Molecular modification of G L P-1R activator and application of dimer thereof in treating metabolic diseases

文档序号：1320466 发布日期：2020-07-14 浏览：8次中文

阅读说明：本技术 Glp-1r激活剂的分子改构及其二聚体在治疗代谢病中的应用 (Molecular modification of G L P-1R activator and application of dimer thereof in treating metabolic diseases ) 是由唐松山张旭东谭宏梅杨莉罗群唐婧晅于 2020-03-18 设计创作，主要内容包括：本发明提供了基于胰高血糖素样肽-1受体(GLP-1R)Exendin-4、葡萄糖依赖性胰岛素营养多肽(GIP)-Exendin 4嵌合多肽、GLP-1的分子变构体和其二聚体在降糖和治疗代谢综合征中的应用。本发明二聚体是两个相同的含有单个半胱氨酸变构的GLP-1、Exendin-4或GIP-Exendin-4嵌合肽单体通过半胱氨酸形成的二硫键连接而成。本发明的H型二聚体(分子内单Ser→Cys替换)在不降低活性的情况下,显著增加了该二聚体的降糖持续时间,本发明的二聚体在体内持续活性长达22天,较阳性对照药Lixisenatide(2天)有明显延长。与Lixisenatide比较,相同摩尔浓度的二聚体2G21产生相似降糖活性和体重降低、诱导产生成倍胰岛素分泌并改善器官毒性的效果,拓展了二聚体的医药用途,如在代谢综合征的治疗,包括糖尿病和肥胖症治疗中的用途,以及用于减少过量食物摄取等。(The invention provides a glucagon-like peptide-1 receptor (G L P-1R) Exendin-4, glucose-dependent insulin nutrition polypeptide (GIP) -Exendin 4 chimeric polypeptide, a molecular variant of G L P-1 and application of a dimer thereof in reducing blood sugar and treating metabolic syndrome.)

1. A glucagon-like peptide-1 receptor (G L P-1R) activator analog peptide is characterized in that the G L P-1R activator analog peptide is formed by allosterically modifying the sequence of Exendin-4, GIP-Exendin-4 chimeric peptide or G L P-1 and modifying a lysine-amino fatty acid chain.

2. The G L P-1R activator analog peptide of claim 1, wherein the analog peptide has the specific sequence of any one of:

(1)(HN₂)H-X₂-EGTFTCDLS-X₁₂-QMEEEAV-X₂₀-LFIEWL-X₂₇-NGGPSSGAPP-X₃₈or;

(2)(HN₂)H-X₂-EGTFTSDLC-X₁₂-QMEEEAV-X₂₀-LFIEWL-X₂₇-NGGPSSGAPP-X₃₈or;

(3)(HN₂)H-X₂-EGTFTSDLS-X₁₂-QMEEEAV-X₂₀-LFIEWL-X₂₇-NGGPCSGAPP-X₃₈or;

(4)(HN₂)H-X₂-EGTFTSDLS-X₁₂-QMEEEAV-X₂₀-LFIEWL-X₂₇-NGGPSCGAPP-X₃₈or;

(5)(HN₂)Y-X₂-EGTFTCDYSI-X₁₃-LDKIAQ-X₂₀-AFVQWLIAGGPSSGAPP-X₃₈or;

(6)(HN₂)Y-X₂-EGTFTSDYCI-X₁₃-LDKIAQ-X₂₀-AFVQWLIAGGPSSGAPP-X₃₈or;

(7)(HN₂)Y-X₂-EGTFTSDYSI-X₁₃-LDKIAQ-X₂₀-AFVQWLIAGGPCSGAPP-X₃₈or;

(8)(HN₂)Y-X₂-EGTFTSDYSI-X₁₃-LDKIAQ-X₂₀-AFVQWLIAGGPSCGAPP-X₃₈or;

(9)(HN₂)H-X₂-EGTFTSDVSCYLEGQAA-X₂₀-EFIAWLV-X₂₈-GRG(NH₂)；

wherein, X₂Or X₁₃Is L- α -glycine or L- α -alanine or α -aminoisobutyric acid (α Aib); X₁₂Or X₂₀Or X₂₇Or X₂₈Is lysine, arginine, or side chain-amino glutamyl fatty acid [ gamma-Glu (N- α -fat acid)]Or glutamyl fatty diacid [ gamma-Glu (N- α -fat diacid)]Modified lysine, or side chain-amino group [2 × AEEAC-gamma-Glu- (N- α -fatty diacid)]A modified lysine; x₃₈Is PS (HN)₂) Or SKKKKKKKK (HN)₂)。

3. The G L P-1R activator-mimetic peptide of claim 2, wherein when X is₁₂Or X₂₀Or X₂₇Or X₂₈The structure of the lysine modified by glutamyl fatty acid gamma-Glu (N- α -fatty acid) or glutamyl fatty diacid gamma-Glu (N- α -fatty diacid) on the side chain-amino is shown in chemical formula 1, when X is₁₂Or X₂₀Or X₂₇Or X₂₈Is a side chain-amino group of [2 × AEEAC-gamma-Glu- (N- α -fatty diacid)]Modified lysine, the structure of which is shown in chemical formula 2:

chemical formula 1:

chemical formula 2:

4. a hypoglycemic peptide mimetic homodimer formed by disulfide bonding of identical monomers as claimed in any one of claims 1 to 3 through cysteine to form a H-form G L P-1R activator peptide mimetic homodimer.

