Peptide modulators of neuroligin 4-neurexin 1-beta axis for treatment of conditions that involve abnormal inflammatory response

Abstract

Provided herein are methods and compositions comprising isolated peptides capable of interfering with neuroligin-4 (NLGn4)-neurexin 1β (Nrx1β) protein-protein interaction (PPI) and uses thereof for treating a condition that involves an abnormal inflammatory response.

Description

[0001] PEPTIDE MODULATORS OF NEUROLIGIN 4-NEUREXIN 1-BETA AXIS FOR TREATMENT OF CONDITIONS THAT INVOLVE ABNORMAL INFLAMMATORY RESPONSE

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to compositions comprising peptides capable of affecting neuroligin-4 (NLGn4)-neurexin i (Nrxip) protein-protein interactions, and uses thereof in treating, preventing and / or attenuating an abnormal inflammatory response.

[0004] BACKGROUND OF THE INVENTION

[0005] Abnormal inflammatory response is associated with diverse pathological states such as autoimmune diseases, acute lung injury (ALI) and cancer.

[0006] Autoimmune diseases are the result of the immune system attacking its own body. Inflammatory bowel disease (IBD) is a group of chronic autoimmune disorders characterized by inflammation of the colon and small intestine. The two main representatives, having increasing incidence and prevalence, are Ulcerative colitis (UC) and Crohn's disease (CD).

[0007] Other forms of IBD, which are not always classified as typical IBD and which include far fewer cases are Collagenous colitis, Lymphocytic colitis, Ischemic colitis, Diversion colitis, Behcet's disease, and Indeterminate colitis. The main difference between CD and UC is the location and nature of the inflammatory changes. CD can affect any part of the gastrointestinal tract, from mouth to anus, although a majority of the cases start in the terminal ileum. UC, in contrast, is restricted to the colon and the rectum. Microscopically, UC is restricted to the mucosa (epithelial lining of the gut), while CD affects the whole bowel wall (transmural lesions). Finally, CD and UC present extra-intestinal manifestations (such as liver complications, arthritis, skin manifestations and eye complications) in different proportions.

[0008] The pathogenesis of IBD is still unclear but in all probability is multifactorial and driven by an exaggerated immune response towards gut microbiome in a genetically susceptible host. Evidence suggests that mis-regulation of leak pathway and intestinal permeability may be crucial for IBD establishment, and that persistent inflammation promotes pathogenesis of IBD, including fibrosis.

[0009] Inflammation results in sustained activation and proliferation of myofibroblasts, leading to extracellular matrix (ECM) over-accumulation. Human intestinal smooth muscle cells (HISMCs) also have a critical role in intestinal fibrosis, responding to chronic inflammation by proliferating and depositing collagen in the ECM. Activated myofibroblasts are considered the final effector cells of intestinal fibrosis. The activated myofibroblasts express high levels of aSMA and therefore represent a markedly increased capability to contract ECM, which in turn contributes to tissue distortion and fibrostenotic stricture.

[0010] In addition, in IBD, imbalance in cholesterol homeostasis and mitochondrial bioenergetics have been identified as essential events for activating the inflammasome- mediated response.

[0011] Surgery is needed in more than 70% of patients with CD within their lifetime to correct severe complications such as intestinal obstruction, perforation, fistula, and abscess. Despite surgery, the rates of postoperative recurrence are high. Endoscopic recurrence by the third postoperative year is reported in 85%— 100% of cases and fibrostenotic stricture is the most common indication for surgery in CD. Surgical resection is related to major morbidity, higher costs, higher risk of unemployment, and poorer quality of life.

[0012] Acute lung injury (ALI) and its most severe manifestation, the acute respiratory distress syndrome (ARDS), is a clinical syndrome defined by acute hypoxemic respiratory failure, bilateral pulmonary infiltrates consistent with edema, and normal cardiac filling pressures. An ALI animal model induced by injection of oleic acid to the animal, reproduces known risk factors for ARDS.

[0013] Neuroligin-4 (NLG4, NLGn4, NLGn4X) is a family member of neuronal cell surface proteins called the Neuroligins. Members of this family are membrane- anchored proteins acting as ligands for P-Neurexin ( -Nrxn), namely neurexin i (Nrxip). P-Nrxn is a member of the presynaptic protein family (Neuroligins ligands) that help to glue together neurons at the synapse. Neurexins are located mostly on the presynaptic membrane and contain a single transmembrane domain.

[0014] Given their amino acid sequence, neurexins are predicted to play a role in extracellular signaling and / or cell adhesion.

[0015] In addition to its role in neurons, the NLGn4X / Nrxip axis has been shown to be related to physiology and pathology of other tissue, for example, the liver.

[0016] International application, publication No. WO 2016 / 157195 related to some of the inventors of the present application is directed to antibodies and recombinant proteins capable of interfering with, inhibiting and / or preventing NLGn4- Nrxip protein-protein interactions (PPI), and to the use thereof for the treatment and / or attenuation of liver disorders.

[0017] Nevertheless, there is an unmet need in the art for improved compositions, which are cost effective, safe and efficient, with reduced side effects, for use in treating at least colon / intestine-related conditions, specifically IBD, by affecting the NLGn4- Nrxip interaction in various cells and tissues including the bowel.

[0018] SUMMARY OF THE INVENTION

[0019] According to some embodiments, herein disclosed is a surprising novel therapeutic target - NLGn4-Nrxip protein-protein interaction (PPI) - for managing the adverse pathological effects associated with conditions in patients that involve abnormal inflammatory response including autoimmune diseases (such as but not limited to Inflammatory bowel disease (IBD)), as well as acute lung injury (ALI) and cancer, through administration of a composition comprising Neuroligin-4 (NLGN4X) inhibitor peptides that, advantageously, is capable of affecting the NLGn4-Nrxip PPI, for example, by interfering or inhibiting the PPI. Each possibility is a separate embodiment.

[0020] Advantageously, according to some embodiments, the NLGN4X inhibitor peptide prevents upregulation of intestinal / colonic P-neurexin (P-Nrxn) expression which is associated with Inflammatory bowel disease (IBD) and / or of lung P-neurexin (P-Nrxn) expression which is associated with acute lung injury (ALI). Each possibility is a separate embodiment.

[0021] Further advantageous, are the disclosed findings that NLGN4X peptides can mitigate the adverse pathological effects associated with IBD, including those related, in some embodiments, to: (i) body weight loss; (ii) morphological changes related to colon size; (iii) severity of inflammation in the bowel; (iv) fibrosis / fibrostenotic strictures in the bowel; and their association with (v) increased paracellular permeability; and (vi) changes in lipid homeostasis, or any combination thereof. Each possibility is a separate embodiment.

[0022] Further advantageous, are the disclosed findings that NLGN4X peptides can mitigate the adverse pathological effects associated with ALI, including those related, in some embodiments, to: (i) increased lung weight; (ii) severity of acute / chronic inflammation in the lung; (iii) fibrosis of the lung; (iv) increment in the number of trNK cells in the lungs; (v) increment in the levels of soluble RAGE; (vi) increment in the levels of surfactant protein D (SP-D), or any combination thereof. Each possibility is a separate embodiment.

[0023] Therefore, it is disclosed that NLGN4X peptides hold a potential as a valuable therapeutic option for alleviating or reducing the pathological effects of conditions in patients that involve abnormal inflammatory response including IBD and / or ALI. Each possibility is a separate embodiment.

[0024] According to one aspect, there is provided a composition comprising an isolated peptide comprising an amino acid sequence as denoted by any one or more of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, for use in treating, attenuating, and / or preventing progression of a condition that involves an abnormal inflammatory response in a subject in need thereof. Each possibility is a separate embodiment.

[0025] In some embodiments, the composition comprises the isolated peptide comprising an amino acid sequence as denoted by SEQ ID NO: 1 and / or SEQ ID NO: 6. Each possibility is a separate embodiment. In some embodiments, the isolated peptide comprises of an amino acid sequence as denoted by SEQ ID NO: 1. In some embodiments, the isolated peptide comprises of an amino acid sequence as denoted by SEQ ID NO: 6.

[0026] In some embodiments, the composition comprises a pharmaceutically acceptable carrier.

[0027] In some embodiments, the composition comprises a plurality of isolated peptides.

[0028] In some embodiments, the condition that involves an abnormal inflammatory response comprises an autoimmune disease.

[0029] In some embodiments, the autoimmune disease is selected from one or more of: Inflammatory bowel disease (IBD), Thyroid diseases including Graves' disease and Hashimoto’s thyroiditis, Adrenal diseases including Addisson disease, Thymus disease including Myasthenia gravis, Multiple sclerosis (MS), Rheumatoid arthritis, Psoriasis and Psoriatic arthritis, Type 1 diabetes, Coeliac disease, Systemic lupus erythematosus, Scleroderma, Polymyositis, Dermatomyositis, and Pernicious anemia, or any combination thereof. Each possibility is a separate embodiment.

[0030] In some embodiments, the IBD is selected from any one or more of: ulcerative colitis (UC), Crohn's disease (CD) Collagenous colitis, Lymphocytic colitis, Ischemic colitis, Diversion colitis, Behcet's disease, and Indeterminate colitis, or any combination thereof. Each possibility is a separate embodiment.

[0031] In some embodiments, the condition that involves an abnormal inflammatory response comprises Acute lung injury (ALI).

[0032] In some embodiments, the ALI comprises acute respiratory distress syndrome (ARDS).

[0033] In some embodiments, the condition that involves an abnormal inflammatory response comprises cancer.

[0034] In some embodiments, administration of the composition comprises systemic and / or localized administration. Each possibility is a separate embodiment.

[0035] In some embodiments, the composition is administered in combination with at least one additional therapeutic agent. In some embodiments, the treatment comprises prevention or reduction of inflammation in the subject.

[0036] In some embodiments, treatment comprises prevention or reduction of fibrosis and / or fibrostenotic strictures in the subject. Each possibility is a separate embodiment.

[0037] In some embodiments, the treatment comprises prevention or reduction of the number of trNK cells in the subject.

[0038] In some embodiments, the treatment comprises prevention or reduction of morphological variations related to colon size of the subject.

[0039] In some embodiments, the treatment comprises prevention or reduction of paracellular permeability and / or susceptibility of the intestine in the subject. Each possibility is a separate embodiment.

[0040] In some embodiments, the treatment comprises prevention or reduction of changes in lipid homeostasis in the subject.

[0041] In some embodiments, the condition involving an abnormal inflammatory response in the subject involves one or more of: inflammation in the subject, fibrosis and / or fibrostenotic strictures in the subject, increased levels of trNK cells in the subject, morphological variations related to colon size or lung weight in the subject, increased paracellular permeability and / or susceptibility of an intestine or lung in the subject, and changes lipid homeostasis in the subject, or any combination thereof. Each possibility is a separate embodiment.

[0042] According to one aspect, there is provided a method of treating, attenuating and / or preventing progression of a condition that involves an abnormal inflammatory response in a subject in need thereof, the method comprising administering a therapeutically effective amount of a composition comprising an isolated peptide consisting of an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6.

[0043] In some embodiments, the method comprises administering a therapeutically effective amount of a composition comprising an isolated peptide consisting of an amino acid sequence as denoted by any one of SEQ ID NO: 1 and / or SEQ ID NO: 6. Each possibility is a separate embodiment. In some embodiments, the method comprises administering a therapeutically effective amount of a composition comprising a plurality of isolated peptide.

[0044] In some embodiments, the composition is administered in combination with at least one additional therapeutic agent.

[0045] According to some aspects, there is provided a composition comprising an isolated peptide including an amino acid sequence as denoted by one or more of any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of autoimmune diseases in a subject in need thereof. Each possibility is a separate embodiment.

[0046] According to some aspects, there is provided a composition comprising an isolated peptide including an amino acid sequence as denoted by one or more of any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of an Inflammatory Bowel Disorder (IBD) in a subject in need thereof. Each possibility is a separate embodiment.

[0047] According to some aspects, there is provided a composition comprising an isolated peptide including an amino acid sequence as denoted by one or more of any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of an Acute Lung Injury (ALI) in a subject in need thereof. Each possibility is a separate embodiment.

[0048] According to some aspects, there is provided a composition comprising an isolated peptide including an amino acid sequence as denoted by one or more of any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of cancer in a subject in need thereof. Each possibility is a separate embodiment.

