Anti-interleukin 36 receptor (IL-36R) therapy for ichthyosis

Abstract

This invention provides a method for treating ichthyosis in the subject. [Solution] A method for treating ichthyosis in a subject, comprising inhibiting IL-36 signaling in the subject, thereby treating the ichthyosis.

Description

[Technical Field]

[0001] Cross-reference of related applications This patent application claims the benefit of U.S. Provisional Patent Application No. 63 / 058,938, filed July 30, 2020, which is incorporated herein by reference in its entirety.

[0002] References to electronically submitted literature The computer-readable nucleotide / amino acid sequence listing submitted concurrently with this specification and identified below is incorporated herein by reference in its entirety: a 76,939-byte ASCII (text) file named "754349_ST25.TXT" created on 26 July 2021. [Background technology]

[0003] Ichthyosis is a family of rare, lifelong genetic disorders that include at least 20 different subtypes. More than 50 gene mutations have been reported to cause ichthyosis, affecting numerous cellular functions, including DNA repair, lipid biosynthesis, adhesion, and desquamation, as well as other pathways. (Marukian, F1000 Research, 5(F1000) Faculty Rev): 1497 (2016). However, all forms of ichthyosis share the characteristic features of systemic or localized scaling, erythema, and underlying skin inflammation. These features are thought to be a compensatory response to the poor epidermal barrier exhibited by all forms of ichthyosis, as evidenced by increased transepidermal water loss (TEWL). Adults and children with ichthyosis may experience highly noticeable and disfiguring skin changes, itching, and functional limitations associated with thickened skin, ultimately leading to a decline in quality of life.

[0004] Currently, there is no known treatment for ichthyosis. Therapies have been limited to ointments and preparations such as keratolytics and retinoids that peel off the thickened scales caused by the disorder. Oral retinoids are considered an effective treatment method for scale removal, but they tend to increase skin inflammation associated with ichthyosis and carry potential side effects. Furthermore, oral retinoids are not etiology-based therapies (i.e., pathway-specific therapies) that generally provide therapeutic, safety, and cost-effective benefits.

[0005] There remains a significant unmet medical need for effective treatments for ichthyosis. [Overview of the project]

[0006] In embodiments, the present invention provides a method for treating ichthyosis in a subject by inhibiting interleukin-36 (IL-36) signaling in the subject, thereby treating ichthyosis.

[0007] Additional embodiments provide a method for selecting subjects having ichthyosis for treatment with an IL-36 pathway inhibitor, which may optionally be used in conjunction with a method for treating ichthyosis in subjects provided herein. In one embodiment, the method includes comparing the expression of at least one of IL-36 cytokine, IL-36R, or the mRNA encoding it in a skin sample from a subject before and after inhibiting IL-36 signaling in the subject, and selecting the subject for treatment if a decrease in the expression of at least one of IL-36 cytokine, IL-36R, or the mRNA encoding it is observed in a skin sample from a subject after inhibiting IL-36 signaling compared to a sample from the subject before inhibiting IL-36 signaling.

[0008] In further embodiments, the present invention provides inhibitors of the IL-36 pathway, such as IL-36 receptor (IL-36R) conjugates and compositions comprising the same, for use in the methods of the present invention. These and other embodiments of the present invention will become apparent to those skilled in the art who have read the embodiments for carrying out the invention. [Brief explanation of the drawing]

[0009] [Figure 1A] Figure 1A shows two bar graphs comparing IL-36α gene expression in skin biopsies from patients with ichthyosis (i.e., Netherton syndrome, phyllodes ichthyosis, congenital ichthyoid erythroderma, epidermolytic ichthyosis, and ichthyosis with confetti) with that of a control. [Figure 1B] Figure 1B shows two bar graphs comparing IL-36β gene expression in skin biopsies from patients with ichthyosis (i.e., Netherton syndrome, phyllodes ichthyosis, congenital ichthyoid erythroderma, epidermolytic ichthyosis, and confetti ichthyosis) with that of a control. [Figure 1C] Figure 1C shows two bar graphs comparing IL-36γ gene expression in skin biopsies from patients with ichthyosis (i.e., Netherton syndrome, phyllodes ichthyosis, congenital ichthyoid erythroderma, epidermolytic ichthyosis, and confetti ichthyosis) with that of a control. [Figure 1D] Figure 1D shows two bar graphs comparing IL-RN gene expression in skin biopsies from patients with ichthyosis (i.e., Netherton syndrome, phyllodes ichthyosis, congenital ichthyoid erythroderma, epidermolytic ichthyosis, and confetti ichthyosis) with that of a control. [Figure 2A] Figure 2A shows IL-36R protein expression in skin biopsies from ichthyosis patients, as determined by immunohistochemical testing (IHC). [Figure 2B] Figure 2B shows IL-36R protein expression in skin biopsies of control patients (i.e., patients without ichthyosis) using IHC. [Modes for carrying out the invention]

[0010] Provided herein is a method of treating ichthyosis in a subject, comprising inhibiting IL-36 signaling in the subject, thereby treating ichthyosis. Inhibition of IL-36 signaling can be achieved by any suitable method, such as preventing binding of IL-36 receptor (IL-36R) to one or more (or all) IL-36 cytokines (e.g., IL-36α, IL-36β, and / or IL-36γ).

[0011] The IL-36 cytokines IL-36α, IL-36β, and IL-36γ (formerly named IL-1F6, IL-1F8, and IL-1F9) are members of the interleukin-1 (IL-1) family that bind to IL-36R (formerly named IL-1Rrp2 or IL-1RL2), a receptor of the IL-1R family, and use IL-1 receptor accessory protein (IL-1RAcP) as a co-receptor to stimulate intracellular signals similar to those induced by IL-1 (Towne et al., J. Biol. Chem., 279(14):13677-13688 (2004)). IL-1F5 is a member of the IL-1 family that has been shown to act as an antagonist of IL-36R, and is currently referred to as IL-36Ra (Dinarello et al., Nat. Immunol., 11(11):973 (2010)).

[0012] The ichthyosis described in the methods of embodiments may be any subtype of ichthyosis, regardless of genotype or phenotype. In some embodiments, ichthyosis may be associated with Th17 activation in the skin and / or consequent induction of IL-17-associated genes or markers synergistically induced by IL-17 and TNF-α. In some embodiments, ichthyosis is associated with upregulation of IL-36γ compared to a normal subject. In some embodiments, ichthyosis is associated with compared to a normal subject or may be associated with an increase in CLA+ (skin-homing) T cells in the IL-17+, IL-22+, and IL-9+ subsets compared to a subject having psoriasis but not ichthyosis. In some embodiments, the ichthyosis is resistant or non-responsive to oral and / or topical retinoids.

[0013] In some embodiments, the ichthyosis is non-syndromic or syndromic ichthyosis. In additional embodiments, the ichthyosis is a common form of ichthyosis (e.g., ichthyosis vulgaris). The ichthyosis may also be an orphan (i.e., rare) form of ichthyosis, including but not limited to congenital ichthyosiform erythroderma, lamellar ichthyosis, epidermolytic ichthyosis, Netherton syndrome, and ichthyosis en confetti.

[0014] The subject may be any subject in need of treatment. In some embodiments, the subject has an IASI score of at least 12 and / or an erythema score of at least 2. IASI quantifies the severity of ichthyosis in a subject based on the severity of erythema or scaling and the percentage of affected BSA, and is a composite score ranging from 0 to 48 that takes into account the degree of erythema and scaling for each of four body regions (each individually scored from 0 to 4), adjusted for the percentage of involved BSA per body region and the ratio of the body region to the whole body (see Paller et al., J Allergy Clin Immunol., 139(1):152-165 (2017)). The subject may be a mammal such as a human or a non-human primate.

[0015] As used herein, terms such as “treat” and “treat” refer to obtaining a desired pharmacological and / or physiological effect. Preferably, the effect is therapeutic, i.e., the treatment reduces the severity of one or more adverse symptoms of ichthyosis, including but not limited to descaling, inhibition of erythema and itching, and reduction of inflammation. The reduction of adverse symptoms may be determined by any preferred technique, such as a reduction in TEWL or a reduction in the clinical severity of ichthyosis as defined by the Ichthyosis Area Severity Index (IASI) described in Paller et al., J Allergy Clin Immunol., 139(1):152-165 (2017) and the Visual Index for Ichthyosis Severity (VISI) described in Marukian et al., J Invest Dermatol, 137:1834-1841 (2017).

[0016] For this purpose, the method of the present invention involves administering a "therapeutably effective amount" of an IL-36 pathway inhibitor described herein, such as an IL-36R conjugate, to the target.

[0017] "Therapeutically effective dose" refers to the amount effective in terms of dosage and duration required to achieve the desired pharmacological and / or physiological effect. The therapeutically effective dose may vary depending on factors such as the disease state, the individual's age, sex, and weight, as well as the ability of the IL-36 pathway inhibitor, such as an IL-36R conjugate, to elicit the desired response in the individual. For example, the therapeutically effective dose of an IL-36 pathway inhibitor is the amount that reduces the biological activity of either the IL-36 cytokine and / or IL-36R, and consequently, the therapeutically effective dose of the IL-36R conjugate described herein is the amount that reduces the biological activity of IL-36R in the subject.

[0018] In some embodiments, the pharmacological and / or physiological effects may be prophylactic, i.e., the effect may completely or partially prevent ichthyosis or its symptoms. In this regard, the method of the present invention involves administering an “anticipatory effective amount” of an IL-36 pathway inhibitor. “An anticipatory effective amount” means an amount that is effective in the dose and duration required to achieve the desired prophylactic outcome (e.g., prevention of the onset of ichthyosis). In some embodiments, the subjects may be subjects with a genetic predisposition to ichthyosis (e.g., a family history or genetic profile of ichthyosis), with or without any clinical symptoms.

[0019] Any suitable dose of IL-36 inhibitor may be used. In some embodiments, the dose is in the range of 1 pg / kg (animal or human body weight) to 20 mg / kg, but doses lower or higher than this exemplary range are also within the scope of the present invention. The parenteral daily dose is approximately 0.00001 μg / kg to 20 mg / kg (total body weight) (for example, approximately 0.001 μg / kg, approximately 0.1 μg / kg, approximately 1 μg / kg, approximately 5 μg / kg, approximately 10 μg / kg, approximately 100 μg / kg, approximately 500 μg / kg, approximately 1 mg / kg, approximately 5 mg / kg, approximately 10 mg / kg, or a range defined by any two of the above values), preferably approximately 0.1 μg / kg to 10 mg / kg (total body weight) (for example, approximately 0.5 μg / kg, approximately 1 μg / kg, approximately 50 μg / kg, approximately 150 μg / kg, approximately 300 μg / kg, approximately 750 μg / kg, approximately 1.5 mg / kg, approximately 5 mg / kg, or the above values). The range may be defined by any two of the above values, more preferably about 1 μg / kg to 5 mg / kg (total body weight) (for example, about 3 μg / kg, about 15 μg / kg, about 75 μg / kg, about 300 μg / kg, about 900 μg / kg, about 2 mg / kg, about 4 mg / kg, or the range defined by any two of the above values), and even more preferably about 0.5 to 15 mg / kg (body weight) / day (for example, about 1 mg / kg, about 2.5 mg / kg, about 3 mg / kg, about 6 mg / kg, about 9 mg / kg, about 11 mg / kg, about 13 mg / kg, or the range defined by any two of the above values).

[0020] In some embodiments, the dose is approximately 20 mg or more (e.g., approximately 30 mg or more, approximately 50 mg or more, approximately 75 mg or more, or approximately 100 mg or more) and approximately 1000 mg or less (e.g., approximately 900 mg or less, approximately 800 mg or less, approximately 700 mg or less, approximately 600 mg or less, approximately 500 mg or less, approximately 400 mg or less, or approximately 300 mg or less) every 1 to 6 weeks (e.g., weekly, every 2 weeks, every 3 weeks, or every 4 weeks). In some embodiments, the dose is approximately 150 to 250 mg (e.g., approximately 200 mg). In some embodiments, the dose is approximately. In some embodiments, the IL-36 inhibitor is administered as a single loading dose of approximately 1.5 to 10 times (e.g., approximately 2 to 8 times) the maintenance dose below. Therefore, for example, an IL-36 pathway inhibitor may be administered as a loading dose of approximately 200 mg to 750 mg, or approximately 300 to 500 mg, or even approximately 350 to 450 mg (e.g., approximately 200 mg, approximately 250 mg, approximately 300 mg, approximately 350 mg, approximately 400 mg, approximately 450 mg, approximately 500 mg, approximately 550 mg, approximately 600 mg, approximately 650 mg, approximately 700 mg, or approximately 750 mg), followed by a maintenance dose of approximately 50 to 250 mg, or approximately 100 to 250 mg, or even approximately 150 to 250 mg (e.g., approximately 50 mg, approximately 100 mg, approximately 150 mg, approximately 200 mg, approximately 250 mg) every two weeks or every one or two months (e.g., every two to eight weeks or every two to four weeks), if necessary for the onset or maintenance of a therapeutic response.

