Il-4 derived peptides for use in the treatment of obesity

Abstract

To provide a peptide or a pharmaceutically acceptable salt thereof for use in the treatment of obesity.SOLUTION: The present disclosure provides a peptide derived from interleukin-4 (IL-4) for the treatment of obesity.SELECTED DRAWING: Figure 1A

Description

[Technical Field]

[0001] The present disclosure relates to peptides derived from IL-4 for the treatment of obesity. In particular, the present disclosure relates to and a method for treating obesity comprising administering to a subject a therapeutically effective amount of an anti-obesity drug comprising the amino acid sequence of SEQ ID NO: 1 or a variant thereof. This relates to peptides. [Background technology]

[0002] (background) Overweight and obesity are defined as abnormal or excessive fat accumulation that presents a risk to health. Body mass index (BMI) is calculated based on weight and height. A BMI above 25 is considered overweight, and a BMI above 30 is considered obese. Obesity has reached epidemic proportions and pandemic levels over the past 50 years. This has nearly tripled since 1975. In 2016, there were more than 1.9 billion adults aged 18 and over. Of these, over 650 million were obese. More than 340 million children and adolescents were overweight or obese in 2016 (Ro glic, G. 2016).

[0003] Both overweight and obesity increase the risk of type 2 diabetes, heart disease, stroke, and some types of cancer. It is a systemic disease associated with the leading causes of morbidity and mortality in the United States, including diabetes (Roglic , G. 2016). Therefore, obesity can also lead to poor mental health and a reduced quality of life. It is a serious condition as it is associated with and contributes to a reduced life expectancy (Abdelaal 201 7).

[0004] Several drugs have been approved for the treatment of obesity. One example is the drug under the trade name Wegovy Semaglutide is marketed in the US and is the first drug for the treatment of type 2 diabetes. It is a glucagon-like peptide-1 (GLP-1) agonist developed and approved in The most notable effect of GLP-1 agonists is the activation of GLP-1 receptors expressed on pancreatic β cells. their ability to stimulate insulin secretion in a glucose-dependent manner by binding to the body Most importantly, GLP-1 agonists increase blood glucose levels above fasting levels. Importantly, this has been shown to inhibit glucagon secretion in response to hypoglycemia. GLP-1 agonists have no effect on the glucagon response compared to insulin. To make it a safe antidiabetic drug with a consistently low incidence of hypoglycemia.

[0005] In June 2021, semaglutide was approved for the treatment of at least one weight-related condition (e.g., high blood pressure, 2 Chronic weight management in obese or overweight adults with type 2 diabetes or high cholesterol It has been approved by the U.S. Food and Drug Administration as a GLP-1 agonist for anti-obesity medication. acts by binding to GLP-1 receptors in the hypothalamus, thereby suppressing appetite Furthermore, GLP-1 agonists bind to GLP-1 receptors in the stomach, inhibiting gastric emptying and acid reflux. GLP-1 inhibits secretion and motility, which collectively promote satiety. Diabetic subjects treated with .beta.-.beta.-.beta.-.beta. receptor agonists also often experience improved control of their blood glucose levels. In addition to the control of obesity, patients experience beneficial weight loss. However, there are no novel or novel methods for the treatment of obesity in the art. There remains a need for more effective treatments.

[0006] Human interleukin-4 (IL-4) consists of the following amino acid sequence (signal peptide, Namely, amino acid AA 1-24It is an anti-inflammatory cytokine that has the following properties: MGLTSQLLPP LFFLLACAGN FVHGHKCDIT LQEIIKT LNS LTEQKTLCTE LTVTDIFAAS KNTTEKETFC RAA TVLRQFY SHHEKDTRCL GATAQQFHRH KQLIRFLKRL DRNLWGLAGL NSCPVKEANQ STLENFLERL KTIMRE KYSK CSS (UniProtKB - P05112; SEQ ID NO: 3)

[0007] Mature IL-4 has 129 amino acid residues and is divided into three components with the following amino acid sequences: Intramolecular disulfide bonds (Cys3-Cys127; Cys46-Cys99; Cys 24-Cys65). HKCDITLQEI IKTLNSLTEQ KTLCTELTVT DIFAASK NTT EKETFCRAAT VLRQFYSHHE KDTRCLGATA QQF HRHKQLI RFLKRLDRNL WGLAGLNSCP VKEANQSTLE NFLERLKTIM REKYSKCSS (SEQ ID NO: 4)

[0008] Like other cytokines, IL-4 acts by binding to receptors on the cell surface. It exerts its biological activity. Two types of receptors for IL-4 are known: type I IL-4 Type I IL-4R is composed of the IL-4Rα chain and the IL-2Rγ chain. (γ c ) chain, whereas type II IL-4R is composed of IL-4Rα and IL-4Rα. IL-4 is composed of the α1 chain and α2 chain of IL-13. It stimulates the proliferation of activated B and T cells, They have many biological roles, including the differentiation of B cells into plasma cells, and in extravascular tissues. The presence of IL-4 in the inflammatory response promotes alternative activation of macrophages into M2 cells and M1 cells. IL-4 inhibits the classical activation of macrophages, resulting in a reduction of pathological inflammation. It is secreted primarily by CD4+ T cells (Th2 cells), which regulate the human immune response. It regulates the coordination of specific cytokine inhibitors and soluble cytokine receptors to promote cytokine production. Maintaining the right balance between pro- and anti-inflammatory activity will help prevent acute and chronic Therefore, modulation of IL-4 signaling plays an important role in the pathogenesis of chronic inflammation. IL-4 has been extensively studied in autoimmune and inflammatory diseases. It has been shown to promote lipolysis through the enhancement of HSL (Hyperlipase) activity ( See Scientific Reports, 2019 9:11974).

[0009] Small peptide fragments derived from IL-4 that act as IL-4 mimetics have previously been reported in macrophages. Inhibits TNF-α release from phage and induces neuritogenic responses from primary neurons It has been shown that (see WO 2010 / 054667). An example is the peptide having the amino acid sequence AQFHRHKQLIRFLKRA (SEQ ID NO: 1). This peptide binds to the IL-4 receptor and inhibits macrophage activation. This can prevent the onset of inflammatory reactions and therefore reduce the risk of rheumatoid arthritis , multiple sclerosis, Alzheimer's disease, Parkinson's disease, and Huntington's disease, among many others. It has been proposed to be useful in inflammatory and autoimmune diseases. Summary of the Invention [Problem to be solved by the invention]

[0010] (Abstract) The present invention relates to mice administered a peptide comprising SEQ ID NO:1 (i.e., treated Regarding the surprising observation that the steroid group (the steroid group) was less sensitive to weight gain compared to the placebo group Therefore, the present invention provides peptides derived from IL-4 that act as IL-4 mimetics. Although previously associated with anti-inflammatory effects, there is little recognition that it can be used to treat obesity. Therefore, peptide fragments derived from IL-4 that act as IL-4 mimetics are The fragment is a peptide fragment of SEQ ID NO: 1 and is a new generation of anti-obesity drug with a novel mode of action. High-affinity partial agonists for type I IL-4R may hold promising promise. It has been shown to be an antagonist of type II IL-4R (Klemen tiv 2013). [Means for solving the problem]

[0011] In a first aspect, the present disclosure provides a peptide or pharmaceutical composition thereof for use in the treatment of obesity. Acceptable salts of the compound of formula (I) are those containing 13 to 19 amino acid residues and further comprising the amino acid sequence AQ A fragment having the sequence AQFHRHKQLIRFLKRA (SEQ ID NO: 1), or 1, 2, or 3 AQFHRHKQLIRFLKRA (SEQ ID NO: 1) having amino acid substitutions and / or deletions of ) or a pharmaceutically acceptable salt thereof.

[0012] In a second aspect, the present disclosure provides a peptide or a pharmaceutically acceptable salt thereof for use in the treatment of obesity. and in the case of an acceptable salt thereof, the peptide may comprise two or more covalently linked peptide fragments according to the first aspect. Includes pieces.

[0013] In a third aspect, the present disclosure provides a pharmaceutical composition for use in treating obesity, comprising: and / or to a pharmaceutical composition comprising a peptide according to the second aspect.

[0014] In a fourth aspect, the present disclosure relates to a method for treating obesity, the method comprising administering to a subject a subject in need thereof a therapeutically effective amount of the compound disclosed herein. According to any of the aspects and embodiments described herein, an effective amount of the peptide or pharmaceutical composition is administered to This includes administering to an individual in need.

[0015] (Definitions and Abbreviations) The term "individual" includes vertebrates, particular members of a mammalian species, preferably humans. As used herein, "subject" and "individual" are used interchangeably. It is possible.

[0016] An "individual in need thereof" refers to an individual who would benefit from the present disclosure. The individual in need thereof is obese (BMI > 30) and preferably has at least one weight and individuals with associated conditions (eg, high blood pressure, type 2 diabetes, or high cholesterol). In another embodiment, the individual in need thereof is overweight (BMI>25), preferably underweight. have at least one weight-related condition (e.g., high blood pressure, type 2 diabetes, or high cholesterol) Therefore, in this context, "obese" preferably refers to an individual who has at least one Overweight (B) with weight-related conditions (e.g., high blood pressure, type 2 diabetes, or high cholesterol) should be understood broadly to include MI > 25.

[0017] An "effective amount" of a peptide may be a single administration or, for example, a total effective amount within a 24-hour period. The dosage can be administered via multiple doses of the same amount. It is important to consider the appropriate amount and timing of administration. An "effective amount" can be determined using standard clinical procedures for determining therapeutic efficacy. Experience and / or individualization (case-by-case) on the part of the treating medical professional and / or individual It is understood that this may be the result of a test.

[0018] In the context of the present invention, it is understood that the present invention provides an amino acid region, e.g., a region in which several consecutive amino acid residues are present. For example, the AA of interleukin-4 (SEQ ID NO: 4) 68 -AA 97 Peptide fragments derived from When referring to fragments, it refers to fragments selected from the respective regions of interleukin-4 (SEQ ID NO: 4). It should be understood as a short stretch (i.e., a continuous sequence) of amino acids. Thus, the cytokine IL-4 (SEQ ID NO: 4) does not form part of the present invention. Amino acid region AA of interleukin-4 (SEQ ID NO: 4) 68 -AA 89 16 derived from A peptide fragment containing consecutive amino acid residues is shown below with the consecutive sequence highlighted in bold. can be obtained from

[0019] HKCDITLQEI IKTLNSLTEQ KTLCTELTVT DIFAASK NTT EKETFCRAAT VLRQFYSHHE KDTRCLGATA QQF HRHKQLI RFLKRLDRNL WGLAGLNSCP VKEANQSTLE NFLERLKTIM REKYSKCSS (SEQ ID NO: 4).

