were referred to an appropriate pulmonologist / specialist if: Signs and / or symptoms suggestive of asthma were observed or reported. AESIs were reported based on the investigator's clinical judgment versus expert reports. Respiratory assessment suggests a decline in the subject's respiratory health. AESIs were reported based on the investigator's clinical judgment versus expert reports. Infection. Severe infection or infection requiring treatment with parenteral antibiotics for more than 2 weeks or oral antibiotics / antivirals / antifungals. Peripheral edema: limbs, both sides Facial edema Elevated ALT or AST (>3×ULN) in combination with elevated bilirubin (>2×ULN)

[0233] Serious Adverse Event (SAE). An SAE is any undesirable medical occurrence or effect, at any dose, that: (1) results in death, (2) is life-threatening, (3) requires or prolongs hospitalization, (4) results in persistent or significant disability / disability, (5) results in congenital anomaly / birth defect, or (6) is a significant medical event that may not result in death, is not life-threatening, or does not require hospitalization and that, based on sound medical judgment, may be considered an SAE, where the event may jeopardize the safety of the subject and may require medical or surgical intervention to prevent one of the outcomes above in this definition (e.g., allergic bronchospasm, hematopoietic dysfunction, or spasms that do not result in hospitalization).

[0234] Unexpected Adverse Reaction. An unexpected adverse reaction is any undesired, unintended response associated with the administration of an investigational product at any dose, the nature or severity of which is inconsistent with the applicable product information (e.g., reference safety information such as the nemolizumab investigator's brochure, study protocol, etc.).

[0235] Clinical Laboratory Evaluations. Hematology, chemistry laboratory analyses, and urinalysis will be performed in a central laboratory as specified in Table 7. Pregnancy testing, tuberculosis screening, complete physical examination (PE), respiratory evaluation, asthma control testing (ACT), respiratory testing, spirometry / peak respiratory flow, and 12-lead electrocardiogram (ECG) were performed. Vital signs, height, and weight were monitored.

[0236] Pharmacokinetics: Blood sample collection. Blood samples were collected according to Table 7 to determine the PK profile of nemolizumab. Serum concentrations of nemolizumab were assessed at baseline, weeks 1-2, 4, 8, 12, 16, and 24, and at any unscheduled visit for safety reasons. At each sample collection time point for PK evaluation, collected blood was allowed to clot at room temperature (within 60 minutes after collection) and then centrifuged at room temperature. Serum was collected into storage tubes. At the study drug injection visits (baseline, weeks 4, 8, and 12), PK samples were collected within 30 minutes prior to study drug injection at weeks 1-2, 16, and 24 (pre-dose samples), with attempts to collect PK samples at approximately the same time of day whenever possible. The date and time of each sample collection was recorded on the CRF along with the time of study drug injection (or missed injection, if applicable) at the same visit. Processing of blood samples for PK evaluation was described in the laboratory manual.

[0237] Quantification of nemolizumab (CD14152) in serum. The concentration of nemolizumab (CD14152) in serum was determined by a designated CRO (Shin Nippon Biomedical Laboratories, Ltd, Japan) using a validated enzyme-linked immunosorbent assay (ELISA) method with a quantification limit of 100 ng / mL.

[0238] Pharmacokinetic Analysis. Pharmacokinetic parameters of nemolizumab in serum were calculated by a designated CRO (Certara) using two analyses: nonlinear mixed-effects modeling and non-compartmental analysis (NCA) analysis.

[0239] Analysis 1: A population PK (popPK) model was used to describe the time course of nemolizumab exposure and to explore sources of variation in subject exposure. A first-order absorption and one-compartment distribution popPK model was developed using nemolizumab serum concentrations from three clinical studies (CIM001JP, CIM003JG, and SPR.114322) conducted in adults and used to derive empirical Bayes estimates in the adolescent population based on their baseline characteristics, their dosing history, and their measured concentrations.

[0240] The suitability of the model to adequately describe the adolescent data was based on the model diagnostic tools described in the dedicated PK modeling plan. The popPK model was used to evaluate the main popPK parameters (CL / F, V d / F,T lag , k a ) was obtained.

[0241] Other individual nemolizumab PK parameters (C max , T max , C trough , AUC 0-4w , AUC 0-8w , AUC 0-12w , AUC 0-16w , AUC inf , and t 1 / 2 ) was derived using the same popPK model.

[0242] Analysis 2: Some individual nemolizumab PK parameters (AUC 0-4w , AUC 0-8w , AUC 0-12w , and AUC 0-16w ) were also calculated using NCA, using only serum concentrations actually observed in the study.

[0243] Pharmacokinetic / pharmacodynamic analysis (PK / PD). A pharmacokinetic / PD model was developed to correlate nemolizumab concentrations to clinical efficacy. The popPK model combined the following pharmacodynamic (PD) endpoints: Eczema Area and Severity Index (EASI), Investigator's Global Assessment of Dermatitis Severity (IGA), and Weekly Average Peak Pruritus Numeric Rating Scale (PP-NRS). For each subject, the concentration time course was predicted based on individual Bayesian posterior estimates of PK parameters up to the time of the last PD assessment and was used as a driver in the PD model.

[0244] Other variables: Immunogenicity. To assess ADA, blood samples were collected according to Table 7. Serum samples were assessed for ADA at baseline, weeks 4, 8, 16, 24, and at any unscheduled visit for safety reasons. ADA was determined at these time points by a designated CRO (LSI Medience Corporation. Japan) using a validated ELISA assay. Serum concentrations were assessed using a multi-tiered approach.

[0245] If serum circulating ADA was detected by the screening assay, the presence was confirmed and characterized (e.g., titer and neutralization potential) using a validated assay. The incidence of positive ADA results was summarized (absolute incidence and percent of subjects).

[0246] For ADA assessment, blood was collected at the same time as PK sampling according to Table 7. Additional visits (considered as unscheduled visits) may be conducted after the last scheduled study visit (12 weeks after the last investigational drug injection) to collect additional samples for further analysis of ADA. Processing of blood samples for PK and ADA assessment was well known in the art.

[0247] Biomarkers. Blood and stratum corneum samples were collected to examine the effect of nemolizumab on selected biomarkers. Unless otherwise specified, samples were shipped to a designated CRO for biomarker evaluation. Biomarker samples (plasma and skin) were destroyed after all testing was completed and when the pharmacodynamic reports were approved.

[0248] Blood samples were collected for evaluation of TARC and Immunoglobulin E (IgE) by a central laboratory. Blood samples were collected for plasma biomarker evaluation according to Table 7. An aliquot of the plasma sample was shipped to a designated CRO for evaluation using a set of biomarkers associated with AD, including but not limited to IL-6, IL-8, and IL-18. Depending on the results of using protein biomarkers, the amount of sample remaining, and the compatibility of the initially collected sample with further investigations, the expression of micro ribonucleic acid (miRNA) and messenger ribonucleic acid (mRNA) markers was subsequently analyzed.

[0249] Stratum corneum (SC) samples were collected using tape strips (D-squames) according to Table 7. At baseline, before administration of the study drug, two lesional and two non-lesional areas were identified and four rectangular D-squames (2.3 cm x 3.5 cm dimensions) were collected from each area, making a total of 16 D-squames (8 from lesional skin and 8 from non-lesional skin). At week 16, four rectangular D-squames were collected from the two lesional areas identified at baseline (8 D-squames total). An aliquot of the sample was shipped to a designated CRO for analysis of selected protein biomarkers related to AD (including TARC, IL-6, IL-8, IL-18, etc.). In addition, at end of study (EOS) ceramide levels were quantified. Depending on the results of protein and ceramide biomarkers, the amount of sample remaining, and the suitability of the initially collected samples for further investigation, lipid expression, as well as miRNA and mRNA marker expression could be subsequently analyzed.

[0250] Data analysis and presentation. Presentation of pharmacokinetic concentrations. Nemolizumab serum concentration data (in ng / mL) were summarized for each visit from baseline to week 24 using the following statistics: n, arithmetic mean, standard deviation, CV%, geometric mean, CV geometric mean%, median, minimum, maximum, 95% CI of the mean, and number of BLQs (below limit of quantification). BLQs were considered missing and excluded from the narrative summary.

[0251] Individual serum concentrations and mean concentrations were plotted by visit on both linear and semi-log scale for the PK population (x-axis units: weeks). For individual concentration plots, BLQ was considered missing. For mean concentration plots, mean concentrations not calculated were considered missing. In addition, individual serum concentrations were plotted over time in different colors for subjects with treatment-related ADA+ (ADA- at baseline and ADA+ after treatment), ADA+ at baseline, and ADA- from baseline during the study). Individual serum concentrations were listed, and non-quantifiable concentrations were listed as BLQ (<100 ng / mL).

[0252] Presentation of pharmacokinetic parameters. Major population PK parameters (Cl / F, Vd / F, T lag Estimates of the mean observed trough (C ) are presented in a dedicated PK report. Individual derived PK parameters at steady state estimated with the popPK model and individual observed PK parameters at steady state calculated with NCA were summarized using the following statistics: n, arithmetic mean, standard deviation, geometric mean, CV%, median, minimum, maximum, and 95% CI. In addition, the mean observed trough (C ) between ADA+ and ADA- samples was analyzed using (analysis of variance) ANOVA. trough ) Nemolizumab concentrations were compared and the comparative results were presented.

[0253] Efficacy variables: All efficacy analyses were based on the intention-to-treat (ITT) population. All efficacy variables were summarized by analysis visit from baseline unless otherwise specified. For efficacy analyses, unscheduled visits or ET visit data were windowed based on the analysis visit window to be considered for visit-by-visit summarization. All efficacy analyses were performed using last observation carried forward (LOCF) or non-responder imputation (unless otherwise specified) and were also performed with OC as a sensitivity analysis.

[0254] Eczema Area and Severity Index (EASI) scores were derived and recorded on the CRF as described in Table 8. EASI was summarized narratively by visit using LOCF and OC. Absolute change and percent change from baseline were included in the analysis. For LOCF analysis, imputation was performed on the EASI overall imputed score. Percent change in EASI using LOCF was illustrated.

[0255] Table 8. Calculation of Eczema Area and Severity Index Score TIFF2024535715000022.tif44167

[0256] Investigator Global Assessment (IGA) is a 5-point scale used by the investigator to assess the overall severity and clinical response of AD, as described in Table 9. IGA scores and changes from baseline were summarized by visit using LOCF and OC. IGA success was defined as 0 (clear) or 1 (almost clear) and at least a 2-grade improvement change from baseline. IGA treatment success rates were obtained as the ratio between the number of subjects who achieved IGA treatment success and the number of subjects with IGA results available at the visit, and were also summarized by visit from baseline using non-responders, LOCF, and OC. The proportion of subjects who achieved IGA success was illustrated using non-responders.

[0257] Table 9. Five-point scale of investigator's global assessment TIFF2024535715000023.tif67167

[0258] Body surface area (BSA) of AD involvement was assessed as a percentage for eight different parts of the body and reported individually and as overall BSA (calculated as the sum of all parts combined) as derived in the CRF. The body parts assessed and their highest possible scores were as follows: head and neck [9%], anterior trunk [18%], back [18%], upper limbs [18%], lower limbs [36%], and genitals [1%]. Overall BSA of AD involvement was summarized descriptively by visit as a continuous variable using LOCF and OC. Absolute change from baseline and percentages were also included in the descriptive analysis. For LOCF analysis, imputation was performed separately for each body part, and the sum of the imputed body domain scores gave the imputed overall BSA.

[0259] Scoring Atopic Dermatitis (SCORAD) is a validated scale commonly used in clinical research and clinical practice to assess the severity and extent of AD signs and symptoms. SCORAD considers the extent and intensity of six basic types of lesions (erythema / darkening, edema / papule, oozing / crusting, peeling, lichenification / prurigo, and dryness) and symptoms (itching and sleep deprivation). SCORAD ranges from 0 to 103 and has three components: extent, intensity, and subjective symptoms. Calculation of SCORAD was performed as follows: the extent of the BSA affected by AD was the "component of the extent of the area involved" (denoted by A). The intensity of all basic types of lesions was then assessed. Depending on the intensity, each of these is assigned a score of 0 (absent), 1 (mild), 2 (moderate), or 3 (severe). The sum of these scores was used to estimate the "intensity component" (denoted by B). Finally, the presence of irritability and poor sleep for the past three nights was assessed based on a VAS (recorded in centimeters) with values ​​from 0 (no symptoms) to 10 (worst symptoms ever had). The sum of the two values ​​was used to estimate the "subjective symptoms component" (denoted C). The SCORAD total score was then calculated as = A / 5 + (7*B) / 2 + C. SCORAD was summarized narratively by visit using LOCF and OC. Absolute change and percent change from baseline were also included in the summary. For LOCF analysis, imputation was performed separately for each SCORAD component, and the imputed components were used to obtain an imputed total score.

[0260] The Pruritus NRS was used by subjects to report the intensity of pruritus (itch) over the past 24 hours (subject-reported ratings of pruritus were collected in a daily diary. The mean and maximum itch intensity scales have values ​​ranging from 0 to 10, with 0 being "no itch" and 10 being "worst itch imaginable." Mean and PP-NRS scores were obtained based on the average and maximum daily scores over the 7 days immediately preceding each scheduled visit (rounding was not allowed) and presented as "weekly" scores (mean week and peak week, respectively). At least four daily scores were required for these calculations. If less than four daily scores are necessary and available, each weekly score is a missing value. Weekly mean peak pruritus (PP) NRS and weekly mean average pruritus (AP) NRS scores were descriptively summarized by visit as continuous variables using LOCF and OC. Absolute values ​​and percent change from baseline were also included in the analysis. Weekly mean peak pruritus NRS and weekly mean average pruritus NRS using LOCF were plotted. For LOCF analysis, imputation was performed separately for each weekly mean or peak pruritus NRS score.

[0261] The Sleep Disorder NRS is a scale used by subjects to report the degree of sleep deprivation associated with AD. The scale has values ​​ranging from 0 to 10, with 0 being "no sleep deprivation associated with AD signs / symptoms" and 10 being "not able to sleep at all due to AD signs / symptoms." Subject-reported ratings of the Sleep Disorder NRS were collected in a daily diary. Weekly Sleep Disorder NRS scores were obtained based on the average daily values ​​for the 7 days immediately preceding each scheduled visit. At least four daily scores were required for this calculation, and if fewer than four daily scores were available, the respective weekly score was a missing value. Weekly mean Sleep Disorder NRS was descriptively summarized by visit as a continuous variable using LOCF and OC. Absolute values ​​and percent change from baseline were also included in the analysis. Absolute values ​​and percent change from baseline of weekly mean Sleep Disorder NRS using LOCF were illustrated. For LOCF analysis, imputation was performed for each weekly mean Sleep Disorder NRS score.

[0262] Application of topical medication for atopic dermatitis. Subject-reported assessments of the use of topical AD medication for eczema were collected daily in a diary. Subjects were instructed to use a licensed basal TCS and were asked to record in an electronic device (i.e., diary) their response (yes / no) to the following question throughout the clinical study (including trial and follow-up periods): "Did you apply the eczema medication given to you by your doctor to your skin today?". A day without topical AD medication was a day without the use of topical AD medication. For each subject, each day was classified as absent or absent. The number of days with the use of topical AD medication was calculated by summing the number of days recorded as yes in the diary. The value of one day in the visit interval was used to calculate the visit information. The number of days without topical AD medication in each visit interval was calculated as follows: days without topical AD medication in visit interval = days in interval - days with the use of topical AD medication in interval. The number of days without topical AD was summarized descriptively by visit interval as a continuous variable. No imputation was used.

[0263] The Dermatology Life Quality Index (DLQI) is a validated 10-item questionnaire for subjects over 16 years of age, covering domains including symptoms / emotions, daily activities, leisure, work / school, personal relationships, and treatment. DLQI data were recorded on a CRF. The pediatric DLQI (cDLQI) is an equivalent validated 10-item questionnaire designed for pediatric subjects, and the cDLQI was used when subjects were 16 years of age or younger. Each question in the DLQI / cDLQI has a value ranging from 0 (not at all) to 3 (very much), as defined in Table 10. The total score of the DLQI / cDLQI was calculated by summing the scores of each question, resulting in a maximum score of 30 and a minimum score of 0. The higher the score, the more impaired the QoL. DLQI and cDLQI scores and their changes from baseline were descriptively summarized by visit as continuous variables.

[0264] Table 1. Questionnaire scores for the Dermatology Quality of Life Index and Pediatric Dermatology Quality of Life Index TIFF2024535715000024.tif31168

[0265] Safety variables: Extent of exposure. Extent of exposure (duration of treatment, total dose administered, planned dose, and treatment compliance percentage) was summarized. Duration of treatment (days) was calculated as follows: [(date of last treatment-date of first treatment)+1]. Treatment compliance was summarized based on the treatment records recorded in the CRF and the medication administration log. Compliance percentage was calculated as follows: (administered dose) / (planned dose)×100, for a planned dose of 60 mg on day 1, followed by 30 mg injections every 4 weeks for 12 weeks. Once the dosing solution was prepared, 1.0 mL was drawn into a syringe and this was administered subcutaneously in the abdomen of the subject as the planned dose.

[0266] Adverse events. Adverse events were coded using the Medical Dictionary for Regulatory Activities (MedDRA) version 21.1. The AE summary included only treatment-emergent AEs (TEAEs), defined as AEs with an onset date / time on or after the date / time of first treatment. AEs with completely missing onset dates / times were considered TEAEs. If an AE's onset date / time was partially missing, the AE was considered a TEAE unless the non-missing portion of the date / time demonstrated onset before the date / time of first treatment.

[0267] Summarization by visit uses analysis visits. Unscheduled and early termination visits are windowed based on the analysis visit windows defined in SAP. Daily diary efficacy data (pruritus and sleep disturbance NRS) were categorized into analysis visits as follows, considering data from the 7 days immediately preceding each scheduled visit. Table 11 shows the weekly diary data windows:

[0268] (Table 11) Weekly diary data window TIFF2024535715000025.tif49168

[0269] Data on daily topical AD medication use were categorized by visit interval as follows (Table 12):

[0270] Table 12. Daily visit intervals for topical atopic dermatitis medication TIFF2024535715000026.tif43168

[0271] result Demographics and other baseline characteristics. As described in Table 13, a total of 31 subjects were screened, and 11 screenings failed due to either not meeting the inclusion / exclusion criteria or subject request. As a result, 20 subjects were enrolled. 18 subjects (90.0% of enrolled subjects) were treated at least once. Two subjects (8238-003 and 8664-001) who did not meet the inclusion criteria and failed screening were enrolled in error. No treatment was administered to any of these subjects. Of the 18 subjects who were treated at least once, 13 subjects (65.0% of enrolled subjects) received all treatment doses and were classified as completers, while 5 subjects (25.0% of enrolled subjects) discontinued treatment early. Premature discontinuation of treatment was due to AEs in 3 subjects (15.0% of enrolled subjects) and protocol deviations in 2 subjects (10.0% of enrolled subjects). Of the subjects who discontinued treatment early, two subjects (8203-005 and 8668-001) completed all study visits up to week 24, and three subjects (8614-001, 8614-002, and 8667-001) terminated the study before the final visit. Overall, 15 subjects (75% of subjects enrolled) completed the study (week 24 visit), whereas five subjects (25.0% of subjects enrolled), including one subject enrolled in error, discontinued the study early.

[0272] (Table 13) Characteristics of the subjects TIFF2024535715000027.tif203167

[0273] There were 12 (60.0%) subjects with at least one major protocol deviation. Most major protocol deviations were reported for study procedures / other (5 [25.0%]) and inclusion or exclusion criteria (4 [20.0%]). Major protocol deviations are summarized in Table 14.

[0274] Table 14. Major protocol deviations (intent-to-treat population) TIFF2024535715000028.tif95167

[0275] Among the ITT population, the mean age of all 20 subjects was 14.8 ± 1.59 years (min-max: 13-17 years), with weight ranging from 49 to 112 kg. Overall, 50% (N = 10) were in the 12-14 age group and 50% were in the 15-17 age group (N = 10). Among the ITT population, 60% of subjects were female. Nearly all subjects were Black or African American (50.0%) or White (35.0), and 80% of subjects were not Hispanic or Latino. Demographic and baseline characteristics of all 20 subjects are shown in Table 15.

[0276] Table 15. Demographic and baseline characteristics (intention-to-treat population) TIFF2024535715000029.tif191167

[0277] As presented in Table 16, at baseline, the mean (SD) baseline EASI score was 25.2 (7.22) for the ITT population. All subjects had moderate or severe IGA scores at baseline, with moderate scores reported for 15 (75%) subjects and severe scores reported for 5 (25%) subjects. The mean percent (SD) of BSA involvement at baseline was 41.3 (12.6). The mean (SD) of the weekly mean of PP NRS and weekly mean of AP NRS at baseline was 6.9 (1.66) and 6.2 (1.7), respectively. The mean (SD) weekly sleep disturbance NRS at baseline was 5.6 (2.27). The mean (SD) SCORAD score was 63.0 (11.34). The mean (SD) scores for DLQI and cDLQI at baseline were 11.8 (3.1) and 10.6 (5.42), respectively.

[0278] Table 16. Summary of baseline disease characteristics (intention-to-treat population) TIFF2024535715000030.tif226167TIFF2024535715000031.tif106167BSA=Body Surface Area, cDLQI=Pediatric Dermatology Life Quality Index, DLQI=Dermatology Life Quality Index, EASI=Eczema Area and Severity Index, IGA=Investigator Global Assessment, Max=Maximum, Min=Minimum, NRS=Numerical Rating Scale, SCORAD=Scoring Atopic Dermatitis Summary, SD=Standard Deviation Note: N = number of subjects in the population, n = number of subjects with available data. Baseline was the last available measurement before or during the first treatment. If the treatment date was missing, the enrollment date was used instead. Percentages (%) were calculated using the number of subjects in the population as the denominator.

[0279] Medical History. All 20 subjects in the ITT population had at least one medical condition recorded in their medical history (Table 14.1.3.2.1). Of these, 10 subjects (50.0%) had asthma. A history of cellulitis and anemia was reported by two subjects, and pneumonia and staphylococcal infection were reported by one subject each. One subject had a history of methicillin-susceptible Staphylococcus aureus (MSSA+), which had resolved before the subject was enrolled in the study. No prior or concomitant medical or surgical procedures were recorded. Nineteen (95%) subjects had used at least one prior medication, all of whom had used a dermatologic corticosteroid. The most commonly previously used dermatologic corticosteroids were triamcinolone (11 [55.0%]) and hydrocortisone (7 [35.0%]). Of the 19 (95%) subjects who used at least one basal TCS, all used mometasone furoate 0.1% cream, 18 (90.0%) subjects used hydrocortisone 1% cream, and 6 (30.0%) subjects used hydrocortisone butyrate 0.1% cream. One subject used at least one rescue medication, which was the systemic corticosteroids betamethasone and triamcinolone and a topical corticosteroid preparation (betamethasone valerate 0.1% lotion).

[0280] Pharmacokinetic results Observed concentrations and noncompartmental analysis. Maximum serum concentrations of nemolizumab (C max) were observed 1 or 2 weeks after the first subcutaneous injection of 60 mg LD (Figure 2 Error! Reference source not found). Maximum concentrations ranged from 2550 ng / mL to 11100 ng / mL with a mean value of 6532.9 ± 2329.9 ng / mL. Steady-state conditions were achieved by week 4 with mean observed trough concentrations ranging from 2935.3 ± 1029.4 ng / mL to 3292.3 ± 2017.8 ng / mL at steady state. Mean observed trough concentrations from weeks 4 to 16 are reported in Table 17 along with the corresponding model predicted values. At the end of the treatment period (week 16), the drug was cleared from serum. NCA was performed on the PK population. The mean (±SD) AUC values ​​calculated from predose samples observed at weeks 0-4, 8, 12, and 16 were 122.3±44.1 μg*day / mL, 233.3±80.1 μg*day / mL, 332.9±121.7 μg*day / mL, and 372.9±149.9 μg*day / mL, respectively. Due to the limited number of PK samples collected at the end of the treatment period, the terminal half-life cannot be accurately calculated from the NCA analysis.

[0281] Table 17. Observed and predicted mean (±SD) nemolizumab serum trough concentrations (ng / mL) in adolescents (PK population, N=18) TIFF2024535715000032.tif41167a. Observed values ​​from the PK population b. Concentrations predicted by the popPK model

[0282] Population Pharmacokinetic Analysis. Analyses were performed on the PK population including all subjects in the safety population who provided at least one post-baseline drug concentration value above the LLOQ (>= 100 ng / mL). Per protocol, at least six subjects with low body weight (i.e., ≤ 61 kg) were enrolled and included in the analysis.

[0283] The popPK model was constructed based on data from 407 adult subjects included in three clinical studies (CIM001JP, CIM003JG, and interim SPR.114322 data), which evaluated exposure to subcutaneous nemolizumab over dose ranges of 0.1-3 mg / kg and 10-90 mg for weight-based and flat dosing, respectively. The effects of several covariates, including age, weight, serum creatinine, estimated glomerular filtration rate, bilirubin, serum albumin, total protein, IgE, sex, and clinical study, were investigated.

[0284] The PK of nemolizumab was described by a one-compartment model with linear elimination and first-order absorption with lag time. A moderate effect of dose on bioavailability was identified, with bioavailability increasing with decreasing dose. Body weight was identified as the main source of variability in the PK profile of nemolizumab, affecting both CL / F and V / F. In fact, increasing body weight led to a decrease in nemolizumab systemic exposure. Albumin plasma concentration also influenced CL / F. Estimates of the main population PK parameters in adults are reported in Table 18.

[0285] No effect of age was observed in estimates of total body clearance, volume of distribution, absorption constant, or terminal half-life obtained with popPK modeling. All summary statistics in the adolescent Bayesian estimates are consistent with adult data either derived from the model (see Tables 17 and 18) or observed in the Phase 2 study, SPR.114322.

[0286] The mean (±SD) model-predicted AUC values ​​calculated at weeks 0 to 4, 8, 12, and 16 were 144.1±50.8 μg*day / mL, 284.4±97.1 μg*day / mL, 411.7±142.9 μg*day / mL, and 549.5±184.5 μg*day / mL, respectively.

[0287] Table 18. Summary of nemolizumab serum pharmacokinetic parameters estimated by popPK analysis (PK population, N=18) TIFF2024535715000033.tif61167NC: not calculated CL / F: apparent clearance, V / F: apparent volume of distribution, Ka: absorption constant, t1 / 2: terminal half-life, Cmax: peak concentration, AUCinf: area under the curve calculated to infinity

[0288] During popPK analysis, the adult popPK model was applied to predict nemolizumab concentration data in adolescents based on the subjects' baseline characteristics and dosing history. The predictions were then compared to the observed data (see Table 17, Figures 2 and 3), showing that the adult model was also able to accurately describe the individual nemolizumab concentration-time profiles in adolescents.

[0289] Pharmacokinetic / pharmacodynamic analysis Individual predictions generated from the popPK model and empirical Bayes inference were used to drive three PK / PD models developed using adult data for the following clinical endpoints: Eczema Area and Severity Index (EASI), Investigator's Global Assessment of Dermatitis Severity (IGA), and weekly mean PP-NRS.

[0290] The EASI model was constructed based on data from 525 subjects with AD from three clinical studies (Studies CIM001JP, CIM003JG, and SPR.114322). It is a turnover model with inhibitory drug effect (sum of placebo effect and nemolizumab effect modeled as Emax model) on the zero-order rate constant for the production of response (Rin), and a model combining proportional and additive residuals. The parameter representing the drug concentration producing half the maximum inhibitory effect (IC50) in the model was replaced with the parameter S0 calculated as Imax / IC50 to stabilize the model. There were no significant covariates that affected the model estimation.

[0291] The PP-NRS model was constructed based on data from 225 subjects with AD from study SPR.114322. It is a turnover model with an inhibitory drug effect (sum of placebo and nemolizumab effects modeled as an Emax model) on the zero-order rate constant for the production of response (Rin), and a model combining proportional and additive residuals. No significant covariates influenced the model estimation.

[0292] The IGA model was constructed based on data from 486 subjects with AD from two clinical studies (Studies CIM003JG and SPR114322). This is a continuous-time Markov model that treats IGA scores as compartments and models the ascending and descending transitions between compartments. A baseline IGA score of 4 or 5 was found to be a significant covariate for all ascending parameters, and therefore the covariate effect was implemented in the model.

[0293] A PK / PD model developed from adult data was applied to simulate the efficacy profile of nemolizumab in adolescents using subject characteristics (e.g., age, weight, body mass index) and dosing history. Predictions were then compared to observed data.

[0294] The EASI PK / PD model showed that the model structure was adequate to describe the adolescent data. The observed bias in predicting observed data to individual predicted scores for higher ESAI scores is explained by data sparseness (see Figure 4).

[0295] The PP-NRS PK / PD model demonstrated that the model structure was appropriate to describe the adolescent data (see Figure 5).

[0296] The IGA PK / PD model was adequate in its predictions and showed some discrepancy only for the aggregated score of 4 / 5, which may result from the lack of a score of 5 in adolescents (see Figure 6).

[0297] Overall, the PK / PD model was able to accurately describe the efficacy profile of nemolizumab in adolescents for the three clinical endpoints, thus supporting the similarity of exposure-response in adults and adolescents.

[0298] Immunogenicity results The overall incidence of ADA formation in the entire ITT population was limited, since only 1 in 20 patients presented treatment-related ADA at week 4. ADA-positive samples did not have neutralizing antibodies. Importantly, as can be observed in Figure 7, serum nemolizumab concentrations in ADA-positive subjects did not differ from serum nemolizumab concentrations in ADA-negative subjects.

[0299] Efficacy analysis Eczema Area and Severity Index (EASI). Using LOCF, the mean (SD) change from baseline in EASI at week 16 was -16.0 (9.9) and the percent change from baseline was -66.5% (32.5) (Table 19). Over the course of the study, EASI scores gradually improved up to week 16. Figure 8 shows a plot of the percent change in EASI scores over time.

[0300] Table 19. Summary of Eczema Area and Severity Index Scores (Last Observation Carried Forward) Intention-to-Treat Population TIFF2024535715000034.tif104167Note: N = number of subjects in the population, n = number of subjects with available data Max=maximum, Min=minimum, SD=standard deviation.

[0301] Investigator's Global Assessment. Improvements were observed from Week 4 to Week 16 (Table 20). At Week 16, 7 (35.0%) subjects achieved IGA success (defined as subjects with 0 [clear] or 1 [almost clear] and at least a 2-grade improvement from baseline data) using non-responder attribution. Over the course of the study, the number of subjects achieving IGA success improved up to Week 16. Figure 9 shows a plot of the percentage of subjects achieving Investigator's Global Assessment Success over time for the ITT population.

[0302] Table 20. Summary of Subjects Who Achieved Investigator Global Assessment of Success (Non-Responders) (Intention-to-Treat Population) TIFF2024535715000035.tif92167N=number of subjects in the population, n=number of subjects with available data, IGA=investigator's global assessment NOTE: Subjects with missing data or data collected after rescue medication intake were considered subjects who did not reach IGA success (non-responders). Percentages (%) were calculated using the number of subjects in the population as the denominator. Baseline was the last available measurement before or during the first treatment. If the treatment date was missing, the date of enrollment was to be used instead. IGA success was defined as subjects with a score of 0 (clear) or 1 (almost clear) and at least a 2-grade improvement from baseline data.

[0303] Body Surface Area (BSA). Using LOCF, the mean (SD) change from baseline in BSA of AD involvement at Week 16 was -19.9 (17.0) and the percent change from baseline was -53.3 (39.8) (Table 21). Over the course of the study, BSA of AD involvement improved up to Week 16.

[0304] Table 21. Summary of Body Surface Area Involved in Atopic Dermatitis (Last Observation Carried Forward) (Intention-to-Treat Population) TIFF2024535715000036.tif110167Note: N = number of subjects in the population, n = number of subjects with available data Max=maximum, Min=minimum, SD=standard deviation. AD=atopic dermatitis, LOCF=last observation carried forward. Baseline was the last available measurement before or during the first treatment. If the treatment date was missing, the enrollment date was used instead. Only missing post-baseline values ​​or values ​​after use of rescue medication were imputed using LOCF; baseline values ​​were not used for imputation.

[0305] Scoring Atopic Dermatitis (SCORAD) Summary. Using LOCF, the mean (SD) change from baseline in SCORAD score at week 16 was -31.2 (19.8) and the percent change from baseline was -50.1 (27.0) (Table 22). Over the course of the study, SCORAD scores improved up to week 16.

[0306] Table 22. Summary of Scoring Atopic Dermatitis Scores (Last Observation Carried Forward) (Intention-to-Treat Population) TIFF2024535715000037.tif110167Note: N = number of subjects in the population, n = number of subjects with available data Max=maximum, Min=minimum, SD=standard deviation, LOCF=last observation carried forward. Baseline was the last available measurement before or during the first treatment. If the treatment date was missing, the enrollment date was used instead. Only missing post-baseline values ​​or values ​​after use of rescue medication were imputed using LOCF; baseline values ​​were not used for imputation.

[0307] Peak pruritus numerical rating scale scores by week. Using LOCF, the mean (SD) change from baseline in the weekly mean of PP NRS at week 16 was -3.1 (2.8) and the percent change from baseline was -43.2 (37.0) (Table 23). Over the course of the study, the weekly mean of PP NRS improved up to week 16. Figure 10 shows a plot of the percent change from baseline in the weekly mean of PP NRS scores over time for the ITT population.

[0308] Table 23. Summary of Weekly Means of Peak Pruritus Numeric Rating Scale Scores (Last Observation Carried Forward) (Intention-to-Treat Population) TIFF2024535715000038.tif99167Note: N = number of subjects in the population, n = number of subjects with available data Max=maximum, Min=minimum, SD=standard deviation, LOCF=last observation carried forward. Baseline was the last available measurement before or during the first treatment. If the treatment date was missing, the enrollment date was used instead. The itch intensity scale ranges from 0 (no itch) to 10 (worst itch imaginable). Missing post-baseline values ​​or values ​​collected after use of rescue medication only were imputed using LOCF; baseline values ​​were not used for imputation.

[0309] Mean Pruritis Numeric Rating Scale Weekly Mean Score. Using LOCF, the mean (SD) change from baseline in the weekly mean of AP NRS at Week 16 was -2.6 (2.6) and the percent change from baseline was -40.9 (37.3) (Table 24). Over the course of the study, the weekly mean of AP NRS improved up to Week 16. Figure 11 shows a plot of the percent change from baseline in the weekly mean of AP NRS score over time for the ITT population.

[0310] Table 24. Summary of Weekly Mean Pruritus Numeric Rating Scale Scores (Last Observation Carried Forward) (Intention-to-Treat Population) TIFF2024535715000039.tif99167Note: N = number of subjects in the population, n = number of subjects with available data Max=maximum, Min=minimum, SD=standard deviation, LOCF=last observation carried forward. Baseline was the last available measurement before or during the first treatment. If the treatment date was missing, the enrollment date was used instead. The itch intensity scale ranges from 0 (no itch) to 10 (worst itch imaginable). Missing post-baseline values ​​or values ​​collected after use of rescue medication only were imputed using LOCF; baseline values ​​were not used for imputation.

[0311] Weekly mean sleep disturbance numeric rating scale scores. Using LOCF, the weekly mean sleep disturbance improved from baseline to week 16 with a mean (SD) change of 3.1 (3.2), a percent change from baseline of -53.5% (47.8) (Table 25). Figure 12 shows a plot of the percent change from baseline in the weekly mean sleep disturbance NRS scores over time for the ITT population.