[0021] In some embodiments, the IL-36R conjugate provides a long-lasting effect against ichthyosis and can be administered relatively infrequently, for example, on a schedule of once every 14, 21, or 30 days or less. In some embodiments, even longer intervals may be used (for example, once every 45, 60, 90, or 120 days or less). Thus, for example, the aforementioned doses described herein may be administered once every 14 days, once every 21 days, once every 30 days, once every 45 days, once every 60 days, once every 90 days, or even once every 120 days. When a maintenance dose is used following a loading dose, the loading dose may be the first dose administered, and the loading dose is administered at the following intervals: In the embodiment, the IL-36R binder is administered as a loading dose (e.g., 200 mg to 750 mg, e.g., about 350 to 450 mg, or even 400 mg), followed by a maintenance dose (e.g., about 100 to 250 mg, or even about 150 to 250 mg, e.g., about 200 mg) at least once every two weeks (e.g., at least once every 21 or 30 days).

[0022] The effectiveness of treatment or prevention is monitored by regularly evaluating the treated patient. It can be administered repeatedly over several days or longer, and depending on the condition, treatment may be repeated until the symptoms of ichthyosis are suppressed as desired, or treatment may be continued for the patient's lifetime. However, other administration regimens may be useful, and this is also within the scope of the present invention. The desired dose can be delivered by a single bolus administration of the IL-36 pathway inhibitor or composition described herein, multiple bolus administrations of the IL-36 pathway inhibitor or composition described herein, or by a series of infusions of the IL-36 pathway or composition described herein.

[0023] IL-36 pathway inhibitors can be formulated as compositions for administration to subjects (e.g., mammals such as humans or non-human primates) by any preferred route of administration, including oral, intravenous, intraperitoneal, subcutaneous, pulmonary, transdermal, intramuscular, intranasal, buccal, sublingual, or suppository administration. Preferably, the compositions are suitable for parenteral administration. As used herein, the term “parenteral” includes administration by intravenous, intramuscular, subcutaneous, rectal, vaginal, and intraperitoneal injection. In some embodiments, the compositions are administered to mammals by peripheral systemic delivery by intravenous, intraperitoneal, or subcutaneous injection.

[0024] The composition may comprise an IL-36 pathway inhibitor and a suitable carrier, such as those well known in the art. The choice of carrier will be determined in part by the specific site where the composition can be administered and the specific method used to administer the composition. The composition may optionally be sterile. The composition may be frozen or lyophilized for storage and reconstituted in a suitable sterile carrier before use. The composition can be produced, for example, according to the prior art described in Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins, Philadelphia, PA (2001).

[0025] Once administered to a mammal (e.g., human or non-human primate), the biological activity of an IL-36 pathway inhibitor, such as an IL-36R conjugate, can be measured by any suitable method known in the art. For example, biological activity can be evaluated by determining the stability of a particular IL-36R conjugate. In one embodiment of the present invention, the IL-36R conjugate (e.g., an antibody) has an in vivo half-life of about 30 minutes to 45 days (e.g., a range defined by about 30 minutes, about 45 minutes, about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 10 hours, about 12 hours, about 1 day, about 5 days, about 10 days, about 15 days, about 25 days, about 35 days, about 40 days, about 45 days, or any two of the above values). In another embodiment, the IL-36R binder has an in vivo half-life of about 2 hours to 20 days (for example, a range defined by about 5 hours, about 10 hours, about 15 hours, about 20 hours, about 2 days, about 3 days, about 7 days, about 12 days, about 14 days, about 17 days, about 19 days, or any two of the aforementioned values). In another embodiment, the IL-36R binder has an in vivo half-life of about 10 days to about 40 days (for example, a range defined by about 10 days, about 13 days, about 16 days, about 18 days, about 20 days, about 23 days, about 26 days, about 29 days, about 30 days, about 33 days, about 37 days, about 38 days, about 39 days, about 40 days, or any two of the aforementioned values).

[0026] The stability of the IL-36R binders described herein is determined by the transition midpoint (T), which is the temperature at which 50% of the amino acid sequence is in its native state and the remaining 50% is denatured. m It can be measured from the perspective of ). Generally, T m The higher the value, the more stable the protein. In one embodiment of the present invention, the IL-36R binder has a transposition midpoint value (T) in vitro at approximately 60-100°C. m ) has. For example, the IL-36R binder has an in vitro T temperature of about 65-80°C (e.g., 66°C, 68°C, 70°C, 71°C, 75°C, or 79°C), about 80-90°C (e.g., about 81°C, 85°C, or 89°C), or about 90-100°C (e.g., about 91°C, about 95°C, or about 99°C). mIt may have.

[0027] The stability of the IL-36R binder can be measured using any other suitable assay known in the art, such as serum half-life measurement, differential scanning calorimetry (DSC), thermal shift assay, and pulse chase assay. Other methods for measuring protein stability in vivo and in vitro that can be used in the context of the present invention include, for example, Protein Stability and Folding, B.A. Shirley (ed.), Human Press, Totowa, New Jersey (1995); Protein Structure, Stability, and This is described in Interactions (Methods in Molecular Biology), Shiver JW (ed.), Humana Press, New York, NY (2010); and Ignatova, Microb. Cell Fact., 4:23 (2005).

[0028] The biological activity of a specific IL-36 pathway inhibitor, such as an IL-36R conjugate, can also be evaluated by determining, for example, its binding affinity to IL-36R or its epitope. The term "affinity" refers to the equilibrium constant for the reversible binding of two agents, and the dissociation constant (K) D ) is expressed as. The affinity of the binder to the ligand, such as the affinity of the antibody for the epitope, may be, for example, in the range of about 1 picomolar concentration (pM) to about 100 micromolar concentration (μM) (for example, about 1 picomolar concentration (pM) to about 1 nanomolar concentration (nM), about 1 nM to about 1 micromolar concentration (μM), or about 1 μM to about 100 μM). In one embodiment, the IL-36R binder is K at a concentration of 1 nanomolar or less (for example, 0.9 nM, 0.8 nM, 0.7 nM, 0.6 nM, 0.5 nM, 0.4 nM, 0.3 nM, 0.2 nM, 0.1 nM, 0.05 nM, 0.025 nM, 0.01 nM, 0.001 nM, or a range defined by any two of the above values). DIt can then bind to the IL-36R protein. In another embodiment, the IL-36R binder is K at a concentration of 200 picomoles or less (e.g., 190 pM, 175 pM, 150 pM, 125 pM, 110 pM, 100 pM, 90 pM, 80 pM, 75 pM, 60 pM, 50 pM, 40 pM, 30 pM, 25 pM, 20 pM, 15 pM, 10 pM, 5 pM, 1 pM, or any two of the above values). D It can bind to IL-36R. The affinity of the immunoglobulin to the target antigen or epitope can be measured using any assay recognized in the art. Such methods include, for example, fluorescence-activated cell sorting (FACS), separable beads (e.g., magnetic beads), surface plasmon resonance (SPR), solution-phase competition (KINEXA®), antigen panning, competitive binding assays, and / or ELISA (see, for example, Janeway et al. (eds.), Immunobiology, 5th ed., Garland Publishing, New York, NY, 2001).

[0029] The IL-36R conjugate of the present invention may be administered alone or in combination with other drugs. For example, the IL-36R conjugate may be administered in combination with other agents for treating or preventing the diseases disclosed herein, such as anti-inflammatory agents including corticosteroids (e.g., prednisone and fluticasone), non-steroidal anti-inflammatory drugs (NSAIDs) (e.g., aspirin, ibuprofen, and naproxen), biological agents (e.g., infliximab (REMICADE®), adalimumab (HUMIRA®), or etanercept (ENBREL®)), methotrexate (MTX), oral retinoids (e.g., acitretin (SORIATANE®)), topical steroids, anti-infective agents and / or antibiotics, or other agents useful for reducing the severity of symptoms of ichthyosis.

[0030] Methods for selecting patient populations and patients suitable for treatment The patient / subject may be any patient / subject with ichthyosis. In one embodiment, the present method The subjects of the method have increased skin expression of at least one of the IL-36 cytokine, IL-36R, or the mRNA encoding it, compared to normal, non-disease subjects. In further embodiments, the IL-36 cytokine is IL-36α, IL-36β, or IL-36γ. In specific embodiments, the IL-36 cytokine is IL-36γ. In some embodiments, the increased skin expression of the IL-36 cytokine (e.g., IL-36γ) is at least about 25% higher (e.g., at least about 30% higher or at least about 50% higher) than the normal baseline expression in healthy subjects.

[0031] In further embodiments, the present invention provides a method for selecting a subject (e.g., a mammal) having ichthyosis for treatment with an IL-36 pathway inhibitor, comprising: comparing the expression of at least one of IL-36 cytokine, IL-36R, or the mRNA encoding it in a skin sample from the subject before and after administration of the IL-36 pathway inhibitor to the subject; and selecting the subject for treatment if a decrease in the expression of at least one of IL-36 cytokine, IL-36R, or the mRNA encoding it is observed in a skin sample from the subject after administration of the IL-36 pathway inhibitor compared to a sample from the subject before administration of the IL-36 pathway inhibitor. In one embodiment, the IL-36 cytokine is IL-36α, IL-36β, or IL-36γ. In further embodiments, the IL-36 pathway inhibitor is the IL-36R conjugate described herein.

[0032] The levels of the target IL-36 cytokine and IL-36R protein can be measured using any suitable method known in the art. Such methods include, for example, radioimmunoassay (RIA) and FACS. Normal or standard expression levels of IL-36 cytokine or IL-36R can be established using any suitable technique, for example, by combining a sample containing or suspected to contain IL-36 cytokine or IL-36R with an IL-36 cytokine-specific or IL-36R-specific antibody under conditions suitable for antigen-antibody complex formation. To facilitate the detection of bound or unbound antibodies, antibodies may be directly or indirectly labeled with a detectable substance. Suitable detectable substances include various enzymes, prosthetic groups, fluorescent substances, luminescent substances, and radioactive substances (see, e.g., Zola, Monoclonal Antibodies: A Manual of Techniques, CRC Press, Inc. (1987)).

[0033] Furthermore, or alternatively, a method for selecting subjects may include determining that a subject has increased Th17 activation in the skin compared to a normal subject, or an increase in CLA+ (skin-homing) T cells of subsets of IL-17+, IL-22+, and IL-9+ compared to a normal subject or compared to a subject with psoriasis but without ichthyosis. Suitable techniques for measuring Th17 activation or an increase in CLA+ T cells are known in the art.

[0034] Furthermore, or alternatively, a method for selecting subjects for treatment may include determining that a subject has at least 12 IASI scores and / or at least 2 erythema scores. The IASI is a composite score ranging from 0 to 48 that quantifies the severity of ichthyosis in a subject based on the severity of erythema or scaling and the percentage of BSA affected, taking into account the degree of erythema and scaling in each of four body regions (each individually scored from 0 to 4), and is adjusted for the percentage of BSA involved in each body region and the proportion of each body region to the whole body (see Paller et al., J Allergy Clin Immunol., 139(1):152-165 (2017)). Methods for making such determinations are known in the art.

[0035] The method for selecting targets for treatment can be used in combination with the methods for treating ichthyosis described herein.

[0036] IL-36 pathway inhibitors None of the methods described herein are limited to the use of any specific IL-36 pathway inhibitor, as long as the inhibitor has a sufficiently rapid and sustained effect to enable therapeutic effects within the dosage parameters described herein. For example, an IL-36 pathway inhibitor may be any inhibitor that inhibits or neutralizes the IL-36 pathway at the level of, for example, cytokines (i.e., inhibiting or neutralizing the biological activity of IL-36α, IL-36β, or IL-36γ), receptors (i.e., inhibiting or neutralizing the biological activity of IL-36R), or inhibits or neutralizes subsequent intracellular signaling induced by IL-36 cytokines or IL-36R. The methods may include, for example, administering an agent that specifically binds to IL-36R; an agent that specifically binds to IL-36 cytokines (e.g., IL-36α, IL-36β, or IL-36γ); or a combination thereof to a subject in need. Examples of IL-36 pathway inhibitors include antibodies or antigen-binding fragments thereof that bind to IL-36 cytokines (e.g., IL-36α, IL-36β, or IL-36γ) or IL-36R, some of which are known in the art and include any of the compounds and compositions disclosed in International Publication Nos. 2016168542A1; 2020018503A2; 2019177883A2; 2018183173A1; U.S. Patent Nos. 10550189B2; 9023995B2; or International Publication No. 2013074569A1 (all incorporated herein by reference).

[0037] In the embodiment, the IL-36 pathway inhibitor is an IL-36R binder such as an antibody or an antigen-binding antibody fragment. The antibody or antigen-binding antibody fragment comprises, consists of, or is essentially composed of, an immunoglobulin heavy chain polypeptide and an immunoglobulin light chain polypeptide, or at least a variable region thereof (e.g., an antigen-binding fragment).

[0038] An entire immunoglobulin typically consists of four polypeptides: two identical copies of a heavy (H) chain polypeptide and two identical copies of a light (L) chain polypeptide. Each heavy chain has one N-terminal variable (V H ) region and three C-terminal constant (C H H1, C H H2, and C H H3) regions, and each light chain contains one N-terminal variable (V L ) region and one C-terminal constant (C L ) region. The light chains of an antibody can be assigned to one of two distinct types, either kappa (κ) or lambda (λ), based on the amino acid sequence of their constant domains. In a typical immunoglobulin, each light chain is linked to a heavy chain by a disulfide bond, and the two heavy chains are linked to each other by disulfide bonds. The light chain variable region aligns with the variable region of the heavy chain, and the light chain constant region aligns with the first constant region of the heavy chain. The remaining constant regions of the heavy chains align with each other.

[0039] The variable regions of each pair of light and heavy chains form the antigen binding site of the antibody. V H L and V L H regions share the same general structure, and each region comprises four framework (FW or FR) regions. As used herein, the term "framework region" refers to a relatively conserved amino acid sequence within the variable region that is located between the hypervariable regions or complementarity determining regions (CDRs). Each variable domain has four framework regions, which are referred to as FR1, FR2, FR3, and FR4. The framework regions form the beta-sheet that provides the structural scaffold for the variable region (see, e.g., C. A. Janeway et al. (eds.), Immunobiology, 5th Ed., Garland Publishing, New York, NY (2001)).

[0040] The framework region is connected by three complementarity-determining regions (CDRs). As previously mentioned, the three CDRs, known as CDR1, CDR2, and CDR3, form the “hypervariable region” of the antibody involved in antigen binding. The CDR regions can also be referred to using “H” or “L” in nomenclature to indicate the heavy or light chain, i.e., CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, or CDRL3. The CDRs of a given Ig sequence can be determined according to one of several conventional numbering schemes such as Kabat, Chothia, Martin (Enhanced Chothia), IGMT, or AHo (e.g., Kabat, et al., Sequences of Proteins of Immunological Interest, USD Department of Health and Human). Services, NIH(1991);Chothia,et al.,Canonical Structures for the Hypervariable Regions of Immunoglobulins,J.Mol.Biol.,196:901-917(1987);Al-Lazikani et al.,Standard Conformations for the Canonical Structures of Immunoglobulins,J.Mol.Biol.,273:927-948(1997);Abhinandan et al.,Analysis and Improvements to Kabat and Structurally Correct Numbering of Antibody Variable Domains,Mol.Immunol.,45:3832-3839(2008);Lefranc et al.,The IMGT unique numbering for immunoglobulins,T cell Receptors and Ig-like domains,The Immunologist,7:132-136(1999);Lefranc et al.,IMGT unique numbering for immunoglobulin and T cell receptor variable domains and I superfamily V-like domains, Dev. Comp. Immunol., 27:55-77 (2003); and Honegger et al., Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool, J. Mol. Biol. 309: 657-670 (2001).

[0041] In one embodiment, the immunoglobulin heavy chain variable region is Gln Val Gln Xaa1 Xaa2 Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Thr Ser Tyr Asp Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Xaa3 Leu Glu Trp Met Gly Trp Ile Tyr Pro Gly Asp Xaa4 Ser Thr Lys Tyr Asn Glu Lys Phe Lys Gly Arg Val Thr Ile Thr Xaa5 Asp Xaa6 Ser Ala Xaa7 Thr Ala Tyr Met Glu Leu Xaa8 Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Xaa9 Cys Thr Arg Ser Phe Tyr Thr Met Asp Tyr Trp Gly Gln Gly Thr Thr Val The amino acid sequence of Thr Val Ser Ser (SEQ ID NO: 56), or at least its CDR, consists of or essentially consists of, where (a) Xaa1 is leucine (Leu) or phenylalanine (Phe), (b) Xaa2 is valine (Val), methionine (Met), or leucine (Leu), (c) Xaa3 is arginine (Arg) or glycine (Gly), and (d) Xaa4 is glycine (Gl (y) is serine (Ser) or alanine (Ala), (e) Xaa5 is arginine (Arg) or alanine (Ala), (f) Xaa6 is threonine (Thr) or lysine (Lys), (g) Xaa7 is serine (Ser) or asparagine (Asn), (h) Xaa8 is serine (Ser) or alanine (Ala), and (i) Xaa9 is tyrosine (Tyr) or phenylalanine (Phe). In some embodiments, the immunoglobulin heavy chain polypeptide is Gln Val Gln Xaa1 Xaa2 Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Thr Ser Tyr Asp Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Xaa3 Leu Glu Trp Met Gly Trp Ile Tyr Pro Gly Asp Xaa4 Ser Thr Lys Tyr Asn Glu Lys Phe Lys Gly Arg Val Thr Ile Thr Xaa5 Asp Xaa6 Ser Ala Ser Thr Ala Tyr Met Glu Leu Xaa7 Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Xaa8 Cys Thr Arg Ser Phe Tyr Thr Met Asp Tyr Trp Gly Gln Gly Thr Thr Val The amino acid sequence of Thr Val Ser Ser (Sequence ID 1), or at least its CDR, consists of or essentially consists of, where (a) Xaa1 is leucine (Leu) or phenylalanine (Phe), (b) Xaa2 is valine (Val), methionine (Met), or leucine (Leu), (c) Xaa3 is arginine (Arg) or glycine (Gly), (d) Xaa4 is glycine (Gly), serine (Ser), or alanine (Ala), (e) Xaa5 is arginine (Arg) or alanine (Ala), (f) Xaa6 is threonine (Thr) or lysine (Lys), (g) Xaa7 is serine (Ser) or alanine (Ala), and (h) Xaa8 is tyrosine (Tyr) or phenylalanine (Phe).

[0042] The heavy chain polypeptide may contain, consist of, or essentially consist of, the amino acid sequence of SEQ ID NO: 56 or SEQ ID NO: 1, or at least its CDR, having any one of the aforementioned amino acid substitutions in any preferred combination. In one embodiment, the immunoglobulin heavy chain polypeptide contains, consists of, or essentially consists of, the amino acid sequence of any one of SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, or SEQ ID NO: 14, or at least its CDR.

[0043] In some embodiments, the IL-36R binder has the amino acid sequence Phe Thr Phe CDR1 (HCDR1) is a heavy chain variable region containing, consisting of, or essentially consisting of Thr Ser Tyr Asp Ile Asn (SEQ ID NO: 59); (a)Trp Ile Tyr Pro Gly Asp Gly Ser Thr Lys Tyr CDR2 (HCDR2) is a heavy chain variable region comprising, consisting of, or essentially comprising the amino acid sequence Asn Glu Lys Phe Lys Gly (SEQ ID NO: 60); (b)Trp Ile Tyr Pro Gly Asp Ser Ser Thr Lys Tyr Asn Glu Lys Phe Lys Gly (SEQ ID NO: 61); or (c)Trp Ile Tyr Pro Gly Asp Ala Ser Thr Lys Tyr Asn Glu Lys Phe Lys Gly (SEQ ID NO: 62); and / or CDR3 (HCDR3) is a heavy chain variable region comprising, consisting of, or essentially comprising the amino acid sequence Ser Phe Tyr Thr Met Asp Tyr (SEQ ID NO: 63).

[0044] In another embodiment, the immunoglobulin heavy chain variable region is an amino acid sequence: Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr Xaa1 Met Xaa2 Trp Val Arg Gln Ala Pro Xaa3 Gln Gly Leu Glu Trp Met Gly Met Phe Xaa4 Pro Xaa5 Xaa6 Xaa7 Val Thr Arg Leu Asn Gln Lys Phe Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Thr Ser Met Ile Ile Gly Gly Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val A compound comprising or essentially comprising Ser Ser (SEQ ID NO: 15), or at least its CDR, wherein (a) Xaa1 is tryptophan (Trp) or tyrosine (Tyr), (b) Xaa2 is histidine (His), asparagine (Asn), or tyrosine (Tyr), (c) Xaa3 is glycine (Gly) or arginine (Arg), (d) Xaa4 is aspartic acid (Asp), glutamic acid (Glu), or histidine (His), (e) Xaa5 is serine (Ser), threonine (Thr), or tyrosine (Tyr), (f) Xaa6 is asparagine (Asn) or glycine (Gly), and (g) Xaa7 is serine (Ser), alanine (Ala), or aspartic acid (Asp).

[0045] The heavy chain variable region may contain, consist of, or essentially consist of, the amino acid sequence of SEQ ID NO: 15 or at least its CDR, having one of the aforementioned amino acid substitutions in any preferred combination. In one embodiment, the immunoglobulin heavy chain polypeptide contains, consists of, or essentially consists of, one of the amino acid sequences of SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, SEQ ID NO: 21, SEQ ID NO: 22, SEQ ID NO: 23, or SEQ ID NO: 24 or at least its CDR.

[0046] In one embodiment, the IL-36 pathway inhibitor is an HCDR1 comprising, consisting of, or essentially comprising an amino acid sequence selected from the group consisting of (a) Tyr Thr Phe Thr Asn Tyr Trp Met His (SEQ ID NO: 64); (b) Tyr Thr Phe Thr Asn Tyr Trp Met Asn (SEQ ID NO: 65); (c) Tyr Thr Phe Thr Asn Tyr Trp Met Tyr (SEQ ID NO: 66); and (d) Tyr Thr Phe Thr Asn Tyr Tyr Met Asn (SEQ ID NO: 67); (a) Met Phe Asp Pro Ser Asn Ser Val Thr Arg Leu Asn Gln Lys Phe Lys Asp (SEQ ID NO: 68); (b) Met Phe Glu Pro Ser Asn Ala Val Thr Arg Leu Asn Gln Lys Phe Lys Asp (SEQ ID NO: 69); (c) Met Phe His Pro Ser Asn Ala Val Thr Arg Leu Asn Gln Lys Phe Lys Asp (SEQ ID NO: 70); and (d) Met Phe His Pro Thr Gly Asp Val Thr Arg Leu Asn Gln Lys Phe Lys HCDR2 comprising, consisting of, or essentially consisting of an amino acid sequence selected from the group consisting of Asp (SEQ ID NO: 71); and / or amino acid sequences Thr Thr Ser It comprises a heavy chain variable region containing HCDR3 which includes, consists of, or is essentially derived from Met Ile Ile Gly Gly Phe Ala Tyr (SEQ ID NO: 72).

[0047] In further embodiments, the immunoglobulin heavy chain variable region is Xaa1 Xaa2 Gln Xaa3 Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Xaa4 Xaa5 Tyr Ser Ile Thr Xaa6 Asp Phe Ala Trp Asn Trp Ile Arg Gln Xaa7 Pro Gly Xaa8 Xaa9 Leu Glu Trp Ile Gly Tyr Ile Ser Tyr Ser Gly Asp Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Xaa10 Xaa11 Asp Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Xaa12 Tyr Xaa13 Cys Ala Ile Arg Gly Pro Tyr Ser Phe Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser The amino acid sequence of Xaa14 (SEQ ID NO: 57), or at least its CDR, consists of or essentially consists of, where Xaa1 is glutamine (Gln) or aspartic acid (Asp); Xaa2 is valine (Val) or leucine (Leu); Xaa3 is leucine (Leu) or phenylalanine (Phe); Xaa4 is threonine (Thr) or serine (Ser); Xaa5 is glycine (Gly) or arginine (Arg); Xaa6 is serine (Ser) or alanine (Ala); Xa a7 is proline (Pro) or phenylalanine (Phe); Xaa8 is lysine (Lys) or asparagine (Asn); Xaa9 is glycine (Gly) or lysine (Lys); Xaa10 is serine (Ser) or threonine (Thr); Xaa11 is valine (Val) or arginine (Arg); Xaa12 is threonine (Thr) or valine (Val); Xaa13 is tyrosine (Tyr) or phenylalanine (Phe); Xaa14 is alanine (Ala) or absent. In some embodiments, the heavy chain variable region is the amino acid sequence Xaa1 Val Gln Xaa2 Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Xaa3 Gly Tyr Ser Ile Thr Ser Asp Phe Ala Trp Asn Trp Ile Arg Gln Xaa4 Pro Gly Xaa5 Xaa6 Leu Glu Trp Ile Gly Tyr Ile Ser Tyr Ser Gly Asp Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Xaa7 Xaa8 Asp Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Xaa9 Cys Ala Ile Arg Gly Pro Tyr Ser Phe Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser (SEQ ID NO: 25), or comprising or essentially comprising at least its CDR, wherein (a) Xaa1 is glutamine (Gln) or aspartic acid (Asp), (b) Xaa2 is leucine (Leu) or phenylalanine (Phe), (c) Xaa3 is threonine (Thr) or serine (Ser), and (d) Xaa4 is proline (Pro) or phenylalanine (e)Xaa5 is lysine (Lys) or asparagine (Asn), (f)Xaa6 is glycine (Gly) or lysine (Lys), (g)Xaa7 is serine (Ser) or threonine (Thr), (h)Xaa8 is valine (Val) or arginine (Arg), and (i)Xaa9 is tyrosine (Tyr) or phenylalanine (Phe).

[0048] In some embodiments, the heavy chain variable region has one or more of the aforementioned amino acid substitutions in any preferred combination, such as the amino acid sequence of SEQ ID NO: 57 or SEQ ID NO: 25, or The immunoglobulin heavy chain variable region may include, consist of, or essentially consist of at least the CDR thereof. In one embodiment, the immunoglobulin heavy chain variable region includes, consists of, or essentially consists of any one amino acid sequence of SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, or SEQ ID NO: 54, or at least the CDR thereof.