[0049] In some embodiments, the isolated peptide includes an amino acid sequence as denoted by SEQ ID NO: 1. In some embodiments, the isolated peptide includes an amino acid sequence as denoted by SEQ ID NO: 6. In some embodiments, the isolated peptide consists of an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof. Each possibility is a separate embodiment.

[0050] In some embodiments, the isolated peptide consists of an amino acid sequence as denoted by SEQ ID NO: 1. In some embodiments, the isolated peptide consists of an amino acid sequence as denoted by SEQ ID NO: 6.

[0051] Further embodiments, features, advantages and the full scope of applicability of the present invention will become apparent from the detailed description and drawings given hereinafter. However, it should be understood that the detailed description, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

[0052] BRIEF DESCRIPTION OF THE FIGURES

[0053] FIGs. 1A-1C show graphs presenting food and water consumption, and body weight, across four experimental groups of mice, including wild type (wt) mice treated with 8pM of either scrambled peptide (group 1), or NLGn4-Nrxip inhibitory peptide 1 (group 2), and an IBD-induced (3% DSS) mice model treated with 8pM of either scrambled peptide (group 3), or NLGn4-Nrxip inhibitory peptide 1 (group 4). Daily food consumption (diet uptake; FIG. 1A) was calculated as gram\day\mice group, while daily drinking (water intake FIG. IB) was calculated as ml\day\mice group. At the culmination of the study, mice were sacrificed, and their weight measured in grams (FIG. 1C). Data are represented as averages ± SD (n = 5 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA); ns= p>0.05, *= p<0.05, **= p<0.01, ***= p<0.001, and ****= pO.OOOl.

[0054] FIGs. 2A-2C show graphs presenting changes in colon size of mice across four experimental groups (groups 1-4 are indicated). Colons from each subject were carefully harvested and their weight in grams were measured (FIG. 2A). Simultaneously, colon lengths in cm were recorded to provide additional insights into potential morphological variations (FIG. 2B). The weight: length ratio, a significant index of colon health, was calculated to ascertain any proportional changes (FIG. 2C). Data are represented as averages ± SD (n = 5 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= p<0.01, ***= p<0.001, and ****= pO.OOOl.

[0055] FIGs. 3A-3D show graphs presenting the quantitation of mRNA expression of Occludin (FIG. 3A), Zo-1 (FIG. 3B) Claudin (FIG. 3C) and Mucin (FIG. 3D) in colon tissues obtained from mice across the four experimental groups (groups 1-4 are indicated). The data are presented as fold change relative to the expression in wt mice treated with 8pM of the scrambled peptide (8pM), with GAPDH serving as the housekeeping gene for normalization. Data are represented as averages ± SD (n = 5 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= pO.Ol, ***= pO.OOl, and ****= pO.OOOl.

[0056] FIG. 4 shows a graph presenting quantification of multiple cytokines, including IL- lb, TNF-a, IL-6 and IL-4, in colon protein extracts (pg / g colon protein) obtained from mice across the four experimental groups (groups 1-4 are indicated). Cytokine concentration was determined simultaneously using a multiplexed sandwich enzyme-linked immunosorbent assay (ELISA). Data are represented as averages ± SD (n = 5 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= pO.Ol, ***= pO.OOl, and ****= pO.OOOL

[0057] FIGs. 5A-5B show a graph presenting quantification of mRNA expression levels of aSMA (green bars), Collagen III (COL III, blue bars), and Collagen I (COL I, yellow bars) (FIG. 5A), and of transforming growth factor-beta (TGF-P) protein (FIG. 5B). Expression levels were measured in colon tissues obtained from mice across four experimental groups (groups 1-4 are indicated). For quantification of mRNA, the data are presented as fold change relative to the expression in naive mice treated with the scrambled peptide (8pM), with GAPDH serving as the housekeeping gene for normalization. For quantification of TGF-b, protein ELISA was performed from colon protein extract (pg\g colon protein). Data are represented as averages ± SD (n = 5 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= pO.Ol, ***= pO.OOl, and ****= pO.OOOL FIGs. 6A-6C show graphs presenting quantification of cholesterol (Choi; FIG. 6A) and triglyceride (TGR; FIG. 6B) concentrations in colon protein extracts (pg / g colon protein) by ELISA, and quantification of SREBP2 (blue bars), HMGCR2 (green bars) and HMGCS III (red bars) mRNA levels in colon tissues (FIG. 6C), in samples obtained from mice across the four experimental groups (groups 1-4 are indicated). For mRNA quantification the data are presented as fold change relative to the expression in naive mice treated with the scrambled (8pM), with GAPDH serving as the housekeeping gene for normalization. Data are represented as averages ± SD (n = 5 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= p<0.01, ***= p<0.001, and ****= pO.OOOl.

[0058] FIG. 7 shows a graph presenting quantification of P-neurexin ( -Nrxn) mRNA expression in colon tissues obtained from mice across four experimental groups (groups 1-4 are indicated). The data are presented as fold change relative to the expression in Naive mice treated with the scrambled (8pM), with GAPDH serving as the housekeeping gene for normalization. Data are represented as averages ± SD (n = 5 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= p<0.01, ***= p<0.001, and ****= pO.OOOL

[0059] FIGs. 8A-8B show graphs presenting changes in lung size / weight of mice across four experimental groups (groups 1-4 are indicated). Lungs from each subject were carefully harvested and their weights in grams measured (FIG. 8A). The ratio of lungs weight : mice weight was calculated to ascertain any proportional changes (FIG. 8B). Data are represented as averages ± SD (n = 4 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= p<0.01, ***= p<0.001, and ****= pO.OOOl.

[0060] FIG. 9 shows a graph presenting quantitation of -Nrxn mRNA expression in lung tissues obtained from mice across four experimental groups (groups 1-4 are indicated,). The data are presented as fold change relative to the expression in naive mice treated with the scrambled peptide (8pM), with GAPDH serving as the housekeeping gene for normalization. Data are represented as averages ± SD (n = 4 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= pO.Ol, ***= pO.OOl, and ****= pO.OOOl.

[0061] FIG. 10 shows a graph presenting quantitation a-SMA (green bars) and GFAP (blue bars) mRNA expression in lung tissues obtained from mice across four experimental groups (groups 1-4 are indicated). The data are presented as fold change relative to the expression in naive mice treated with the scrambled (8pM), with GAPDH serving as the housekeeping gene for normalization. Data are represented as averages ± SD (n = 4 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= pO.Ol, ***= pO.OOl, and ****= p .OOOL

[0062] FIG. 11 shows a graph presenting quantitation of multiple cytokines, including IFN-y, IL-6 and IL-4, in bronchoalveolar lavage fluid (BALF) extract (pg / ml BALF) obtained from mice across four experimental groups (groups 1-4 are indicated). Cytokine concentration was determined simultaneously using a multiplexed sandwich ELISA. Data are represented as averages ± SD (n = 4 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= pO.Ol, ***= pO.OOl, and ****= pO.OOOl.

[0063] FIG. 12 shows a graph presenting quantitation of trNK cells count in the lungs of mice from across the same four experimental groups (groups 1-4 are indicated) and calculated as (cells\mg lung tissue weight). Data are represented as averages ± SD (n = 4 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= pO.05, **= pO.Ol, ***= pO.OOl, and ****= pO.OOOl.

[0064] FIG. 13 shows a graph presenting quantitation of soluble RAGE (sRAGE), in BALF extract (pg / ml BALF) obtained from mice across the four experimental groups (groups 1-4 are indicated). sRAGE concentration was determined simultaneously using a multiplexed sandwich ELISA. Data are represented as averages ± SD (n = 4 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= pO.05, **= pO.Ol, ***= pO.OOl, and ****= pO.OOOl.

[0065] FIG. 14 shows a graph presenting quantitation of surface protein-D (SP-D) mRNA expression in lung tissues obtained from mice across four experimental groups (groups 1-4 are indicated). The data are presented as fold change relative to the expression in naive mice treated with the scrambled peptide (8pM), with GAPDH serving as the housekeeping gene for normalization. Data are represented as averages ± SD (n = 4 per group). Significance was determined using Newman-Keuls two-way analysis of variance (ANOVA), ns= p>0.05, *= p<0.05, **= p<0.01, ***= p<0.001, and ****= pO.OOOl.

[0066] DETAILED DESCRIPTION

[0067] In the following description, various aspects of the disclosure will be described. For the purpose of explanation, specific configurations and details are set forth in order to provide a thorough understanding of the different aspects of the disclosure. However, it will also be apparent to one skilled in the art that the disclosure may be practiced without specific details being presented herein. Furthermore, well-known features may be omitted or simplified in order not to obscure the disclosure.

[0068] As used herein, the term "about" when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20% or in some instances ±10%, or in some instances ±5%, or in some instances ±1%, or in some instances ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

[0069] As used herein, the terms “prevent”, “reduce”, “attenuate”, “ameliorate”, “inhibit” may be used interchangeably.

[0070] As used herein, the terms “enhanced”, “increased”, “elevated” may be used interchangeably.

[0071] As used herein, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise.

[0072] As used herein, the term “comprising” is synonymous with the terms "including," "containing," or "characterized by," and is inclusive or open-ended i.e. does not exclude additional, unrecited elements. According to some embodiments, the term comprising may be replaced with the term with the term “consisting of’ which excludes any element, step, or ingredient not specified in the claim. According to some embodiments, the term comprising may be replaced with the term “consisting essentially of’ which limits the scope of a claim to the specified materials or steps "and those that do not materially affect the basic and novel characteristics" of the claimed invention.

[0073] As used herein, the terms “essentially” and “substantially” are synonymous and when referring to a stated material such as a composition, a substance, and the like, is meant to encompass variations of in some embodiments, ±0.1%, or in some embodiments, ±1%, or in some embodiments, ±2%, or in some embodiments, ±5% or in some embodiments, ±10% from a stated amount, as such variations / deviations are appropriate to perform the disclosed methods. Each possibility is a separate embodiment.

[0074] Unless otherwise noted, technical terms are used according to conventional usage. Definitions of common terms in molecular biology may be found in Benjamin Lewin, Genes V, published by Oxford University Press, 1994 (ISBN 0-19-854287-9); Kendrew et al. (eds.), The Encyclopedia of Molecular Biology, published by Blackwell Science Ltd., 1994 (ISBN 0-632-02182-9); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995 (ISBN 1-56081-569-8).

[0075] The terms that are used throughout the description, should be understood in accordance with the various embodiments to mean as follows:

[0076] According to some embodiments, the present invention stems, in part, from the surprising finding that specific isolated peptides capable of affecting (for example, by interfering or inhibiting) NLGn4X-Nrxip protein-protein interactions can modulate the NLGn4X-Nrxip axis in a manner that mitigates adverse pathological effects associated with Inflammatory bowel disease (IBD)

[0077] According to some embodiments, provided herein are advantageous compositions comprising isolated inhibitory peptides, capable of effectively and safely affecting NLGn4X-Nrxip interactions, for treating colon / intestine-related conditions, namely IBD. According to some embodiments, the isolated peptide includes or consists of an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, wherein the isolated peptide is capable of affecting interactions between neuroligin 4 (NLGn4X) and Neurexin ip (Nrxip). Each possibility is a separate embodiment.

[0078] According to some embodiments, the isolated peptide includes or consists of an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 or any combination thereof. Each possibility is a separate embodiment.

[0079] In some embodiments, the isolated peptide includes or consists of an amino acid sequence as denoted by SEQ ID NO: 1. Each possibility is a separate embodiment.

[0080] According to some embodiments, the isolated peptide includes or consists of an amino acid sequence as denoted by SEQ ID NO: 6. Each possibility is a separate embodiment.

[0081] As used herein, the terms "isolated peptide", "modulator peptide", "inhibitory peptide", NLGN4X inhibitor peptide, “NLGn4-Nrxip protein-protein interaction (PPI) inhibitor peptide”, “NLGn4-Nrxip protein-protein interaction (PPI) modulator peptide” and “NLGn4-Nrxip inhibitory peptide” may be used interchangeably to refer to the peptide(s) of the present invention that may interact with and / or bind to a corresponding site / region in NLGn4 and / or Nrx 1 p, to thereby modulate and / or interfere with the interaction (for example, protein-protein interaction) between NLGn4 and Nrxip. In some embodiments, the binding and / or modulation / interference is transient. In some embodiments, the binding and / or modulation / interference is reversible. In some embodiments, the binding and / or modulation / interference is permanent (irreversible) . The binding of the inhibitory peptide to NLGn4 and / or Nrxl p may occur both in-vivo or in-vitro. For example, in some embodiments, binding of the inhibitory peptide to NLGn4 and / or Nrxip may occur in-vivo, in tissue / cells of a subject (for example, in the colon).