[0020] Thus, the amino acid region AA of interleukin-4 (SEQ ID NO: 4) 68 -AA 89 The 16 consecutive amino acid residues derived from the fragment / sequence ATAQQFHRHKQLI RFL;TAQQFHRHKQLIRFLK;AQQFHRHKQLIRFLKR,QQ FHRHKQLIRFLKRL, QFHRHKQLIRFLKRLD, FHRHKQLI RFLKRLDR, HRHKQLI RFLKRLDRN, and RHKQLI RFLK The peptide contains a fragment derived from interleukin-4 (SEQ ID NO: 4) / sequence variants. Non-limiting examples include AQFHRHKQLIRFL KRA (SEQ ID NO: 1) is a variant of the fragment / sequence QQFHRHKQLIRFLKRL where the terminal amino acids Q and L are replaced with A (i.e., AQFHRHKQ LIRFLKRA (SEQ ID NO: 1) has two amino acid substitutions). In this context, the present disclosure provides, for example, the amino acid region AA of interleukin-4 (SEQ ID NO: 4). 68 -AA 97 A peptide containing a fragment of 13 to 19 consecutive amino acid residues derived from When referring to a single amino acid sequence, the number of consecutive amino acids must not be less than 13. 19 in the fragment (i.e., without considering any linker, if any) The peptide may consist of two or more peptides linked via a linker (spacer). It should be understood that such fragments may be included. Non-limiting examples include those of formula (I): The following peptides each consist of 16 consecutive amino acids linked via a linker at their C-terminus: It contains two peptide fragments of amino acids. [ka]

[0021] Peptide fragments can also be lipidated to alter the pharmacokinetic properties of the peptide. .

[0022] As used herein, "sequence identity" refers to an exact match between two different sequences being compared. The number of amino acid residues that the variants share is 90% of the sequence with 10 amino acid residues. If identical, the variant may differ from the sequence by a single amino acid substitution.

[0023] As used herein, the terms "treat," "treatment," and "therapy" refer to curative therapy. The term "treatment," "prophylactic treatment," "preventative treatment," or "ameliorative treatment" refers equally to a beneficial or desirable treatment. This includes approaches to achieve physiological outcomes such as weight loss or fat percentage. This can be clinically established, such as a decrease in

[0024] The term "covalently bonded" refers to molecules covalently linked to each other via a linker or spacer. , should be understood as at least two peptides / fragments according to the present disclosure. The peptides can have the same or different sequences. The peptides can be linked together using a dendrimer, for example. , fusion peptides / proteins, or hybrid peptides are commonly known in the art. For example, the linker (spacer) can be a peptide. Linkers, e.g., lysine-beta alanine, a single amino acid, as exemplified herein , for example lysine, or a small stretch of amino acids (e.g., Adv Drug Deliv Rev. 2013 Oct 15; 65(10): 1357-136 9). The linker may also be a non-peptide linker, such as a PEG chain, or an ethylenediamine diamine diamine. Those skilled in the art will appreciate that the linker (spacer) used may be a diamine such as methylamine. It is understood that other linkers (spacers) may be used without departing from the concept of the invention. Most preferably, the peptide / fragment is a peptide fragment thereof, as exemplified herein. Covalently bonded via their N- or C-terminus, most preferably via their C-terminus Most preferably, the two peptides / fragments are lysine-coated, as exemplified herein. The beta-alanine in the lysine linker reduces steric hindrance during synthesis. Acts as a spacer between the linkers used in SPPS to Therefore, the beta-alanine moiety is used in the field of SPPS. Other common spacers may be substituted. Various branched lysine-based dendrimers For example, (Proc. Natl. Acad. Sci. USA, Vol. 85 , pp. 5409-5413, August 1988) The dimers disclosed herein have been shown to perform better than the monomers. was discovered.

[0025] The term "variant" refers to a peptide sequence that has, for example, one or more amino acid residue substitutions. Both L- and D-amino acids can be used. Alternatively, unnatural amino acids, preferably L-amino acids, may be used. When referring to variants having one, two, or three amino acid substitutions and / or deletions, It is understood that the total number of substitutions and / or deletions may be up to three. For example, a variant may have two substitutions and one deletion (total = 3). A variant may have three deletions and a zero substitution (total = 3), or a variant may have three deletions and a zero substitution. As an example, the present disclosure may be used in the treatment of obesity. For example, the peptide has 13 amino acid residues, and The peptide is a variant of AQFHRHKQLIRFLKRA (SEQ ID NO: 1, i.e., 16 amino acids). If it contains 3 amino acids, it is a variant with 3 deletions (16-3 = 13 amino acids). Preferably, the substitutions in the variant are conservative amino acids. Conservative amino acid groups are as follows: A, G (neutral, weakly hydrophobic), Q, N, S, T (hydrophilic, uncharged) E, D (hydrophilic, acidic) H, K, R (hydrophilic, basic) A, L, P, I, V, M, F, Y, W (hydrophobic, aromatic) C (crosslink formation)

[0026] Conservative substitutions may be introduced at any position in a peptide for use according to the present disclosure. While non-conservative substitutions may be introduced at any position in a peptide for use according to the present disclosure, It may also be desirable.

[0027] The peptide sequences of the present disclosure can be synthesized by any conventional synthetic method, such as solid phase peptide synthesis (SPPS). It can be prepared by

[0028] The peptides according to the present invention may be in the form of pharmaceutically acceptable salts. Pharmaceutically acceptable salts include any salt commonly used in peptide formulations. Such salts include both acid addition and base salts, e.g. Remington's Pharmaceutical Sciences, 1 It can be found in the 7th edition. [Brief explanation of the drawings]

[0029] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1A] FIG. 1A shows the crystal structure of IL-4 (PDB identifier: 1CYL) with the amino acid residues shown in the region AA71-AA86 forming part of α-helix C (AA71-AA94). [Figure 1B] FIG. 1B shows the crystal structure of IL-4 (PDB identifier: 1CYL) with the amino acid residues shown in the region AA71-AA86 that constitute part of α-helix C (AA71-AA94). DETAILED DESCRIPTION OF THE INVENTION

[0030] (Detailed explanation) The present invention relates to small peptide fragments derived from IL-4, more specifically those associated with anti-inflammatory effects. The α-helix C of IL-4 (i.e., IL-4R type I agonist) ) was found to significantly reduce the body weight of animals upon administration. do.

[0031] Accordingly, the present disclosure provides a peptide for use in the treatment of obesity, comprising IL-4 Peptides that are IL-4 receptor type I agonists, preferably IL-4 receptor type II antagonists Regarding.

[0032] Mature IL-4 has 129 amino acid residues and three intramolecular disulfide bonds (Cys 3-Cys127; Cys46-Cys99; Cys24-Cys65) The peptides used in the experimental section of this specification do not contain a linker (spacer) (i.e., lysine -beta alanine) through AQFHRHKQLIRFLKRA (SEQ ID NO: 1) (i.e. AQFHRHKQ covalently linked to another peptide having the amino acid sequence of It has the amino acid sequence of LIRFLKRA (SEQ ID NO: 1) (i.e., the first fragment). The peptide in column number 1 is amino acid AA of SEQ ID NO: 4 71 -AA 86 (Contains signal sequence It is a variant of a peptide derived from IL-4 (not IL-4) (highlighted in bold below).

[0033] HKCDITLQEI IKTLNSLTEQ KTLCTELTVT DIFAASK NTT EKETFCRAAT VLRQFYSHHE KDTRCLGATA QQF HRHKQLI RFLKRLDRNL WGLAGLNSCP VKEANQSTLE NFLERLKTIM REKYSKCSS (SEQ ID NO: 4)

[0034] Amino acid AA of SEQ ID NO: 1 and SEQ ID NO: 4 71 -AA 86 It can be seen from the alignment with Thus, SEQ ID NO: 1 represents a peptide fragment derived from this site with two amino acid substitutions. is a variant of (highlighted in bold).

[0035] QQFHRHKQLIRFLKRL IL-4 AA 71 -AA 86 (SEQ ID NO: 4) AQFHRHKQLIRFLKRA (SEQ ID NO: 1).

[0036] Region AA of SEQ ID NO: 4 71 -AA 86 As shown in the crystal structure of Figure 1A / B, IL -4 α-helix C (AA 71 -AA 94 ) (PDB identifier: 1CY L). This α-helix C interacts with the ectodomain of IL-4Rα1, Therefore, α-helix C(AA 71 -AA 94 ) is a peptide fragment derived from SEQ ID NO: It is likely to have similar binding characteristics and pharmacology to No. 1. In other words, AA 71 -AA 94 , preferably AA of SEQ ID NO: 4 71 -AA 86 Peptides derived from or near the region of The compound is suitable for use in the treatment of obesity. 71 -AA 94 ) The peptide fragment derived from the present invention has an α-helix for at least a part of the amino acid sequence. The tendency of short peptides to form α-helices is Predictions are made using software such as Arture Prediction Server. or can be determined experimentally using, for example, X-ray crystallography. PROTEUS Structure Prediction Server The amino acids highlighted in bold AQFHRHKQLIRFLKRA (SEQ ID NO: 1) are α-heterocyclic amino acids. Predicts that a person has a high tendency to form dyslexia (on a scale ranging from 0 to 9) (Confidence score 9, 9 being the highest confidence level). Therefore, most preferably, IL-4 α- Helix C (AA 71 -AA 94 ) at least a portion of the fragment derived from Form.

[0037] In particular, the present invention provides a method for the preparation of a polypeptide having the amino acid sequence AQFHRHKQLIRFLKRA (SEQ ID NO: 1). As shown in the experimental section of the present specification, a peptide containing a fragment of the The findings are related to a significant decrease in

[0038] Thus, in a first aspect, the present disclosure provides a peptide for use in the treatment of obesity. The peptide comprises 13 to 19 amino acid residues, and further comprises: a fragment having the amino acid sequence AQFHRHKQLIRFLKRA (SEQ ID NO: 1), or 1, AQFHRHKQLIRF with 2 or 3 amino acid substitutions and / or deletions Includes variants of LKRA (SEQ ID NO: 1).

[0039] In a preferred embodiment, the present disclosure provides a peptide for use in the treatment of obesity, comprising: It contains 14 to 18 amino acid residues and further contains the amino acid sequence AQFHRHKQLIRFL A fragment having KRA (SEQ ID NO: 1) or one, two, or three amino acid substitutions and / or deletions A peptide containing a variant of AQFHRHKQLIRFLKRA (SEQ ID NO: 1) with a deletion Regarding do.

[0040] In a more preferred embodiment, the present disclosure relates to peptides for use in the treatment of obesity. and contains 15 to 17 amino acid residues, and further contains the amino acid sequence AQFHRHKQLIR A fragment having FLKRA (SEQ ID NO: 1) or a fragment having 1, 2, or 3 amino acid substitutions and / or The gene encoding the mutant AQFHRHKQLIRFLKRA (SEQ ID NO: 1) contains a deletion. Regarding Petite.

