Compound containing triazole group as well as application and preparation method thereof

Abstract

The invention discloses a triazole group-containing compound as well as application and a preparation method thereof. The invention specifically discloses a compound I. The compound I has one or more advantages as follows: the compound I is novel in structure, good in stabilizing effect on membrane protein with a complex structure, easy to synthesize, capable of being prepared on a large scale, excellent in character and easy in quality control.

Description

Technical Field

[0001] This invention relates to a compound containing a triazole group, its uses, and its preparation method. Background Technology

[0002] Membrane proteins (MPs) are the main carriers of biological membrane function. They play a crucial role in intracellular and intercellular signal transduction and the exchange of metabolites. Due to their diverse biological functions, MPs are essential in drug design, accounting for approximately 70% of all current drug targets. In recent years, with the development of X-ray diffraction (X-ray), nuclear magnetic resonance (NMR), and cryo-electron microscopy (EM), more and more protein structures have been resolved. However, membrane proteins are unstable outside biological membranes and are prone to denaturation. Membrane proteins (MPs) are more difficult to study in vitro than soluble proteins. Therefore, currently, membrane protein structures account for only about 1.5% of those in protein databases (PDB).

[0003] In vitro studies of membrane proteins require an amphiphilic system that mimics natural biological membranes to stabilize the proteins. However, this involves a time-consuming process of selecting a suitable membrane simulation environment to stabilize specific protein molecules (MPs) and ensure that their structure and function are not disrupted. To stabilize membrane proteins, many amphiphilic systems have been developed in recent years, including detergent micelles and other membrane-like systems such as bicelles, nanodiscs, and liposomes. In a hydrophilic environment, membrane proteins need to be dissolved and purified using the amphiphilic properties of detergents, thereby extracting the proteins from the phospholipid bilayer.

[0004] Detergents, as common amphiphilic systems, are frequently used as membrane mimics in membrane protein (MP) preparation and structural studies. They can form membrane protein-detergent complexes (PDCs) with membrane proteins, which can be directly used in crystallography or NMR experiments. Even when proteins are recombined in other amphiphilic systems for crystallization experiments or electron microscopy, detergents are almost always necessary in intermediate steps involving protein purification. Therefore, detergents are important tools for the biochemical and structural studies of membrane proteins. An ideal detergent can maintain membrane proteins in a soluble state without having a measurable effect on their function, structure, and thermodynamic properties. Among them, n-octyl-β-D-glucopyranoside (OG) and n-dodecyl-β-D-maltoside (DDM) are currently the most widely used traditional detergents. However, these detergents have poor stabilizing effects on complex membrane proteins, such as those of the GPCR family.

[0005] Currently, many novel detergents reported in the literature employ pre-assembly strategies and have successfully achieved good solubilization and stabilization effects on membrane proteins, such as lauryl maltose neopentyl glycol (LMNG). However, most of these detergents use inefficient polysaccharide glycosylation reactions to link hydrophilic and hydrophobic groups, making them difficult to prepare on a large scale. This limits the widespread application of novel detergents in membrane protein research. Summary of the Invention

[0006] The technical problem to be solved by this invention is to overcome the shortcomings of existing detergents, such as their relatively simple structure, low synthesis efficiency, low accessibility, difficulty in large-scale preparation, poor properties, and inconvenience in quality control. To this end, this invention provides a compound containing a triazole group, which has one or more of the following advantages: novel structure, good stabilizing effect on complex membrane proteins, easy synthesis, large-scale preparation, excellent properties, and easy quality control.

[0007] This invention provides a compound I,

[0008] ;

[0009] Among them, R 3 for Or H;

[0010] A 1 and A 2 The monosaccharide is independently a monosaccharide glycosyl or oligosaccharide glycosyl; the monosaccharide is a furanose or pyranose of a pentose, or a furanose or pyranose of a hexose; the oligosaccharide is obtained by dehydration polymerization of 2-10 of the monosaccharides;

[0011] L 1 L 2 L 3 and L 4 Independently for C 1-6 Alkylene;

[0012] R 1 C 1-25 Hydrophobic aliphatic groups or H;

[0013] R 2 C 1-25 Hydrophobic aliphatic groups.

[0014] In certain preferred embodiments of the present invention, certain groups of compound I are defined as follows, and groups not mentioned are as described in any embodiment of the present invention (hereinafter referred to as "in a certain embodiment of the present invention"). Other aspects of the present invention are referred to using the same abbreviations as this invention and will not be described further.

[0015] In one aspect of the present invention, A1 and A 2 In this context, the pentose is an aldose.

[0016] In one aspect of the present invention, A 1 and A 2 In this context, the hexose is a ketose or an aldose, preferably an aldose;

[0017] The preferred ketose is allulose, fructose, sorbitol, or tagatose;

[0018] The aldose is preferably allose, adroose, glucose, mannose, gulose, idose, galactose or tarose, more preferably glucose or galactose;

[0019] The glucose is preferably D-glucose; the galactose is preferably D-galactose.

[0020] In one aspect of the present invention, A 1 and A 2 In this context, the monosaccharide is a furanose or pyranose of a hexose, preferably a pyranose of a hexose.

[0021] In one aspect of the present invention, A 1 and A 2 In this context, the furanose of the hexose is a furanose of D-glucose, such as α-D-furanose or β-D-furanose.

[0022] In one aspect of the present invention, A 1 and A 2 In this context, the pyranose of the hexose is either the pyranose of glucose or the pyranose of galactose;

[0023] The pyranose of the glucose is preferably a pyranose of D-glucose, such as α-D-glucose pyranose or β-D-glucose pyranose;

[0024] The pyranose of the galactose is preferably a pyranose of D-galactose, such as α-D-galactopyranose or β-D-galactopyranose.

[0025] In one aspect of the present invention, A 1 and A 2 In this context, the monosaccharide is... .

[0026] In one aspect of the present invention, A 1 and A 2 In this context, the monosaccharide glycosyl group is... .

[0027] In one aspect of the present invention, A 1 and A 2In this process, the oligosaccharide is obtained by dehydration polymerization of 2-5 monosaccharides, preferably by dehydration polymerization of 2 monosaccharides.

[0028] In one aspect of the present invention, A 1 and A 2 In the oligosaccharide, the "monosaccharide dehydration polymerization" is the "dehydration polymerization" of a hemiacetal or hemiketal hydroxyl group of a monosaccharide with any hydroxyl group of another monosaccharide.

[0029] The "monosaccharide dehydration polymerization" is preferably a process in which the hemiacetal or hemiketal hydroxyl group of one monosaccharide undergoes "dehydration polymerization" with the 4- or 6-position hydroxyl group of another monosaccharide.

[0030] The "monosaccharide dehydration polymerization" is preferably a process in which the α-hydroxyl or β-hydroxyl group of a hemiacetal of one monosaccharide undergoes "dehydration polymerization" with the 4- or 6-hydroxyl group of another monosaccharide.

[0031] The above description of "monosaccharide dehydration polymerization" is not a limitation on the preparation process of "oligosaccharides", but only to indicate the structure of "oligosaccharides", that is, the connection mode between the monosaccharides of each monomer in "oligosaccharides"; and the connection mode between the monosaccharides of each monomer in "oligosaccharides" is independent of each other.

[0032] In one aspect of the present invention, A 1 and A 2 In this process, the oligosaccharide is obtained by dehydration polymerization of 2-10 monosaccharides; the monosaccharide is a furanose or pyranose of a hexose, preferably a pyranose of a hexose; the oligosaccharide is preferably obtained by dehydration polymerization of 2-5 monosaccharides.

[0033] The pyranose of the hexose is preferably D-glucose pyranose or D-galactopyranose;

[0034] The D-glucose pyranose is preferably α-D-glucose pyranose or β-D-glucose pyranose;

[0035] The D-galactopyranose is preferably α-D-galactopyranose or β-D-galactopyranose;

[0036] In the oligosaccharide, the linkages between each monosaccharide are preferably α-1,4 glycosidic bonds, β-1,4 glycosidic bonds, or α-1,6 glycosidic bonds.