[0049] In additional embodiments, the IL-36 pathway inhibitor is an HCDR1 comprising, consisting of, or essentially comprising an amino acid sequence selected from the group consisting of (a) Tyr Ser Ile Thr Ser Asp Phe Ala Trp Asn (SEQ ID NO: 73); and (b) Tyr Ser Ile Thr Ala Asp Phe Ala Trp Asn (SEQ ID NO: 74); amino acid sequence Tyr Ile Ser Tyr Ser Gly HCDR2 may contain, consist of, or be essentially derived from the amino acid sequence Asp Thr Asn Tyr Asn Pro Ser Leu Lys Ser (SEQ ID NO: 75); and / or HCDR3 may contain, consist of, or be essentially derived from the amino acid sequence Arg Gly Pro Tyr Ser Phe Thr Tyr (SEQ ID NO: 76).

[0050] In another embodiment, the IL-36 pathway inhibitor comprises an immunoglobulin heavy chain polypeptide comprising, or essentially comprising, the amino acid sequence of SEQ ID NO: 33, SEQ ID NO: 34, or SEQ ID NO: 35, or at least its CDR.

[0051] In some embodiments, the immunoglobulin heavy chain polypeptide includes an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to any of the aforementioned variable region sequences. As described herein, nucleic acid or amino acid sequence "identity" can be determined by comparing the nucleic acid or amino acid sequence of interest with a reference nucleic acid or amino acid sequence. The identity percentage is obtained by dividing the number of nucleotide or amino acid residues that are the same (i.e., identical) between the sequence of interest and the reference sequence by the length of the longest sequence (i.e., the length of the longer of the sequence of interest or the reference sequence). Numerous mathematical algorithms are known and incorporated into many available software programs for obtaining optimal alignment and calculating identity between two or more sequences. Examples of such programs include CLUSTAL-W, T-Coffee, and ALIGN (for aligning nucleic acid and amino acid sequences), the BLAST program (e.g., BLAST2.1, BL2SEQ, and newer versions thereof), and the FASTA program (e.g., FASTA3x, FASTM, and SSEARCH) (for sequence alignment and sequence similarity searches).Furthermore, sequence alignment algorithms can be found in, for example, Altschul et al., J. Molecular Biol., 215(3):403-410 (1990), Beigert et al., Proc. Natl. Acad. Sci. USA, 106(10):3770-3775 (2009), Durbin et al., eds., Biological Sequence Analysis: Probabalistic Models of Proteins and Nucleic Acids, Cambridge University Press, Cambridge, UK (2009), Soding, Bioinformatics, 21(7):951-960 (2005), Altschul et al., Nucleic Acids Res., 25(17):3389-3402 (1997), and Gusfield, Algorithms on Strings, Trees and Sequences, Cambridge University Press, Cambridge. It is disclosed in UK(1997)).

[0052] In addition to the heavy chain variable region described herein, the IL-36R binder is an amino acid sequence Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu His Ser Asn Xaa1 Asn Thr Ty Leu Tyr Trp Tyr Pro Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile The immunoglobulin light chain variable region comprises Lys (SEQ ID NO: 36), or at least its CDR, consisting of, or essentially consisting of, (a) Xaa1 is glycine (Gly) or alanine (Ala), (b) Xaa2 is phenylalanine (Phe) or tyrosine (Tyr), and (c) Xaa3 is tyrosine (Tyr) or serine (Ser).

[0053] The light chain variable region may contain, consist of, or essentially consist of, the amino acid sequence of SEQ ID NO: 36 or at least its CDR, having one or more of the aforementioned amino acid substitutions in any preferred combination. In one embodiment, the isolated immunoglobulin light chain variable region contains, consists of, or essentially consists of, the amino acid sequence of SEQ ID NO: 37, SEQ ID NO: 38, or SEQ ID NO: 39 or at least its CDR.

[0054] In some embodiments, the IL-36R binder is (a) Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly Asn Thr Tyr Leu Tyr (SEQ ID NO: 77); or (b) Arg Ser Ser Lys Ser Leu Leu His Ser Asn Ala Asn Thr Tyr CDR1 (LCDR1) is a light chain variable region comprising, consisting of, or essentially comprising an amino acid sequence selected from Leu Tyr (SEQ ID NO: 78); CDR2 (LCDR2) is a light chain variable region comprising, consisting of, or essentially comprising the amino acid sequence Arg Met Ser Asn Leu Ala Ser (SEQ ID NO: 79); and CDR3 (LCDR3) is a light chain variable region comprising, consisting of, or essentially comprising the amino acid sequence Met Gln His Leu Glu Tyr Pro Phe Thr (SEQ ID NO: 80).

[0055] In some embodiments, the immunoglobulin light chain variable region is the amino acid sequence Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu His Xaa1 Asn Xaa2 Ile Thr Tyr Phe Tyr Trp Tyr Leu Xaa3 Lys Pro Gly Gln Pro Pro Gln Leu Leu Ile Tyr Gln Met Ser Asn Leu Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ala Gln Asn Leu Glu Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys (Sequence ID 40), or comprising or essentially comprising at least its CDR, wherein (a)Xaa1 is serine (Ser) or arginine (Arg) (b) Xaa2 is glycine (Gly) or alanine (Ala), and (c) Xaa3 is glutamine (Gln) or histidine (His).

[0056] The light chain variable region may include, consist of, or essentially consist of, the amino acid sequence of SEQ ID NO: 40 or at least its CDR having any combination of the aforementioned amino acid substitutions. In one embodiment, the immunoglobulin light chain polypeptide includes, consists of, or essentially consists of, one of the amino acid sequences of SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, or SEQ ID NO: 44; or at least its CDR.

[0057] In some embodiments, the light chain variable region is an amino acid sequence (a) Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly Ile Thr Tyr Phe Tyr (SEQ ID NO: 81); (b) Arg Ser Ser Lys Ser Leu Leu His Ser Asn Ala Ile Thr Tyr Phe Tyr (SEQ ID NO: 82); or (c) Arg Ser Ser Lys Ser Leu Leu His Arg Asn Ala Ile Thr Tyr Phe LCDR1 contains, consists of, or is essentially derived from Tyr (SEQ ID NO: 83); LCDR2 contains, consists of, or is essentially derived from the amino acid sequence Gln Met Ser Asn Leu Ala Ser (SEQ ID NO: 84); and LCDR3 contains, consists of, or is essentially derived from the amino acid sequence Ala Gln Asn Leu Glu Leu Pro Leu Thr (SEQ ID NO: 85).

[0058] In further embodiments, the immunoglobulin light chain variable region is: Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Xaa1 Ile Asn Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Tyr Thr Ser Xaa2 Leu His Ser Gly Val Pro Ser Arg Phe Ser Xaa3 Ser Gly Ser Gly Xaa4 Asp Xaa5 Thr Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly His Thr Leu Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Xaa6 The amino acid sequence of Xaa7 (SEQ ID NO: 58), or comprising or essentially comprising at least its CDR, wherein (a) Xaa1 is aspartic acid (Asp) or tryptophan (Trp), (b) Xaa2 is arginine (Arg) or methionine (Met), (c) Xaa3 is glycine (Gly), serine (Ser), or proline (Pro), (d) Xaa4 is threonine (Thr) or asparagine (Asn), (e) Xaa5 is phenylalanine (Phe) or tyrosine (Tyr), (f) Xaa6 is arginine (Arg) or absent, and (g) Xaa7 is threonine (Thr) or absent.In some embodiments, the immunoglobulin light chain variable region is an amino acid sequence Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Xaa1 Ser Gly Ser Gly Thr. Asp Xaa2 Thr Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gl n Gln Gly His Thr Leu Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys (SEQ ID NO: 45), or comprising or essentially comprising at least the CDR thereof, wherein (a) Xaa1 is serine (Ser) or proline (Pro), and (b) Xaa2 is phenylalanine (Phe) or tyrosine (Tyr).

[0059] The light chain variable region may contain, consist of, or essentially consist of, the amino acid sequence of SEQ ID NO: 58 or SEQ ID NO: 45, or at least its CDR, having one or more of the aforementioned amino acid substitutions in any preferred combination. In one embodiment, the immunoglobulin light chain polypeptide contains, consists of, or essentially consists of, the amino acid sequence of SEQ ID NO: 46, SEQ ID NO: 47, or SEQ ID NO: 55, or at least its CDR.

[0060] In some embodiments, the light chain variable region is the amino acid sequence (a)Arg Ala Ser LCDR1 comprises, consists of, or essentially consists of, the amino acid sequence (a) Tyr Thr Ser Arg Leu His Ser (SEQ ID NO: 88) or (b) Tyr Thr Ser Met Leu His Ser (SEQ ID NO: 89); and LCDR2 comprises, consists of, or essentially consists of, the amino acid sequence (a) Tyr Thr Ser Arg Leu His Ser (SEQ ID NO: 88) or (b) Tyr Thr Ser Met Leu His Ser (SEQ ID NO: 89); and LCDR3 comprises, consists of, or essentially consists of, the amino acid sequence Gln Gln Gly His Thr Leu Pro Trp Thr (SEQ ID NO: 90).

[0061] In another embodiment, the immunoglobulin light chain variable region comprises or essentially comprises the amino acid sequence of SEQ ID NO: 48, SEQ ID NO: 49, or SEQ ID NO: 50, or at least its CDR.

[0062] In some embodiments, the immunoglobulin light chain variable region includes an amino acid sequence that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to any of the immunoglobulin light chain variable region sequences described herein. The "identity" of nucleic acid or amino acid sequences can be determined using the methods described herein.

[0063] As described above, the IL-36R binder may include either of the heavy chain and light chain variable region sequences described above, or the heavy chain and light chain variable region of an immunoglobulin having the CDR thereof. The CDR sequence may be one of the CDR sequences described herein, or one of several known methods (e.g., Kabat, Chothia, Martin (Enhanced Chothia), IGMT, or AHo).

[0064] In one embodiment, the IL-36R binder has an immunoglobulin heavy chain variable region including SEQ ID NO: 15 or SEQ ID NO: 22, or at least its CDR region; and an immunoglobulin light chain variable region including SEQ ID NO: 40 or SEQ ID NO: 44, or at least its CDR sequence, wherein the CDR is as determined according to any of the various known immunoglobulin numbering schemes, particularly Kabat, Chothia, Martin (Enhanced Chothia), IGMT, or AHo. For example, in some embodiments, the antibody includes the heavy chain variable region of SEQ ID NO: 22 and the light chain variable region of SEQ ID NO: 44, or at least its CDR as determined by Kabat. In some embodiments, the antibody includes the heavy chain variable region of SEQ ID NO: 22 and the light chain variable region of SEQ ID NO: 44, or at least its CDR as determined by Chothia. In some embodiments, the antibody includes the heavy chain variable region of SEQ ID NO: 22 The antibody includes the variable region and the light chain variable region of SEQ ID NO: 44, or its CDR as determined by Martin. In some embodiments, the antibody includes the heavy chain variable region of SEQ ID NO: 22 and the light chain variable region of SEQ ID NO: 44, or its CDR as determined by IGMT. In some embodiments, the antibody includes the heavy chain variable region of SEQ ID NO: 22 and the light chain variable region of SEQ ID NO: 44, or its CDR as determined by AHo.

[0065] In some embodiments, the IL-36R binder is (a) Tyr Thr Phe Thr Asn Tyr Trp Met His (SEQ ID NO: 64); (b) Tyr Thr Phe Thr Asn Tyr Trp Met Asn (SEQ ID NO: 65); (c) Tyr Thr Phe Thr Asn Tyr Trp Met Tyr (SEQ ID NO: 66); and (d) Tyr Thr Phe Thr Asn Tyr Tyr Met HCDR1 comprises, consists of, or is essentially derived from an amino acid sequence selected from the group consisting of Asn (SEQ ID NO: 67); (a) Met Phe Asp Pro Ser Asn Ser Val Thr Arg Leu Asn Gln Lys Phe Lys Asp (SEQ ID NO: 68); (b) Met Phe Glu Pro Ser Asn Ala Val Thr Arg Leu Asn Gln Lys Phe Lys Asp (SEQ ID NO: 69); (c) Met Phe His Pro Ser Asn Ala Val Thr Arg Leu Asn Gln Lys Phe Lys Asp (SEQ ID NO: 70); and (d) Met Phe His Pro Thr Gly Asp Val Thr Arg Leu Asn Gln Lys Phe Lys Asp (SEQ ID NO: 71); and / or the amino acid sequence Thr Thr Ser Met Ile Ile Gly Gly Phe Ala The heavy chain variable region comprises HCDR3 containing, consisting of, or essentially consisting of Tyr (SEQ ID NO: 72); amino acid sequence (a) Arg Ser Ser Lys Ser Leu Leu His Ser Asn Gly Ile Thr Tyr Phe Tyr (SEQ ID NO: 81); (b) Arg Ser Ser Lys Ser Leu Leu His Ser The light chain variable region comprises LCDR1 containing, consisting of, or essentially consisting of the amino acid sequence Asn Ala Ile Thr Tyr Phe Tyr (SEQ ID NO: 82); or (c)Arg Ser Ser Lys Ser Leu Leu His Arg Asn Ala Ile Thr Tyr Phe Tyr (SEQ ID NO: 83); LCDR2 containing, consisting of, or essentially consisting of the amino acid sequence Gln Met Ser Asn Leu Ala Ser (SEQ ID NO: 84); and LCDR3 containing, consisting of, or essentially consisting of the amino acid sequence Ala Gln Asn Leu Glu Leu Pro Leu Thr (SEQ ID NO: 85).