[0041] In an even more preferred embodiment, the present disclosure provides a peptide for use in the treatment of obesity. comprising 16 amino acid residues and further having 1, 2 or 3 amino acid substitutions. SEQ ID NO: 1 (AQFHRHKQLIRFLKRA) or SEQ ID NO: 1 (AQFHRHKQ This invention relates to peptides containing variants of LIRFLKRA.

[0042] In another embodiment, the present invention provides a peptide or pharmaceutical composition thereof for use in the treatment of obesity. Regarding acceptable salts, the peptide is 68 -AA 97 , preferably AA 71 -AA 94 , more preferably AA 71 -AA 89 , most preferably interloy AA of Kin-4 (SEQ ID NO: 4) 71 -AA 86 13 to 19 consecutive amino acids derived from a variant thereof containing an acid residue or having 1, 2, 3, or 4 amino acid substitutions In a more preferred embodiment, the peptide has the amino acid region AA 68 -AA 97 , preferred Or AA 71 -AA 94 , preferably AA 71 -AA 89 , most preferably AA 71 - AA 86 14 to 18 consecutive amino acids derived from interleukin-4 (SEQ ID NO: 4) a barrier having one, two, three, or four amino acid residues or having one, two, three, or four amino acid substitutions In a more preferred embodiment, the peptide is 68 -AA 97 , Preferably AA 71 -AA 94 , preferably AA 71 -AA 89 , most preferably Inter -Leukin-4 AA 71 -AA 86 15 to 17 consecutive amino acids derived from (SEQ ID NO: 4) or a variant thereof having one, two, three, or four amino acid substitutions. In an even more preferred embodiment, the peptide is a variant of the amino acid region AA 68 - AA 97 , preferably AA 71 -AA 94 , more preferably AA 71 -AA 89 , most preferred Preferably, the AA of interleukin-4 (SEQ ID NO: 4) 71 -AA 86 16 derived from Consecutive amino acid residues or 1, 2, 3, or 4 amino acid substitutions In the above embodiment, the peptide has the amino acid sequence AQFHRHK QLIRFLKRA (SEQ ID NO: 1), or AQFHRHKQ with one or two substitutions It is highly preferred to include variants of LIRFLKRA (SEQ ID NO: 1).

[0043] In any of the above embodiments, the variant preferably has two amino acid substitutions and and / or deletions, even more preferably one amino acid substitution or deletion.

[0044] Peptides for use according to the invention are amidated at the C-terminus (-CONH2). It can also be expressed as a free carboxylic acid (-COOH) or a methyl ester (-COOMe). The peptides according to the invention may have a free amine (-NH2), N-acylation (-NHCOR), N-methylation (-NHCH3 or -N(CH3)2), The peptide may also be deaminated at the N-terminus, which alters the PK properties of the peptide. For example, the N-terminal end and / or one or more lysine residues may be lipidated to achieve the desired effect. It is possible.

[0045] Thus, in one embodiment of the present disclosure, the C-terminal amino acid is a free carboxylic acid ("-COO In another embodiment, the C-terminal amino acid is present as an amidated derivative (" In another embodiment, the N-terminal amino acid is a free amino group (“NH In another embodiment, the N-terminal amino acid comprises an acetylated derivative (COCH Most preferably, the C-terminus is connected to another peptide / fragment via a linker. Unless linked to a fragment, it is amidated ("-CONH2"). Most preferably The N-terminus contains a free amino group ("NH2").

[0046] In a highly preferred embodiment, the present disclosure relates to peptides for use in the treatment of obesity. The peptide comprises 16 amino acid residues, and further comprises a fragment of SEQ ID NO: 1. SEQ ID NO: 1 contains 16 amino acids. In view of the fact that peptides can be lipidated and / or modified in a manner known in the art, The term "comprising" refers to the possibility that the term may include post-translational modifications commonly used in This should be understood from the wording.

[0047] In another highly preferred embodiment, the present disclosure relates to peptides for use in the treatment of obesity. The peptide has one, two or three amino acid substitutions. KQLIRFLKRA) or a variant of SEQ ID NO: 1 (AQFHRHKQLIRFLKR A).

[0048] In a further highly preferred embodiment, the present disclosure provides a peptide for use in the treatment of obesity. wherein the peptide consists of Ac-AQFHRHKQLIRFLKRA (SEQ ID NO: 2). It is a peptide.

[0049] In a most preferred embodiment, the present disclosure relates to peptides for use in the treatment of obesity. and a peptide consisting of AQFHRHKQLIRFLKRA (SEQ ID NO: 1). Regarding.

[0050] The peptides according to the present invention are agonists for type I IL-4R, preferably type II The agonists are superagonists and complete IL-4R antagonists. It can be either a full agonist or a partial agonist.

[0051] The peptides of the invention may form part of a multimer, such as a dimer, trimer, or tetramer. Multimers incorporating peptides of the present disclosure, such as peptide dendrimers, can also be used. Most preferably, the peptide is in the form of a monomer or a dimer, most preferably a dimer. The peptides according to the present disclosure are those commonly used in the art for fusion peptides or proteins. Covalently linking peptides via various linkers (spacers), such as those used in can be covalently linked to form multimers (e.g., dimers) by

[0052] A second aspect of the present disclosure relates to a peptide or a pharmaceutically acceptable salt thereof for use in the treatment of obesity. In relation to possible salts, the peptide comprises two or more covalently linked peptides derived from α-helix C of IL-4. It contains covalently linked peptide fragments.