[0082] In some embodiments, the isolated peptide is an isolated modulator peptide.

[0083] As used herein, the terms “modulate”, “affect”, “alter” and “interfere”, may be used interchangeably. The phrase “capable of affecting / modulating / interfering” as used herein refers to the capability of the isolated peptide(s) of the invention to inhibit or prevent interaction or binding between NLGn4 and Nrxip or to interact with a target of NLGn4 or Nrxip, such that the target becomes less accessible, preferably inaccessible, to binding by its corresponding binding partner (for example, Nrxip or NLGn4), for example the receptor's natural antigen. In some embodiments, the isolated peptide is capable of inhibiting binding between NLGn4 and Nrxip.

[0084] The term “binding site” or “epitope” as used herein refers to the region of NLGn4 and / or Nrxip that specifically reacts with a particular modulator peptide.

[0085] The terms “Neuroligin 4”, “Neuroligin 4X”, “NLGn4” and “NLG4” are interchangeable and as used herein refer to the protein product of the NLGn4 gene e.g., NP 00I269075.I, NP_001269074.1, NP_851849.1 and NP_065793.1. In some embodiments, the amino acid sequence of NLGn4 protein is denoted by SEQ ID NO: 4.

[0086] The terms “P-Nrxn” “neurexin-l-beta”, “P-neurexin” “Nrxip” and “Nrxnl” are interchangeable and as used herein refer to the protein product of the Nrxip gene e.g., NP_001129131.1 NP_004792.1 NP_620072.1. In some embodiments, the amino acid sequence of Nrxip protein is denoted by SEQ ID NO: 5.

[0087] The terms “peptide” and “polypeptide” as used herein are intended to encompass any amino acid sequence including modified sequences. The terms “peptide” and “polypeptide” are specifically intended to cover naturally occurring proteins, as well as those, which are recombinantly or synthetically produced. The inhibitory peptides of the invention also include modified peptides (with amino acid substitutions, both conservative and non-conservative as described below) that have the same or improved activity as a wild-type or unmodified peptide. “Salts” of the peptides of the invention contemplated by the invention are physiologically and pharmaceutically acceptable organic and inorganic salts. The invention also provides conservative amino acid variants of the peptides according to the invention. Variants according to the invention also may be made that conserve the overall molecular structure of the encoded peptides. Given the properties of the individual amino acids comprising the disclosed peptide products, some rational substitutions will be recognized by the skilled worker. Amino acid substitutions, i.e., “conservative substitutions,” may be made, for instance, on the basis of similarity in polarity, charge, solubility, hydrophobicity, hydrophilicity, and / or the amphipathic nature of the residues involved. The amino acids used in this invention are those, which are available commercially or are available by routine synthetic methods. Certain residues may require special methods for incorporation into the peptide, and either sequential, divergent or convergent synthetic approaches to the peptide sequence are useful in this invention. Natural coded amino acids and their derivatives are represented by three- letter codes according to IUPAC conventions. When there is no indication, the L isomer was used. The D isomers are indicated by “D” before the residue abbreviation.

[0088] Conservative substitutions of amino acids as known to those skilled in the art are within the scope of the present invention. Conservative amino acid substitutions include replacement of one amino acid with another having the same type of functional group or side chain, e.g., aliphatic, aromatic, positively charged, negatively charged. These substitutions may enhance oral bioavailability, penetration into the islets, targeting to specific beta cell populations, immunogenicity, and the like. One of skill will recognize that individual substitutions, deletions or additions to a peptide, polypeptide, or protein sequence which alters, adds or deletes a single amino acid or a small percentage of amino acids in the encoded sequence is a “conservatively modified variant” where the alteration results in the substitution of an amino acid with a chemically similar amino acid. Conservative substitution tables providing functionally similar amino acids are well known in the art.

[0089] The following six groups each contain amino acids that are conservative substitutions for one another:

[0090] 1( - Alanine (A), Serine (S), Threonine (T);

[0091] 2- Aspartic acid (D), Glutamic acid (E);

[0092] 3- Asparagine (N), Glutamine (Q);

[0093] 4- Arginine (R), Lysine (K);

[0094] 5- Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and

[0095] 6- Phenylalanine (L), Tyrosine (Y), Tryptophan (W).

[0096] The peptides of the present invention may be produced by any method known in the art, including recombinant and synthetic methods. Synthetic methods include exclusive solid phase synthesis, partial solid phase synthesis, fragment condensation, or classical solution synthesis. Solid phase peptide synthesis procedures are well known to one skilled in the art and described, for example by John Morrow Stewart and Janis Dillaha Young, Solid Phase Polypeptide Syntheses (2nd Ed., Pierce Chemical Company, 1984). In some embodiments, synthetic peptides are purified by preparative high-performance liquid chromatography (Creighton T. (1983) Proteins, structures and molecular principles. WH Freeman and Co. N.Y.) The peptide sequence may be confirmed by amino acid sequencing using methods known to one skilled in the art.

[0097] As used herein, the term “isolated" means either: 1) separated from at least some of the components with which it is usually associated in nature with respect of the source protein; 2) prepared or purified by a process that involves the hand of man; 3) not occurring in nature.

[0098] As used herein, the term "isolated peptide” refers to the peptides of the present invention produced by any method known in the art, including recombinant and synthetic methods and thereafter purified using any method known in the art so as to become at least a major component of the solution, mixture, or extract and dry forms thereof (i.e., powder), which may also include smaller amounts of other molecules; or so as to become essentially the only component of the solution, mixture, or extract and dry forms thereof (i.e., powder) (i.e. other components are present in residual amounts only).

[0099] In some embodiments, the isolated peptide refers to solution, mixture, or extract and dry forms thereof (i.e., powder) containing at least 60% (w / w or w / v), at least 70% at least 80%, or at least 90%, or at least about 95%, or at least about 96%, or at least about 97% or at least about 98%, or at least about 99% of the isolated peptide. Each possibility is a separate embodiment.

[0100] In some embodiments, the solution, mixture, or extract and dry forms thereof (i.e., powder) containing essentially the isolated peptide contains less than about 10% (w / w or w / v) or less than 5%, or less than 4%, or less than 3%, or less than 2%, or less than 1%, or less than 0.1% of other molecules than the isolated peptide. Each possibility is a separate embodiment.

[0101] In some embodiments, recombinant protein techniques are used to generate the peptide of the present invention. In some embodiments, recombinant protein techniques are used for generation of polypeptides or nucleic acid sequences or viral or bacterial vectors for vaccine formulation. Recombinant techniques are described for example by Bitter et al., (1987) Methods in Enzymol. 153:516-544, Studier et al. (1990) Methods in Enzymol. 185:60-89, Brisson et al. (1984) Nature 310:511-514, Takamatsu et al. (1987) EMBO J. 6:307-311, Coruzzi et al. (1984) EMBO J. 3: 1671-1680 and Brogli et al., (1984) Science 224:838-843, Gurley et al. (1986) Mol. Cell. Biol. 6:559-565 and Weissbach & Weissbach, 1988, Methods for Plant Molecular Biology, Academic Press, NY, Section VIII, pp 421-463.

[0102] Isolated polynucleotide sequences comprising at least one sequence encoding a peptide, peptide analog, peptide homolog conjugate or fusion protein of a modulator peptide are also included in the scope of the present invention. According to some embodiments, the polynucleotide sequence encoding the peptide or peptide analog, peptide homolog, is translationally linked to another polynucleotide sequence such as an RNA or DNA molecule and is recombinantly expressed within target cells. According to other embodiments, the polynucleotide sequence is part of a recombinant viral or bacterial vector.

[0103] The term “analog” refers to a molecule, which has the amino acid sequence according to the invention except for one or more amino acid changes. Analogs according to the present invention may include peptidomimetics. “Peptidomimetic” refers to a peptide modified in such a way that it includes at least one non-coded residue or non-peptidic bond. Such modifications include, e.g., alkylation and more specific methylation of one or more residues, insertion of or replacement of natural amino acid by non-natural amino acids, replacement of an amide bond with another covalent bond. A peptidomimetic according to the present invention may optionally comprise at least one bond, which is an amide -replacement bond such as urea bond, carbamate bond, sulfonamide bond, hydrazine bond, or any other covalent bond. The design of appropriate “analogs” may be computer assisted. Analogs are included in the invention as long as they remain pharmaceutically acceptable.

[0104] In some embodiment, the polypeptide comprises one or more amino acid substitutions, additions, or deletions. In some embodiment, the polypeptide comprises one or more substitutions corresponding to a conservative variant of the amino acid. According to some embodiments, there is provided a composition comprising an isolated peptide comprising an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of an Inflammatory Bowel Disorder (IBD) in a subject in need thereof. Each possibility is a separate embodiment.

[0105] In some embodiments, the isolated peptide comprises an amino acid sequence as denoted by SEQ ID NO: 1 and / or SEQ ID NO: 6. Each possibility is a separate embodiment.

[0106] In some embodiments, the isolated peptide comprises an amino acid sequence as denoted by SEQ ID NO: 1. In some embodiments, the isolated peptide comprises an amino acid sequence as denoted by SEQ ID NO: 6.

[0107] In some embodiments, the isolated peptide consists of an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof. Each possibility is a separate embodiment.

[0108] In some embodiments, the isolated peptide consists of an amino acid sequence as denoted by SEQ ID NO: 1 and / or SEQ ID NO: 6. Each possibility is a separate embodiment.

[0109] According to some embodiments, the amino acid sequence of the peptide has at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or at least 99.9% sequence homology / identity / similarity to the amino acid sequence as set forth in any one of SEQ ID NO: 1, SEQ ID NO: 2 SEQ ID NO: 3 and / or SEQ ID NO: 6. Each possibility is a separate embodiment.

[0110] The terms “similarity”, "homology" and “sequence similarity / homology” may be interchangeably used and refer hereinafter to the level of identities between two homologous sequences when they are compared by aligning them using an alignment tool. As used herein, the sequence similarity is with respect to any one of the amino acid sequences disclosed herein and denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2 SEQ ID NO: 3 and / or SEQ ID NO: 6.

[0111] The level of similarity or homology is determined by the number of identities in the amino acid sequence when they are aligned. In some embodiments, the composition includes a plurality of isolated peptides.

[0112] The terms “plurality” or “plurality of isolated peptides” refer to a combination / mixture including two or more of the isolated peptides of the invention as set forth in any one of SEQ ID NO: 1, SEQ ID NO: 2 SEQ ID NO: 3 and / or SEQ ID NO: 6 and / or variants or homologs thereof including peptides having conservative amino acid substitutions and / or peptide analog / modified peptide thereof.

[0113] In some embodiments, the composition includes a combination of two or more isolated peptides selected from SEQ ID NO: 1, SEQ ID NO: 2 SEQ ID NO: 3 and SEQ ID NO: 6, or any combination thereof.

[0114] According to some embodiments, there is provided a method of treating, attenuating and / or preventing progression of Inflammatory Bowel Disorder (IBD) in a subject in need thereof, the method comprising administering a therapeutically effective amount of a composition comprising an isolated peptide comprising an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6.

[0115] In some embodiments, the administration includes systemic and / or localized administration. Each possibility is a separate embodiment.

[0116] In some embodiments, the composition comprising the isolated peptide is administered in combination with at least one additional therapeutic agent.

[0117] The terms “subject” and “patient” are interchangeable and as used herein refer to any individual suffering from IBD.

[0118] The term “treatment” as used herein refers to both therapeutic treatment and prophylactic or preventative measures. In some embodiments, those in need of treatment include those already having a disorder. In some embodiments, those in need of treatment include those in which the progression of the disorder is to be prevented. In some embodiments, those in need of treatment include those in which the disorder is to be prevented. As used herein, the terms “prevent”, “reduce”, “attenuate”, “ameliorate”, “inhibit”, “mitigate”, “alleviate” may be used interchangeably.

[0119] The terms “administration” and “administering” refer to providing or giving a subject a therapeutic agent (e.g. an isolated peptide or composition comprising the same), by any effective route. Exemplary routes of administration include, but are not limited to, injection or infusion (such as subcutaneous, intramuscular, intradermal, intraperitoneal, intrathecal, intravenous, intracerebroventricular, intrastriatal, intracranial and into the spinal cord), oral, intraductal, sublingual, rectal, transdermal, intranasal, vaginal and inhalation routes. Each possibility is a separate embodiment.