[0066] In certain embodiments, the IL-36R binder comprises, consists of, or is essentially derived from an amino acid sequence selected from the group consisting of Tyr Thr Phe Thr Asn Tyr Trp Met Asn (SEQ ID NO: 65) as HCDR1; comprises, consists of, or is essentially derived from an amino acid sequence selected from the group consisting of Met Phe His Pro Thr Gly Asp Val Thr Arg Leu Asn Gln Lys Phe Lys Asp (SEQ ID NO: 71) as HCDR2; and / or the amino acid sequence Thr Thr Ser Met Ile Ile Gly Gly Phe Ala The heavy chain variable region comprises HCDR3 containing, consisting of, or essentially consisting of Tyr (SEQ ID NO: 72); amino acid sequence Arg Ser Ser Lys Ser Leu LCDR1 contains, consists of, or is essentially derived from the amino acid sequence Leu His Arg Asn Ala Ile Thr Tyr Phe Tyr (SEQ ID NO: 83); LCDR2 contains, consists of, or is essentially derived from the amino acid sequence Gln Met Ser Asn Leu Ala Ser (SEQ ID NO: 84); and contains, or is essentially derived from the amino acid sequence Ala Gln Asn Leu Glu Leu Pro Leu Thr (SEQ ID NO: 85). It includes a light chain variable region comprising an LCDR3 which is essentially derived from or from.

[0067] Furthermore, the IL-36R binder may include heavy chain and light chain variable regions of immunoglobulins that have a specified percentage of identity with respect to the heavy chain and light chain variable region sequences, for example, at least 90% identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity). In some embodiments, the variation in the sequence occurs outside the CDR (determined by any known method including Kabat, Chothia, Martin (Enhanced Chothia), IGMT, or AHo) such that the heavy chain and light chain sequences having specified sequence identity with respect to the specified sequences described herein retain the CDR of such sequences. In embodiments, the IL-36R binder includes an immunoglobulin heavy chain variable region that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 15 or SEQ ID NO: 22, and optionally the sequence holds the CDR of SEQ ID NO: 15 or SEQ ID NO: 22; and also includes an immunoglobulin heavy chain variable region that is at least 90% identical (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical) to SEQ ID NO: 40 or SEQ ID NO: 44, and optionally the sequence holds the CDR of SEQ ID NO: 40 or SEQ ID NO: 44, and the CDR is according to any of the various known immunoglobulin numbering schemes, particularly Kabat, Chothia, Martin (Enhanced As determined in accordance with Chothia, IGMT, or AHo.In certain embodiments, the IL-36R conjugate comprises an immunoglobulin heavy chain variable region that is at least 90% identical to (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to) SEQ ID NO: 22, and optionally the sequence retains the CDR of SEQ ID NO: 22; and comprises an immunoglobulin heavy chain variable region that is at least 90% identical to (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to) SEQ ID NO: 44, and optionally the sequence retains the CDR of SEQ ID NO: 44, the CDR being determined according to any of the various known immunoglobulin numbering schemes, particularly according to Kabat, Chothia, Martin (Enhanced Chothia), IGMT, or AHo.

[0068] Differences in sequence identity can result from the addition, substitution, or deletion of one or more amino acid residues. Amino acid "substitution" or "replacement" refers to the replacement of an amino acid at a given position or residue with another amino acid at the same position or residue within the polypeptide sequence. Amino acid substitutions or replacements can be conserved, semi-conservative, or non-conservative, depending on whether the substitution is made by an amino acid residue with similar properties to the residue being replaced. A functional method for defining common properties between individual amino acids is to analyze the normalized frequency of amino acid changes between corresponding proteins in homologous organisms (Schulz and Schirmer, Principles of Protein Structure, Springer-Verlag, New York (1979)). According to such analysis, groups of amino acids can be defined as those in which amino acids preferentially exchange with each other and are therefore most similar in their influence on the overall protein structure (Schulz and Schirmer, cited above).

[0069] Amino acids can be broadly classified into "aromatic" or "aliphatic." Aromatic amino acids contain an aromatic ring. Examples of aromatic amino acids include histidine (H or His) and phenylalanine. Examples include glycine (F or Phe), tyrosine (Y or Tyr), and tryptophan (W or Trp). Non-aromatic amino acids are broadly classified as "aliphatic." Examples of "aliphatic" amino acids include glycine (G or Gly), alanine (A or Ala), valine (V or Val), leucine (L or Leu), isoleucine (I or Ile), methionine (M or Met), serine (S or Ser), threonine (T or Thr), cysteine ​​(C or Cys), proline (P or Pro), glutamic acid (E or Glu), aspartic acid (A or Asp), asparagine (N or Asn), glutamine (Q or Gln), lysine (K or Lys), and arginine (R or Arg).

[0070] Aliphatic amino acids can be subdivided into four subgroups. The "major aliphatic nonpolar subgroup" consists of valine, leucine, and isoleucine. The "aliphatic micropolar subgroup" consists of methionine, serine, threonine, and cysteine. The "aliphatic polar / charged subgroup" consists of glutamic acid, aspartic acid, asparagine, glutamine, lysine, and arginine. The "small residue subgroup" consists of glycine and alanine. The charged / polar amino acid group can be subdivided into three subgroups: the "positively charged subgroup" consisting of lysine and arginine, the "charged subgroup" consisting of glutamic acid and aspartic acid, and the "polar subgroup" consisting of asparagine and glutamine.

[0071] Aromatic amino acids can be further subdivided into two subgroups: the "nitrogen ring subgroup" consisting of histidine and tryptophan, and the "phenyl subgroup" consisting of phenylalanine and tyrosine.

[0072] Examples of conservative amino acid substitutions include substitutions of amino acids within the above subgroups, such as the substitution of arginine with lysine (and vice versa) in which a positive charge can be maintained, the substitution of aspartic acid with glutamic acid (and vice versa) in which a negative charge can be maintained, the substitution of threonine with serine in which a free -OH can be maintained, and the substitution of asparagine with glutamine in which a free -NH2 can be maintained. "Semi-conservative mutations" include amino acid substitutions of amino acids within the same group listed herein but not within the same subgroup. For example, the substitution of asparagine with aspartic acid or lysine with asparagine involves amino acids within the same group but from different subgroups. "Non-conservative mutations" include amino acid substitutions between different groups, such as the substitution of tryptophan with lysine or serine with phenylalanine.

[0073] If the variable region of the light or heavy chain of an immunoglobulin "essentially consists of" any of the aforementioned heavy or light chain variable region amino acid sequences, the polypeptide may contain additional components that do not significantly affect the polypeptide, such as those described herein. If the variable region of the light or heavy chain of an immunoglobulin "consists of," the polypeptide does not contain any additional components.

[0074] IL-36R conjugates (e.g., antibodies or antibody fragments) may be conjugates that compete with IL-36R conjugates containing heavy-chain or light-chain polypeptides of immunoglobulins described herein for binding to IL-36R, for example, by binding to the same epitope or overlapping epitopes. Antibody competition can be assayed using routine peptide competition assays utilizing ELISA, Western blotting, or immunohistochemical methods (see, for example, U.S. Patents 4,828,981 and 8,568,992; and Braitbard et al., Proteome Sci., 4:12 (2006)).

[0075] The "biological activity" of an IL-36R conjugate refers, for example, to its binding affinity for a specific IL-36R epitope, and to the neutralization or inhibition of IL-36R's binding to its receptor(s). Neutralization or inhibition of IL-36R activity in vivo (e.g., IC 50 This refers to the pharmacokinetics and cross-reactivity (e.g., with non-human homologs or orthologs of the IL-36R protein, or with other proteins or tissues). In certain embodiments, the IL-36R conjugate preferably exhibits one or more of the following biological activities: (a) inhibiting the interaction between IL-36R and IL-36α, IL-36β, and / or IL-36γ; (b) inhibiting intracellular signaling mediated by IL-36R; and / or (c) cross-reacting with and inhibiting the activity of human and non-human primate (e.g., cynomolgus monkey) IL-36R. Other biological properties or features of antigen conjugates recognized in the art include, for example, binding affinity, selectivity, solubility, folding, immunotoxicity, expression, and formulation. The aforementioned properties or characteristics can be observed, measured, and / or evaluated using standard techniques including, but not limited to, ELISA, competitive ELISA, surface plasmon resonance analysis (BIACORE®), or KINEXA®, in vitro or in vivo neutralization assays, receptor-ligand binding assays, cytokine or growth factor production and / or secretion assays, and signal transduction and immunohistochemical assays.

[0076] When used herein with respect to the activity of IL-36 pathway inhibitors, such as IL-36R conjugates, the terms “inhibit” or “neutralize” refer to the ability to substantially antagonize, inhibit, prevent, suppress, slow, disrupt, modify, eliminate, halt, or reverse the biological activity of IL-36 cytokines or IL-36R, or the progression or severity of diseases or conditions associated with IL-36 cytokines or IL-36R, such as ichthyosis. For example, an IL-36R conjugate preferably inhibits or neutralizes the activity of IL-36R to the extent defined by at least about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 100%, or any two of the aforementioned values.

[0077] The IL-36R conjugate of the method of the present invention may be an entire antibody as described herein, or an antibody fragment. The terms “antibody fragment,” “antibody fragment,” and “functional fragment of antibody” are used interchangeably herein and mean one or more fragments of an antibody that retain the ability to specifically bind to an antigen (see Holliger et al., Nat. Biotech., 23(9):1126-1129 (2005) in general). The IL-36R conjugate may contain any IL-36R-conjugated antibody fragment. The antibody fragment preferably includes, for example, one or more CDRs, variable regions (or parts thereof), constant regions (or parts thereof), or a combination thereof. Examples of antibody fragments include (i)V L , V H , C L (ii) a monovalent fragment consisting of the CH1 domain, (ii) a Fab fragment, which is a bivalent fragment containing two Fab fragments linked by disulfide crosslinks in the hinge region, and (iii) a single arm of the antibody V L and V HExamples include, but are not limited to, (iv) an Fv fragment consisting of the domain, (iv) a Fab' fragment resulting from disrupting the disulfide crosslinks of the F(ab')2 fragment using mild reducing conditions, (v) a disulfide-stabilized Fv fragment (dsFv), and (vi) a domain antibody (dAb) which is an antibody single variable region domain (VH or VL) polypeptide that specifically binds to the antigen.

[0078] If the IL-36R binder is an antibody or antibody fragment, the antibody or antibody fragment has any suitable class of heavy chain constant region (F c ) may include. In some embodiments, the antibody or antibody fragment comprises a heavy chain constant region based on a wild-type IgG1, IgG2, or IgG4 antibody, or a variant thereof. It will be understood that each antibody class or isotype, once recognized, associates with a different set of effector mechanisms for disposing of or neutralizing the antigen. Thus, in some embodiments, when the IL-36R conjugate is an antibody or antibody fragment, it may exhibit one or more effector functions, such as involvement in antibody-dependent complement-mediated lysis or antibody-dependent cytotoxicity via interaction with effector molecules and cells (e.g., activation of the complement system).

[0079] Furthermore, the IL-36R binder may also be a single-chain antibody fragment. An example of a single-chain antibody fragment is (i) two domains of an Fv fragment joined by a synthetic linker that allows the two domains to be synthesized as a single polypeptide chain (i.e., V L and V H (ii) a monovalent molecule consisting of a single-chain Fv (scFv) (see, for example, Bird et al., Science, 242:423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA, 85:5879-5883 (1988); and Osbourn et al., Nat. Biotechnol., 16:778 (1998)), and (ii) V that is too short to be found in the same polypeptide chain H and V L V is formed by a peptide linker that cannot pair with it. L V connectedH Each includes, and thereby, different V H -V L Examples include, but are not limited to, dimers of polypeptide chains, called diabodies, which drive the pairing of complementary domains in the polypeptide chain to produce a dimeric molecule having two functional antigen-binding sites. Antibody fragments are known in the art and are described in detail, for example, in U.S. Patent Application Publication 2009 / 0093024 A1.

[0080] Furthermore, the IL-36R conjugate may be an intrabody or a fragment thereof. An intrabody is an antibody that is expressed and functions within a cell. Intrabodies typically lack disulfide bonds and can regulate the expression or activity of target genes through their specific binding activity. Intrabodies are isolated from V H and V L The intrabody includes domains and single-domain fragments such as scFv. The intrabody may include an intracellular transport signal attached to its N-terminus or C-terminus so that it can be expressed at high concentrations in the intracellular compartment where the target protein is located. Upon interaction with the target gene, the intrabody modulates the function of the target protein and / or achieves phenotypic / functional knockout by mechanisms such as accelerating the degradation of the target protein and sequestrating the target protein to a non-physiological intracellular compartment. Other mechanisms of gene inactivation mediated by the intrabody, such as binding to a catalytic site on the target protein or binding to an epitope involved in protein-protein, protein-DNA, or protein-RNA interactions, may depend on the epitope targeted by the intrabody.