[0053] Thus, in a second aspect, the present invention provides a peptide or peptides thereof for use in the treatment of obesity. With respect to pharmaceutically acceptable salts of the peptide, the peptide comprises two or more covalently linked fragments, Each fragment consists of the amino acid sequence AA68 -AA 97 , preferably AA 68 -AA 89 ,twist Preferably AA 71 -AA 94 (i.e., α-helix C of IL-4), and even more Preferably AA 71 -AA 89 , most preferably AA 71 -AA 86 Interleukin -4 (SEQ ID NO: 4), or a variant thereof having 1, 2, 3 or 4 amino acid substitutions The preferred embodiment of the second aspect is a polypeptide comprising 13 to 19 consecutive amino acid residues derived from a polypeptide of the present invention. In the form, each fragment is a fragment of the amino acid region AA of interleukin-4 (SEQ ID NO: 4). 68 -AA 97 , preferably AA 68 -AA 89 , more preferably AA 71 -AA 94 ,Sara By AA 71 -AA 89 , most preferably AA 71 -AA 86 1 derived from Consisting of 4 to 18 consecutive amino acid residues, or 1, 2, 3 or 4 amino acids In a more preferred embodiment of the second aspect, each fragment is a variant thereof having a substitution. The fragment is the amino acid region AA of interleukin-4 (SEQ ID NO: 4). 68 -AA 97 , preferred Kuha AA 68 -AA 89 , more preferably AA 71 -AA 94 , and even more preferably A A 71 -AA 89 , most preferably AA 71 -AA 86 15 to 17 consecutive lines derived from or a variant thereof with 1, 2, 3 or 4 amino acid substitutions. In an even more preferred embodiment of the second aspect, each fragment is an interleukin. Amino acid region AA of Kin-4 (SEQ ID NO: 4) 68 -AA 89 , more preferably AA 71 - AA 94 , and even more preferably AA 71 -AA 89 , most preferably AA 71 -AA8 6, or 1, 2, 3 or 4 consecutive amino acid residues In the above embodiment of the second aspect, each fragment is a variant thereof having an amino acid substitution. The amino acid sequence AQFHRHKQLIRFLKRA (SEQ ID NO: 1) has one or two substitutions. No. 1), or a variant of AQFHRHKQLIRFLKRA (SEQ ID NO: 1) is highly preferred. Most preferably, the covalently linked fragments are identical. Most preferably, The peptide is a dimer (ie, two covalently linked fragments).

[0054] In a preferred embodiment of the second aspect, the present disclosure provides a peptide for use in the treatment of obesity. wherein the peptide comprises two or more covalently linked fragments, each fragment having 13 to 19 amino acids. and wherein the peptide comprises the amino acid sequence AQFHRHKQLIRF LKRA (SEQ ID NO: 1), or a variant having one, two, or three amino acid substitutions and / or deletions. A peptide comprising a variant of AQFHRHKQLIRFLKRA (SEQ ID NO: 1) Regarding.

[0055] In a more preferred embodiment of the second aspect, the present disclosure provides a peptide for use in the treatment of obesity. For peptides, the peptide comprises two or more covalently linked fragments, each fragment having 14 to 18 amino acids. and each fragment has the amino acid sequence AQFHRHKQLIRFLK RA (SEQ ID NO: 1), or one, two, or three amino acid substitutions and / or deletions The variant of AQFHRHKQLIRFLKRA (SEQ ID NO: 1) having the following structure:

[0056] In an even more preferred embodiment of the second aspect, the present disclosure provides a method for treating obesity. With respect to the peptide of claim 1, said peptide comprises two or more covalently linked fragments, each fragment being 15 to 20% by weight. It consists of 17 amino acid residues, and each fragment has the amino acid sequence AQFHRHKQLIR FLKRA (SEQ ID NO: 1), or one, two, or three amino acid substitutions and / or deletions This includes variants of AQFHRHKQLIRFLKRA (SEQ ID NO: 1) having the amino acid sequence

[0057] In an even more preferred embodiment of the second aspect, the present disclosure provides a method for treating obesity. 1. The peptide of claim 1, wherein the peptide comprises two or more covalently linked fragments, each fragment having 16 amino acids. and the peptide comprises one, two, or three amino acid substitutions. SEQ ID NO: 1 (AQFHRHKQLIRFLKRA) or a variant of SEQ ID NO: 1 (A QFHRHKQLIRFLKRA).

[0058] In any of the above-described embodiments of the second aspect, the variant preferably comprises two amino acids amino acid substitutions and / or deletions, even more preferably one amino acid substitution or deletion In any of the above-described embodiments of the second aspect, the peptide most preferably comprises two covalent It comprises linked fragments (i.e., dimers), each of which consists of the same sequence.

[0059] In a most preferred embodiment of the second aspect, the present invention provides a method for treating obesity comprising administering to a subject a compound of formula (I) comprising administering to said subject a compound of formula (I) ... Regarding peptides, said peptides have the formula (I): [ka] or its barrier having one or two substitutions in each peptide fragment. It is a nt.

[0060] Each fragment of SEQ ID NO: 1 in the dimeric peptide of formula (I) (i.e., AQFHRHKQLI RFLKRA) are covalently linked at their C-termini via a linker.