[0312] Table 25. Summary of Weekly Mean Sleep Disorder Numeric Rating Scale Scores (Last Observation Carried Forward) (Intention-to-Treat Population) TIFF2024535715000040.tif99167Note: N = number of subjects in the population, n = number of subjects with available data Max=maximum, Min=minimum, SD=standard deviation, LOCF=last observation carried forward. Baseline was the last available measurement before or during the first treatment. If the treatment date was missing, the enrollment date was used instead. Missing post-baseline values ​​or values ​​collected after use of rescue medication only were imputed using LOCF; baseline values ​​were not used for imputation.

[0313] Number of days free of topical atopic dermatitis medication. The mean (SD) number of medication-free days improved from 3.5 (2.25) days at baseline to 10.7 (5.82) days at weeks 12-16, and further increased to 23.4 (11.88) days at weeks 16-24.

[0314] Dermatology Life Quality Index. Both DLQI and cDLQI mean (SD) scores showed improvement over the course of the study. DLQI improved from 11.8 (3.11) to 3.0 (2.58) at baseline and week 16, respectively. cDLQI improved from 10.6 (5.42) to 6.2 (5.29) at baseline and week 16, respectively.

[0315] Biomarker analysis Approach: Biomarker samples were available from 19 AD subjects aged 13-17 years treated with nemolizumab. For plasma analysis, blood samples were collected at baseline, 8, and 16 weeks after treatment. For SC analysis, lesioned and nonlesional samples were collected at baseline, whereas only lesioned samples were collected after 16 weeks of treatment. For biomarker evaluation, 30 proteins were measured in plasma and SC samples. These included CCL13, CCL2, CCL20, CCL22, CCL26, CCL27, CX3CL1, CXCL6, EGF, FAS L, galectin-1, IL-1RA, CXCL8 / IL-8, IL-11, IL-21, PLA2G7, CCL17 / TARC, CCL21, IL-1α, IL-2, oncostatin M, SP-D, VEGF-A, CCL5, cystatin C, CCL18, adiponectin, TGFb1, IL-31, and IgE (plasma only).

[0316] IL-6 and IL-18 analysis was not performed in plasma or SC samples because the above 30 biomarkers were considered more relevant to AD, but the two cytokines are more considered as markers of general inflammation. Furthermore, preliminary analysis from a previous study in atopic dermatitis (SPR114322, phase 2b) revealed that these cytokines do not predict responsiveness. Finally, due to the limited amount of SC samples, only protein biomarker analysis could be performed, and EOS ceramide, or mRNA and miRNA analysis could not be performed.

[0317] Clinical parameters - EASI-75, IGA, and PP-NRS at week 16 were considered as the primary outcomes of the biomarker substudy. Clinical responders using these parameters were defined as follows: ● EASI-75 Responder: At least 75% reduction in EASI score from baseline. ●IGA-01 responders: IGA≦1. ● PP-NRS Responder: At least a 4-point reduction in Peak Pruritus Maximum Itch NRS from baseline.

[0318] Stratum corneum: Supervised analysis - mixed model repeated measures (MMRM). Six proteins significantly associated with EASI-75 outcome: CCL18, CCL20, CCL22, CCL27, IL1RA with false discovery rate (FDR) < 0.05, and VEGF with FDR < 0.10 (Figure 13), the FDR considered is the minimum adjusted p-value (interaction, main effect). Although not significant with respect to the FDR criterion, CCL17 and IL31 are also presented due to the particular interest in these proteins (Figure 13). Nevertheless, the expression of CCL17 was notably different between lesional and non-lesional skin in EASI75 responders and was simultaneously reduced after treatment in lesional skin at week 16. The fold change between lesional and non-lesional samples at baseline was 1.9-3.5 higher in responders than in non-responders for four of the six proteins, namely CCL20, CCL22, CCL27, and VEGF. This difference is statistically significant (shown in FIG. 13). These data suggest that these four proteins can be used as potential biomarkers of clinical response (FIG. 14). Changes in the measured protein biomarkers were not found to be associated with IGA and PP-NRS at week 16. Since only 10 subjects were documented at week 16, PP-NRS was also considered at week 12, with 14 subjects documented. Significant associations with CCL17 and CCL20 were observed.

[0319] Unsupervised analysis, non-negative tensor factorization. Unsupervised analysis was performed on 15 subjects with at least one documented outcome (EASI-75, IGA, PP-NRS) and proteomic profile in at least one condition (non-lesional at baseline, lesion at baseline, or lesion at week 16). The association between subject ranking provided by non-negative tensor factorization (NTF) and EASI-75 was significant (p=0.0224). Fifteen proteins (including 6 proteins found by supervised analysis) contributed most to the achieved ranking: CCL18, CCL20, CCL22, CCL27, CX3CL1, CXCL6, CXCL8, cystatin, FASL, galectin, IL11, IL21, IL1RA, SPD, VEGF (Error! Reference source not found). When the analysis was restricted to these 15 proteins, the association between subject ranking provided by NTF and EASI-75 was even more significant (p=0.0041). As in the supervised analysis, IGA-01 score was not associated with subject ranking (p=0.5493). The association p-value with PP-NRS could not be calculated due to unstable estimation caused by the small sample size (only 10 subjects were documented). When using PP-NRS at week 12, the association remained non-significant. The results of the analysis are shown in the table below.

[0320] TIFF2024535715000041.tif120167

[0321] Plasma: Neither supervised nor unsupervised analyses showed significant correlations between measured proteins and clinical scores.

[0322] Safety evaluation results. Extent of Exposure. The mean duration of study treatment was 68.7 days, with a median of 85.0 days. The mean total dose planned and administered during the study was 131.7 mg (100% compliance). A summary of exposure to study drug can be found in Table 26.

[0323] Table 26. Summary of Exposure Levels (Safety Population) TIFF2024535715000042.tif185167N=number of subjects in the population, n=number of subjects with available data, SD=standard deviation. The study duration was calculated as the date of the last visit minus the date of the first visit plus one.

[0324] Adverse Events. A total of 6 (33.3%) subjects experienced 14 TEAEs. Of these subjects, 3 (16.7%) were reported to have 10 TEAEs related to the study drug. The majority of reported TEAEs by maximum severity were either mild (6 events in 3 [16.7%] subjects) or moderate (4 events in 1 [5.6%] subject). Two (11.1%) subjects had 4 TEAEs that were considered severe. Two of these TEAEs were serious (i.e., infectious eczematous dermatitis with staphylococcal skin infection and right leg edema). The four treatment-emergent AESIs reported in this study were experienced by the same two subjects. Each of these two subjects (8667-001 and 8668-001) had one serious AESI (infectious eczematous dermatitis with staphylococcal skin infection and right leg edema, respectively) and one non-serious AESI (bilateral leg edema and bilateral distal leg edema, respectively). All AESIs were considered study drug related. These same two subjects also had seven related AEs that led to permanent discontinuation of study drug (subject 8667-001 [atopic dermatitis, bilateral leg edema, bilateral leg pain, and pustular rash] and subject 8668-001 [bilateral leg edema, right leg edema, and staphylococcal skin infection]). No TEAES led to discontinuation of the study.

[0325] As shown in Table 27, 6 of 18 subjects (33.3%) experienced at least one TEAE. One subject had two SAEs and one subject had one SAE for a total of three SAEs. The same two subjects had three TEAEs rated as severe. Seven TEAEs reported for these two subjects led to permanent discontinuation of study drug. Two subjects reported four AESIs. No deaths were reported during the study.