[0081] The IL-36R conjugate may be an antibody conjugate. In this regard, the IL-36R conjugate may be a conjugate of (1) an antibody, an alternative scaffold, or a fragment thereof, and (2) a protein or non-protein portion containing the IL-36R conjugate. For example, the IL-36R conjugate may be all or part of an antibody conjugated to a peptide, a fluorescent molecule, or a chemotherapeutic agent.

[0082] The IL-36R conjugate may be a human antibody, a non-human antibody, or a chimeric antibody, or may be derived from them. A "chimeric" antibody is an antibody or fragment thereof that contains both a human region and a non-human region. Preferably, the IL-36R conjugate is a humanized antibody. A "humanized" antibody is a monoclonal antibody comprising a human antibody scaffold and at least one CDR obtained from or derived from a non-human antibody. Examples of non-human antibodies include antibodies isolated from any non-human animal, such as rodents (e.g., mice or rats). The humanized antibody may contain one, two, or three CDRs obtained from or derived from a non-human antibody. In one embodiment of the present invention, the CDRH3 of the IL-36R conjugate is obtained from or derived from a mouse monoclonal antibody, while the remaining variable and constant regions of the IL-36R conjugate are obtained from or derived from a human monoclonal antibody.

[0083] Human antibodies, non-human antibodies, chimeric antibodies, or humanized antibodies are supplied from in vitro sources (e.g., hybridomas or recombinant antibody-producing cell lines) and in vivo sources (e.g., It can be obtained by any means, including via rodents. Methods for generating antibodies are known in the art, for example, Kohler and Milstein. This is described in Eur.J.Immunol., 5:511-519 (1976); Harlow and Lane (eds.), Antibodies: A Laboratory Manual, CSH Press (1988); and Janeway et al. (eds.), Immunobiology, 5th Ed., Garland Publishing, New York, NY (2001). In certain embodiments, human antibodies or chimeric antibodies can be generated using transgenic animals (e.g., mice) in which one or more endogenous immunoglobulin genes are replaced with one or more human immunoglobulin genes. Examples of transgenic mice in which the endogenous antibody gene is effectively replaced with a human antibody gene include, but are not limited to, Medarex HUMAB-MOUSE®, Kirin TC MOUSE®, and Kyowa Kirin KM-MOUSE® (see, for example, Lonberg, Nat. Biotechnol., 23(9):1117-25 (2005) and Lonberg, Handb. Exp. Pharmacol., 181:69-97 (2008)). Humanized antibodies can be produced using any suitable method known in the art, for example, grafting non-human CDRs onto a human antibody scaffold (see, for example, Kashmiri et al., Methods, 36(1):25-34 (2005); and Hou et al., J. Biochem., 144(1):115-120 (2008) (see, for example, An, Z. (ed.), Therapeutic Monoclonal Antibodies: From Bench to Clinic, John Wiley & Sons, Inc., Hoboken, New Jersey (2009)). In one embodiment, humanized antibodies can be produced using, for example, the method described in U.S. Patent Application Publication 2011 / 0287485 A1.

[0084] In one embodiment, the CDRs (e.g., CDR1, CDR2, or CDR3) or variable regions of the immunoglobulin heavy chain polypeptide and / or immunoglobulin light chain polypeptide described herein can be transplanted (i.e., grafted) onto another molecule, such as an antibody or non-antibody polypeptide, using either protein chemistry or recombinant DNA technology. In this regard, the present invention encompasses an IL-36R binder comprising at least one CDR of the immunoglobulin heavy chain and / or light chain polypeptide described herein. The IL-36R binder may comprise one, two, or three CDRs of the immunoglobulin heavy chain and / or light chain variable regions described herein.

[0085] In a preferred embodiment, the IL-36R conjugate binds to an IL-36R epitope, blocking the binding of IL-36R to any of its ligands (e.g., IL-36α, IL-36β, and IL-36γ), thereby inhibiting IL-36R-mediated signaling. The methods of the present invention also encompass isolated or purified epitopes of IL-36R that indirectly or allosterically block the binding of IL-36R to any of its ligands.

[0086] Furthermore, nucleic acids encoding at least the variable region of the heavy or light chain of an IL-36R binder are provided herein. In one embodiment, the nucleic acid encodes the immunoglobulin light chain variable region or the entire immunoglobulin light chain of an IL-36R binder. In another embodiment, the nucleic acid encodes the immunoglobulin heavy chain variable region or the entire immunoglobulin heavy chain of an IL-36R binder. In yet another embodiment, the nucleic acid encodes both the immunoglobulin light chain variable region or the entire immunoglobulin light chain and the immunoglobulin heavy chain variable region or the entire immunoglobulin heavy chain provided herein.

[0087] The terms "nucleic acid" and "nucleic acid sequence" refer to polymers of DNA or RNA, which may be single-stranded or double-stranded and may contain unnatural or altered nucleotides. That is, it is intended to encompass polynucleotides. The terms “nucleic acid” and “polynucleotide,” as used herein, refer to polymeric forms of nucleotides of any length, which are either ribonucleotides (RNA) or deoxyribonucleotides (DNA). These terms refer to the primary structure of molecules and therefore include double-stranded and single-stranded DNA and double-stranded and single-stranded RNA. The terms include, as equivalents, analogs of either RNA or DNA made from modified polynucleotides, such as nucleotide analogs and methylated and / or capped polynucleotides, etc. Nucleic acids are typically linked via phosphate bonds to form nucleic acid sequences or polynucleotides, although many other linkages are also known in the art (e.g., phosphorothioates, boranophosphates, etc.).

[0088] Nucleic acids may be part of the vector. The vector may be, for example, a plasmid, episome, cosmid, viral vector (e.g., retrovirus or adenovirus), or phage. Suitable vectors and methods for vector preparation are well known in the art (see, for example, Sambrook et al., Molecular Cloning, a Laboratory Manual, 3rd edition, Cold Spring Harbor Press, Cold Spring Harbor, NY (2001) and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and John Wiley & Sons, New York, NY (1994)).

[0089] In addition to nucleic acid sequences encoding the heavy and / or light chains of immunoglobulins, vectors may include expression regulatory sequences that cause the coding sequence to be expressed in host cells, such as promoters, enhancers, polyadenylation signals, transcription terminators, and intra-sequence ribosome entry sites (IRESs). Exemplary expression regulatory sequences are known in the art and are described, for example, in Goeddel, Gene Expression Technology: Methods in Enzymology, Vol. 185, Academic Press, San Diego, Calif. (1990).

[0090] Numerous promoters, including constitutive, inducible, and repressive promoters from various different sources, are well known in the art. Typical sources of promoters include, for example, viruses, mammals, insects, plants, yeasts, and bacteria. Suitable promoters from these sources are readily available or can be synthesized based on sequences publicly available from depositaries such as ATCC and other commercial or private sources. Promoters may be unidirectional (i.e., initiate transcription in one direction) or bidirectional (i.e., initiate transcription in either the 3' or 5' direction). Non-limiting examples of promoters include, for example, the T7 bacterial expression system, the pBAD(araA) bacterial expression system, the cytomegalovirus (CMV) promoter, the SV40 promoter, and the RSV promoter. Examples of inducible promoters include the Tet system (US Patent Nos. 5,464,758 and 5,814,618), the Ecdysone-inducible system (No et al., Proc. Natl. Acad. Sci., 93:3346-3351 (1996)), the T-REX (Trademark) system (Invitrogen, Carlsbad, CA), the LACSWITCH (Trademark) system (Stratagene, San Diego, CA), and the Cre-ERT tamoxifen-inducible recombinase system (Indra et al., Nuc. Acid. Res., 27:4324-4327 (1999); Nuc. Acid. Res., 28:e99 (2000); US Patent No. 7,112,715; and Kramer & Fussenegger, Methods Mol. Biol., 308:123- 144 (2005) is one example.

[0091] As used herein, the term “enhancer” refers, for example, to a DNA sequence that increases the transcription of a nucleic acid sequence to which it is operably ligated. Enhancers may be located several kilobases away from the coding region of the nucleic acid sequence and can mediate the binding of regulatory factors, the pattern of DNA methylation, or changes in DNA structure. Numerous enhancers from various different sources are well known in the art and are available as cloned polynucleotides or within such polynucleotides (e.g., from depositary institutions such as ATCC, and other commercial or private sources). Many polynucleotides containing promoters (such as the commonly used CMV promoter) also contain enhancer sequences. Enhancers may be located upstream, within, or downstream of the coding sequence.

[0092] The vector may also contain a selection marker gene. As used herein, the term “selection marker gene” means a nucleic acid sequence that, in the presence of a corresponding selector, enables cells expressing that nucleic acid sequence to be positively or negatively specifically selected. Suitable selection marker genes are known in the art, for example, in International Publication Nos. 1992 / 008796 and 1994 / 028143; Wigler et al, Proc. Natl. Acad. Sci. USA, 77:3567-3570 (1980); O'Hare et al., Proc. Natl. Acad. Sci. USA, 78:1527-1531 (1981); Mulligan & Berg, Proc. Natl. Acad. Sci. USA, 78:2072-2076 (1981); Colberre-Garapin et al., J. Mol. Biol., 150:1-14 (1981); Santerre et al., Gene, 30:147-156 (1984); Kent et al. This information is also found in: al., Science, 237:901-903 (1987); Wigler et al., Cell, 11:223-232 (1977); Szybalska & Szybalski, Proc. Natl. Acad. Sci. USA, 48:2026-2034 (1962); Lowy et al., Cell, 22:817-823 (1980); and U.S. Patent Nos. 5,122,464 and 5,770,359.

[0093] In some embodiments, the vector is an “episomal expression vector” or “episome” that is replicable within the host cell and persists as an extrachromosomal segment of DNA within the host cell in the presence of appropriate selective pressure (see, e.g., Conese et al., Gene Therapy, 11:1735-1742 (2004)). Representative commercially available episomal expression vectors include, but are not limited to, episomal plasmids utilizing Epstein-Barr virus nuclear antigen 1 (EBNA1) and Epstein-Barr virus (EBV) origin of replication (oriP). Vectors pREP4, pCEP4, pREP7, and pcDNA3.1 from Invitrogen (Carlsbad, CA), and pBK-CMV from Stratagene (La Jolla, CA) represent non-exclusive examples of episomal vectors that use the T-antigen and SV40 origin of replication instead of EBNA1 and orip.

[0094] Other suitable vectors include embedded expression vectors that may be randomly integrated into the host cell's DNA or may contain recombination sites that enable specific recombination between the expression vector and the host cell's chromosomes. Such embedded expression vectors can express desired proteins by utilizing endogenous expression regulatory sequences on the host cell's chromosomes. Examples of site-specific embedded vectors include, for example, the flp-in system from Invitrogen (Carlsbad, CA) (e.g., pcDNA® 5 / FRT) or components of the cre-lox system found in, for example, the pExchange-6 core vector from Stratagene (La Jolla, CA). Examples of vectors that are randomly incorporated into chromosomes include pcDNA3.3 from ThermoFisher (Carlsbad, CA) (when introduced in the absence of the T-antigen), UCOE from Millipore (Billerica, MA), and pCI or pFN10A(ACT)FLEXI® from Promega (Madison, WI).

[0095] Viral vectors may also be used. Representative commercially available viral expression vectors include the adenovirus-based Per.C6 series from Crucell, Inc. (Leiden, The Netherlands), the lentivirus-based pLP1 from ThermoFisher (Carlsbad, CA), and the retroviral vector pFB-RV from Agilent (Stratagene, La Jolla, CA). Examples include, but are not limited to, pCFB-EGSH.

[0096] The nucleic acid sequences encoding the amino acid sequences of the present invention can be supplied to cells on the same vector (i.e., in cis). The expression of each nucleic acid sequence can be controlled using a unidirectional promoter. In another embodiment, the expression of multiple nucleic acid sequences can be controlled using a combination of bidirectional and unidirectional promoters. Alternatively, the nucleic acid sequences encoding the amino acid sequences of the present invention can be supplied to a population of cells on separate vectors (i.e., in trans). Each nucleic acid sequence in each separate vector may contain the same or different expression regulatory sequences. The separate vectors can be supplied to cells simultaneously.

[0097] A vector(s) containing nucleic acids(s) encoding the amino acid sequence of the present invention may be introduced into a host cell(s) (including any suitable prokaryotic or eukaryotic cell) capable of expressing the polypeptide encoded thereby. Thus, the present invention provides in vitro cells or cell lines containing the vector of the present invention. The present invention also provides in vitro cells or cell lines expressing immunoglobulin heavy chain and / or light chain polypeptides or IL-36R conjugates. Preferred host cells are those that can be easily and reliably grown, have a moderately fast growth rate, have a well-characterized expression system, and can be easily and efficiently transformed or transfected.

[0098] Suitable prokaryotic cells include, but are not limited to, cells of the genera Bacillus (such as Bacillus subtilis and Bacillus brevis), Escherichia (such as Escherichia coli), Pseudomonas, Streptomyces, Salmonella, and Erwinia. Particularly useful prokaryotic cells include various strains of Escherichia coli (e.g., K12, HB101 (ATCC number 33694), DH5α, DH10, MC1061 (ATCC number 53338), and CC102).

[0099] In some embodiments, the vector is introduced into eukaryotic cells. Suitable eukaryotic cells are known in the art and include, for example, yeast cells, insect cells, and mammalian cells. Examples of suitable yeast cells include those from the genera Kluyveromyces, Pichia, Rhino-sporidium, Saccharomyces, and Schizosaccharomyces. Preferred yeast cells include, for example, Saccharomyces cerivisae and Pichia pastoris.

[0100] Suitable insect cells are, for example, Kitts et al., Biotechniques This is described in 14:810-817 (1993); Lucklow, Curr. Opin. Biotechnol., 4:564-572 (1993); and Lucklow et al., J. Virol., 67:4566-4579 (1993). Preferred insect cells include Sf-9 and HI5 (Invitrogen, Carlsbad, CA).

[0101] In some embodiments, the present invention utilizes mammalian cells. Numerous suitable mammalian host cells are known in the art, many of which are available from the American Type Culture Collection (ATCC, Manassas, VA). Examples of suitable mammalian cells include, but are not limited to, Chinese hamster ovary cells (CHO) (e.g., ATCC number CCL61), CHO DHFR- cells (e.g., Urlaub et al., Proc. Natl. Acad. Sci. USA, 97:4216-4220 (1980)), human embryonic kidney (HEK) 293 or 293T cells (e.g., ATCC number CRL1573), and 3T3 cells (e.g., ATCC number CCL92). Other suitable mammalian cell lines include monkey COS-1 (e.g., ATCC number CRL1650) and COS-7 cell lines (e.g., ATCC number CRL1651), and CV-1 cell line (e.g., ATCC number CCL70). Further exemplary mammalian host cells include primate and rodent cell lines, including mouse cell line NS0, derivatives of mouse myeloma cell line MOPC21 (e.g., Tysabri), and transformed cell lines. In addition to conventional diploid cells and cell lines obtained from in vitro culture of primary tissues, primary explants are also suitable. Other suitable mammalian cell lines include, but are not limited to, mouse neuroblastoma N2A cells, HeLa, mouse L-929 cells, and BHK or HaK hamster cell lines, all of which are available from ATCC. Methods for selecting suitable mammalian host cells, as well as methods for cell transformation, culture, amplification, screening, and purification, are known in the art.

[0102] In some embodiments, the mammalian cells are human cells. For example, the mammalian cells may be human lymphocytes or lymphocyte-derived cell lines, such as cell lines of pre-B lymphocyte origin. Examples of human lymphocyte cell lines include, but are not limited to, RAMOS (e.g., CRL-1596), Daudi (e.g., CCL-213), EB-3 (e.g., CCL-85), Raji cells (e.g., CCL-86), and their derivatives. The nucleic acid sequence encoding the amino acid sequence of the present invention can be introduced into cells by any suitable technique such as "transfection," "transformation," or "transduction." "Transfection," "transformation," or "transduction," as used herein, refers to the introduction of one or more exogenous polynucleotides into a host cell by physical or chemical methods. Many transfection techniques are known in the art, including, for example, calcium phosphate DNA coprecipitation (e.g., Murray EJ (ed.), Methods in Molecular Biology, Vol. 7, Gene Transfer and Expression Protocols, Humana Press (1991)); DEAE-dextran; electroporation; cationic liposome-mediated transfection; tungsten particle-enhanced microparticle gun (Johnston, Nature, 346:776-777 (1990)); and strontium phosphate DNA coprecipitation (Brash et al., Mol. Cell Biol., 7:2031-2034 (1987)). Infection particles may be grown in suitable packaging cells, many of which are commercially available, and then the phage or viral vector may be introduced into the host cell.

[0103] Nucleic acids and cells can be used for any purpose, such as the production of IL-36R binders as described herein. In this regard, the present invention includes culturing cells containing nucleic acids encoding heavy and / or light immunoglobulin polypeptides of IL-36R binders, A method for preparing IL-36R conjugates is provided. In other words, the method involves expressing in cells the nucleic acids encoding the heavy and / or light chains of the immunoglobulin of the IL-36R conjugate. It will be understood that the heavy and light chains of the immunoglobulin may be expressed from a single nucleic acid in a given cell, or the heavy and light chains of the immunoglobulin may be expressed from separate nucleic acids in the same cell. The method may further include collecting and / or purifying the IL-36R conjugate from cells or cell culture media using known techniques.

[0104] The present invention further encompasses compositions comprising an effective amount of an IL-36 pathway inhibitor (e.g., an IL-36R conjugate) or a nucleic acid sequence encoding it, and a pharmaceutically acceptable (e.g., physiologically acceptable) carrier. The choice of carrier will be determined in part by the specific site to which the composition can be administered and the specific method used to administer the composition. The composition may optionally be sterile. The composition may be frozen or lyophilized for storage and reconstituted in a suitable sterile carrier before use. The composition may be produced according to the prior art described, for example, Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins, Philadelphia, PA (2001).

[0105] The following examples further illustrate the present invention, but should not be construed as limiting its scope in any way. The compositions can be used in any of the methods described herein. [Examples]

[0106] Example 1 This embodiment demonstrates that patients with ichthyosis show increased expression of the IL-36 cytokine and IL-36R gene and protein in the skin.

[0107] Skin biopsies were collected from 60 patients of all ages with at least 15 different genotypes underlying orphan ichthyosis (e.g., congenital erythrodermic ichthyosis, phyllodes ichthyosis, epidermolytic ichthyosis, Netherton syndrome, and confetti ichthyosis) and age-matched controls (measured age 0 to <6 years, 6 to <12 years, 12 to <18 years, and adults). Buccal swabs or saliva samples from patients were collected to confirm genotype, and if unknown, the appropriate subtype of ichthyosis was identified.

[0108] A biopsy was performed from the upper arm for gene expression analysis and extraction. Blood was collected for serum proteomics analysis using the OLINK proseek proteomics platform, and nearly 300 inflammatory biomarkers were detected in the serum.

[0109] Gene expression data obtained from RNA-seq experiments were preprocessed using Harshlight for quality control, the GCRMA algorithm for normalization, and GC content background correction. Expression values, converted to Log2 for microarrays, were modeled using a mixed-effects linear model with severity as a fixed factor and patient as a random effect, allowing for consideration of within-patient correlation structures. Hypothesis testing of target comparisons was performed for specified contrasts using the R limma framework. P-values ​​were adjusted for multiplicity using the Benjamini and Hochberg approaches. Genes with differing expression (DEGs) were identified using FCH>2 and FDR<0.05 as cutoff values. Downstream analysis included extensive use of bioinformatics tools providing biological insights, including Ingenuity Pathway Analysis (IPA). Gene Set Enrichment Analysis (GSEA) and Gene Set Variation Analysis (GSEA) were performed to obtain enrichment scores for AD-related pathways. We used on Analysis (GSVA).

[0110] As is evident from Figures 1A-D and 2A and 2B, patients with ichthyosis showed significantly higher expression levels of the IL-36 cytokine and IL-36R gene and protein compared to controls.

[0111] Example 2 This example demonstrates the effect of an IL-36R conjugate (e.g., an anti-IL-36R antibody) on ichthyosis.

[0112] Adolescent and adult males and females with a clinically confirmed diagnosis of ichthyosis were selected for treatment with either placebo or an IL-36R conjugate. Participants may have a diagnosis of a subtype of ichthyosis confirmed by genetic testing, and on day 1, may have a total Ichthyosis Area Severity Index (IASI) score of at least 18, an erythema score of at least 2 (moderate severity) in at least one body area as assessed by IASI, a scaling score of at least 2 (moderate severity) in at least one body area, and a BSI related to ichthyosis of at least 50%.

[0113] Subjects may be randomized (e.g., 2:1) to receive either an IL-36R conjugate (e.g., heavy chain variable region SEQ ID NO: 22, light chain variable region SEQ ID NO: 44, or an antibody containing at least its CDR) or a placebo subcutaneously. For example, four doses may be administered. For example, on day 1, subjects receive a 400 mg dose of either the IL-36R conjugate or placebo. On days 29, 57, and 85, subjects receive a 200 mg dose of either the IL-36R conjugate or placebo.

[0114] Disease activity can be assessed using the IASI, IASI erythema (IASI-E) subscore, IASI scaling (IASI-S) subscore, Netherton area and severity assessment (NASA) for subjects with Netherton syndrome only, physician's general assessment (IGA), and body surface area (BSA) associated with ichthyosis. Quality of life can be assessed using the iQoL-32 (Ichthyosis Quality of Life-32) for subjects 15 years of age and older, the Dermatological Quality of Life Index (DLQI) for subjects 16 years of age and older, and the Pediatric Dermatological Quality of Life Index (CDLQI) for subjects under 16 years of age. Subjects are also assessed for disease-related characteristics such as worst and mean pruritus and pain using a numerical rating scale (NRS), patient's general impression of severity using the Patient General Impression of Severity-S, patient's general impression of change using the Patient General Impression of Change (PGI-C), and changes in transepidermal water loss (TEWL).

[0115] Administration of IL-36R conjugates treats ichthyosis compared to placebo administration by reducing disease activity, improving quality of life, and / or decreasing disease-related features in subjects.

[0116] All references cited herein, including publications, patent applications, and patents, are incorporated herein by reference to the same extent as if they were included herein, each reference being indicated as being incorporated by reference individually and specifically.

[0117] In connection with the description of the present invention (particularly in connection with the following claims), the use of the terms “a,” “an,” “the,” and “at least one,” as well as similar references, should be construed to encompass both singular and plural forms unless otherwise specified herein or unless clearly contradicted by the context. The term “at least one” following a list of one or more items The use of "at least one" (e.g., "at least one of A and B") should be interpreted as meaning one item (A or B) selected from the enumerated items or any combination of two or more items (A and B) unless otherwise specified herein or unless clearly contradicted by the context. The terms "comprising," "having," "including," and "containing" should be interpreted as open-ended terms (i.e., meaning "including but not limited to") unless otherwise specified herein. Enumerations of value ranges herein are intended, unless otherwise specified herein, simply to function as an abbreviation for each distinct value within the range individually, and each distinct value is incorporated into the specification as if it were individually enumerated herein. All methods described herein may be performed in any preferred order unless otherwise specified herein or unless clearly contradicted by the context. Any and all examples or illustrative expressions provided herein (e.g., "etc.") are intended, unless otherwise specifically asserted, merely to further illustrate the invention and not to propose any limitation of its scope. No expression in the specification should be construed as indicating that any unclaimed element is essential for the practice of the invention.

[0118] Preferred embodiments of the Invention, including the best mode known to the inventors for carrying out the Invention, are described herein. Variations of the preferred embodiments may become apparent to those skilled in the art upon reading the foregoing description. The inventors anticipate that those skilled in the art will use such variations as appropriate, and they intend that the Invention may be carried out in ways other than those specifically described herein. Accordingly, the Invention includes all variations and equivalents of the subject matter of the Invention enumerated in the claims appended herein, as permitted by applicable law. Furthermore, any combination of the above elements in all possible variations is encompassed by the Invention unless otherwise specified herein or is clearly inconsistent with the context.

Claims

1. A method for treating ichthyosis in a subject, comprising inhibiting IL-36 signaling in the subject, thereby treating the ichthyosis.

2. The method according to claim 1, wherein the ichthyosis is selected from congenital ichthyoid erythroderma, phyllodes ichthyosis, epidermolytic ichthyosis, Netherton syndrome, and confetti ichthyosis.

3. The method according to claim 1 or 2, wherein the subject is a mammal.

4. The method according to any one of claims 1 to 3, wherein the mammal is a human.

5. The method according to any one of claims 1 to 4, wherein in the subject, the expression of at least one of the interleukin-36 (IL-36) cytokine, interleukin-36 receptor (IL-36R), or mRNA encoding it is increased in the skin.

6. The method according to claim 5, wherein the IL-36 cytokine is IL-36α, IL-36β, or IL-36γ.

7. The method according to any one of claims 1 to 6, wherein IL-36 signaling is inhibited by administering an IL-36 pathway inhibitor, or optionally an IL-36R conjugate, to the subject.