[0037] In one aspect of the present invention, A 1 and A 2 In this context, the oligosaccharide is cellobiose, maltose, lactose, melibiose, or sucrose, preferably maltose, cellobiose, or lactose, and more preferably maltose;

[0038] The maltose is preferably α-maltose or β-maltose; the β-maltose is preferably β-D-maltose.

[0039] The cellobiose is preferably α-cellobiose or β-cellobiose; the β-cellobiose is preferably β-D-cellobiose.

[0040] The lactose is preferably α-lactose or β-lactose; the β-lactose is preferably β-D-lactose.

[0041] The melibiose is preferably α-melibiose or β-melibiose; the β-melibiose is preferably β-D-melibiose.

[0042] In one aspect of the present invention, A 1 and A 2 In the text, the oligosaccharide glycosyl group is... , , or .

[0043] In one aspect of the present invention, A 1 and A 2 Independently , , , , or .

[0044] In one aspect of the present invention, R 1 and R 2 In, the C 1-25 The hydrophobic aliphatic group is independently C 1-25 Alkyl, C 1-25 Heteroalkyl or -C 1-20 Alkylene-C 3-8 cycloalkyl;

[0045] The C 1-25 The heteroalkyl group contains 1-5 heteroatoms, wherein each heteroatom is independently O or S;

[0046] The C 3-8 Cycloalkyl groups are preferably C 4-6 Cycloalkyl groups, such as cyclohexyl.

[0047] In one aspect of the present invention, R 1 and R 2 In, the C 1-25 Heteroalkyl , or .

[0048] In one aspect of the present invention, R 1 and R 2 In the context, the -C 1-20 Alkylene-C 3-8Cycloalkyl group is -C 1-10 Alkylene-C 3-8 cycloalkyl, preferably -C 1-6 Alkylene-C 3-8 cycloalkyl, for example .

[0049] In one aspect of the present invention, R 1 and R 2 In, the C 1-25 Alkyl group is C 4-20 Alkyl, for example , , , , , , , , or ;

[0050] The C 1-25 Alkyl groups are preferably C 5-10 Alkyl, for example , , or .

[0051] In one aspect of the present invention, R 1 C 1-25 Hydrophobic aliphatic groups.

[0052] In one aspect of the present invention, R 1 and R 2 Independently for 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.

[0053] In one aspect of the present invention, R 1 and R 2 Independently , , , , , , , , , or .

[0054] In one aspect of the present invention, R 3 for .

[0055] In one aspect of the present invention, L 1 L 2 L 3 and L 4 Independently for C 1-3 Alkylene.

[0056] In one aspect of the present invention, L 1 It is a methylene group.

[0057] In one aspect of the present invention, L 2 It is a methylene group.

[0058] In one aspect of the present invention, L 3 It is a methylene group.

[0059] In one aspect of the present invention, L 4 It is a methylene group.

[0060] In one embodiment of the present invention, the structure of compound I is as follows:

[0061] ,

[0062] Among them, A 1 A 2 L 1 L 2 L 3 L 4 R 1 and R 2 The definition is as described in any of the previous schemes;

[0063] Preferably, compound I has any of the following structures:

[0064] or ;

[0065] Among them, A 1 A 2 L 1 L 2 L 3 L 4 R 1 and R 2 The definition is as described in any of the previous schemes;

[0066] More preferably, the structure of compound I is as follows:

[0067]

[0068] Among them, A 1 A 2 R1 and R 2 The definition is as described in the previous scheme.

[0069] In one embodiment of the present invention, the structure of compound I is as follows:

[0070]

[0071] Among them, A 1 and A 2 The oligosaccharide is independently a monosaccharide glycosyl or an oligosaccharide glycosyl; the monosaccharide is a pyranose of a hexose; the oligosaccharide is obtained by dehydration polymerization of 2-10 of the monosaccharides; preferably, the oligosaccharide is obtained by dehydration polymerization of 2-5 of the monosaccharides.

[0072] In the oligosaccharide, the linkage between each monosaccharide is preferably an α-1,4 glycosidic bond, a β-1,4 glycosidic bond, or an α-1,6 glycosidic bond;

[0073] L 1 L 2 L 3 and L 4 Independently for C 1-3 Alkylene;

[0074] R 1 C 4-20 alkyl;

[0075] R 2 C 4-20 alkyl.

[0076] In one embodiment of the present invention, the structure of compound I is as follows:

[0077]

[0078] Among them, A 1 and A 2 Independently, it is an oligosaccharide glycosyl group; the oligosaccharide is obtained by dehydration polymerization of 2-5 monosaccharides; the monosaccharide is a pyranose of a hexose, such as α-D-glucopyranose, β-D-glucopyranose, α-D-galactopyranose or β-D-galactopyranose;

[0079] In the oligosaccharide, the linkage between each monosaccharide is preferably an α-1,4 glycosidic bond or a β-1,4 glycosidic bond;

[0080] L 1 L 2 L 3 and L 4 Independently for C 1-3 Alkyl groups, such as methylene groups;

[0081] R 1 C 5-10alkyl;

[0082] R 2 C 5-10 alkyl;

[0083] Preferably, the oligosaccharide is maltose, cellobiose, or lactose, such as β-D-maltose, β-D-cellobiose, or β-D-lactose.

[0084] In one embodiment of the present invention, compound I is any of the following compounds:

[0085] , , , or Each n is independently 2-18, where n is an integer.

[0086] In one embodiment of the present invention, compound I is any of the following compounds:

[0087] , , , , , , , , , , , , , , , , , , or .

[0088] The present invention also provides the use of the above-mentioned compound I in the preparation of a detergent formulation, or as a detergent.

[0089] In one aspect of the present invention, the detergent preparation or the detergent is used to stabilize membrane proteins, or to improve the stability of membrane proteins, or to improve the thermal stability of membrane proteins.

[0090] The membrane protein is preferably a G protein-coupled receptor or a GLP-1 receptor; the G protein-coupled receptor is preferably an A2a receptor.

[0091] In one aspect of the present invention, the detergent or the detergent works by the following steps: causing the biomembrane to disintegrate and release membrane proteins, and providing a hydrophobic environment for the membrane proteins in the demembrane state in the solution, thereby extracting the membrane proteins on the biomembrane into the solution;

[0092] The solution is preferably an aqueous solution; the membrane protein is preferably a G protein-coupled receptor, such as the A2a receptor.

[0093] In one aspect of the present invention, the detergent formulation or the detergent forms micelles in the solution, wherein the diameter of the micelles is preferably 4-50 nm, for example 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;

[0094] The micelles are preferably amphiphilic micelles; the solution is preferably an aqueous solution.

[0095] The present invention also provides a method for preparing the above-mentioned compound I, which includes the following steps:

[0096] 1. Preparation of hydrophobic fragments:

[0097] (1) In a solvent (e.g., THF), dimethyl malonate, a base (e.g., NaH), and a monohalogenated product (R-Br, where R is C) with a hydrophobic aliphatic group are added. 1-25 An alkylation reaction is carried out using a hydrophobic aliphatic group (e.g., R-Br is 1-bromodecane). After the reaction is completed, the first intermediate is separated. The first intermediate refers to the intermediate compound obtained by di-substituting the active methylene group of dimethyl malonate with a hydrophobic aliphatic group (e.g., n-decyl).

[0098] (2) In a solvent (e.g., THF), the first intermediate and a reducing agent (e.g., LiAlH4) are reduced to obtain the second intermediate; the substance obtained in this step is the intermediate compound obtained after both ester groups on the first intermediate are reduced to primary alcohols.