[0061] In another most preferred embodiment of the second aspect, the present invention provides a method for treating obesity comprising administering to a subject a Regarding the peptide, the peptide is linked to a linker (lysine- beta-alanine): [ka]

[0062] In another embodiment of the second aspect the invention relates to a peptide for use in the treatment of obesity. and the peptide is a tetrameric peptide containing four copies of SEQ ID NO: 1 and / or SEQ ID NO: 2. a portion of a nucleotide sequence of a nucleotide sequence containing one, two, or three amino acid substitutions, or a variant thereof In another preferred embodiment, the peptide is a dimer comprising two copies of SEQ ID NO: 1. A dendrimer or variant thereof containing one, two, or three amino acid substitutions of SEQ ID NO:2. or two copies of SEQ ID NO: 2, or one, two, or three amino acid substitutions In another embodiment, the peptide comprises 4 copies of SEQ ID NO: 1. or a variant thereof containing one, two, or three amino acid substitutions. or 4 copies of SEQ ID NO: 2, or variants thereof containing 1, 2, or 3 amino acid substitutions It is a riant.

[0063] In a third aspect, the present disclosure relates to a pharmaceutical composition for use in the treatment of obesity, said pharmaceutical The composition comprises one or more peptides according to the aspects and embodiments disclosed herein. A pharmaceutical composition may include an acceptable pharmaceutical carrier and, optionally, one or more excipients. The pharmaceutical composition (i.e., formulation) is a tablet, pill, or capsule intended to be dissolved prior to administration. These formulations include, but are not limited to, pharmaceutical preparations, emulsions, suspensions, sustained release formulations, solutions, or lyophilized powders. In some embodiments, the formulation may be a depot formulation that provides sustained release. Depending on the choice of formulation and chemical and / or metabolic stability of the polypeptide, different dosages may be obtained. It is understood that any route of administration may be used. Such routes of administration include oral administration, parenteral administration, and the like. administration (intravenous (IV), subcutaneous (SC), intradermal (ID) and intramuscular (IM)), or inhalation In a preferred embodiment of the present invention, the route of administration is parenteral. Oral administration. In an even more preferred embodiment, the route of administration is subcutaneous.

[0064] In a fourth aspect, the present disclosure provides a method for the treatment of obesity, comprising administering to a subject a therapeutically effective amount of a compound according to any one of the aspects disclosed herein. According to any of the methods and embodiments, an effective amount of the peptide is administered to an individual in need thereof. The present invention relates to a method comprising: [Example]

[0065] Example Example 1: Effect of IL-4 derived peptides on body weight Materials and Methods (animal) Thirty female mice (BKS.Cg-Dock7m + / + Leptin db / db) were BLKS, 5 weeks old) were transferred to the Gubra Research Animal Unit in Denmark. Throughout the acclimation and testing periods, animals were allowed free access to food and water, and were kept in a well-ventilated area. All animal experiments were conducted in accordance with the Care and Use of Laboratory Animals Act. According to the Gubra Bioethical Guidelines, which are fully in accordance with internationally recognized principles regarding At the start of the study (baseline), on days 13, 27, 34, 43, and 60 and on day 77 (end of study), animal weights were recorded.

[0066] (compound) Compound A was prepared using standard solid phase peptide synthesis (SPPS) procedures. was purchased from Phlogo ApS, Copenhagen, Denmark (10 mg / ml miliQ The vehicle was provided by a 1:1 mixture of miliQ:PBS and 1:1 dilution of the dose formulation with water. It was 5.

[0067] In vivo procedures Mice were randomized to vehicle group (n=15) and 10 mg / kg Compound A (n=15). The compound and vehicle were administered subcutaneously once daily. On day 27, the dose was increased to 5 mg / kg. On day 43, the dose was reduced to 2.5 mg / kg of Compound A (n=15). n=15).

[0068] (result) Compared to vehicle, the tested Compound A significantly increased weight gain in the treatment group receiving Compound A. The weight results (average values) are shown in Table 1 below.

[0069] [Table 1]

[0070] (Consideration) Interestingly, weight gain was significantly less in the treatment group compared to the placebo group. The average weight of animals in the placebo group increased by 5.76 g (12.8%) over the course of the study. In contrast, the treated group only increased their weight by 3.51 g (7.8%). Thus, Compound A (SEQ ID NO: 1) reduced weight gain.

[0071] (Array Overview) SEQ ID NO: 1 AQFHRHKQLIRFLKRA

[0072] SEQ ID NO: 2 Ac-AQFHRHKQLIRFLKRA

[0073] SEQ ID NO: 3 (signal peptide, IL-4 peptide containing 153 amino acids) MGLTSQLLPP LFFLLACAGN FVHGHKCDIT LQEIIKT LNS LTEQKTLCTE LTVTDIFAAS KNTTEKETFC RAA TVLRQFY SHHEKDTRCL GATAQQFHRH KQLIRFLKRL DRNLWGLAGL NSCPVKEANQ STLENFLERL KTIMRE KYSK CSS

[0074] SEQ ID NO: 4 (IL-4 peptide without signal peptide, 129 amino acids) HKCDIT LQEIIKTLNS LTEQKTLCTE LTVTDIFAAS KNTTEKETFC RAATVLRQFY SHHEKDTRCL GATAQQF HRH KQLIRFLKRL DRNLWGLAGL NSCPVKEANQ STL ENFLERL KTIMREKYSK CSS

[0075] (References) Roglic, G. World Health Organization. (2 016). Global report on diabetes. ISBN 97 89241565257. Abdelaal M. et al. Morbidity and mortali ty associated with obesity. Ann Transl M ed. 2017 Apr;5(7):161, p.1-12. Klementiev B. et al. Antiinflammatory pr operations of a peptide derived from inter leukin-4. Cytokine, Vol. 64, No. 1, 10.2 013, pp. 112-21.

[0076] (item) 1. AA 68 -AA 97 , preferably AA of interleukin-4 (SEQ ID NO: 4) 68 -AA 89 a fragment consisting of 13 to 19 adjacent amino acid residues derived from or a variant thereof having four amino acid substitutions for use in the treatment of obesity. A peptide or a pharmaceutically acceptable salt thereof for the purpose of

[0077] 2. The peptide is preferably an AA of interleukin-4. 68 -AA 89 (Array number No. 4) amino acid region AA68 -AA 97 14 to 18 consecutive amino acid residues derived from or a variant thereof having 1, 2, 3 or 4 amino acid substitutions. 4. A peptide for use according to any of the preceding paragraphs, comprising:

[0078] 3. The peptide is preferably an AA of interleukin-4. 68 -AA 89 (Array number No. 4) amino acid region AA 68 -AA 97 15-17 consecutive amino acid residues derived from A fragment consisting of the group or a variant thereof having 1, 2, 3, or 4 amino acid substitutions. 4. A peptide for use according to any of the preceding claims, comprising:

[0079] 4. The peptide is preferably an AA of interleukin-4. 68 -AA 89 (array No. 4) amino acid region AA 68 -AA 97 16 consecutive amino acid residues derived from or a variant thereof having 1, 2, 3, or 4 amino acid substitutions. 2. A peptide for use according to any of the preceding claims.

[0080] 5. The amino acid residue is in the amino acid region AA 71 -AA 94 (i.e., IL-4 α- Helix C), preferably AA 71 -AA 89 , most preferably AA 71 -AA 86 to 2. A peptide for use according to any of the preceding paragraphs, derived from

[0081] 6. The peptide has the amino acid sequence AQFHRHKQLIRFLKRA (SEQ ID NO: 1). or a variant thereof having one or two substitutions. A peptide for use as described in any one of the preceding claims.

[0082] 7. The peptide comprises two or more identical fragments covalently linked via a linker; A peptide for use according to any of the preceding paragraphs.

[0083] 8. A peptide or a pharmaceutically acceptable salt thereof for use in the treatment of obesity. , wherein the peptide is a dimer comprising two fragments, each fragment containing interleukin-4 (sequence No. 4) amino acid region AA 71 -AA 94 13 to 19 consecutive amino acids derived from a peptide or a pharmaceutically acceptable salt thereof consisting of the residues (i.e., the α-helix of IL-4) or a variant thereof having 1, 2, 3, or 4 amino acid substitutions; A peptide or a pharmaceutically acceptable salt thereof, wherein each fragment is covalently linked via a linker. Useful salt.

[0084] 9. The amino acid residue is in the amino acid region AA 71 -AA 89 most preferably AA7 1-AA 86 9. The peptide for use according to item 8, wherein

[0085] 10. Each fragment is composed of 14 to 18 consecutive amino acid residues, preferably 15 to 17 consecutive amino acid residues. Any of items 8 and 9, consisting of consecutive amino acid residues, most preferably 16 amino acid residues. A peptide for use as described in any one of the preceding claims.

[0086] 11. A peptide for use according to any one of items 8 to 10, comprising two identical fragments. Petite.

[0087] 12. A peptide for use according to any of the preceding claims, having the structure of formula (I): [ka] or its barrier having one or two substitutions in each peptide fragment. It is a nt.

[0088] 13. The peptide is an agonist for type I IL-4R and an agonist for type II IL-4R. 4. The peptide according to any one of the preceding paragraphs, which is an antagonist for

Claims

1. 1. A peptide, or a pharmaceutically acceptable salt thereof, for use in the treatment of obesity, comprising: The peptide is a dimer containing two fragments, each fragment containing interleukin-4 (sequence Amino acid region AA of column number 4 ７１ -AA ９４ 13 to 19 consecutive amino acids derived from a fragment consisting of 1, 2, 3, or a variant thereof having four amino acid substitutions, each fragment being covalently linked via a linker; A peptide or a pharmaceutically acceptable salt thereof, wherein the peptide is synthetically linked to the amino acid sequence of ...

2. Each fragment is composed of 14 to 18 consecutive amino acid residues, preferably 15 to 17 consecutive amino acid residues.

2. The method of claim 1, comprising a sequence of amino acid residues, most preferably 16 consecutive amino acid residues. Peptides for use in

3. The amino acid residues are in the amino acid region AA ７１ -AA ８９ , most preferably AA ７１ -A A ８６ 10. A peptide for use according to any one of the preceding claims, derived from

4. The peptide is an agonist for type I IL-4R and an agonist for type II IL-4R.

10. A peptide according to any preceding claim which is an antagonist of

5. 10. Any of the preceding claims, wherein at least a portion of the peptide forms an α-helix. The peptide according to claim 1.

6. The peptide is a dimer containing two fragments, each fragment containing interleukin-4 ( Amino acid region AA of SEQ ID NO: 4 ７１ -AA ９４ 13 to 19 consecutive ami derived from a fragment consisting of 1, 2, 3, or 4 amino acid residues (i.e., the α-C helix of IL-4) is a variant thereof having four amino acid substitutions, and each fragment is shared by a linker. and wherein the peptide is an agonist for type I IL-4R and a type II IL-4R. and an antagonist for L-4R, wherein at least a portion of the peptide is 2. The peptide for use according to claim 1, which forms an α-helix.

7. The peptide has the formula (I): 【Chemistry 1】 or a barrier thereof having one or two substitutions in each peptide fragment.

10. A peptide for use according to any of the preceding claims, which is a compound.

8. The peptide has the formula (II): 【Chemistry 2】 or a barrier thereof having one or two substitutions in each peptide fragment.

10. A peptide for use according to any of the preceding claims, which is a compound.

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