8. The IL-36R binder comprises an immunoglobulin heavy chain polypeptide and an immunoglobulin light chain polypeptide, wherein the immunoglobulin heavy chain polypeptide is Gln Val Gln Xaa1 Xaa2 Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Thr Ser Tyr Asp Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Xaa3 Leu Glu Trp Met Gly Trp Ile Tyr Pro Gly Asp Xaa4 Ser Thr Lys Tyr Asn Glu Lys Phe Lys Gly Arg Val Thr Ile Thr Xaa5 Asp Xaa6 Ser Ala Xaa7 Thr Ala Tyr Met Glu Leu Xaa8 Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Xaa9 Cys Thr Arg Ser Phe Tyr Thr Met Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser (Sequence ID 56) (In the formula, (a) Xaa1 is leucine (Leu) or phenylalanine (Phe), (b) Xaa2 is valine (Val), methionine (Met), or leucine (Leu), (c) Xaa3 is arginine (Arg) or glycine (Gly), (d) Xaa4 is glycine (Gly), serine (Ser), or alanine (Ala), (e) Xaa5 is arginine (Arg) or alanine (Ala), (f) Xaa6 is threonine (Thr) or lysine (Lys), (g) Xaa7 is serine (Ser) or asparagine (Asn), (h) Xaa8 is serine (Ser) or alanine (Ala), (i) Xaa9 is tyrosine (Tyr) or phenylalanine (Phe). or Gln Val Gln Xaa1 Xaa2 Gln Ser Gly Ala Gl u Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Phe Thr Phe Thr Ser Tyr Asp Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Xaa3 Leu Glu Trp Met Gly Trp Ile Tyr Pro Gly Asp Xaa4 Ser Thr Lys Tyr Asn Glu Lys Phe Lys Gly Arg Val Thr Ile Thr Xaa5 Asp Xaa6 Ser Ala Ser Thr Ala Tyr Met Glu Leu Xaa7 Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Xaa8 Cys Thr Arg Ser Phe Tyr Thr Met Asp Tyr Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser (SEQ ID NO: 1) (in the formula, (a) Xaa1 is leucine (Leu) or phenylalanine (Phe), (b) Xaa2 is valine (Val), methionine (Met), or leucine (Leu), (c) Xaa3 is arginine (Arg) or glycine (Gly), (d) Xaa4 is glycine (Gly), serine (Ser), or alanine (Ala), (e) Xaa5 is arginine (Arg) or alanine (Ala), (f) Xaa6 is threonine (Thr) or lysine (Lys), (g) Xaa7 is serine (Ser) or alanine (Ala), (h) Xaa8 is tyrosine (Tyr) or phenylalanine (Phe). or Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr Xaa1 Met Xaa2 Trp Val Arg Gln Ala Pro Xaa3 Gln Gly Leu Glu Trp Met Gly Met Phe Xaa4 Pro Xaa5 Xaa6 Xaa7 Val Thr Arg Leu Asn Gln Lys Phe Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Thr Ser Met Ile Ile Gly Gly Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser (SEQ ID NO: 15) (in the formula, (a) Xaa1 is tryptophan (Trp) or tyrosine (Tyr), (b) Xaa2 is histidine (His), asparagine (Asn), or tyrosine (Tyr), (c) Xaa3 is glycine (Gly) or arginine (Arg), (d) Xaa4 is aspartic acid (Asp), glutamic acid (Glu), or histidine (His), (e) Xaa5 is serine (Ser), threonine (Thr), or tyrosine (Tyr), (f) Xaa6 is asparagine (Asn) or glycine (Gly), (g) Xaa7 is serine (Ser), alanine (Ala), or aspartic acid (Asp). or Xaa1 Xaa2 Gln Xaa3 Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser L eu Thr Cys Thr Val Xaa4 Xaa5 Tyr Ser Ile Thr Xaa6 Asp Phe Ala Trp Asn Trp Ile Arg Gln Xaa7 Pro Gly Xaa8 Xaa9 Leu Glu Trp Ile Gly Tyr Ile Ser Tyr Ser Gly Asp Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Xaa10 Xaa11 Asp Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Ser Val Thr Ala Ala Asp Thr Ala Xaa12 Tyr Xaa13 Cys Ala Ile Arg Gly Pro Tyr Ser Phe Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Xaa14 (Sequence No. 57) (In the formula, (a) Xaa1 is glutamine (Gln) or aspartic acid (Asp), (b) Xaa2 is valine (Val) or leucine (Leu), (c) Xaa3 is leucine (Leu) or phenylalanine (Phe), (d) Xaa4 is threonine (Thr) or serine (Ser), (e) Xaa5 is glycine (Gly) or arginine (Arg), (f) Xaa6 is serine (Ser) or alanine (Ala), (g) Xaa7 is proline (Pro) or phenylalanine (Phe), (h) Xaa8 is lysine (Lys) or asparagine (Asn), (i) Xaa9 is glycine (Gly) or lysine (Lys), (j) Xaa10 is serine (Ser) or threonine (Thr), (k) Xaa11 is valine (Val) or arginine (Arg), (l) Xaa12 is threonine (Thr) or valine (Val), (m)Xaa13 is tyrosine (Tyr) or phenylalanine (Phe), (n) Xaa14 is either alanine (Ala) or absent. or Xaa1 Val Gln Xaa2 Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Xaa3 Gly Tyr Ser Ile Thr Ser Asp Phe Ala Trp Asn Trp Ile Arg Gln Xaa4 Pro Gly Xaa5 Xaa6 Leu Glu Trp Ile Gly Tyr Ile Ser Tyr Ser Gly Asp Thr Asn Tyr Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Xaa7 Xaa8 Asp Thr Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Xaa9 Cys Ala Ile Arg Gly Pro Tyr Ser Phe Thr Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser (Sequence ID 25) (In the formula, (a) Xaa1 is glutamine (Gln) or aspartic acid (Asp), (b) Xaa2 is leucine (Leu) or phenylalanine (Phe), (c) Xaa3 is threonine (Thr) or serine (Ser), (d) Xaa4 is proline (Pro) or phenylalanine (Phe), (e) Xaa5 is lysine (Lys) or asparagine (Asn), (f) Xaa6 is glycine (Gly) or lysine (Lys), (g) Xaa7 is serine (Ser) or threonine (Thr), (h) Xaa8 is valine (Val) or arginine (Arg), (i) Xaa9 is tyrosine (Tyr) or phenylalanine (Phe). or Any one of sequence numbers 2-14, 16-24, 26-35, or 51-54 Tsu; or at least CDR1, CDR2, and CDR3 Including; The immunoglobulin light chain polypeptide is Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Ser Lys Ser Leu Leu His Ser Asn Xaa1 Asn Thr Tyr Leu Tyr Trp Xaa2 Leu Gln Lys Pro Gly Gln Ser Pro Gln Leu Leu Ile Xaa3 Arg Met Ser Asn Leu Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln His Leu Glu Tyr Pro Phe Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys (Sequence No. 36) (In the formula, (a) Xaa1 is glycine (Gly) or alanine (Ala), (b) Xaa2 is phenylalanine (Phe) or tyrosine (Tyr), (c) Xaa3 is tyrosine (Tyr) or serine (Ser) or Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu His Xaa1 Asn Xaa2 Ile Thr Tyr Phe Tyr Trp Tyr Leu Xaa3 Lys Pro Gly Gln Pro Pro Gln Leu Leu Ile Tyr Gln Met Ser Asn Leu Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ala Gln Asn Leu Glu Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys (SEQ ID NO: 40) (a) Xaa1 is serine (Ser) or arginine (Arg), (b) Xaa2 is glycine (Gly) or alanine (Ala), (c) Xaa3 is glutamine (Gln) or histidine (His)) or Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Xaa1 Ile Asn Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Tyr Thr Ser Xaa2 Leu His Ser Gly Val Pro Ser Arg Phe Ser Xaa3 Ser Gly Ser Gly Xaa4 Asp Xaa5 Thr Phe Thr Ile Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly His Thr Leu Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Xaa6 Xaa7 (Sequence No. 58) (In the formula, (a) Xaa1 is aspartic acid (Asp) or tryptophan (Trp), (b) Xaa2 is arginine (Arg) or methionine (Met), (c) Xaa3 is glycine (Gly), serine (Ser), or proline (Pro), (d) Xaa4 is threonine (Thr) or asparagine (Asn), (e) Xaa5 is phenylalanine (Phe) or tyrosine (Tyr), (f) Xaa6 is either arginine (Arg) or absent. (g) Xaa7 is either threonine (Thr) or absent. or Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Ser Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Asn Asn Tyr Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Xaa1 Ser Gly Ser Gly Thr Asp Xaa2 Thr Phe Thr Ile Ser Ser Ser Leu Gln Pro Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Gly His Thr Leu Pro Trp Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys (SEQ ID NO: 45) (in the formula, (a) Xaa1 is serine (Ser) or proline (Pro), (b) Xaa2 is phenylalanine (Phe) or tyrosine (Tyr). Does it include; Or any one of sequence numbers 37-39, 41-44, 46-50, or 55; or at least CDR1, CDR2, and CDR3 The method according to claim 7, including the method described in claim 7.

9. The immunoglobulin heavy chain polypeptide of the IL-36R binder is Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr Xaa1 Met Xaa2 Trp Val Arg Gln Ala Pro Xaa3 Gln Gly Leu Glu Trp Met Gly Met Phe Xaa4 Pro Xaa5 Xaa6 Xaa7 Val Thr Arg Leu Asn Gln Lys Phe Lys Asp Arg Val Thr Met Thr Arg Asp Thr Ser Thr Ser Thr Val Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Thr Thr Ser Met Ile Ile Gly Gly Phe Ala Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser (SEQ ID NO: 15) (in the formula, (a) Xaa1 is tryptophan (Trp) or tyrosine (Tyr), (b) Xaa2 is histidine (His), asparagine (Asn), or tyrosine (Tyr), (c) Xaa3 is glycine (Gly) or arginine (Arg), (d) Xaa4 is aspartic acid (Asp), glutamic acid (Glu), or histidine (His), (e) Xaa5 is serine (Ser), threonine (Thr), or tyrosine (Tyr), (f) Xaa6 is asparagine (Asn) or glycine (Gly), (g) Xaa7 is serine (Ser), alanine (Ala), or aspartic acid (Asp). or at least CDR1, CDR2, and CDR3 Including; The immunoglobulin light chain of the IL-36R binder is Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Ser Val Thr Pro Gly Gln Pro Ala Ser Ser Ile Ser Cys Arg Ser Ser Ser Lys Ser Leu Leu His Xaa1 Asn Xaa2 Ile Thr Tyr Phe Tyr Trp Tyr Leu Xaa3 Lys Pro Gly Gln Pro Pro Gln Leu Leu Ile Tyr Gln Met Ser Asn Leu Ala Ser Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ala Gln Asn Leu Glu Leu Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys (SEQ ID NO: 40) (a) Xaa1 is serine (Ser) or arginine (Arg), (b) Xaa2 is glycine (Gly) or alanine (Ala), (c) Xaa3 is glutamine (Gln) or histidine (His) or at least CDR1, CDR2, and CDR3 The method according to claim 8, including the method described in claim 8.

10. The method according to claim 8 or 9, wherein the immunoglobulin heavy chain polypeptide of the IL-36R binder comprises SEQ ID NO: 22 or at least its CDR.

11. The method according to any one of claims 8 to 10, wherein the immunoglobulin light chain polypeptide of the IL-36-binding agent comprises SEQ ID NO: 44 or at least its CDR.

12. The method according to any one of claims 7 to 11, wherein the IL-36R binder exhibits one or more of the following biological activities: (a) Inhibit the interaction between IL-36R and IL-36α, IL-36β, and / or IL-36γ, (b) Inhibit intracellular signaling mediated by IL-36R, (c) It cross-reacts with human IL-36R, cynomolgus monkey IL-36R, and non-human primate IL-36R, inhibiting their activity.

13. The method according to any one of claims 7 to 12, wherein the IL-36R binder is an antibody, an antibody conjugate, or an antigen-binding fragment thereof.

14. The method according to claim 13, wherein the IL-36R binder is an F(ab')2 fragment, a Fab' fragment, a Fab fragment, an Fv fragment, an scFv fragment, a dsFv fragment, a dAb fragment, or a single-strand linked polypeptide.

15. The method according to claim 7, wherein the IL-36R binder competes with the IL-36R binder described in any one of claims 8 to 14 for binding to IL-36R.

16. The method according to any one of claims 7 to 15, wherein the half-life of the IL-36R binder in mammals is 30 minutes to 45 days.

17. The method according to any one of claims 7 to 16, wherein the IL-36R binder binds to IL-36R at a KD of about 1 picomolar concentration (pM) to about 100 micromolar concentration (μM).

18. The method according to any one of claims 7 to 17, comprising administering an effective amount of a composition comprising (a) an IL-36R binder according to any one of claims 8 to 17 and (b) a pharmaceutically acceptable carrier.

19. An IL-36R conjugate for treating ichthyosis in a subject according to any one of claims 7 to 18.

20. Use of an IL-36R binder for preparing a medicament for treating ichthyosis in a subject according to any one of claims 7 to 18.

21. A method for selecting subjects with ichthyosis for treatment with IL-36 pathway inhibitors, The expression of at least one of the IL-36 cytokine, IL-36R, or mRNA encoding it in skin samples from subjects before and after administration of the IL-36 pathway inhibitor, If a decrease in the expression of at least one of the IL-36 cytokine, IL-36R, or the mRNA encoding it is observed in a skin sample from a subject after administration of the IL-36 pathway inhibitor, compared to a sample from the subject before administration of the IL-36 pathway inhibitor, the subject should be selected for treatment. A method that includes this.

22. The method according to claim 21, wherein the ichthyosis is selected from congenital ichthyoid erythroderma, phyllodes ichthyosis, epidermolytic ichthyosis, Netherton syndrome, and confetti ichthyosis.

23. The method according to claim 21 or 22, wherein the subject is a mammal.

24. The method according to any one of claims 21 to 23, wherein the mammal is a human.

25. The method according to any one of claims 21 to 24, wherein the IL-36 cytokine is IL-36α, IL-36β, or IL-36γ.

26. The method according to any one of claims 21 to 25, wherein the inhibitor of the IL-36 pathway is the IL-36R binder according to any one of claims 8 to 17.

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