[0099] (3) In a solvent (e.g., DMF), the second intermediate, a base (e.g., NaOH) and 3-bromopropyne are subjected to an etherification reaction. After the reaction is completed, a hydrophobic fragment is separated. The hydrophobic fragment refers to the third intermediate compound obtained after both primary hydroxyl groups of the second intermediate are etherified.

[0100]

[0101] 2. Preparation of hydrophilic fragments:

[0102] (4) In a solvent (e.g., DCM), a hydroxyl-protected monosaccharide or oligosaccharide (preferably a hydroxyl-protected monosaccharide or oligosaccharide, such as β-D-maltose octaethyl ester), an azide reagent (e.g., trimethyl azidosilane), and a Lewis acid (e.g., FeCl3) are subjected to a terminal azide reaction. After the reaction is completed, a hydrophilic fragment is separated. The hydrophilic fragment refers to the fourth intermediate compound obtained by replacing the terminal of the hydroxyl-protected monosaccharide or oligosaccharide with an azide group.

[0103]

[0104] 3. Preparation of the detergent:

[0105] (5) In a solvent (e.g., DCM and / or MeOH), the third intermediate and the fourth intermediate, sodium ascorbate and copper sulfate are subjected to a Click reaction. After the reaction is completed, the fifth intermediate is separated. The fifth intermediate obtained in this step is an intermediate compound obtained by linking the hydrophobic fragment and the hydrophilic fragment through a triazole group using a Click reaction.

[0106] (6) The fifth intermediate and sodium methoxide are subjected to methanololysis in a solvent (e.g., MeOH). After the reaction is completed, compound I is separated.

[0107] .

[0108] The present invention also provides a compound II, which has any of the following structures:

[0109] , , , , , , , , , , , , , , , , , , or .

[0110] Unless otherwise specified, the terms used in this invention may be defined as follows:

[0111] The term "alkyl" refers to a straight-chain or branched alkyl group having a specified number of carbon atoms (e.g., C1-C6). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, etc.

[0112] The term "alkylene" refers to a straight-chain divalent hydrocarbon group or a branched divalent hydrocarbon group having a specified number of carbon atoms (e.g., C1-C6). Examples of alkylene groups include, but are not limited to, methylene, ethylene, propylene, 1-methylpropylene, butylene, etc.

[0113] The term "heteroalkyl" refers to a substance having a specified number of carbon atoms (e.g., C1-C1). 20 A straight-chain or branched monovalent group having a specified number of heteroatoms (e.g., 1-5) and a specified type of heteroatoms (e.g., one or two of O and S); the term "heteroalkyl" can be understood as a group obtained by independently replacing one or more carbon-carbon single bonds in an "alkyl" having the same number of carbon atoms with -O- or -S-, for example or .

[0114] In this invention, "pentose" is understood as a monosaccharide with a chain-like (straight-chain or branched-chain) structure containing five carbon atoms, such as aldoses or ketoses, or, for example, D-ribose. ;

[0115] "Pentose furanose or pyranose" can be understood as the carbonyl group and hydroxyl group of a chain-like pentose forming an intramolecular cyclic hemiacetal or hemiketal. If the hemiacetal or hemiketal is a five-membered ring, it is "pentose furanose"; if the hemiacetal or hemiketal is a six-membered ring, it is "pentose pyranose".

[0116] For example, a furanose of a pentose is a furanose of a D-ribose. .

[0117] In this invention, "hexose" is understood as a monosaccharide containing six carbon atoms with a chain-like (straight-chain or branched-chain) structure, such as aldoses or ketoses, for example, D-glucose. ;

[0118] "Hexose furanose or pyranose" can be understood as the carbonyl group and hydroxyl group of a chain-like hexose forming an intramolecular cyclic hemiacetal or hemiketal. If the hemiacetal or hemiketal is a five-membered ring, it is "hexose furanose"; if the hemiacetal or hemiketal is a six-membered ring, it is "hexose pyranose".

[0119] For example, a pyranose of a hexose is a pyranose of D-glucose. or .

[0120] In this invention, although "oligosaccharide" is defined as "obtained by dehydration polymerization of 2-10 monosaccharides," those skilled in the art will understand that the expression "dehydration polymerization" in this definition is not a limitation on its preparation process, but only to indicate the structure of the "oligosaccharide," that is, the connection mode between the monosaccharides of each monomer in the "oligosaccharide." Moreover, the connection mode between the monosaccharides of each monomer in the "oligosaccharide" is independent of each other.

[0121] In this invention, "glycosyl" refers to a group formed by losing a hemiacetal hydroxyl or hemiketal hydroxyl group from the corresponding prototype sugar; for example, β-D-glucose pyranopyran. The sugar group is For example, β-D-maltose The sugar group is .

[0122] "Hydrophobic aliphatic groups" refer to saturated or unsaturated chain or cyclic hydrocarbon groups that do not contain aryl or heteroaryl groups; they have no affinity for water, are insoluble in water or have very low solubility, and can generate hydrophobic interactions.

[0123] Detergents are a class of amphiphilic molecules that, in a hydrophilic environment, can extract membrane proteins from the phospholipid bilayer. In solution, detergents mimic the amphiphilic system of natural biological membranes to stabilize membrane proteins, thus ensuring that their structure and function are not disrupted. Detergents can form membrane protein-detergent complexes (PDCs) with membrane proteins, which can be directly used in crystallography or NMR experiments. An ideal detergent can maintain membrane proteins in a soluble state without having a measurable effect on their function, structure, and thermodynamic properties.

[0124] Biological membranes: Except for some viruses, all living organisms possess biological membranes. Eukaryotic cells, in addition to the plasma membrane (also known as the cell membrane), have a membrane system separating various organelles, including the nuclear membrane, mitochondrial membrane, endoplasmic reticulum membrane, lysosomal membrane, Golgi apparatus membrane, chloroplast membrane, vacuoles, and peroxisomal membranes. The internal membrane system includes the nuclear membrane, endoplasmic reticulum membrane, lysosomal membrane, Golgi apparatus membrane, and vacuoles (including endosomes and secretory vacuoles). Morphologically, biological membranes are all bilayered sheet structures, approximately 5–10 nanometers thick. Their main components are lipids and proteins, with a small amount of carbohydrates covalently bonded to lipids or proteins. Different biological membranes have different functions.

[0125] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0126] The reagents and raw materials used in this invention are all commercially available.

[0127] The positive and progressive effects of this invention are as follows:

[0128] (1) The compound has a novel structure, better stabilizing effect on complex membrane proteins, is easy to synthesize, can be prepared on a large scale, has excellent properties, and is easy to control in terms of quality;

[0129] (2) The compounds provided by this invention can cause lipid membrane disintegration and release membrane proteins, and provide a hydrophobic environment for the membrane proteins in the demembrane state in solution, thereby extracting membrane proteins from biological membranes into aqueous solutions. In addition, the above compounds can form micelles of suitable diameter in water, and the thermal stability of membrane proteins can be significantly improved in the presence of the above compounds;

[0130] (3) G protein-coupled receptors have relatively poor stability, and some traditional detergents have difficulty effectively stabilizing such membrane proteins. However, the compounds provided in this application have a better stabilizing effect on these proteins. In a specific embodiment of the present invention, the membrane protein may be selected from A2a receptors;

[0131] (4) The compounds provided by this invention are dimer compounds with “pre-assembled” characteristics in molecular structure. These compounds can significantly improve the stability of membrane proteins in the thermal stability experiment of membrane proteins, and the effect is significantly higher than that of commonly used commercial detergents.

[0132] (5) The compounds provided by this invention link hydrophilic and hydrophobic fragments through triazole groups, which can replace the inefficient polysaccharide glycosylation reaction with a highly efficient click chemistry reaction in the preparation process, greatly improving the accessibility of detergents, and can be synthesized efficiently, with great commercial prospects.

[0133] (6) The detergent compound provided by the present invention is superior to traditional detergents in terms of separation and quality control. (Traditional detergents require the precursors without the removal of the protecting group to be synthesized by glycosylation reaction under harsh reaction conditions and using metal catalysts. The detergent precursors themselves are oily and difficult to separate and purify, which makes it difficult to control the quality of the detergent precursors and ultimately affects the separation and quality control of the detergent. The detergent precursors of the present application, namely M-CuAAc compounds, are solid and have better properties than traditional oily detergent precursors, which is more conducive to the quality control of the detergent.) Attached Figure Description

[0134] Figure 1 The particle size distribution of the membrane protein-detergent complex at 1 min and 120 min is shown in Example 4 when the detergent is DDM.

[0135] Figure 2 The particle size distribution of the membrane protein-detergent complex at 1 min and 120 min is shown in Example 4 when the detergent is LMNG.

[0136] Figure 3The particle size distribution of the membrane protein-detergent complex at 1 min and 120 min is shown in Example 4 when the detergent is DTG-0101.

[0137] Figure 4 The particle size distribution of the membrane protein-detergent complex at 1 min and 120 min is shown in Example 4 when the detergent is DTG-0102.

[0138] Figure 5 The particle size distribution of the membrane protein-detergent complex at 1 min and 120 min is shown in Example 4 when the detergent is DTG-0103.

[0139] Figure 6 The particle size distribution of the membrane protein-detergent complex at 1 min and 120 min is shown in Example 4 when the detergent is DTG-0104. Detailed Implementation

[0140] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0141] Example 1: Triazole-linked dimer detergent molecule and its synthesis:

[0142] Preparation of hydrophobic fragments:

[0143]

[0144] 1.1 Preparation method of intermediate M-Dialkyl-C10C10 (n=8):

[0145]

[0146] Add 10.00 g (75.70 mmol) of dimethyl malonate and 100 ml of THF, and heat to 0°C on ice. Add 2.00 g (83.27 mmol) of NaH and 18.42 g (83.27 mmol) of 1-bromodecane, and heat to 25-30°C with stirring for 12 h. Heat to 0°C again, add 2.00 g (83.27 mmol) of NaH and 18.42 g (83.27 mmol) of 1-bromodecane, and continue stirring at 25-30°C for 12 h. The reaction was monitored by TLC and found to be complete. Quench the reaction solution with 100 ml of saturated NH4Cl solution, and then extract the aqueous phase with EA (100 ml x 2). Combine the organic phases, dry to anhydrous NaSO4, filter, and concentrate to obtain a crude, golden-yellow oil. The crude product was then purified by silica gel column chromatography (eluent PE:EA = 100:1), and the collected solution was concentrated to obtain 21.50 g of pure colorless oil (molar yield: 68.8%).

[0147] By following the above method and only changing the type of haloalkane, the following M-Dialkyl series intermediates can be obtained.

[0148] 1.2 The structural characterization of the intermediate M-Dialkyl is as follows:

[0149] Structural characterization of M-Dialkyl-C3C3-Cy:

[0150]

[0151] A colorless oily substance with a separation yield of 72.0%. 1 H NMR (300 MHz, Chloroform-d)δ3.70 (s,6H), 1.93-1.81 (m, 4H), 1.78-1.53 ​​(m, 6H), 1.30-1.03 (m, 20H), 0.95-0.78 (m,4H). ESI-MS(m / z):381.30[M+H] +

[0152] Structural characterization of M-Dialkyl-C4C4 (n=2):

[0153]

[0154] A colorless oily substance with a separation yield of 72.0%. 1H NMR (600 MHz, Chloroform-d)δ3.70 (s,6H), 1.93-1.81 (m, 4H), 1.32-1.23 (m, 4H), 1.17-1.06 (m, 4H), 0.87 (t, J =6.8 Hz, 6H). ESI-MS(m / z):245.18[M+H] +

[0155] Structural characterization of M-Dialkyl-C6C6 (n=4):

[0156]

[0157] A colorless oily substance with a separation yield of 68.5%. 1 H NMR (600 MHz, Chloroform-d)δ3.70 (s,6H), 1.93-1.81 (m, 4H), 1.32-1.23 (m, 12H), 1.17-1.06 (m, 4H), 0.87 (t, J =6.8 Hz, 6H). ESI-MS(m / z):301.24[M+H] +

[0158] Structural characterization of M-Dialkyl-C7C7 (n=5):

[0159]

[0160] A colorless oily substance with a separation yield of 68.7%. 1 H NMR (600 MHz, Chloroform-d)δ3.70 (s,6H), 1.99-1.74 (m, 4H), 1.33-1.20 (m, 16H), 1.13 (dq, J = 14.7, 6.7, 6.2 Hz,4H), 0.88 (t, J = 7.0 Hz, 6H). ESI-MS(m / z):329.27[M+H] +

[0161] Structural characterization of M-Dialkyl-C8C8 (n=6):

[0162]

[0163] A colorless oily substance with a separation yield of 71.5%. 1H NMR (600 MHz, Chloroform-d)δ3.70 (s,6H), 1.99-1.74 (m, 4H), 1.33-1.20 (m, 20H), 1.13 (dq, J = 14.7, 6.7, 6.2 Hz,4H), 0.88 (t, J = 7.0 Hz, 6H). ESI-MS(m / z):357.30[M+H]+

[0164] Structural characterization of M-Dialkyl-C9C9 (n=7):

[0165]

[0166] A colorless oily substance with a separation yield of 69.5%. 1 H NMR (400 MHz, Chloroform-d) δ 3.70 (s,6H), 1.90 -1.82 (m, 4H), 1.33-1.20 (m, 24H), 1.11 (dq, J = 11.6, 6.6, 5.2 Hz,5H), 0.88 (t, J = 6.8 Hz, 7H). ESI-MS(m / z):385.33[M+H] +

[0167] Structural characterization of M-Dialkyl-C10C10 (n=8):

[0168]

[0169] A colorless oily substance with a separation yield of 68.8%. 1 H NMR (400 MHz, Chloroform-d) δ3.70 (s,6H), 1.92-1.79 (m, 4H), 1.33-1.20 (m, 28H), 1.12 (dd, J = 10.4, 6.3 Hz, 4H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):413.36[M+H] +

[0170] Structural characterization of M-Dialkyl-C12C12 (n=10):

[0171]

[0172] A colorless oily substance with a separation yield of 71.2%. 1H NMR (600 MHz, Chloroform-d)δ3.70 (s,6H), 1.90-1.80 (m, 4H), 1.33-1.20 (m, 36H), 1.12 (td, J = 11.5, 9.6, 5.5 Hz,4H), 0.88 (t, J = 7.0 Hz, 6H). ESI-MS(m / z):469.42[M+H] +

[0173] Structural characterization of M-Dialkyl-C14C14 (n=12):

[0174]

[0175] A colorless oily substance with a separation yield of 71.8%. 1 H NMR (400 MHz, Chloroform-d)δ3.70 (s,6H), 1.94-1.79 (m, 4H), 1.31-1.21 (m, 44H), 1.11 (td, J = 11.2, 9.6, 5.2 Hz,4H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):525.49[M+H] +

[0176] Structural characterization of M-Dialkyl-C16C16 (n=14):

[0177]

[0178] Colorless oily substance, separation yield 68.9%. ¹H NMR (400 MHz, Chloroform-d) δ 3.70 (s, 6H), 1.94–1.81 (m, 4H), 1.30–1.25 (m, 52H), 1.15–1.07 (m, 4H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS (m / z): 581.55 [M+H]+

[0179] Structural characterization of M-Dialkyl-C20C20 (n=18):

[0180]

[0181] Colorless oily substance, separation yield 68.0%. ¹H NMR (400 MHz, Chloroform-d) δ 3.70 (s, 6H), 1.94–1.81 (m, 4H), 1.30–1.25 (m, 68H), 1.15–1.07 (m, 4H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS (m / z): 693.685 [M+H]+

[0182] 2.1 Preparation method of intermediate M-Reduc-C10C10:

[0183]

[0184] 10.00 g (24.24 mmol) of M-Dialkyl-C10C10 and 100 ml of THF were added, and the mixture was kept in an ice bath at 0 °C. 2.02 g (53.33 mmol) of LiAlH4 was added in portions, with the temperature controlled below 10 °C. After the addition was complete, the mixture was stirred at 25-30 °C for 4 h, and TLC was monitored until the M-Dialkyl-C10C10 was completely converted. The reaction mixture was cooled to 0 °C in an ice bath, and 2.00 ml of water was slowly added dropwise to quench the reaction. Then, 2.00 ml of 15% NaOH solution was slowly added dropwise. 6.00 ml of water was added, and the mixture was heated to 25 °C and stirred for 15 min (a white granular precipitate formed, indicating a solid-liquid two-phase system). An appropriate amount of anhydrous MgSO4 was added, and stirring was continued for 5 min. The mixture was filtered, and the mother liquor was concentrated to obtain a colorless oily substance. The crude product was purified by silica gel column chromatography to obtain 7.65 g of pure product (molar yield 88.5%).

[0185] By following the above method, and only changing the types of M-Dialkyl series compounds in the raw materials, the following M-Reduc series intermediates can be obtained.

[0186] 2.2 The structural characterization of the intermediate M-Reduc is as follows:

[0187] Structural characterization of M-Reduc-C3C3-Cy:

[0188]

[0189] A colorless oily substance with a separation yield of 98.9%. 1H NMR (300 MHz, Chloroform-d): δ 3.56 (d,J= 5.2 Hz, 4H), 2.43 (s, 2H), 1.78-1.53 ​​(m, 10H), 1.30-1.03 (m, 20H), 0.95-0.78 (m, 4H). ESI-MS(m / z):347.29[M+Na] +

[0190] Structural characterization of M-Reduc-C4C4 (n=2):

[0191]

[0192] A colorless oily substance with a separation yield of 80.2%. 1 H NMR (400 MHz, Chloroform-d) δ3.46 (s,4H), 2.94 (s, 2H), 1.28-1.06 (m, 12H), 0.88 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):189.19[M+H] +

[0193] Structural characterization of M-Reduc-C6C6 (n=4):

[0194]

[0195] A colorless oily substance with a separation yield of 79.2%. 1 H NMR (400 MHz, Chloroform-d) δ3.46 (s,4H), 2.94 (s, 2H), 1.28-1.06 (m, 20H),0.88 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):489.49[2M+H] +

[0196] Structural characterization of M-Reduc-C7C7 (n=5):

[0197]

[0198] A colorless oily substance with a separation yield of 78.4%. 1H NMR (400 MHz, Chloroform-d) δ3.56 (m,4H), 2.37 (s, 2H), 1.32-1.19 (m, 24H), 0.88 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):271.26[MH] -

[0199] Structural characterization of M-Reduc-C8C8 (n=6):

[0200]

[0201] A colorless oily substance with a separation yield of 82.3%. 1 H NMR (400 MHz, Chloroform-d) δ3.56 (s,4H), 2.37 (s, 2H), 1.32-1.19 (m, 28H), 0.88 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):301.31[M+H] +

[0202] Structural characterization of M-Reduc-C9C9 (n=7):

[0203]

[0204] White paste-like substance, separation yield 86.2%. 1 H NMR (600 MHz, Chloroform-d) δ3.56 (s,4H), 2.23 (s, 2H), 1.31-1.18(m, 32H), 0.88 (t, J = 7.0 Hz, 6H). ESI-MS(m / z):351.32[M+Na] +

[0205] Structural characterization of M-Reduc-C10C10 (n=8):

[0206]

[0207] White paste-like substance, separation yield 88.5%. 1 H NMR (600 MHz, Chloroform-d) δ3.56 (s,4H), 2.22 (s, 2H), 1.31-1.18(m, 36H), 0.88 (t, J = 7.0 Hz, 6H). ESI-MS(m / z):357.38[M+H] +

[0208] Structural characterization of M-Reduc-C12C12 (n=10):

[0209]

[0210] White paste-like substance, separation yield 88.1%. 1 H NMR (400 MHz, Chloroform-d) δ3.56 (s,4H), 2.09 (s, 2H), 1.32-1.20 (m, 44H), 0.88 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):435.42[M+Na] +

[0211] Structural characterization of M-Reduc-C14C14 (n=12):

[0212]

[0213] White paste-like substance, with a separation yield of 89.1%. 1 H NMR (400 MHz, Chloroform-d) δ3.50 (s,4H), 1.94 (s, 2H), 1.23-1.14 (m, 52H), 0.81 (t, J = 6.7 Hz, 6H). ESI-MS(m / z): / [M+H] +

[0214] Structural characterization of M-Reduc-C16C16 (n=14):

[0215]

[0216] White paste-like substance, with a separation yield of 88.9%. 1 H NMR (400 MHz, Chloroform-d) δ3.70 (s,4H), 1.28-1.20 (m, 60H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z): / [M+H] +

[0217] Structural characterization of M-Reduc-C20C20 (n=18):

[0218]

[0219] White paste-like substance, with a separation yield of 89.9%. 1H NMR (400 MHz, Chloroform-d) δ3.70 (s,4H), 1.28-1.20 (m, 76H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):636.69[M+H] +

[0220] 3.1 Preparation method of intermediate M-William-C10C10:

[0221]

[0222] 6.00 g (16.82 mmol) of M-Reduc-C10C10 sample was dissolved in 20 mL of DMF, and 1.68 g (42.05 mmol) of NaOH and 5.00 g (42.05 mmol) of 3-bromopropyne were added. The mixture was stirred at 25-30 °C for 24 h, and TLC monitoring showed no further reaction. The above reaction solution was diluted with 200 mL of water and extracted with n-hexane (100 mL x 3). The organic phases were combined. The mixture was then washed successively with 100 mL of saturated NH4Cl solution and 100 mL of saturated NaCl solution to separate the aqueous phase. The organic phase was dried over anhydrous MgSO4, filtered, and the mother liquor was concentrated to obtain a crude golden-yellow oily product. The crude product was then purified by column chromatography (eluent PE→P:E=30:1), and the collected solution was concentrated to obtain 4.12 g of pure product (molar yield 56.7%).

[0223] By following the above method, and only changing the types of M-Reduc series compounds in the raw materials, the following M-William series intermediates can be obtained.

[0224] 3.2 The structural characterization of the intermediate M-William is as follows:

[0225] Structural characterization of M-William-C3C3-Cy:

[0226]

[0227] Yellow oily substance, separation yield 72.3%. 1 H NMR (400 MHz, Chloroform-d) δ4.11 (d, J= 2.4 Hz, 4H), 3.31 (s, 4H), 2.38 (t, J = 2.4 Hz, 2H), 1.78-1.53 ​​(m, 10H), 1.30-1.03 (m, 20H), 0.95-0.78 (m, 4H). ESI-MS(m / z):401.34[M+H] +

[0228] Structural characterization of M-William-C4C4 (n=2):

[0229]

[0230] Yellow oily substance, separation yield 65.4%. 1 H NMR (400 MHz, Chloroform-d) δ4.11 (d, J= 2.4 Hz, 4H), 3.31 (s, 4H), 2.38 (t, J = 2.4 Hz, 2H), 1.29-1.19 (m,12H),0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):265.22[M+H] +

[0231] Structural characterization of M-William-C6C6 (n=4):

[0232]

[0233] Yellow oily substance, separation yield 64.4%. 1 H NMR (400 MHz, Chloroform-d) δ4.11 (d, J= 2.4 Hz, 4H), 3.31 (s, 4H), 2.38 (t, J = 2.4 Hz, 2H), 1.29-1.19 (m, 20H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):321.28[M+H] +

[0234] Structural characterization of M-William-C7C7 (n=5):

[0235]

[0236] Yellow oily substance, separation yield 63.2%. 1 H NMR (400 MHz, Chloroform-d) δ4.11 (d, J= 2.4 Hz, 4H), 3.31 (s, 4H), 2.38 (t, J = 2.4 Hz, 2H), 1.30-1.20 (m, 24H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):349.31[M+H] +

[0237] Structural characterization of M-William-C8C8 (n=6):

[0238]

[0239] Yellow oily substance, separation yield 60.0%. 1 H NMR (400 MHz, Chloroform-d) δ4.11 (d, J= 2.4 Hz, 4H), 3.30 (s, 4H), 2.38 (t, J = 2.4 Hz, 2H), 1.30-1.20 (m, 28H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):377.34[M+H] +

[0240] Structural characterization of M-William-C9C9 (n=7):

[0241]

[0242] Yellow oily substance, separation yield 58.5%. 1 H NMR (400 MHz, Chloroform-d) δ4.11 (d, J= 2.3 Hz, 4H), 3.30 (s, 4H), 2.38 (t, J = 2.3 Hz, 2H), 1.30-1.22(m, 32H), 0.88 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):405.37[M+H] +

[0243] Structural characterization of M-William-C10C10 (n=8):

[0244]

[0245] Yellow oily substance, separation yield 56.7%. 1 H NMR (400 MHz, Chloroform-d)δ4.04 (d, J =2.4 Hz, 4H), 3.23 (s, 4H), 2.31 (t, J = 2.4 Hz, 2H), 1.24-1.19 (m, 36H), 0.81(t, J = 6.8 Hz, 6H). ESI-MS(m / z):471.38[M+K] +

[0246] Structural characterization of M-William-C12C12 (n=10):

[0247]

[0248] Yellow oily substance, separation yield 56.3%. 1 H NMR (400 MHz, Chloroform-d)δ4.11 (d, J =2.4 Hz, 4H), 3.30 (s, 4H), 2.38 (t, J = 2.4 Hz, 2H), 1.30-1.22 (m, 44H), 0.88(t, J = 6.8 Hz, 6H). ESI-MS(m / z):489.47[M+H] +

[0249] Structural characterization of M-William-C14C14 (n=12):

[0250]

[0251] Yellow oily substance, separation yield 56.5%. 1 H NMR (400 MHz, Chloroform-d)δ4.11 (d, J =2.4 Hz, 4H), 3.30 (s, 4H), 2.38 (t, J = 2.4 Hz, 2H), 1.30-1.22 (m, 52H), 0.88(t, J = 6.8 Hz, 6H). ESI-MS(m / z):545.53[M+H] +

[0252] Structural characterization of M-William-C16C16 (n=14):

[0253]

[0254] Yellow oily substance, separation yield 55.8%. 1 H NMR (400 MHz, Chloroform-d)δ4.11 (d, J =2.4 Hz, 4H), 3.30 (s, 4H), 2.38 (t, J = 2.3 Hz, 2H), 1.30-1.22 (m, 60H), 0.88(t, J = 6.8 Hz, 6H). ESI-MS(m / z): 601.59[M+H] +

[0255] Structural characterization of M-William-C20C20 (n=20):

[0256]

[0257] Yellow oily substance, separation yield 56.7%. 1 H NMR (400 MHz, Chloroform-d)δ4.11 (d, J =2.4 Hz, 4H), 3.30 (s, 4H), 2.38 (t, J = 2.3 Hz, 2H), 1.30-1.22 (m, 76H), 0.88(t, J = 6.8 Hz, 6H). ESI-MS(m / z): 713.71[M+H] +

[0258] 4. Preparation of hydrophilic glycosyl azides:

[0259] 4.1 Preparation method of terminal azidoglycoester:

[0260] Preparation of β-D-maltose azide heptaacetate:

[0261]

[0262] 30.00 g (44.21 mmol) of β-D-maltose octaethyl ester was dissolved in 300 mL of DCM, and 7.64 g (66.31 mmol) of azidotrimethylsilane and 360 mg (2.21 mmol) of FeCl3 were added. The mixture was stirred at 25-30 °C. TLC monitoring showed that the β-D-maltose octaethyl ester was completely converted after 12 h of reaction. 240 mg of triethylamine was added to quench the reaction. The reaction solution was washed successively with saturated NaHCO3 solution (150 mL x 2) and saturated NaCl solution (150 mL) to separate the aqueous phase. The resulting organic phase was dried over anhydrous MgSO4, filtered, and the mother liquor was concentrated to obtain a foamy colloidal crude product. This was then subjected to silica gel column chromatography (eluent: PE:EA = 2:1), and the collected solution was concentrated to give 28.50 g of a white powdery solid (molar yield 97.4%).

[0263] By following the above method and only changing the type of sugar ester in the raw materials, the following terminal azidoglycoesters can be obtained.

[0264] 4.2 Structural characterization of terminal azidoglycoesters:

[0265] β-D-glucosyl azide tetraacetate:

[0266]

[0267] White foamy solid, separation yield 97.0%. 1H NMR (400 MHz, Chloroform-d) δ5.22(t, J = 9.5 Hz, 1H), 5.10 (t, J = 9.7 Hz, 1H), 4.95 (t, J = 9.2 Hz, 1H), 4.64(d, J = 8.9 Hz, 1H), 4.27 (dd, J = 12.5, 4.8 Hz, 1H), 4.17 (dd, J = 12.5, 2.3Hz, 1H), 3.79 (ddd, J = 10.0, 4.8, 2.3 Hz, 1H), 2.10 (s, 3H), 2.08 (s, 3H), 2.03 (s, 3H), 2.01 (s, 3H). ESI-MS(m / z): 396.10 [M+Na] +

[0268] β-D-maltose azide heptaacetate:

[0269]

[0270] White foamy solid, separation yield 96.9%. 1 H NMR (400 MHz, Chloroform-d) δ5.34(d, J = 4.0 Hz, 1H), 5.29 (dd, J = 10.5, 9.5 Hz, 1H), 5.21 (dd, J = 17.9, 9.0Hz, 1H), 4.99 (t, J = 9.9 Hz, 1H), 4.79 (dd, J = 10.5, 4.0 Hz, 1H), 4.72 (t,J = 8.9 Hz, 1H), 4.65 (d, J = 8.7 Hz, 1H), 4.45 (dd, J = 12.3, 2.4 Hz, 1H), 4.26-4.11 (m, 2H), 4.03-3.92 (m, 2H), 3.89 (ddd, J = 10.3, 4.0, 2.3 Hz, 1H), 3.72 (ddd, J = 9.9, 4.5, 2.5 Hz, 1H), 2.09 (s, 3H), 2.04 (s, 3H), 1.99 (s,3H), 1.98 (s, 3H), 1.96 (s, 3H), 1.95 (s, 3H), 1.94 (s, 3H). ESI-MS(m / z):684.18[M+Na] +9

[0271] β-D-Cellobiose azide heptaacetate:

[0272]

[0273] White foamy solid, with a separation yield of 97.8%. 1 H NMR (400 MHz, Chloroform-d) δ5.20 -5.08 (m, 2H), 5.04 (t, J = 9.6 Hz, 1H), 4.93 - 4.81 (m, 2H), 4.60 (d, J = 8.8Hz, 1H), 4.54-4.47 (m, 2H), 4.36 (dd, J = 12.5, 4.4 Hz, 1H), 4.10 (dd, J =12.2, 5.0 Hz, 1H), 4.02 (dd, J = 12.5, 2.1 Hz, 1H), 3.82 - 3.74 (m, 1H), 3.72- 3.61 (m, 2H), 2.12 (s, 3H), 2.07 (s, 3H), 2.05 (s, 3H), 2.01 (s, 3H), 2.00(s, 3H), 1.99 (s, 3H), 1.96 (s, 3H). ESI-MS(m / z): 684.18[M+Na] +

[0274] β-D-lactosylazidoheptaacetate:

[0275]

[0276] White foamy solid, separation yield 98.2%. 1H NMR (400 MHz, Chloroform-d) δ5.34(d, J= 3.6, 1H), 5.20 (t, J= 9.4, 1H), 5.09 (dd, J= 10.4, 8, 1H), 4.94 (dd, J= 10.4, 3.6, 1H), 4.85 (t, J= 9.6, 1H), 4.62 (d, J= 8.8, 1H), 4.52-4.46 (m,2H), 4.14-4.05 (m, 3H), 3.87 (t, J = 6.8, 1H), 3.81 (t, J= 9.6, 1H), 3.70-3.71 (m, 1H), 2.14 (s, 3H), 2.13 (s, 3H), 2.08 (s, 3H), 2.06 (s, 3H), 2.05(s, 3H), 2.04 (s, 3H), 1.95 (s, 3H). ESI-MS(m / z): 684.18[M+Na] +

[0277] β-D-Melbiose-syl azide heptaacetate:

[0278]

[0279] White foamy solid, separation yield 98.0%. 1 H NMR (400 MHz, Chloroform-d) 5.44 (d,J = 3.8, 1H), 5.35 (dd, J = 10.8, 3.8, 1H), 5.21 (t, J = 9.0, 1H), 5.15 (d, J =3.8, 1H), 5.10 (dd, J = 10.8, 3.8, 1H), 5.08 (t, J = 9.0, 1H), 4.90 (t, J =9.0, 1H), 4.60 (d, J = 9.0, 1H), 4.21 (t, J = 6.8, 1H), 4.07 (d, J = 6.8,2H), 3.70-3.80 (m, 2H), 3.55-3.60 (m, 1H), 2.12 (s, 3H), 2.10 (s, 3H), 2.06(s, 3H), 2.03 (s, 6H), 2.00 (s, 3H), 1.97 (s, 3H). ESI-MS(m / z): 684.18[M+Na] +

[0280] Preparation of detergent:

[0281]

[0282] 5.1 Preparation method of M-CuAAc:

[0283] Taking the preparation of M-Maltose-CuAAc-C10C10 as an example:

[0284]

[0285] 1.00 g (2.31 mmol) of M-William-C10C10 and 3.21 g (2.10 eq) of β-D-maltose azide heptaacetate were dissolved in a mixed solvent of DCM and MeOH (DCM:MeOH = 1:1). Then, 362 mg (1.85 mmol) of sodium ascorbate and 274 mg (1.16 mmol) of CuSO4 were added sequentially, and the mixture was stirred at 25-30 °C. TLC monitoring showed that the starting material M-William-C10C10 was completely converted after 12 h of reaction. The reaction solution was concentrated, diluted with 20 ml of saturated NH4Cl solution, and then extracted with DCM (20 ml x 3). The organic phases were combined. The solution was washed with saturated NH4Cl solution (20 ml x 2) and the aqueous phase was separated. The organic phase was dried with anhydrous MgSO4, filtered, and the mother liquor was concentrated to obtain a yellow-green foamy solid. The crude product was purified by column chromatography (eluent P:E = 1:1 → 1:2) to obtain 3.45 g of white powdery solid (molar yield 85.1%).

[0286] Following the method described above, the following M-CuAAc series compounds can be obtained.

[0287] 5.2 Structural characterization of M-CuAAc:

[0288] Structural characterization of M-Maltose-CuAAc-C3C3-Cy:

[0289]

[0290] White foamy solid, separation yield 94.9%. 1H NMR (400 MHz, Chloroform-d) δ7.72(s, 2H), 5.96 (d, J = 9.1 Hz, 2H), 5.50-5.36 (m, 8H), 5.08 (t, J = 9.9 Hz,2H), 4.89 (dd, J = 10.5, 4.0 Hz, 2H), 4.64-4.54 (m, 4H), 4.52 (dd, J = 12.8,2.7 Hz, 2H), 4.27 (dt, J = 12.5, 3.8 Hz, 4H), 4.19 (t, J = 9.1 Hz, 2H), 4.06(ddd, J = 12.6, 11.1, 2.2 Hz, 4H), 4.00 (dd, J = 10.3, 3.2 Hz, 2H), 3.41 -3.16 (m, 4H), 2.12 (s, 6H), 2.11 (s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03(s, 6H), 2.02 (s, 6H), 1.81 (s, 6H), 1.78-1.53 ​​(m, 10H), 1.30-1.03 (m, 20H), 0.95-0.78 (m, 4H). ESI-MS(m / z):1723.74[M+H] +

[0291] Structural characterization of M-Glucose-CuAAc-C4C4 (n=2):

[0292]

[0293] White foamy solid, separation yield 96.8%. 1H NMR (400 MHz, Chloroform-d) δ7.96 (2H, s), 6.00-5.93 (m, 2H), 5.56-5.47 (m, 2H), 5.44 (ddd,J= 11.2, 6.6, 2.9 Hz,2H), 5.31-5.23 (m, 2H), 4.64-4.54 (m, 4H), 4.32 (dt,J= 13.1, 4.0 Hz, 2H), 4.15(d,J= 12.7 Hz, 2H), 4.06 (dt, J = 8.3, 3.3 Hz, 2H), 3.39-3.16 (m, 4H), 2.10(s, 6H), 2.07 (s, 3H ), 2.03 (s, 3 H), 1.85 ( s, 3 H), 1.28-1.16 (m, 12H), 0.87(d, J = 7.2 Hz, 6H). ESI-MS(m / z):1011.44[M+H] +

[0294] Structural characterization of M-GlucoseCuAAc-C6C6 (n=4):

[0295]

[0296] White foamy solid, separation yield 96.1%. 1 H NMR (400 MHz, Chloroform-d) δ 7.96 (2H, s), 6.00–5.93 (m, 2H), 5.56–5.47 (m, 2H), 5.44 (ddd,J= 11.2, 6.6, 2.9 Hz,2H), 5.31–5.23 (m, 2H), 4.64-4.54 (m, 4H), 4.32 (dt,J= 13.1, 4.0 Hz, 2H), 4.15(d,J= 12.7 Hz, 2H), 4.06 (dt, J = 8.3, 3.3 Hz, 2H), 3.39-3.16 (m, 4H), 2.10(s, 6H), 2.07 (s, 3H ), 2.03 (s, 3 H), 1.85 ( s, 3 H), 1.28 -1.16 (m, 20H), 0.87(d, J = 7.2 Hz, 6H). ESI-MS(m / z):1067.51[M+H] +

[0297] Structural characterization of M-Maltose-CuAAc-C6C6 (n=4):

[0298]

[0299] White foamy solid, separation yield 94.7%. 1 H NMR (400 MHz, Chloroform-d) δ7.72(s, 2H), 5.96 (d, J = 9.1 Hz, 2H), 5.50-5.36 (m, 8H), 5.08 (t, J = 9.9 Hz,2H), 4.89 (dd, J = 10.5, 4.0 Hz, 2H), 4.64-4.54 (m, 4H), 4.52 (dd, J = 12.8,2.7 Hz, 2H), 4.27 (dt, J = 12.5, 3.8 Hz, 4H), 4.19 (t, J = 9.1 Hz, 2H), 4.06(ddd, J = 12.6, 11.1, 2.2 Hz, 4H), 4.00 (dd, J = 10.3, 3.2 Hz, 2H), 3.41 -3.16 (m, 4H), 2.12 (s, 6H), 2.11 (s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03(s, 6H), 2.02 (s, 6H), 1.81 (s, 6H), 1.28 -1.16 (m, 20H), 0.87 (d, J = 7.2Hz, 6H). ESI-MS(m / z):1643.66[M+H] +

[0300] Structural characterization of M-Cellobiose-CuAAc-C6C6 (n=4):

[0301]