5. The dimer of claim 4, wherein the amino acid sequence is any one of:

6. The dimer of claim 5, wherein when X is₁₂Or X₂₀Or X₂₇Or X₂₈Is side chain-amino glutamyl fatty acid [ gamma-Glu (N- α -fat acid)]Or glutamyl fatty diacid [ gamma-Glu (N- α -fattydiacid)]Modified lysine has a structure shown in chemical formula 1; when X is present₁₂Or X₂₀Or X₂₇Or X₂₈Is a side chain-amino group of [2 × AEEAC-gamma-Glu- (N- α -fatty diacid)]The structure of modified lysine is shown in chemical formula 2.

7. Use of a G L P-1R activator analogue peptide according to any one of claims 1-3, or a homodimer according to any one of claims 4-6, for the manufacture of a medicament for the treatment of a metabolic syndrome disorder.

8. The use of claim 7, wherein the metabolic syndrome disorder comprises hyperglycemia, diabetes, and obesity.

9. A medicament for treating a metabolic syndrome disorder, comprising a G L P-1R activator-analogous peptide according to any one of claims 1 to 3, or a homodimer according to any one of claims 4 to 6 and pharmaceutically acceptable salts thereof as an active ingredient.

Technical Field

The invention belongs to the field of medical biology, and particularly relates to molecular modification of G L P-1R activator similar peptide and application of homodimer thereof in treating metabolic diseases.

Background

Exendin-4 is an incretin analogue isolated from Heloderma subspecium saliva, has 39 amino acids and has 53% sequence homology with G L P-1. GIP is a gastrointestinal regulatory peptide of 42 amino acids, which has the functions of regulating body sugar metabolism, promoting insulin release from islet β cells and reducing body weight. G β P-1 is an incretin-like peptide of 30 amino acid residues, which is released by intestinal β cells upon nutrient intake. Exendin-4 and G β P-1 are two G β P-1R activators found so far, and two G β P-1R activators are obtained based on the three active polypeptide amino acid sequences regulating sugar metabolism, and through the significant structural change of last decade, the FDA obtains sugar-reducing G β P-1R activators which are available in the U.S. or Chinese SFDA or are clinically approved twice per time, such as once daily administration of IRE 25, and twice daily administration of bifunctional G β P-1R activators (Clitin) which are produced by the methods of the polypeptide of Tailite L, Taizeitanium 7, the Tai II, the polypeptide synthesized by the Tailite-8, the Tailite III, the Tai-8, the polypeptide can be synthesized by the Tailite III, the Tailite, the polypeptide of the Tailite, the Tailite III, the Tailite can be synthesized by the Tailite III, the Tailite III, the polypeptide of the Tailite, the Tailite can be synthesized by the Tailite III, the Tailite III, the Tailite can be synthesized by the Tailite III, the Tailite of the Tailite III, the Tailite of the Tailite III, the Tailite of the Tailite, the Tailite of the Tailite, the Tailite of the Tailite.

Because The hypothalamic-pituitary-adrenal axis (HPA or HTPA) is part of The physiological stress response, G L P-1R activators stimulate The HPA axis leading to an increase in corticosterone, resulting in a partial arrhythmia, therefore, there remains a need for ① to simultaneously antagonize The activation of glucagon and G L P-2 receptors while activating GIP receptors and/or G L P-1 receptor effects, ② to provide weight loss, antagonizing DPP-4 and other forms of degradation mechanisms while maintaining low immunogenicity by activating GIP receptors and/or G L P-1 receptor effects, and ③ G L P-1R activators remain to be optimized because current long-acting activators provide less effective activity than G1-26 or Exendin for activation of specific glucose-lowering effects (glucose lowering effect per unit mass), dosage, weight reduction, and side effects, and natural G L P-1R activators.

Disclosure of Invention

The invention also provides a homodimer formed by the G L P-1R activator similar peptide, and finds that H-type structure dimers (intramolecular single Cys → Ser substitutions) formed by different positions of cysteine generate different activities and last 22 days, and the clinical medicine can be obviously increased compared with the current 1-7 days clinical medicine.

In order to achieve the purpose, the technical scheme includes that the glucagon-like peptide-1 receptor (G L P-1R) activator similar peptide is formed by allosterizing a sequence of Exendin-4, GIP-Exendin-4 chimeric peptide or G L P-1 and modifying a lysine-amino fatty acid chain, the G L P-1R activator similar peptide is formed by replacing an amino acid sequence of the Exendin-4, GIP-Exendin-4 or G L P-1 peptide with Cys as a main chain, replacing Ser on the main chain with Cys, and only containing one Cys in the amino acid sequence of the main chain, wherein the side chain is the amino-fatty acid chain of one lysine of the main chain.

Preferably, the G L P-1R activator analogue peptide has a specific sequence of any one of the following: