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CN-122011068-A - Triazole group-containing compound, application and preparation method thereof

CN122011068ACN 122011068 ACN122011068 ACN 122011068ACN-122011068-A

Abstract

The invention discloses a compound containing triazole groups, and application and a preparation method thereof. The invention specifically discloses a compound I, which has one or more of the following advantages of novel structure, good stabilizing effect on complex structure membrane proteins, easy synthesis, large-scale preparation, excellent properties and easy quality control.

Inventors

  • CAI HONGMING
  • LIN KUAILE
  • CAI ZHENGYAN
  • LIU YUBIN
  • JIN SHIXIN
  • WU XINYU
  • DONG JUNJUN

Assignees

  • 上海医药工业研究院有限公司
  • 中国医药工业研究总院有限公司

Dates

Publication Date
20260512
Application Date
20251111
Priority Date
20241111

Claims (11)

  1. 1. A compound of formula (I), ; Wherein R 3 is Or H; A 1 and A 2 are independently monosaccharide glycosyl or oligosaccharide glycosyl, wherein the monosaccharide is pentose furanose or pyranose, or hexose furanose or pyranose, and the oligosaccharide is obtained by dehydrating and polymerizing 2-10 monosaccharides; L 1 、L 2 、L 3 and L 4 are independently C 1-6 alkylene; R 1 is a C 1-25 hydrophobic aliphatic group or H; R 2 is a C 1-25 hydrophobic aliphatic group.
  2. 2. Compound I according to claim 1, characterized in that it satisfies one or more of the following conditions: (1) In a 1 and a 2 , the pentose is an aldose; (2) In a 1 and a 2 , the hexose is a hexose or aldohexose, preferably aldohexose; (3) In a 1 and a 2 , the monosaccharide is a furanose or a pyranose of a hexose, preferably a pyranose of a hexose; (4) In A 1 and A 2 , the furanose of the hexose is the furanose of D-glucose; (5) In a 1 and a 2 , the pyranose of the hexose is a glucopyranose of glucose or a galactopyranose of galactose; (6) In A 1 and A 2 , the oligosaccharide is obtained by water-loss polymerization of 2-5 monosaccharides, preferably 2 monosaccharides; (7) In a 1 and a 2 , the "monosaccharide dehydration polymerization" is a hemiacetal or hemiketal hydroxyl group of one monosaccharide, and is performed with any hydroxyl group of another monosaccharide; (8) In R 1 and R 2 , the C 1-25 hydrophobic aliphatic groups are independently C 1-25 alkyl, C 1-25 heteroalkyl, or-C 1-20 alkylene-C 3-8 cycloalkyl; The C 1-25 heteroalkyl group contains 1-5 heteroatoms, and the heteroatoms are independently O or S; The C 3-8 cycloalkyl is preferably C 4-6 cycloalkyl, such as cyclohexyl; (9) R 1 is a C 1-25 hydrophobic aliphatic group; (10) R 3 is 。
  3. 3. Compound I according to claim 2, characterized in that it satisfies one or more of the following conditions: (1) In a 1 and a 2 , when the hexose is ketohexose, the ketohexose is psicose, fructose, sorbose or tagatose; (2) In a 1 and a 2 , when the hexose is aldohexose, the aldohexose is allose, altrose, glucose, mannose, gulose, idose, galactose, or talose; (3) In A 1 and A 2 , when the furanose of the hexose is the furanose of D-glucose, the furanose of D-glucose is alpha-D-furanose or beta-D-furanose; (4) In A 1 and A 2 , when the pyranose of the hexose is a glucopyranose, the glucopyranose is a D-glucopyranose, e.g., α -D-glucopyranose or β -D-glucopyranose; (5) In A 1 and A 2 , when the pyranose of the hexose is a galactopyranose, the galactopyranose is a D-galactose pyranose, e.g., a-D-galactopyranose or β -D-galactopyranose; (6) In A 1 and A 2 , the "monosaccharide dehydration polymerization" is a hemiacetal or hemiketal hydroxyl group of one monosaccharide, and the "dehydration polymerization" is performed with the hydroxyl group at the 4-position or the 6-position of the other monosaccharide; (7) R 1 and R 2 , the C 1-25 heteroalkyl radical is 、 Or (b) ; (8) In R 1 and R 2 , the-C 1-20 alkylene-C 3-8 cycloalkyl is-C 1-10 alkylene-C 3-8 cycloalkyl, preferably-C 1-6 alkylene-C 3-8 cycloalkyl, for example ; (9) In R 1 and R 2 , the C 1-25 alkyl is C 4-20 alkyl, e.g 、 、 、 、 、 、 、 、 Or (b) ; The C 1-25 alkyl is preferably C 5-10 alkyl, for example 、 、 Or (b) ; (10) L 1 is methylene; (11) L 2 is methylene; (12) L 3 is methylene; (13) L 4 is methylene; (14) R 1 and R 2 are independently C 1-25 alkyl or-C 1-20 alkylene-C 3-8 cycloalkyl, preferably C 4-20 alkyl or-C 1-6 alkylene-C 3-8 cycloalkyl.
  4. 4. A compound I according to claim 3, characterized in that it satisfies one or more of the following conditions: (1) In A 1 and A 2 , when the hexose is aldohexose, the aldohexose is glucose or galactose, the glucose is preferably D-glucose, and the galactose is preferably D-galactose; (2) In A 1 and A 2 , the "monosaccharide dehydration polymerization" is a hemiacetal alpha-type hydroxyl group or beta-type hydroxyl group of one monosaccharide, and the "dehydration polymerization" is performed on the hydroxyl group at the 4-position or the 6-position of the other monosaccharide.
  5. 5. Compound I according to claim 1, characterized in that it satisfies one or more of the following conditions: (1) In A 1 and A 2 , the monosaccharide is ; (2) In A 1 and A 2 , the monosaccharide sugar group is ; (3) In a 1 and a 2 , the oligosaccharide is cellobiose, maltose, lactose, melibiose or sucrose, preferably maltose, cellobiose or lactose, more preferably maltose; The maltose is preferably alpha-maltose or beta-maltose, and the beta-maltose is preferably beta-D-maltose; The cellobiose is preferably alpha-cellobiose or beta-cellobiose, and the beta-cellobiose is preferably beta-D-cellobiose; The lactose is preferably alpha-lactose or beta-lactose, and the beta-lactose is preferably beta-D-lactose; The melibiose is preferably alpha-melibiose or beta-melibiose, and the beta-melibiose is preferably beta-D-melibiose; (4) In A 1 and A 2 , the oligosaccharide glycosyl is 、 、 Or (b) ; (5) A 1 and A 2 are independently 、 、 、 、 Or (b) ; (6) R 1 and R 2 are independently 、 、 、 、 、 、 、 、 、 Or (b) 。
  6. 6. Compound I according to claim 1, characterized in that it satisfies any of the following schemes: In schemes (1) A 1 and A 2 , the oligosaccharides are obtained by water-loss polymerization of 2-10 monosaccharides, the monosaccharides are furanose or pyranose of hexoses, preferably pyranose of hexoses, and the oligosaccharides are preferably obtained by water-loss polymerization of 2-5 monosaccharides; The pyranose of the hexose is preferably D-glucopyranose or D-galactopyranose; the D-glucopyranose is preferably alpha-D-glucopyranose or beta-D-glucopyranose; the D-galactopyranose is preferably alpha-D-galactopyranose or beta-D-galactopyranose; in the oligosaccharide, the connecting bond between the monosaccharides is independently preferably an alpha-1, 4 glycosidic bond, a beta-1, 4 glycosidic bond or an alpha-1, 6 glycosidic bond; Scheme (2) the compound I has the structure: , wherein a 1 、A 2 、L 1 、L 2 、L 3 、L 4 、R 1 and R 2 are as defined in any one of claims 1 to 5; Scheme (3) the compound I structure is as follows: ; Wherein A 1 and A 2 are independently a monosaccharide or oligosaccharide glycosyl, the monosaccharide is a pyranose of hexose, the oligosaccharide is obtained by water-loss polymerization of 2-10 monosaccharides, and the oligosaccharide is preferably obtained by water-loss polymerization of 2-5 monosaccharides; In the oligosaccharide, the connecting bond between the monosaccharides is preferably alpha-1, 4 glycosidic bond, beta-1, 4 glycosidic bond or alpha-1, 6 glycosidic bond; l 1 、L 2 、L 3 and L 4 are independently C 1-3 alkylene; R 1 is C 4-20 alkyl; R 2 is C 4-20 alkyl; scheme (4) the compound I structure is as follows: ; Wherein A 1 and A 2 are independently oligosaccharyl, the oligosaccharyl is obtained by water-loss polymerization of 2-5 monosaccharides, and the monosaccharides are hexose glucopyranoses, such as alpha-D-glucopyranose, beta-D-glucopyranose, alpha-D-galactopyranose or beta-D-galactopyranose; in the oligosaccharide, the connecting bond between the monosaccharides is preferably alpha-1, 4 glycosidic bond or beta-1, 4 glycosidic bond; L 1 、L 2 、L 3 and L 4 are independently C 1-3 alkylene, e.g., methylene; R 1 is C 5-10 alkyl; r 2 is C 5-10 alkyl; Preferably, the oligosaccharide is maltose, cellobiose or lactose, e.g. β -D-maltose, β -D-cellobiose or β -D-lactose.
  7. 7. Compound I according to claim 1, characterized in that it is any one of the following compounds: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 Or (b) 。
  8. 8. Use of a compound I as claimed in any of claims 1 to 7 for the preparation of detergent formulations or as a detergent.
  9. 9. Use according to claim 8, characterized in that it fulfils one or more of the following conditions: (1) The detergent preparation or the detergent is used for stabilizing or improving the stability or the thermal stability of membrane proteins, wherein the membrane proteins are preferably G protein coupled receptors or GLP-1 receptors, and the G protein coupled receptors are preferably A2a receptors; (2) The detergent formulation or the detergent acts by disrupting the biofilm to release membrane proteins and providing a hydrophobic environment in solution for membrane proteins in a de-membraned state, thereby extracting membrane proteins on the biofilm into solution; The membrane protein is preferably a G protein coupled receptor, such as an A2a receptor; (3) The detergent formulation or the detergent forms micelles in a solution, preferably 4-50 nm a diameter, such as 5.8 nm、5.9 nm、6.1 nm、6.4 nm、7.6 nm、7.7 nm、7.8 nm、8.0 nm、10.2 nm、28.4 nm、29.8 nm、31.2 nm、32.7 nm、34.1 nm or 47.4 nm a micelle is preferably an amphiphilic micelle and the solution is preferably an aqueous solution.
  10. 10. A process for the preparation of compound I according to any one of claims 1 to 7, comprising the steps of: (1) In a solvent, carrying out alkylation reaction on dimethyl malonate, alkali and monohalogenide with a hydrophobic aliphatic group, and separating to obtain a first intermediate after the reaction is finished; the first intermediate refers to an intermediate compound obtained by disubstituted active methylene of dimethyl malonate by a hydrophobic aliphatic group; (2) In a solvent, carrying out reduction reaction on the first intermediate and a reducing agent, and separating to obtain a second intermediate; the substance obtained in the step is an intermediate compound obtained after two ester groups on the first intermediate are reduced to primary alcohol; (3) In a solvent, carrying out etherification reaction on a second intermediate, alkali and 3-bromopropyne, and separating to obtain a hydrophobic segment after the reaction is finished, wherein the hydrophobic segment refers to a third intermediate compound obtained after all 2 primary hydroxyl groups of the second intermediate are etherified; ; (4) Performing terminal azide reaction on hydroxyl-protected monosaccharide or oligosaccharide, an azide reagent and Lewis acid in a solvent, and separating to obtain a hydrophilic fragment after the reaction is finished, wherein the hydrophilic fragment refers to a fourth intermediate compound obtained after terminal azide of the hydroxyl-protected monosaccharide or oligosaccharide is substituted; ; (5) In a solvent, performing a Click reaction on the third intermediate, the fourth intermediate, sodium ascorbate and copper sulfate, and separating to obtain a fifth intermediate after the reaction is finished, wherein the fifth intermediate obtained in the step is an intermediate compound obtained by connecting the hydrophobic segment and the hydrophilic segment through a triazole group by adopting the Click reaction; (6) In a solvent, carrying out methanolysis reaction on the fifth intermediate and sodium methoxide, and separating to obtain the compound I after the reaction is finished; 。
  11. 11. a compound II, which is any one of the following structures: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 Or (b) 。

Description

Triazole group-containing compound, application and preparation method thereof Technical Field The invention relates to a compound containing triazole groups, and application and a preparation method thereof. Background Membrane Proteins (MPs) are the main contributors to the function of biological membranes. Plays an important role in intracellular or intercellular signal transduction and the exchange of metabolites. Since MPs have a wide variety of biological functions, they are critical in drug design, accounting for about 70% of all drug targets at present. With the development of X-Ray diffraction (X-Ray), nuclear Magnetic Resonance (NMR) and electron-freezing (EM) technologies, more and more protein structures have been resolved in recent years. However, membrane proteins are unstable outside the biological membrane and are easily denatured. Membrane Proteins (MPs) are more difficult to study in vitro than soluble proteins. Thus, in the Protein Database (PDB), the structure of membrane proteins is only about 1.5%. Membrane protein in vitro studies require an amphiphilic system that mimics the natural biological membrane to stabilize the membrane protein, but this requires a relatively time-consuming process to select an appropriate membrane-mimic environment to stabilize a particular MP, thereby ensuring that its membrane protein structure and function is not compromised. In order to stabilize membrane proteins, a number of amphiphilic systems have been developed in recent years, with other membrane-like systems such as Bicelles, nanodiscs, liposomes, etc., in addition to detergent micelles. In a hydrophilic environment, membrane proteins need to be solubilized and purified by means of the amphiphilicity of detergents, so that the membrane proteins are extracted from the phospholipid bilayer. Detergents are commonly used as membrane mimics for MP preparation and structural studies as a common amphiphilic system. Can form a complex (PDC) of membrane protein-detergent with membrane protein, and is directly used for crystallography or nuclear magnetic resonance experiments. Even in the case where proteins are recombined in other two parents for crystallization experiments or electron microscopy, detergents are almost always necessary in intermediate steps involving protein purification, and thus detergents are important tools for membrane protein biochemistry and structural studies. Ideal detergents would be able to maintain the membrane protein in a soluble state without a measurable effect on the function, structure and thermodynamic properties of the protein. Among them, n-Octyl- β -D-glucopyranoside (n-Octyl- β -D-Glucopyranoside, OG), dodecyl- β -D-maltoside (n-Dodecyl- β -D-maltoside, DDM) and the like are the most widely used conventional detergents at present, but these detergents have poor stability to complex structural membrane proteins, for example, membrane proteins of GPCR family. At present, many novel detergents reported in the literature successfully achieve better solubilization and stabilization of membrane proteins by adopting a pre-assembly strategy, such as dodecyl neopentyl glycol maltoside (Lauryl Maltose Neopentyl Glycol, LMNG), but most of the detergents adopt low-efficiency polysaccharide glycosylation reaction to link hydrophilic and hydrophobic groups, so that the novel detergents are difficult to prepare in a large scale, and the popularization and application of the novel detergents in membrane protein research are limited. Disclosure of Invention The invention aims to overcome the defects of single structure, low synthesis efficiency, low accessibility, difficult large-scale preparation, poor properties and inconvenient quality control of the detergent in the prior art. Therefore, the compound containing the triazole group has one or more advantages of novel structure, good stabilizing effect on complex structure membrane proteins, easiness in synthesis, large-scale preparation, excellent properties and easiness in quality control. The present invention provides a compound of formula I, ; Wherein R 3 isOr H; A 1 and A 2 are independently monosaccharide glycosyl or oligosaccharide glycosyl, wherein the monosaccharide is pentose furanose or pyranose, or hexose furanose or pyranose, and the oligosaccharide is obtained by dehydrating and polymerizing 2-10 monosaccharides; L 1、L2、L3 and L 4 are independently C 1-6 alkylene; R 1 is a C 1-25 hydrophobic aliphatic group or H; R 2 is a C 1-25 hydrophobic aliphatic group. In certain preferred embodiments of the invention, certain groups of the compound I are defined below, and the unrecited groups are as described in any of the embodiments of the invention (abbreviated as "in a certain embodiment of the invention"). For other invention items, the same shorthand manner as the invention is adopted, and no further description is given. In one embodiment of the invention, in A 1 and A 2, the pentose is an aldose. In one embodiment of th