[0120] According to some embodiments, any suitable route of administration to a subj ect may be used for the compositions comprising the isolated peptides of the present invention, including but not limited to, local and systemic routes. Exemplary suitable routes of administration include, but are not limited to: orally, intra-nasally, parenterally, intravenously, topically, enema or by inhalation. According to another embodiment, systemic administration of the composition is via an injection. For administration via injection, the composition may be formulated in an aqueous solution, for example in a physiologically compatible buffer including, but not limited, to Hank’s solution, Ringer’s solution, or physiological salt buffer. Formulations for injection may be presented in unit dosage forms, for example, in ampoules, or in multi -dose containers with, optionally, an added preservative.

[0121] According to another embodiment, administration systemically is through a parenteral route. According to some embodiments, parenteral administration is administration intravenously, intra-arterially, intramuscularly, intraperitoneally, intradermally, intravitreally, or subcutaneously. Each of the abovementioned administration routes represents a separate embodiment of the present invention. According to another embodiment, parenteral administration is performed by bolus injection. According to another embodiment, parenteral administration is performed by continuous infusion. According to some embodiments, preparations of the composition of the invention for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, or emulsions, each representing a separate embodiment of the present invention.

[0122] According to some embodiments, the administration may include any suitable administration regime, depending, inter aha, on the medical condition, patient characteristics, administration route, and the like. In some embodiments, administration may include administration twice daily, every day, every other day, every third day, every fourth day, every fifth day, once a week, once every second week, once every third week, once every month, and the like.

[0123] The terms “IBD” and “colon / intestinal-related disorders” may be used interchangeably, and refer to pathologies that cause the bowel / colon / intestine to function improperly as generally known in the art.

[0124] In some embodiments, IBD pathologies include at least one of any one of: (i) body weight loss; (ii) morphological changes related to colon size; (iii) chronic inflammation; (iv) fibrosis and / or fibrostenotic strictures; (v) increased paracellular permeability or colon susceptibility; and (vi) changes in lipid homeostasis, or any combination thereof. Each possibility is a separate embodiment.

[0125] In some embodiments, IBD pathologies may also include loss of body weight.

[0126] In some embodiments, the colon / intestinal-related disorders / IBD includes but is not limited to one or more of: ulcerative colitis (UC), Crohn's disease (CD) Collagenous colitis, Lymphocytic colitis, Ischemic colitis, Diversion colitis, Behcet' s disease, and Indeterminate colitis, or any combination thereof. Each possibility is a separate embodiment.

[0127] In some embodiments, the IBD is selected from any one or more of: ulcerative colitis (UC), Crohn's disease (CD) Collagenous colitis, Lymphocytic colitis, Ischemic colitis, Diversion colitis, Behcet's disease, and Indeterminate colitis, or any combination thereof.

[0128] The terms “composition” and “pharmaceutical composition” are used interchangeably and as used herein refers to any composition comprising at least one isolated peptide of the invention., or a composition which comprises a mixture / plurality of isolated peptides. In some embodiments, the composition can include a plurality of peptides. In some embodiments, the plurality of peptides includes a combination of several peptides. As a non-limiting example, in some embodiments, the composition may include a combination of peptide 1, peptide 2 and / or peptide 3, and in other embodiments, the composition may include a mixture of peptide 1, peptide 3 and / or peptide 6. In some embodiments, the composition includes the modulator peptide and a pharmaceutically acceptable carrier.

[0129] The term “pharmaceutically acceptable carrier” refers to any carrier conventional used in the production of pharmaceutical compositions. Remington's Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, Pa., 15th Edition, 1975, describes compositions and formulations suitable for pharmaceutical delivery of the compositions disclosed herein.

[0130] According to some aspects, the present invention provides compositions including novel isolated peptides, and uses thereof for treating IBD by modulating, interfering with, inhibiting and / or preventing NLGn4-Nrxip interaction, in particular, by preventing protein-protein interaction ("PPI") thereof.

[0131] In some embodiments, the isolated peptides may be derived from NLGN4X. The peptides may include different amino acid sequences that can vary in length, and may be derived from the same binding site as that of p-neurexin (i.e., the ligand of the NLG4NX receptor). In some embodiments, the active binding site is in a domain containing amino acids 359-364 in NLGN4X protein. In some embodiments, epitopes involved in binding may include, E361, L363, H267, Y463 and E270. In some embodiments, the isolated peptides are derived from E361 site epitope.

[0132] According to some embodiments the isolated peptide is used in a method of treating, attenuating and / or preventing progression of a bowel disorder, the method includes administering a therapeutically effective amount of peptide (or a composition including the same), the peptide having an amino acid sequence as denoted by any one of: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, wherein said peptide is capable of interfering with, inhibiting and / or preventing neuroligin 4 (NLGn4X) - Neurexin ip (Nrxip) interaction.

[0133] According to some embodiments, the peptide, may be used in combination with at least one additional therapeutic agent, such as, but not limited to PD1 and PDL1 inhibitors.

[0134] In some embodiments, the additional therapeutic agent comprises a classical therapeutic agent. In some embodiments, the isolated peptide may be comprised in a single composition with the additional therapeutic agent. In some embodiments, the peptide may be comprised in a separate composition than the additional therapeutic agent.

[0135] According to some embodiments, the isolated peptide may be a synthetic peptide or a recombinant peptide. Each possibility is a separate embodiment.

[0136] According to some embodiments, there are provided isolated peptides for affecting the NLGn4-Nrxip interaction, the peptides have an amino acid sequence as set forth in SEQ ID NO: 6: EQGEFLNY. In some embodiments, the isolated peptide having amino acid sequence as denoted by SEQ ID NO: 6 is designated "peptide 6". In some embodiments, the isolated peptide may consist or comprise of amino acid sequence as denoted by SEQ ID NO: 6.

[0137] According to some embodiments, the isolated peptide sequence of NLGn4- Nrxip PPI may have an amino acid sequence as denoted by SEQ ID NO: 1: MEQGEFLNYD. In some embodiments, the isolated peptide having amino acid sequence as denoted by SEQ ID NO: 1 is designated "peptide 1". In some embodiments, peptide 1 may comprise or consist of amino acid sequence as denoted by SEQ ID NO: 1.

[0138] According to some embodiments, the isolated peptide sequence may have an amino acid sequence as denoted by SEQ ID NO: 2: QILMEQGEFLNYDIM. In some embodiments, the isolated peptide having amino acid sequence as denoted by SEQ ID NO: 2 is designated "peptide 2". In some embodiments, peptide 2 may comprise or consist of amino acid sequence as denoted by SEQ ID NO: 2.

[0139] According to some embodiments, the isolated peptide sequence may have an amino acid sequence as denoted by SEQ ID NO: 3: DPQILMEQGEFLNYDIMLGV. In some embodiments, the isolated peptide having amino acid sequence as denoted by SEQ ID NO: 3 is designated "peptide 3". In some embodiments, peptide 3 may comprise or consist of amino acid sequence as denoted by SEQ ID NO: 3.

[0140] According to some embodiments, there is provided a composition comprising an isolated peptide capable of affecting interactions between neuroligin 4 (NLGn4X) and Neurexin ip (Nrxip), and wherein the isolated peptide comprises or consist of an amino acid sequence as denoted by SEQ ID NO: 6.

[0141] According to some embodiments, the isolated peptide comprises or consist an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3. Each possibility is a separate embodiment.

[0142] According to some embodiments, the isolated peptide comprises or consist of an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6. Each possibility is a separate embodiment.

[0143] In some embodiments, the isolated peptide comprises an amino acid sequence as denoted by SEQ ID NO: 1. In some embodiments, the isolated peptide comprises an amino acid sequence as denoted by SEQ ID NO: 6.

[0144] According to some embodiments, the present invention provides methods and compositions for treating, attenuating, and / or preventing progression of IBD.

[0145] According to some embodiments, the present invention provides methods and compositions for treating, attenuating, and / or preventing progression of autoimmune disease.

[0146] According to some embodiments, the inhibitory peptide can modulate / disrupt / block the NLGn4X / p-neurexin axis.

[0147] According to some embodiments, the isolated peptide can ameliorate or reduce the severity of one or more pathological characteristics associated with IBD.

[0148] According to some embodiments, the isolated peptide can ameliorate or reduce the severity of one or more pathological characteristics associated with autoimmune disease.

[0149] In some embodiments, pathological characteristics associated with IBD may be evident through examination of serum / blood, and / or colon / inte stine samples.

[0150] In some embodiments, pathological characteristics associated with IBD may be evident through examination of mRNA and / or protein levels biomarkers.

[0151] Advantageously, in some embodiments, the pathological characteristics associated with IBD includes, but are not limited to: weight loss, morphological variations related to colon size, persistent inflammation, fibrotic tissue / stricture, gut barrier permeability, and changes in lipid homeostasis, or any combination thereof.

[0152] Advantageously, according to some embodiments, treatment of IBD patients using the isolated peptide or composition including the same, includes prevention of or protection from body weight loss. Each possibility is a separate embodiment.

[0153] According to some embodiments, the isolated peptide does not affect diet / food uptake and water intake.

[0154] In accordance, in some embodiments a isolated peptide has no adverse effects and does not negatively affect the well-being of a treated subject, estimated by food intake and / or water intake.

[0155] Advantageously, according to some embodiments, treatment of IBD patients using the isolated peptide or composition including the same, includes prevention of or protection from morphological variations related to colon size. Each possibility is a separate embodiment.

[0156] In some embodiments, morphological variations related to colon size include, one or more of colon weight, colon length and / or ratio thereof, or any combination thereof. Each possibility is a separate embodiment.

[0157] In some embodiments, prevention of or protection from morphological variations related to colon size includes prevention of an increase in colon weight and / or prevention of reduction in colon length. Each possibility is a separate embodiment.

[0158] In some embodiments, prevention of or protection from morphological variations related to colon size includes prevention of an increase in the ratio of colon weight / colon length. Each possibility is a separate embodiment.

[0159] Advantageously, according to some embodiments, treatment of IBD patients using the isolated peptide or composition including the same, includes prevention of or protection from increased paracellular permeability and / or susceptibility of the intestine thereby reducing leakiness through the gut barrier. Each possibility is a separate embodiment.

[0160] In some embodiments, prevention of or protection from paracellular permeability and / or susceptibility of the intestine includes prevention of increased expression of one or more of: Occludin, ZO-1, Claudin and / or Mucin, or any combination thereof. Each possibility is a separate embodiment.

[0161] Advantageously, according to some embodiments, treatment of IBD patients using the isolated peptide or composition including the same, includes prevention of or protection from inflammation. Each possibility is a separate embodiment.

[0162] In some embodiments, prevention of or protection from inflammation includes prevention of increased expression of one or more of: IL-ip, TNF-a, IL-6 and / or IL- 4, or any combination thereof. Each possibility is a separate embodiment.

[0163] Advantageously, according to some embodiments, treatment of IBD patients using the isolated peptide or composition including the same, includes prevention of or protection from fibrosis / fibrostenotic strictures. Each possibility is a separate embodiment.

[0164] In some embodiments, prevention of or protection from fibrosis / fibrostenotic strictures includes prevention of increased expression of one or more of: aSMA, collagen I, and collagen III expression, or any combination thereof. Each possibility is a separate embodiment.

[0165] In some embodiments, prevention of or protection from fibrosis / fibrostenotic strictures includes prevention of upregulation of one or more of: TGF-P expression. Each possibility is a separate embodiment.

[0166] According to some embodiments, the isolated peptide can prevent or reduce P- neurexin expression and / or activity in fibrotic colon / intestine.

[0167] According to some embodiments, the isolated peptide of the invention can inhibit fibrostenotic strictures in vivo.

[0168] According to some embodiments, the NLGn4-Nrxip inhibitory peptides can advantageously promote anti-fibrotic effects, by modulating NLGN4X / p-neurexin axis.

[0169] According to some embodiments, there is provided a method of preventing or protecting from fibrostenotic strictures in the colon / intestine, the method including administering to a subject in need thereof the isolated peptide of the invention, or compositions including the same; in some embodiments the subject in need thereof suffers from IBD. According to some embodiments, there is provided a method of inhibiting or ameliorating fibrosis in a subject in the need thereof, the method includes administrating to a subject in need thereof an inhibitory peptide of the invention, or a composition including the same; in some embodiments the subject in need thereof suffers from IBD.

[0170] Advantageously, according to some embodiments, treatment of IBD patients using the isolated peptide or composition including the same, includes prevention of or protection from changes in lipid homeostasis. Each possibility is a separate embodiment.

[0171] In some embodiments, prevention of or protection from changes in lipid homeostasis includes prevention of increased expression and / or variability in expression of one or more of: Cholesterol (Choi), Triglycerides (TRG), SREBP2, HMGCR, and HMGCS, or any combination thereof. Each possibility is a separate embodiment.

[0172] Advantageously, according to some embodiments, treatment of IBD patients using the isolated peptide or composition including the same, includes prevention of or protection from increase in P-Nrxn expression.

[0173] According to some embodiments, the present invention provides methods and compositions for treating, attenuating, and / or preventing progression of acute lung injury (ALI).

[0174] According to some embodiments, the inhibitory peptide can modulate / disrupt / block the NLGn4X / p-neurexin axis.

[0175] According to some embodiments, the isolated peptide can ameliorate or reduce the severity of one or more pathological characteristics associated with ALI.

[0176] In some embodiments, pathological characteristics associated with ALI may be evident through examination of serum / blood, and / or lung samples.

[0177] In some embodiments, pathological characteristics associated with ALI may be evident through examination of mRNA and / or protein levels biomarkers.

[0178] Advantageously, in some embodiments, the pathological characteristics associated with ALI includes, but are not limited to: increased lung weight, acute / chronic inflammation in the lung, fibrosis, increment in the number of trNK cells in the lungs, increment in the levels of soluble RAGE, increment in the levels of surfactant protein D (SP-D), or any combination thereof. Each possibility is a separate embodiment.

[0179] Advantageously, according to some embodiments, treatment of ALI patients using the isolated peptide or composition including the same, includes prevention of or protection from increased lung weight. Each possibility is a separate embodiment.

[0180] According to some embodiments, the isolated peptide does not affect diet / food uptake and water intake.

[0181] In accordance, in some embodiments a isolated peptide has no adverse effects and does not negatively affect the well-being of a treated subject, estimated by food intake and / or water intake.

[0182] Advantageously, according to some embodiments, treatment of ALI patients using the isolated peptide or composition including the same, includes prevention of or protection from inflammation. Each possibility is a separate embodiment.

[0183] In some embodiments, prevention of or protection from inflammation includes prevention of increased expression of one or more of: IFN-y, IL-6 and IL-4, or any combination thereof. Each possibility is a separate embodiment.

[0184] Advantageously, according to some embodiments, treatment of ALI patients using the isolated peptide or composition including the same, includes prevention of or protection from fibrosis / fibrostenotic strictures. Each possibility is a separate embodiment.

[0185] In some embodiments, prevention of or protection from fibrosis / fibrostenotic strictures includes prevention of increased expression of: aSMA and / or GFAP. Each possibility is a separate embodiment.

[0186] According to some embodiments, the isolated peptide can prevent or reduce - neurexin expression and / or activity in fibrotic lung.

[0187] According to some embodiments, the isolated peptide of the invention can inhibit fibrostenotic strictures in vivo. In some embodiments, treatment comprises prevention or reduction of number of trNK cells in the lungs.

[0188] According to some embodiments, the NLGn4-Nrxip inhibitory peptides can advantageously promote anti-fibrotic effects, by modulating NLGN4X / p-neurexin axis.

[0189] According to some embodiments, there is provided a method of preventing or protecting from fibrosis / fibrostenotic strictures in the lung, the method including administering to a subject in need thereof the isolated peptide of the invention, or compositions including the same; in some embodiments the subject in need thereof suffers from IBD.

[0190] According to some embodiments, there is provided a method of inhibiting or ameliorating fibrosis in a subject in the need thereof, the method includes administrating to a subject in need thereof an inhibitory peptide of the invention, or a composition including the same; in some embodiments the subject in need thereof suffers from ALL

[0191] Advantageously, according to some embodiments, treatment of ALI patients using the isolated peptide or composition including the same, includes prevention of or protection from increase in P-Nrxn expression.

[0192] According to some aspects, there is provided a composition comprising an isolated peptide including an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of one or more conditions that involve an abnormal inflammatory response in a subject in need thereof. Each possibility is a separate embodiment.

[0193] In some embodiments, the isolated peptide comprises an amino acid sequence as denoted by SEQ ID NO: 1. In some embodiments, the isolated modulator peptide comprises an amino acid sequence as denoted by SEQ ID NO: 6.

[0194] In some embodiments, the abnormal inflammatory response is acute and / or chronic.

[0195] In some embodiments, the condition that involves an abnormal inflammatory response is localized and / or systemic. In some embodiments, the inflammatory response is localized and / or systemic. In some embodiments, administration of the composition is localized and / or systemic.

[0196] In some embodiments, the one or more conditions that involve an abnormal inflammatory response are selected from one or more of an autoimmune disease, Acute lung injury (ALI), and cancer, or any combination thereof. Each possibility is a separate embodiment.

[0197] In some embodiments, the conditions at involve an abnormal inflammatory response include an autoimmune disease.

[0198] In some embodiments, the autoimmune disease is selected from one or more of: inflammatory bowel disorder (IBD), Thyroid diseases including Graves' disease and Hashimoto’s thyroiditis, Adrenal diseases including Addisson disease, Thymus disease including Myasthenia gravis, Multiple sclerosis (MS), Rheumatoid arthritis, Psoriasis and Psoriatic arthritis, Type 1 diabetes, Coeliac disease, Systemic lupus erythematosus, Scleroderma, Polymyositis, Dermatomyositis, and Pernicious anemia, or any combination thereof. Each possibility is a separate embodiment.

[0199] In some embodiments, the autoimmune disease includes inflammatory bowel disorder (IBD).

[0200] In some embodiments, the IBD is selected from any one or more of: ulcerative colitis (UC), Crohn's disease (CD) Collagenous colitis, Lymphocytic colitis, Ischemic colitis, Diversion colitis, Behcet's disease, and Indeterminate colitis, or any combination thereof. Each possibility is a separate embodiment.

[0201] In some embodiments, the conditions that involve an abnormal inflammatory response include acute lung injury (ALI). In some embodiments, the Acute lung injury (ALI) includes acute respiratory distress syndrome (ARDS).

[0202] In some embodiments, the conditions that involve an abnormal inflammatory response include cancer. In some embodiments, the cancer includes lung cancer.

[0203] In some embodiments, the cancer is not liver cancer, colorectal cancer, breast cancer, and prostate cancer, or any combination thereof. Each possibility is a separate embodiment.

[0204] According to some aspects, there is provided a composition comprising an isolated peptide including an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of an autoimmune disease in a subject in need thereof. Each possibility is a separate embodiment.

[0205] In some embodiments, the autoimmune disease includes abnormal inflammatory response.

[0206] In some embodiments, the abnormal inflammatory response is acute and / or chronic.

[0207] In some embodiments, the autoimmune disease is associated with cancer. In some embodiments, the cancer includes lung cancer.

[0208] In some embodiments, the association of autoimmunity with cancer comprises a higher risk of developing the cancer.

[0209] In some embodiments, the autoimmune disease is associated with Acute lung injury (ALI). In some embodiments, the autoimmune disease is associated with acute respiratory distress syndrome (ARDS).

[0210] In some embodiments, the association of autoimmunity with ALI or ARDS comprises a higher risk of developing ALI or ARDS.

[0211] In some embodiments, the Acute lung injury (ALI) includes acute respiratory distress syndrome (ARDS).

[0212] According to some aspects, there is provided a composition comprising an isolated peptide including an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of Acute lung injury (ALI) in a subject in need thereof. Each possibility is a separate embodiment.

[0213] In some embodiments, the Acute lung injury (ALI) includes abnormal inflammatory response.

[0214] In some embodiments, the abnormal inflammatory response is acute and / or chronic.

[0215] In some embodiments, the Acute lung injury (ALI) includes acute respiratory distress syndrome (ARDS). According to some aspects, there is provided a composition comprising an isolated peptide including an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of cancer in a subject in need thereof. Each possibility is a separate embodiment.

[0216] In some embodiments, the cancer includes abnormal inflammatory response.

[0217] According to some embodiments, the isolated peptide of the invention inhibits / attenuates proliferation and / or oncogenicity of cancer cells.

[0218] In some embodiments, the cancer includes lung cancer.

[0219] According to some embodiments, there is provided a method of promoting anti- cancerous effects in the lung, the method including administering to a subject in need thereof a composition comprising isolated peptide of the invention, or compositions including the same.

[0220] According to some aspects, there is provided a composition comprising an isolated peptide including an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, or any combination thereof, for use in treating, attenuating, and / or preventing progression of one or more immune associated disease / condition in a subject in need thereof. Each possibility is a separate embodiment.

[0221] In some embodiments, immune associated disease / condition is localized and / or systemic.

[0222] In some embodiments, the inflammatory response is acute and / or chronic.

[0223] In some embodiments, the one or more immune associated disease / conditions are selected from one or more of an autoimmune disease, Acute lung injury (ALI), and cancer, or any combination thereof. Each possibility is a separate embodiment.

[0224] In some embodiments, the immune associated disease / condition includes an abnormal inflammatory response.

[0225] In some embodiments, the immune associated disease / condition that involves an abnormal inflammatory response include an autoimmune disease. In some embodiments, the autoimmune disease is selected from one or more of: inflammatory bowel disorder (IBD), Thyroid diseases including Graves' disease and Hashimoto’s thyroiditis, Adrenal diseases including Addisson disease, Thymus disease including Myasthenia gravis, Multiple sclerosis (MS), Rheumatoid arthritis, Psoriasis and Psoriatic arthritis, Type 1 diabetes, Coeliac disease, Systemic lupus erythematosus, Scleroderma, Polymyositis, Dermatomyositis, and Pernicious anemia, or any combination thereof. Each possibility is a separate embodiment.

[0226] In some embodiments, the autoimmune disease includes inflammatory bowel disorder (IBD).

[0227] In some embodiments, the IBD is selected from any one or more of: ulcerative colitis (UC), Crohn's disease (CD) Collagenous colitis, Lymphocytic colitis, Ischemic colitis, Diversion colitis, Behcet's disease, and Indeterminate colitis, or any combination thereof.

[0228] In some embodiments, the immune associated disease / condition that involves an abnormal inflammatory response includes acute lung injury (ALI). In some embodiments, the Acute lung injury (ALI) includes acute respiratory distress syndrome (ARDS).

[0229] In some embodiments, the immune associated disease / condition that involves an abnormal inflammatory response includes cancer. In some embodiments, the cancer includes lung cancer.

[0230] In some embodiments, the cancer is not liver cancer, colorectal cancer, breast cancer, and prostate cancer, or any combination thereof. Each possibility is a separate embodiment.

[0231] In some embodiments, the abnormal inflammatory response related to IBD is associated with one or more of (i) morphological changes related to colon size; (ii) acute / chronic inflammation in the bowel; (iii) fibrosis / fibrostenotic strictures in the bowel; and their association with (iv) increased paracellular permeability; and (v) changes in lipid homeostasis, or any combination thereof. Each possibility is a separate embodiment.

[0232] In some embodiments, the abnormal inflammatory response related to ALI is associated with one or more of (i) acute / chronic inflammation in the lung; (ii) fibrosis of the lung; (iii) increment in the number of trNK cells in the lungs; (iv) increment in the levels of soluble RAGE; (v) increment in the levels of surfactant protein D (SP-D), or any combination thereof. Each possibility is a separate embodiment.

[0233] In some embodiments, the condition involving an abnormal inflammatory response in the subject involves one or more of: inflammation in the subject, fibrosis and / or fibrostenotic strictures in the subject, increased levels of trNK cells in the subject, morphological variations related to colon size or lung weight in the subject, increased paracellular permeability and / or susceptibility of an intestine or lung in the subject, and changes lipid homeostasis in the subject, or any combination thereof. Each possibility is a separate embodiment.

[0234] In some embodiments, the fibrosis and / or fibrostenotic strictures are not liver fibrosis and / or liver fibrostenotic strictures. Each possibility is a separate embodiment.

[0235] According to some embodiments, there is provided a nucleic acid encoding for the isolated peptides of the invention. In some embodiments, there is provided a DNA construct / vector (such as, an expression vector) harboring or comprising a nucleic acid encoding for the peptides (optionally in addition to one or more regulatory sequences, non-coding sequences, and the like). In some embodiments, various suitable vectors are known to those skilled in art, and the choice of which depends on the function desired. Such vectors include, for example, plasmids, cosmids, viruses, bacteriophages and other vectors. In some embodiments, the polynucleotides and / or vectors harboring the same can be reconstituted into vehicles, such as, for example, liposomes for delivery to target cells. Any cloning vector and / or expression vector known in the art may be used, depending on the purpose, the host cell, and the like. Such vectors may be used for in- vitro and / or in-vivo introduction / expression.

[0236] According to some embodiments, the encoding nucleic acid molecules and / or the vectors disclosed herein may be designed for direct introduction or for introduction via carrier, such as, liposomes, viral vectors (adenoviral, retroviral) into target cells / tissue.

[0237] According to some embodiments, there are provided kits comprising the isolated peptides or composition comprising the same, as disclosed herein. In some embodiments, such kits / compositions can be used, for example, in the treatment of IBD or ALL Each possibility is a separate embodiment.

[0238] In some embodiments, the kit may further include one or more reagents, buffers and / or instructions for using the same. In some embodiments, the composition may further include a pharmaceutically acceptable carrier.

[0239] The following examples are presented to provide a more complete understanding of the invention. The specific techniques, conditions, materials, proportions and reported data set forth to illustrate the principles of the invention are exemplary and should not be construed as limiting the scope of the invention.

[0240] EXAMPLES:

[0241] Materials and Methods:

[0242] In-vivo experiments:

[0243] NLGn4-Nrxip Inhibitory Peptide synthesis - inhibitory peptide 1: MEQGEFLNYD (SEQ ID NO: 1), inhibitory peptide 2: QILMEQGEFLNYDIM (SEQ ID NO: 2), inhibitory peptide 3: DPQILMEQGEFLNYDIMLGV (SEQ ID NO: 3), and a control peptide having a scrambled sequence: LEGEDQNFYM (SEQ ID NO: 7) were synthesized by Pepscan Ltd. at a purity of 98.3%.

[0244] Animal ethics - the mice used in the Examples described hereinbelow were housed in a barrier facility and received care according to the Hebrew University ethical regulations and NIH guidelines. The institutional animal care ethics committee of the Hebrew University approved all animal protocols under the ethics number of MD-18- 15494-3.

[0245] Experimental design of in-vivo experiments with Inflammatory Bowel Disease (IBD) model mice - Twelve-week-old male weight 24±1.2 g wild-type C57BL / 6J obtained from Envigo (Harlan, Israel). IBD administration was performed by providing the animals with water containing 3% Dextran sulfate sodium salt (DSS 3%; Merck, 67578-5G >95% purity, as determined by LC / MS analyses). Treatment with scrambled peptide and NLGn4-Nrxip inhibitor peptide 1 was prepared fresh daily in phosphate buffered saline (PBS) at a concentration of 8pM at the volume of 100ml.

[0246] The prepared peptides were administered i.p. once daily for 5 days, n=5 mice per group.

[0247] Groupl: wild-type mice drinking regular water without DSS with scrambled peptide (8pM), i.p. injection daily for five days.

[0248] Group : wild-type mice drinking regular water without DSS with NLGn4-Nrxip inhibitor peptide 1 (8pM), i.p. injection daily for five days.

[0249] Group3: wild-type mice drinking regular water with DSS 3% with scrambled peptide (8pM), i.p. injection daily for five days.

[0250] Group4: wild-type mice drinking regular water with DSS 3% with NLGn4-Nrxip inhibitor peptide 1 (8pM), i.p. injection daily for five days.

[0251] Additional procedures - Mice were weighed, and water intake and diet uptake were measured daily. Prior to sacrifice, animals were anesthetized by 5% isoflurane inhalation for 10 seconds before cervical dislocation. Colon protein extract were assessed for pro-inflammatory cytokine profile assessments by ELISA, and mRNA was obtained for RT-PCR as described below.

[0252] RNA isolation, cDNA preparation, and real-time PCR - Total cellular of 2mg / ml RNA (purity 98%) determined using a NanoDrop ND- 1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE) was isolated from all mice groups (n=5) using 2 ml of TRI reagent (Bio Lab; Cat# 90102331). The samples were centrifuged (1,400 rpm) for 15 minutes at 4°C to collected RNA-supematant. For RNA precipitation, the supernatant of each sample was transferred to a new microcentrifuge tube, and 0.5 ml of isopropanol (Bio Lab; Cat# 16260521) was added and incubated at 25°C for 10 minutes. The tubes were then centrifuged (12,000 rpm) for 10 minutes at 4°C, the supernatants were removed, and one milliliter of 75% ethanol was added to the pellets prior to centrifugation (7,500 rpm) for 5 minutes. The pellets were air-dried at room temperature for 15 minutes, 50 pl of DEPC was added, and the samples were heated for ten minutes at 55 °C. cDNA was prepared with a High-Capacity cDNA Isolation Kit (R&D; Cat# 1406197). Real-time PCR was performed with TaqMan Fast Advanced Master Mix (Cat# 4371130, Applied Biosystems) to quantify b-Nrxn, aSMA, Col I, Col III, SREBP2, HMGCR, HMGCS gene expression with normalization to the expression of the housekeeping gene GAPDH. Cycling condition for Thermo Fisher One Step RT-PCR kit involved RT step for 30 min at 50 °C and denaturation for 15 min at 95 °C. Further the reaction mixture was incubated for 40 cycles of 94 °C for 30 s, 60 °C for 30 s, and 72 °C for 1 min followed by 72 °C for 10 min. Data analysis were analyzed using QuantStudio ™ 5 Real-Time PCR System (Cat# A34322, Applied Biosystem).

[0253] Cytokines and lipids assessment in colon protein lysate - Protein was extracted from colon tissues by lysis in a buffer (50 mmol / L Tris-HCl (pH 7.6), 0.25% Triton X-100, 0. 15 M NaCl, 10 mM CaCh) and cOmplete™ Mini EDTA-Free Protease Inhibitor Cocktail (Roche Diagnostics, Mannheim, Germany). Protein concentrations were determined using BCA protein assay kit (Beyotime, Shanghai, China), and the proteins were mixed with 5 x loading buffer (protein: 5 x loading buffer = 4: 1) and boiled at 95 °C for 10 min. A multiplexed sandwich enzyme-linked immunosorbent assay -based technology (Cat# MHSTCMAG-70K; R&D Systems) was used to simultaneously determine the concentration of multiple cytokines of IL- lb, IL-6, IL-4, TNF-a, TGF-b. Moreover, cholesterol (Abeam; ab285242), triglyceride (Abeam; ab65336), were performed using ELISA kits according to the manufacture protocols. Briefly, all reagents and samples were brought to room temperature (18 - 25 °C) before use. A volume of 100 pL of each standard and protein sample were added into appropriate wells and incubate for 2.5 hours at room temperature with gentle shaking. The solution was discarded, wells were wash 4 times with IX Wash Solution, of note; washing was done by filling each well with Wash Buffer (300 pL) using a multichannel Pipette or auto washer. Following the washing, the liquid was complete removal at each step is essential to good performance. A volume of 100 pL of lx prepared detection antibody were added to each well for 1 hour at room temperature with gentle shaking. A volume of 100 pL of prepared streptavidin solution to each well for 45 minutes at room temperature with gentle shaking. A volume of 100 pL of TMB One-Step Substrate Reagent (Item H) were added to each well for 30 minutes at room temperature in the dark with gentle shaking. Finally, 50 pL of Stop Solution (Item I) were added to each well. Absorbance was read at 450 nm immediately using ELISA reader (Tecan Ml 00 Plate Reader). Statistical analysis - Statistical differences were analyzed with a two-tailed unpaired Student’ s t test (for comparisons between two groups) or one-way or two-way analysis of variance (ANOVA; one-way ANOVA with the Newman-Keuls post hoc test for comparisons among multiple groups) with GraphPad Prism 9.0 (GraphPad Software, La Jolla, CA). A t-test of p value <0.05 is considered statistically significant and was calculated as the difference in means between two variables. Results are presented as mean ± SD or as average means of experimental replicates ± SD.

[0254] Example 1 - NLGn4-NrxlB inhibitory peptide protected IBP model mice from weight loss

[0255] In-vivo experiments were performed in a mouse model of Inflammatory Bowel Disease (IBD) generated by exposing the mice to 3% Dextran sulfate sodium salt (DSS 3%) that were added to their drinking water in order to induce colitis. The experiments were carried out by comparing between four groups of mice, according to the experimental procedure of IBD-animal model described in detail in the material and methods section and summarized herein, as follows: group 1 - wt mice administered with 8pM of scrambled peptide (SEQ ID NO: 7); group 2 - wt mice administered with 8pM of peptide 1 (SEQ ID NO: 1); group 3 - IBD-model mice (3% DSS) administered with 8pM of scrambled peptide (SEQ ID NO: 7); and group 4 - IBD-model mice (3% DSS) administered with 8pM of peptide 1 (SEQ ID NO: 1).

[0256] Initially, the general well-being of the mice participating in these experiments was assessed. Assessment was performed by daily measurements of food and water consumed by the mice, as well as their body weight (FIGs. 1A-1C).

[0257] The mice's food and water consumption were meticulously recorded throughout the study, with daily measurements taken to capture precise intake levels. At the culmination of the study, mice were sacrificed, and their body weights were measured.

[0258] Analysis of the data revealed no significant disparities in either food (FIG. 1A) consumption or water intake (FIG. IB) among experimental groups 1-4, indicating consistent consumption patterns emphasizing the meticulous nature of the research design and indicate that factors beyond food and water availability might be influential in any observed effects. One such observed effect is related to a reduction of about 2g in the measured body weight exhibited by mice induced with IBD using 3% DSS and receiving treatment with 8pM scrambled peptide relative to wt mice (FIG. 1C; compare group 3 with groups 1-2).

[0259] Advantageously and surprisingly, treatment of mice induced with an IBD model with 8pM peptide 1 improved this effect, by preventing about 50% (about 1g) of the loss of weight (compare group 3 with group 4).

[0260] Example 2 - NLGn4-NrxlB inhibitory peptide protected IBD model mice from morphological variations related to colon size

[0261] Next, morphological variations related to the size of the colon were assessed in mice belonging to same experimental groups 1-4 presented in the previous example.

[0262] Changes in colon length, for example, may provide additional insights into other potential morphological variations, and its proportion with respect to the weight of the colon constitutes a predictive measure / index indicative of colon health.

[0263] Therefore, following the experimental timeline, the size of the colon of the mice participated in these experiments was assessed by measurements of colon weight and colon length, and by calculating the ratio between them (FIGs. 2A-2C).

[0264] Analysis of the data uncovered statistically significant differences in morphology / size of the colon of the IBD-induced mice model and treated with 8pM scrambled peptide relative to wt mice. These differences include an increase in colon weight (FIG. 2A; compare group 3 with groups 1-2) in parallel to a decrease in colon length (FIG. 2B; compare group 3 with groups 1-2).

[0265] In accordance with these differences, the calculated ratio of colon weight / length was about 2.5-fold higher in mice induced with IBD model and treated with 8pM scrambled peptide relative to wt mice (FIG. 2C; compare group 3 with groups 1-2).

[0266] Advantageously and surprisingly, treatment of IBD model mice with peptide 1 improved this effect, by preventing increase in weight as well as the potential reduction in length (FIGs 2A-2B, respectively; compare groups 3 with groups 4). Accordingly, these effects also improved the calculated ratio of colon weight / length by about 2-fold (FIG. 2C; compare group 4 with groups 3), thereby bringing the value down and close to its normal levels (compare group 4 with groups 1-2).

[0267] Example 3 - NLGn4-NrxlB inhibitory peptide protected IBP model mice from changes in gene expression related to paracellular permeability and colon susceptibility

[0268] To gain further insight into other potential morphological variations that may be manifested by colons of mice induced with 3% DSS - such as changes in paracellular permeability, and to assess the potential of positively affecting them by treatment with NLGn4-Nrxip inhibitory peptide 1 - changes in gene expression of specific genes related to leakiness in the gut barrier, namely, Occludin, ZO-1, and Claudin was evaluated by quantifying mRNA levels of these genes in mice belonging to all experimental groups 1-4, as in previous examples (FIGs. 3A-3C).

[0269] In addition, the expression of Mucin mRNA levels, which relates to colon susceptibility was also evaluated using RT-PCR (FIG. 3D).

[0270] Results of the quantification of the expression of Occludin, ZO-1, Claudin, and Mucin show the levels of the mRNAs are increased by 2-fold to 4-fold in the IBD- induced mice model using 3% DSS and treated with 8pM scrambled peptide relative to wt mice (FIG. 3A, FIG.3B FIG.3C and FIG.3D; compare group 3 with groups 1-2).

[0271] Advantageously and surprisingly, treatment of IBD-induced mice with peptide 1 improved this effect, by preventing the potential increase in mRNA expression, keeping it at the range of about 1-fold to about 3 -fold for Occludin, ZO-1 and Mucin (FIG. 3A, FIG. 3B, and FIG. 3D, respectively; compare groups 4 with groups 3, while completely diminishing the potential increase in Claudin keeping it close to wt levels (FIG. 3C, respectively; compare group 4 with groups 1-2)

[0272] Therefore, it can be concluded that NLGN4X peptide administration to IBD- induced mice led to substantial enhancements in intestinal and colonic permeability through inhibitions in down expression of Occludin, Zo-1 and claudin protein levels indicating reduced leakiness in the gut barrier, which is often compromised in IBD.

[0273] Example 4 - NLGn4-NrxlB inhibitory peptide protected IBP model mice from colonic inflammation Colonic inflammation, i.e., colitis was induced by exposing mice to drinking water containing 3% DSS. The severity of inflammation was assessed in mice belonging to all experimental groups 1-4, as described in any of the previous examples, by measuring the level of the pro-inflammatory cytokines IL- lb, TNF-a, IL-6 and IL-4 in the colon, using a multiplexed sandwich enzyme-linked immunosorbent assay (ELISA).

[0274] As can be seen in FIG. 4, mice exposed to 3% DSS and treated with 8pM scrambled peptide exhibit a strong and highly significant increment in the levels of all four cytokines tested, compared to wt mice (compare group 3 with groups 1-2).

[0275] Nevertheless, administration of 8pM NLGn4-Nrxip inhibitory peptide 1 as a treatment to IBD model mice, was very potent with preventing the potential increase in the levels of all four inflammatory cytokines.

[0276] The analysis of the data showed that peptide 1 exerted a positive effect on the severity of the inflammation, as suggested from the reduction of about 25%-70% in the expression of IL- lb, TNF-a, IL-6 and IL-4 in IBD model mice relative to IBD model mice treated with 8pM scrambled peptide. (FIG. 4; compare group 4 with group 3).

[0277] Advantageously and surprisingly, it is disclosed that treatment with NLGn4- Nrxip inhibitory peptide 1 alleviated colonic inflammation in IBD-induced mice model.

[0278] Therefore, it can be concluded that the observed remarkable decrease in inflammation levels echoed by IL- lb, IL4, TNF-a, and IL-6 in the gut tissues of DSS mice treated with NLGN4X peptide, showcase its potent anti-inflammatory properties.

[0279] Example 5 - NLGn4-NrxlB inhibitory peptide protected IBD model mice from colon fibrosis

[0280] Next, extracellular matrix (ECM) markers of collagen I, collagen III and aSMA that characterize fibrosis / fibrostenotic strictures were evaluated in mice belonging to all experimental groups 1-4, as in any of the previous examples, by measuring mRNA expression levels of aSMA, Collagen III and Collagen I, as well as expression levels TGF-P protein, evaluated by ELISA (FIGs 5A-5B, respectively). Activated myofibroblasts express high levels of aSMA and therefore represent a markedly increased capability to contract ECM, which in turn contributes to tissue distortion and fibrostenotic stricture.

[0281] The analysis of the data was indicative of about 2-4 fold increase in expression of aSMA, Collagen I, Collagen III and TGF-P, in IBD model mice treated with 8pM scrambled peptide relative to wt mice (FIG. 5A and FIG. 5B; compare group 3 and groups 1-2).

[0282] However, when IBD-induced mice were treated with 8pM peptide 1 this effect was either completely diminished with a reduction of about 90%-100% for aSMA, Collagen I, Collagen III (FIG. 5A; compare group 4 and group 3) bringing their levels of expression to levels comparable to those of wt mice (compare group 4 and groups 1- 2), or that the effect was significantly reduced by about 20-40% for TGF-P (FIG. 5B; compare group 4 and group 3).

[0283] Advantageously and surprisingly, it is disclosed that treatment with peptide 1 alleviated colon fibrosis in mice induced with IBD model.

[0284] Therefore, it can be concluded that NLGN4X peptide may prevent fibrostenotic strictures associated with IBD.

[0285] Example 6 - NLGn4-NrxlB inhibitory peptide protected IBD-induced mice by preserving lipid homeostasis

[0286] Evaluation of lipid homeostasis was performed in mice belonging to all experimental groups 1-4, as in any of the previous examples, by measuring levels of cholesterol (Choi), triglyceride (TGR), and related genes SREBP2, HMGCR2 and HMGCS III.

[0287] The analysis of the data is indicative of increased levels and / or variability in each of the above-mentioned molecules and protein when measured in colon extract of mice induced for IBD and treated with 8pM scrambled peptide relative to wt mice (FIGs. 6A-6C; compare group 3 with groups 1-2).

[0288] Contrary, it was observed that treatment of IBD-induced mice with 8 pM peptide 1 reduced the potential increment observed in IBD-induced mice treated with scrambled peptide by preventing any increase in cholesterol levels, keeping its levels similar to those of wt (FIG. 6A; compare group 4 with groups 1-2), and by significantly inhibiting by more than 50% the potential induction of triglycerides (FIG. 6B; compare group 4 with group 3).

[0289] Similarly, treatment of IBD-induced mice with peptide 1 protected mice from potential increment in levels and / or variability of expression of SREBP2, HMGCR2 or HMGCS III (FIG. 6C; compare group 4 with group 3) also keeping the level of expression of these genes at about the same levels and / or variability of wt.

[0290] Advantageously and surprisingly, it is disclosed that peptide 1 protected IBD- induced mice from changes in lipid homeostasis, preserving the level of homoeostasis comparable to that of wt.

[0291] Therefore, it can be concluded that therapeutic intervention with NLGN4X peptide show a marked reduction in lipidemia reproduced by cholesterol and triglycerides levels in intestinal and colon tissues, and is indicative of its beneficial impact on lipid metabolism in the context of IBD.

[0292] Example 7 - NLGn4-NrxlB inhibitor peptide modulates B-neurexin expression in-vivo

[0293] P-Nrxn expression was evaluated in mice belonging to all experimental groups 1-4, as in any of the previous examples, by qPCR (FIG. 7).

[0294] More than about 2-fold increase in mRNA levels was observed in mice induced with 3% DSS and treated with 8pM scramble peptide, relative to wt mice (compare group 3 with groups 1-2).

[0295] This upregulation of P-neurexin expression in whole mice tissue of intestine and colon in mice model of DSS, is associated with any of the established effects of the IBD model, exemplified throughout any one of previous Examples 1-6, including for example, increased extracellular matrix (ECM) markers of collagen I, collagen III and aSMA that characterize fibrostenotic strictures, as was exemplified in Example 5.

[0296] Advantageously, as can be seen in FIG. 7, NLGn4-Nrxip inhibitor peptide 1 prevented any increase in the levels of P-Nrxn mRNA expression, keeping it at about same or even below wt levels (compare group 4 with groups 1-2). This is in accordance with its demonstrated ability to prevent any of the established effects of the IBD model, exemplified throughout any one of previous examples 1-6, including for example, preventing potential establishment of fibrostenotic strictures.

[0297] Therefore, it can be concluded that NLGN4X peptides holds a potential as a valuable therapeutic option for mitigating the pathological effects of IBD, in particular fibrostenotic strictures.

[0298] Example 8 - NLGn4-NrxlB inhibitory peptide protected mice from Acute Lung Injury (ALI)

[0299] Materials and Methods:

[0300] In-vivo experiments:

[0301] NLGn4-Nrxip Inhibitory Peptide synthesis - inhibitory peptide 1: MEQGEFLNYD (SEQ ID NO: 1), inhibitory peptide 2: QILMEQGEFLNYDIM (SEQ ID NO: 2), inhibitory peptide 3: DPQILMEQGEFLNYDIMLGV (SEQ ID NO: 3), and a control peptide having a scrambled sequence: LEGEDQNFYM (SEQ ID NO: 7) were synthesized by Pepscan Ltd. at a purity of 98.3%.

[0302] Animal ethics - The animals used were housed in a barrier facility and received care according to the Hebrew University ethical regulations and NIH guidelines. The institutional animal care ethics committee of the Hebrew University approved all animal protocols under the ethics number of MD- 18- 15494-3.

[0303] Experimental design of in-vivo experiments with Acute Lung Injury (ALI) mice - Twelve-week-old male weight 25±1.5 g wild-type C57BL / 6J obtained from Envigo (Harlan, Israel). ALI administration / induction was performed by i.p injection of the mice with Oleic acid (OA; Merck, O1008-1G >99% purity, as determined by LC / MS analyses) in the concentration of 100 mg / kg twice a week for two weeks. Treatment with scrambled peptide and NLGn4-Nrxip inhibitor peptide 1 was prepared fresh daily in phosphate buffered saline (PBS) at a concentration of 8pM at the volume of lOOpl day following OA administration. The prepared peptides were administered i.p. of twice a week for two weeks, n=4 mice per group.

[0304] Acute lung injury (ALI) and its most severe manifestation, the acute respiratory distress syndrome (ARDS), is a clinical syndrome defined by acute hypoxemic respiratory failure, bilateral pulmonary infiltrates consistent with edema, and normal cardiac filling pressures. An ALI-induced animal model was generated by injection of oleic acid to the animal, reproducing known risk factors for ARDS.

[0305] Groupl: wild-type mice without OA with scrambled peptide (8pM), i.p. injection twice a week for two weeks.

[0306] Group : wild-type mice without OA with NLGn4-Nrxip inhibitor peptide 1 (8pM), i.p injection twice a week for two weeks.

[0307] Group3: wild-type with OA (=ALI -induced mouse model) (100 mg / kg) with scrambled peptide (8pM), i.p. injection twice a week for two weeks.

[0308] Group4: wild-type mice with OA (= ALI-induced mouse model) (100 mg / kg) with NLGn4-Nrxip inhibitor peptide 1 (8pM), i.p injection twice a week for two weeks.

[0309] Additional procedures - Mice and diet were weighed, and water intake was measured daily. Prior to sacrifice, animals were anesthetized by 5% isoflurane inhalation for 10 seconds before cervical dislocation. On the sacrificing day whole blood was withdrawn through cardiac puncture collected in heparinized tubes for studying immune cells. Lung BALF extract were assessed for pro-inflammatory cytokine profile assessments by the ELISA and white blood cells (WBCs) count, mRNA was obtained from lung for gene expressions assessment by RT-PCR as described below.

[0310] RNA isolation, cDNA preparation, and real-time PCR - Total cellular of lOpg / pl RNA (purity 98%) determined using a Nanodrop ND- 1000 spectrophotometer (Nanodrop Technologies, Wilmington, DE) was isolated from all mice groups (n=5) using 2 ml of TRI reagent (Bio Lab; Cat# 90102331). The samples were centrifuged (1,400 rpm) for 15 minutes at 4°C to collected RNA-supematant. For RNA precipitation, the supernatant of each sample was transferred to a new microcentrifuge tube, and 0.5 ml of isopropanol (Bio Lab; Cat# 16260521) was added and incubated at 25°C for 10 minutes. The tubes were then centrifuged (12,000 rpm) for 10 minutes at 4°C, the supernatants were removed, and one milliliter of 75% ethanol was added to the pellets prior to centrifugation (7,500 rpm) for 5 minutes. The pellets were air-dried at room temperature for 15 minutes, 50 pl of DEPC was added, and the samples were heated for ten minutes at 55 °C. cDNA was prepared with a High-Capacity cDNA Isolation Kit (R&D; Cat# 1406197). Real-time PCR was performed with TaqMan Fast Advanced Master Mix (Cat# 4371130, Applied Biosystems) to quantify P-Nrxn, aSMA, GFAP and SP-D gene expressions with normalization to the expression of the housekeeping gene GAPDH. Cycling condition for Thermofisher one step RT-PCRkit involved RT step for 30 min at 50 °C and denaturation for 15 min at 95 °C. Further the reaction mixture was incubated for 40 cycles of 94 °C for 30 s, 60 °C for 30 s, and 72 °C for 1 min followed by 72 °C for 10 min. Data analysis were analyzed using QuantStudio™ 5 Real-Time PCR System (Cat# A34322, Applied biosystem).

[0311] Cytokines assessment in BALF - A multiplexed sandwich enzyme-linked immunosorbent assay -based technology (Cat# MHSTCMAG-70K; R&D Systems) was used to simultaneously determine the concentration of multiple cytokines of IL-6, IL- 4, IFN-y and sRAGE concentrations. Briefly, all reagents and samples were brought to room temperature (18 - 25 °C) before use. A volume of 100 pL of each standard and BALF sample were added into appropriate wells and incubate for 2.5 hours at room temperature with gentle shaking. The solution was discarded, wells were wash 4 times with IX Wash Solution, of note; washing was done by filling each well with Wash Buffer (300 pL) using a multi-channel Pipette or auto washer. Following the washing, the liquid was complete removal at each step is essential to good performance. A volume of 100 pL of lx prepared detection antibody were added to each well for 1 hour at room temperature with gentle shaking. A volume of 100 pL of prepared streptavidin solution to each well for 45 minutes at room temperature with gentle shaking. A volume of 100 pL of TMB One-Step Substrate Reagent (Item H) were added to each well for 30 minutes at room temperature in the dark with gentle shaking. Finally, 50 pL of Stop Solution (Item I) were added to each well. Absorbance was read at 450 nm immediately using ELISA reader (Tecan Ml 00 Plate Reader). trNK cells isolation from the lung - Isolation of lung trNK cells were prepared by digesting the lung tissue samples at 37°C for 40 minutes with 20 ml of GBSS solution containing 0.1% (wt / vol) pronase, 0.1% (wt / vol) collagenase, and 0.01% (wt / vol) deoxyribonuclease I (DNase I) (all enzymes from Roche Diagnostics GmbH, Mannheim, Germany). The cell suspension was fdtered through a 150-mm steel mesh and centrifuged on a FICOLL (Sigma, St. Louis, USA) at 1,400 x g for 20 minutes at 25 °C to produce a WBCs-enriched fraction in the middle opaque layer. The whole opaque layer was collected and washed by PBS. trNK purification was performed using EasySep™ Mouse NK Cell Isolation Kit. Statistical analysis - Statistical differences were analyzed with a two-tailed unpaired Student’s t test (for comparisons between two groups) or one-way or two-way analysis of variance (ANOVA; one-way ANOVA with the Newman-Keuls post hoc test for comparisons among multiple groups) with GraphPad Prism 9.0 (GraphPad Software, La Jolla, CA). A t-test of p value <0.05 is considered statistically significant and was calculated as the difference in means between two variables. Results are presented as mean ± SD or as average means of experimental replicates ± SD.

[0312] Results:

[0313] In-vivo experiments including mice induced with Acute Lung Injury (ALI) were carried out by comparing between four groups of mice, according to the experimental procedure of ALI-induced animal model described hereinabove and summarized herein below, as follows: group 1 - wt mice administered with 8pM of scrambled peptide (SEQ ID NO: 7); group 2 - wt mice administered with 8pM of peptide 1 (SEQ ID NO: 1); group 3 - ALI-induced mice model (Oleic acid-treated) administered with 8pM of scrambled peptide (SEQ ID NO: 7); and group 4 - ALI-induced mice model (Oleic acid-treated) administered with 8pM of peptide 1 (SEQ ID NO: 1).

[0314] Assessment of the general well-being of mice induced with ALI model - The mice's food and water consumption were meticulously recorded throughout the study, with daily measurements taken to capture precise intake levels. Analysis of the data revealed no significant disparities in either food consumption, or water intake, among experimental groups 1-4, indicating consistent consumption patterns emphasize the meticulous nature of the research design and indicate that factors beyond food and water availability might be influential in any observed effects (Data not shown).

[0315] At the culmination of the study, mice were sacrificed, and their weights were measured in grams. No significant disparities were observed in mice body weight among experimental groups 1-4 (Data not shown).

[0316] NLGn4-NrxlB inhibitory peptide protected ALI-induced mice from increased lung weight - Next, variations related to the weight of the lung were assessed in mice belonging to same experimental groups 1-4.

[0317] Therefore, following the experimental timeline, lungs from each subject were carefully harvested, the weight of the lungs was measured, and the ratio between lungs weight and mice weight (lungs weight: mice weight ratio) was also calculated to ascertain any proportional changes.

[0318] The analysis of the data suggests that NLGn4-Nrxip inhibitory peptide 1 protected the lungs of ALI mice from increased weight (FIG. 8A; compare group 4 with groups 3 relative to groups 1-2), and from increment in the calculated ration of lungs weight : mice weight (FIG. 8B; compare group 4 with groups 3 relative to groups 1-2).

[0319] NLGn4-NrxlB inhibitory peptide protected ALI-induced mice from increased B- Nrxn expression in the lungs - Quantification of B-Nrxn mRNA expression was performed in lung tissues obtained from mice belonging to experimental groups 1-4. The analysis of the data suggests that B-Nrxn is important for mediating lung injury in ALI mice of the current model (FIG. 9; compare group 4 with groups 3 relative to groups 1-2).

[0320] NLGn4-NrxlB inhibitory peptide protected ALI-induced mice from fibrosis in the lungs - Lung fibrosis was assessed by measuring mRNA expression levels of aSMA and GFAP in mice belonging to all experimental groups 1-4. The analysis of the data suggests that NLGn4-NrxlB inhibitory peptide 1 protected the lungs of ALI mice from increased expression of fibrosis markers aSMA and GFAP (FIG. 10; compare group 4 with groups 3 relative to groups 1-2).

[0321] NLGn4-NrxlB inhibitory peptide protected ALI-induced mice from increment in inflammation in the lungs - lung inflammation was assessed by measuring the levels of pro-inflammatory cytokines, including IFN-y, IL-6 and IL-4, in fluid recovered by bronchoalveolar lavage (BAL fluid or BALF) from mice belonging to all experimental groups 1-4. The analysis of the data suggests that NLGn4-NrxlB inhibitory peptide 1 protected the lungs of ALI mice from increased inflammation (FIG. 11; compare group 4 with groups 3 relative to groups 1-2).

[0322] NLGn4-NrxlB inhibitory peptide protected ALI-induced mice from increment in the number of trNK cells in the lungs - the number of trNK cells purified / isolated from the lungs was assessed in mice belonging to all experimental groups 1-4, and was calculated as (cells\mg lung tissue weight). The analysis of the data suggests that NLGn4-NrxlB inhibitory peptide 1 protected the lungs of ALI mice from increased amount of trNK cells (FIG. 12; compare group 4 with groups 3 relative to groups 1-2). NLGn4-NrxlB inhibitory peptide protected ALI-induced mice model from increment in the levels of soluble RAGE associated with lung injury - the soluble form of the receptor for advanced glycation end products (sRAGE) was assessed by measuring its levels in fluid recovered by bronchoalveolar lavage (BAL fluid or BALF) from mice belonging to all experimental groups 1-4. The analysis of the data suggests that NLGn4-Nrxl P inhibitory peptide 1 protected the lungs of ALI mice from increment in levels of sRAGE associated with severe injury (FIG. 13; compare group 4 with groups 3 relative to groups 1-2).

[0323] NLGn4-NrxlB inhibitory peptide protected ALI-induced mice from increment in the levels of surfactant protein D (SP-D) - the level of SP-D was assessed in mice belonging to all experimental groups 1 -4. The analysis of the data suggests that NLGn4- Nrxi inhibitory peptide 1 protected the lungs of ALI mice from increment in levels of SP-D (FIG. 14; compare group 4 with groups 3 relative to groups 1-2).

[0324] To conclude, NLGN4X peptides hold a potential as a valuable therapeutic option for mitigating the pathological effects of ALI, in particular those effects related to fibrosis and inflammation of lung tissue.

[0325] The foregoing description of the specific embodiments will so fully reveal the general nature of the invention that others can, by applying current knowledge, readily modify and / or adapt for various applications such specific embodiments without undue experimentation and without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. The means, materials, and steps for carrying out various disclosed functions may take a variety of alternative forms without departing from the invention. It is to be understood that further trials are being conducted to establish clinical effects.

Claims

CLAIMS:

1. A composition comprising an isolated peptide comprising an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6, for use in treating, attenuating, and / or preventing progression of a condition that involves an abnormal inflammatory response in a subject in need thereof.

2. The composition for use according to claim 1, wherein the isolated peptide comprises the amino acid sequence as denoted by SEQ ID NO: 1.

3. The composition for use according to any one of claims 1 or 2, comprising a pharmaceutically acceptable carrier.

4. The composition for use according to any one of claims 1-3, comprising a plurality of isolated peptides.

5. The composition for use according to any one of claims 1-4, wherein the condition that involves an abnormal inflammatory response comprises an autoimmune disease.

6. The composition for use according to claim 5, wherein the autoimmune disease is selected from one or more of: Inflammatory bowel disease (IBD), Thyroid diseases including Graves' disease and Hashimoto’s thyroiditis, Adrenal diseases including Addisson disease, Thymus disease including Myasthenia gravis, Multiple sclerosis (MS), Rheumatoid arthritis, Psoriasis and Psoriatic arthritis, Type 1 diabetes, Coeliac disease, Systemic lupus erythematosus, Scleroderma, Polymyositis, Dermatomyositis, and Pernicious anemia, or any combination thereof.

7. The composition for use according to claim 6, wherein the IBD is selected from any one or more of: ulcerative colitis (UC), Crohn's disease (CD) Collagenous colitis, Lymphocytic colitis, Ischemic colitis, Diversion colitis, Behcet's disease, and Indeterminate colitis, or any combination thereof.

8. The composition for use according to any one of claims 1-4, wherein the condition that involves an abnormal inflammatory response comprises Acute lung injury (ALI).

9. The composition for use according to claim 8, wherein the ALI comprises acute respiratory distress syndrome (ARDS).

10. The composition for use according to any one of claims 1-4, wherein the condition that involves an abnormal inflammatory response comprises cancer.

11. The composition for use according to any one of claims 1-10, wherein administration of the composition comprises systemic and / or localized administration.

12. The composition for use according to any one of claims 1-11, administered in combination with at least one additional therapeutic agent.

13. The composition for use according to any one of claims 1-12, wherein the treatment comprises prevention or reduction of inflammation in the subject.

14. The composition for use according to any one of claims 1-13, wherein the treatment comprises prevention or reduction of fibrosis and / or fibrostenotic strictures in the subject.

15. The composition for use according to any one of claims 1-14, wherein the treatment comprises prevention or reduction of number of trNK cells in the subject.

16. The composition for use according to any one of claims 1-7, wherein the treatment comprises prevention or reduction of morphological variations related to colon size in the subject.

17. The composition for use according to any one of claims 1-7, wherein the treatment comprises prevention or reduction of paracellular permeability and / or susceptibility of an intestine of the subject.

18. The composition for use according to any one of claims 1-7, wherein the treatment comprises prevention or reduction of changes in lipid homeostasis in the subject.

19. The composition for use according to any one of claims 1-4, wherein the condition that involves an abnormal inflammatory response in the subject involves one or more of: inflammation in the subject, fibrosis and / or fibrostenotic strictures in the subject, increased levels of trNK cells in the subject, morphological variations related to colon size or lung weight in the subject, increased paracellular permeability and / or susceptibility of an intestine or lung in the subject, and changes lipid homeostasis in the subject, or any combination thereof.

20. A method of treating, attenuating and / or preventing progression of a condition that involves an abnormal inflammatory response in a subject in need thereof, the method comprising administering a therapeutically effective amount of a composition comprising an isolated peptide consisting of an amino acid sequence as denoted by any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and / or SEQ ID NO: 6.

21. The method of claim 20, comprising administering a therapeutically effective amount of a composition comprising an isolated peptide consisting of an amino acid sequence as denoted by any one of SEQ ID NO: 1 and / or SEQ ID NO: 6.

22. The method of claim 20 or 21, comprising administering a therapeutically effective amount of a composition comprising a plurality of isolated peptide.

23. The method according to any one of claims 1-10, wherein the administration of the composition comprises systemic and / or localized administration.

24. The method according to any one of claims 20-23, wherein the composition is administered in combination with at least one additional therapeutic agent.

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