[0326] Table 27. Overall summary of treatment-emergent adverse events, all causalities (safety population) TIFF2024535715000043.tif82167AESI=Adverse Events of Special Interest, TEAE=Treatment-Emergent Adverse Events

[0327] As shown in Table 28, the system organ class (SOC) with the most reported TEAES was infections and infestations (5 events in 4 subjects). With the exception of peripheral edema, which was experienced by 2 subjects, all other AEs were experienced by 1 subject each.

[0328] Table 28. Treatment-emergent adverse events by system organ class, preferred term, and all causalities (safety population) TIFF2024535715000044.tif168167

[0329] Adverse events related to the study drug. Three subjects experienced 10 TEAEs that were deemed related to the study drug by the investigator. Serious TEAEs, severe TEAEs, TEAEs of special interest, and TEAEs leading to permanent discontinuation of study drug considered related to the study drug were each experienced by two (11.1%) subjects. No subjects experienced TEAEs considered related to study procedures.

[0330] Adverse events by severity. The majority of reported TEAEs by maximum severity were mild (six events in three [16.7%] subjects), severe (four events in two [11.1%] subjects), or moderate (four events in one [5.6%] subject). Among the four moderate events, the most frequently reported TEAE was peripheral edema (two events in one [5.6%] subject) in the SOC for general disorders and administration site conditions. Four severe events (peripheral edema, eczema infection, staphylococcal skin infection, and atopic dermatitis) were reported in two subjects. One subject (8667-001) experienced eczema infection and atopic dermatitis in the Infections and Infestations and Skin and Subcutaneous Skin Disorders SOCs, respectively, and the other subject (8668-001) experienced staphylococcal skin infection and peripheral edema in the Infections and Infestations and Systemic Disorders and Administration Site Conditions SOCs, respectively.

[0331] Deaths, other serious AEs, and other significant AEs. No deaths occurred during the study. Two subjects experienced three SAEs (eczema infection, peripheral edema, and staphylococcus skin infection), all considered drug-related. A summary of serious TEAEs is shown in Table 29.

[0332] Table 29. Serious treatment-emergent adverse events of special interest by organ system class and preferred term (safety population) TIFF2024535715000045.tif69167N = number of subjects in the population, n = number of subjects with available data. Note: Percentages (%) are calculated using the number of subjects in the population as the denominator. Adverse events are coded using MedDRA version 21.1. Treatment-emergent adverse events (TEAEs) are defined as events with an onset date on or after the date of study drug administration.

[0333] Adverse Events of Particular Interest. The four treatment-emergent AESIs reported in this study were experienced by the same two subjects. Each of these two subjects (8667-001 and 8668-001) had one serious AESI (infectious eczematous dermatitis with staphylococcal skin infection and right leg edema, respectively) and one non-serious AESI (bilateral leg edema and bilateral distal leg edema, respectively). All AESIs were considered to be study drug-related. A summary of TEAEs of particular interest is provided in Table 30.

[0334] Table 30. Treatment-emergent adverse events of special interest by system organ class and preferred term, all causalities (safety population) TIFF2024535715000046.tif69167N = number of subjects in the population, n = number of subjects with available data. Note: Percentages (%) are calculated using the number of subjects in the population as the denominator. Adverse events are coded using MedDRA version 21.1. Treatment-emergent adverse events (TEAEs) are defined as events with an onset date on or after the date of study drug administration.

[0335] Treatment-emergent adverse events leading to discontinuation. The seven TEAEs leading to permanent study drug discontinuation were experienced by the same two subjects as the SAEs and AESIs (subject 8667 001 [atopic dermatitis, bilateral leg edema, bilateral leg pain, and pustular rash] and subject 8668 001 [bilateral leg edema, right leg edema, and staphylococcal skin infection]). No TEAEs led to subject discontinuation from the study.

[0336] Clinical laboratory evaluation results: Hematology. Small changes from baseline in hematology parameters were observed during the study. No significant trends in shifts from baseline among hematology laboratory values ​​were observed. For hematology values ​​recorded after baseline, nearly all that were normal at baseline remained normal at the post-baseline time points.

[0337] Chemistry. Small changes from baseline in chemistry parameters were observed during the study. No significant trends in shifts from baseline among chemistry laboratory values ​​were observed. For chemistry values ​​recorded after baseline, nearly all that were normal at baseline remained normal at the post-baseline time points.

[0338] Urinalysis. No PCS urinalysis laboratory results were observed.

[0339] Vital signs, physical examination, and other observations relevant to safety: Vital Signs. Five subjects had potentially clinically significant vital signs during the study. One subject had a PCS diastolic blood pressure value at week 12 (>90 mmHg, increase of >10 mmHg from baseline), body temperature at weeks 1-2 (<35°C), and weight change at weeks 16 and 24 (<95% change from baseline). Two other subjects had PCS changes in weight (>95% change from baseline) from weeks 8-24 and beginning at week 12, respectively. One subject had a PCS change in weight (>105% change from baseline) at week 8, and four subjects had PCS changes in weight (>105% change from baseline) at weeks 12, 16, and 24, respectively. No significant trends were observed in vital sign values.

[0340] Electrocardiogram Data. No significant trends were observed in electrocardiogram parameters, and no clinically significant abnormalities were reported at screening or week 16.

[0341] Physical Examination Findings. Clinically significant skin abnormalities (atopic dermatitis) were reported in one subject at week 4, seven subjects at week 8, and one subject at week 16. No significant trends were observed in physical examination values.

[0342] Respiratory evaluation results. One subject (8614-001) with a history of asthma had an abnormal PEF (77.6% of predicted) at screening, which was later identified as a major protocol deviation and the site was instructed to immediately discontinue administration of any study drug. At screening, the subject's ACT score was normal (25). The subject returned for all subsequent safety procedures as required by the sponsor. PEF values ​​remained abnormal until the early termination visit, at which point the PEF was 104.8% of predicted. A summary of the abnormal PEF is shown in Table 31.

[0343] Table 31. Summary of abnormal peak expiratory flows (safety population) TIFF2024535715000047.tif71167PEF=Peak expiratory flow

[0344] One subject (8668-001) had an abnormal Asthma Control Test (ACT) score of 13 at screening, which was later identified as a major protocol deviation. An ACT score ≦19 suggests that asthma may not be adequately controlled. However, the baseline loading dose was administered in error. The subject's PEF was normal at screening (87.7% of predicted). The subject returned for all subsequent safety procedures as required by the sponsor. ACT scores were normal at all visits except for Week 12, where the ACT score was 15. ACT scores were normal at all time points for all other subjects. No notable trends were observed in respiratory values. A summary of ACT scores ≦19 is shown in Table 32.

[0345] Table 32. Summary of Asthma Control Test Score ≦19 (Safety Population) TIFF2024535715000048.tif106167ACT=Asthma Control Test

[0346] Safety Conclusions: Six (33.3%) subjects experienced 14 TEAEs, most of which were mild or moderate in maximum severity. Two subjects experienced three SAEs (eczema infection, peripheral edema, and staphylococcal skin infection), all considered drug-related. The same two subjects who experienced SAEs experienced four AESIs, with each subject having one serious and one non-serious AESI. Seven TEAEs led to permanent study drug discontinuation, experienced by the same two subjects as SAEs and AESIs. No TEAEs led to study discontinuation. No deaths occurred during the study. No notable trends were observed between laboratory, vital sign, and respiratory assessments over time.

Claims

1. A medicament comprising an anti-IL-31RA antibody for use in a method of treating or preventing atopic dermatitis (AD) in a subject, wherein the method comprises subcutaneously administering an anti-IL-31RA antibody to a subject having AD, wherein the subject has skin lesions in which the expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is upregulated compared to non-lesional skin of an individual without AD or non-lesional skin of the subject, and wherein administration of the anti-IL-31RA antibody reduces the expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in the skin lesions of the subject, said medicament.

2. A medicament comprising an anti-IL-31RA antibody for use in a method of altering or reducing the expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in skin lesions of a subject having atopic dermatitis (AD), wherein the method comprises subcutaneously administering an anti-IL-31RA antibody to a subject having AD, wherein at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 is upregulated in the skin lesions of the subject compared to non-lesional skin of an individual without AD or non-lesional skin of the subject, wherein administration of the anti-IL-31RA antibody reduces said expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in the skin lesions, said medicament.

3. The medicament according to claim 1 or 2, wherein the expression of CCL20, CCL22, CCL27, VEGF, IL1RA, and / or CCL18 is determined by RT-qPCR, RT-PCR, RNA-seq, Northern blotting, serial analysis of gene expression (SAGE), DNA or RNA microarray, Western blotting, ELISA, surface plasmon resonance, or mass spectrometry.

4. The medicament according to claim 1 or 2, wherein 1, 2, 3, 4, 5, or 6 of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 are upregulated in the skin lesions of the subject compared to non-lesional skin of an individual without AD or non-lesional skin of the subject.

5. The expression of at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 changes or decreases within 2 weeks, 4 weeks, 8 weeks, 10 weeks, 12 weeks, 14 weeks, 16 weeks, 18 weeks, or 20 weeks from the administration of the anti-IL-31RA antibody as compared to the baseline expression level in the skin lesions of the subject before the administration of the anti-IL-31RA antibody. The pharmaceutical according to claim 1 or 2.

6. The expression of CCL20 is reduced by at least about one-third, the expression of CCL22 is reduced by at least about one-eleventh, the expression of CCL27 is reduced by at least about one-third, the expression of VEGF is reduced by at least about one-sixteenth, the expression of CCL18 is reduced by at least about one-eighth, and / or the expression of IL1RA is increased by at least about 1.1-fold. The pharmaceutical according to claim 1 or 2.

7. A pharmaceutical comprising an anti-IL-31RA antibody for use in a method of reducing the expression of inflammatory biomarkers in the skin of a subject having atopic dermatitis (AD), The method comprising administering an anti-IL-31RA antibody to a subject having AD, thereby reducing the inflammatory response in the skin, and The inflammatory biomarker is selected from at least one of CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18. The pharmaceutical.

8. The expression of 1, 2, 3, or all 4 of CCL20, CCL22, CCL27, and VEGF is reduced. The pharmaceutical according to claim 7.

9. The subject achieves a reduction of at least 66.5% in the eczema area and severity index (EASI) scoring after administration of the anti-IL-31RA antibody. The pharmaceutical according to any one of claims 1, 2, and 7.

10. The subject achieves a reduction of at least 43.2% in the peak pruritus numerical rating scale (PP-NRS) scoring after administration of the anti-IL-31RA antibody. The pharmaceutical according to any one of claims 1, 2, and 7.

11. The subject experiences improved sleep after administration of the anti-IL-31RA antibody. The pharmaceutical according to any one of claims 1, 2, and 7.

12. The subject is an adult. The pharmaceutical according to any one of claims 1, 2, and 7.

13. The pharmaceutical according to any one of claims 1, 2, and 7, wherein in the method, the anti-IL-31RA antibody is administered once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, or once every eight weeks.

14. The pharmaceutical according to any one of claims 1, 2, and 7, wherein in the method, the anti-IL-31RA antibody is administered at a dose of about 0.01 mg / kg to about 0.1 mg / kg, about 0.1 mg / kg to about 0.5 mg / kg, about 0.5 mg / kg to about 1.5 mg / kg, about 1.5 mg / kg to about 2.5 mg / kg, or about 2.5 mg / kg to about 10 mg / kg.

15. The pharmaceutical according to any one of claims 1, 2, and 7, wherein in the method, the anti-IL-31RA antibody is administered at a dose of about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg.

16. The pharmaceutical according to claim 15, wherein in the method, the anti-IL-31RA antibody is administered at a dose of 30 mg or 60 mg.

17. The pharmaceutical according to any one of claims 1, 2, and 7, wherein the anti-IL-31RA antibody comprises a heavy chain variable region comprising an HCDR1 comprising SEQ ID NO: 8, an HCDR2 comprising SEQ ID NO: 9, and an HCDR3 comprising SEQ ID NO: 10, and a light chain variable region comprising an LCDR1 comprising SEQ ID NO: 12, an LCDR2 comprising SEQ ID NO: 13, and an LCDR3 comprising SEQ ID NO:

14.

18. The pharmaceutical according to any one of claims 1, 2, and 7, wherein the anti-IL-31RA antibody is nemolizumab or a fragment or variant thereof.

19. The pharmaceutical according to any one of claims 1, 2, and 7, wherein in the method, the anti-IL-31RA antibody is administered subcutaneously at a loading dose of 60 mg, followed by a dose of 30 mg every four weeks for at least 12, 14, 16, or 18 weeks.

20. The pharmaceutical according to any one of claims 1, 2, and 7, wherein in the method, the anti-IL-31RA antibody is administered according to a flat dosing regimen.

21. The pharmaceutical according to any one of claims 1, 2 and 7, wherein in the method, the anti-IL-31RA antibody is administered according to a loading dose regimen.

22. A method of testing for suspected atopic dermatitis (AD), comprising: detecting the expression level of at least 1, at least 2, at least 3, at least 4, at least 5, or all 6 biomarkers selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a sample obtained from a subject suspected of having AD; comparing the expression level of the biomarker with a reference level; wherein the reference level is the expression level of each corresponding biomarker in a skin sample from an individual without AD or a sample of non-lesional skin from the subject. Method.

23. A method of testing whether a subject having atopic dermatitis (AD) responds to treatment with an anti-IL-31RA antibody, comprising: detecting the expression level of at least 1, at least 2, at least 3, at least 4, at least 5, or all 6 biomarkers selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a sample obtained from a subject suspected of having AD; comparing the expression level of the biomarker with a reference level; wherein if the expression level of the biomarker is higher than the reference level, the subject responds to treatment, and the reference level is the expression level of each corresponding biomarker in a skin sample from an individual without AD or a sample of non-lesional skin from the subject. Method.

24. A method of testing whether a subject having atopic dermatitis (AD) responds to treatment with an anti-IL-31RA antibody, comprising: detecting the expression level of at least 1, at least 2, at least 3, at least 4, at least 5, or all 6 biomarkers selected from CCL20, CCL22, CCL27, VEGF, IL1RA, and CCL18 in a post-treatment sample obtained from a subject having AD who has received at least one dose of an anti-IL-31RA antibody; comparing the expression level of the biomarker with a baseline expression level from a sample obtained from the same subject before treatment was initiated; wherein A decrease in the expression level of CCL20, CCL22, CCL27, VEGF, and / or CCL18 and / or an increase in the expression level of IL1RA indicating that the subject responds to treatment Method.

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