[0302] White foamy solid, separation yield 93.5%. 1H NMR (400 MHz, Chloroform-d) δ7.70(s,2H), 5.80 (d, J = 9.1 Hz, 2H), 5.34-5.32 (m, 4H), 5.12 (s, 4H), 5.01 (t, J=10.5 Hz, 2H) 4.87 (t, J= 10.2 Hz, 2H), 4.64-4.54 (m, 4H), 4.53 (d, J= 8.2Hz, 2H), 4.44 (d, J= 11.5 Hz, 2H), 4.30 (dd, J= 3.3, 10.5 Hz, 2H), 4.07 (d, J= 10.5 Hz, 2H), 3.98 (d, J= 11.4 Hz, 2H), 3.92-3.89 (m, 4H), 3.41-3.16 (m, 4H), 2.01 (s, 6H), 2.00 (s, 6H), 1.96 (s, 6H), 1.95 (s, 6H), 1.92 (s, 6H), 190 (s, 6H), 1.81 (s, 6H), 1.28 -1.16 (m, 20H), 0.87 (d, J = 7.2 Hz, 6H). ESI-MS(m / z):1643.66[M+H] +

[0303] Structural characterization of M-Lactose-CuAAc-C6C6 (n=4):

[0304]

[0305] White foamy solid, separation yield 92.7%. 1H NMR (400 MHz, Chloroform-d) δ7.82(s, 2H), 5.90-5.74 (m, 2H), 5.49-5.28(m, 10H), 4.89 (dd, J = 10.5, 4.0 Hz,2H), 4.97 (dd, J = 10.4, 3.4 Hz, 2H), 4.53-4.46 (m, 4H), 4.19-4.04 (m, 6H), 4.00 (ddd, J = 12.6, 11.1, 2.2 Hz, 4H), 3.84 (dd, J = 10.3, 3.2 Hz, 2H), 3.41-3.16 (m, 4H) 2.12 (s, 6H), 2.11 (s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03(s, 6H), 2.02 (s, 6H), 1.81 (s, 6H), 1.28 -1.16 (m, 20H), 0.87 (d, J = 7.2Hz, 6H). ESI-MS(m / z):1643.66[M+H] +

[0306] Structural characterization of M-Melibiose-CuAAc-C6C6 (n=4):

[0307]

[0308] White foamy solid, separation yield 92.7%. 1H NMR (400 MHz, Chloroform-d) δ7.73(s, 2H), 5.44 (d, J = 3.8Hz, 2H), 5.36(dd, J = 10.8, 3.8Hz, 2H), 5.20 (t, J =9.0Hz, 2H), 5.16 (d, J = 3.8Hz, 2H), 5.09 (dd, J = 10.8, 3.8Hz, 2H), 5.08 (t,J = 9.0Hz, 2H), 4.90 (t, J = 9.0Hz, 2H), 4.66-4.56 (m, 4H), 4.62 (d, J =9.0Hz, 2H), 4.27 (dt, J = 12.5, 3.8 Hz, 4H), 4.21 (t, J = 6.8Hz, 2H), 4.07(d, J = 6.8Hz, 4H), 3.40-3.15 (m, 4H), 2.13 (s, 6H), 2.11 (s, 6H), 2.07 (s,6H), 2.03 (s, 6H), 2.02 (s, 6H),2.00 (s, 6H),1.81 (s, 6H), 1.28 -1.16 (m,20H), 0.87 (d, J = 7.2 Hz, 6H). ESI-MS(m / z):1643.66[M+H] +

[0309] Structural characterization of M-Maltose-CuAAc-C7C7 (n=5):

[0310]

[0311] White foamy solid, separation yield 94.5%. 1H NMR (400 MHz, Chloroform-d) δ7.72(s, 2H), 5.96 (d, J = 9.0 Hz, 2H), 5.51-5.36 (m, 8H), 5.09 (t, J = 9.9 Hz,2H), 4.89 (dd, J = 10.5, 4.0 Hz, 2H), 4.64-4.54 (m, 4H), 4.54-4.46 (m, 2H),4.27 (dt, J = 12.5, 3.9 Hz, 4H), 4.19 (t, J = 9.1 Hz, 2H), 4.12-4.03 (m, 4H),4.00 (dt, J = 10.3, 3.4 Hz, 2H), 3.39-3.16 (m, 4H), 2.12 (s, 6H), 2.11 (s,6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.81 (s, 6H),1.29-1.15 (m, 24H), 0.87 (d, J = 7.3 Hz, 6H). ESI-MS(m / z):1671.70[M+H] +

[0312] Structural characterization of M-Maltose-CuAAc-C8C8 (n=6):

[0313]

[0314] White foamy solid, with a separation yield of 89.4%. 1H NMR (400 MHz, Chloroform-d) δ7.71(s, 2H), 5.95 (d, J = 9.0 Hz, 2H), 5.51-5.36 (m, 8H), 5.08 (t, J = 9.9 Hz,2H), 4.90 (dd, J = 10.5, 4.0 Hz, 2H), 4.66-4.54 (m, 4H), 4.52 (dd, J = 12.3,2.3 Hz, 2H), 4.27 (dt, J = 12.5, 4.0 Hz, 4H), 4.19 (t, J = 9.1 Hz, 2H), 4.13-4.03 (m, 4H), 4.00 (dt, J = 10.3, 3.2 Hz, 2H), 3.39-3.12 (m, 4H), 2.12 (s, 6H), 2.11 (s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.01 (s, 6H), 1.81 (s, 6H), 1.29-1.16 (m, 28H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):1699.73[M+H] +

[0315] Structural characterization of M-Cellobiose-CuAAc-C8C8 (n=6):

[0316]

[0317] White, foamy solid, with a separation yield of 88.5%. 1H NMR (400 MHz, Chloroform-d) δ7.70(s,2H), 5.80 (d, J = 9.1 Hz, 2H), 5.34-5.32 (m, 4H), 5.12 (s, 4H), 5.01 (t, J=10.5 Hz, 2H) 4.87 (t, J= 10.2 Hz, 2H), 4.64-4.54 (m, 4H), 4.53 (d, J= 8.2Hz, 2H), 4.44 (d, J= 11.5 Hz, 2H), 4.30 (dd, J= 3.3, 10.5 Hz, 2H), 4.07 (d, J= 10.5 Hz, 2H), 3.98 (d, J= 11.4 Hz, 2H), 3.92-3.89 (m, 4H), 3.39-3.12 (m, 4H), 2.01 (s, 6H), 2.00 (s, 6H), 1.96 (s, 6H), 1.95 (s, 6H), 1.92 (s, 6H), 190 (s, 6H), 1.81 (s, 6H), 1.28 -1.16 (m, 28H), 0.87 (d, J = 7.2 Hz, 6H). ESI-MS(m / z):1699.73[M+H] +

[0318] Structural characterization of M-Lactose-CuAAc-C8C8 (n=6):

[0319]

[0320] White foamy solid, separation yield 91.7%. 1H NMR (400 MHz, Chloroform-d) δ7.82(s, 2H), 5.90-5.74 (m, 2H), 5.49-5.28(m, 10H), 4.89 (dd, J = 10.5, 4.0 Hz,2H), 4.97 (dd, J = 10.4, 3.4 Hz, 2H),4.53-4.46 (m, 4H), 4.19-4.04 (m, 6H),4.00 (ddd, J = 12.6, 11.1, 2.2 Hz, 4H), 3.84 (dd, J = 10.3, 3.2 Hz, 2H),3.39-3.12 (m, 4H), 2.12 (s, 6H), 2.11 (s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.81 (s, 6H), 1.28 -1.16 (m, 28H), 0.87 (d, J =7.2 Hz, 6H). ESI-MS(m / z):1699.73[M+H] +

[0321] Structural characterization of M-Melibiose-CuAAc-C8C8 (n=6):

[0322]

[0323] White foamy solid, separation yield 91.7%. 1H NMR (400 MHz, Chloroform-d) δ7.73 (s,2H), 5.44 (d, J = 3.8Hz, 2H), 5.36 (dd, J = 10.8, 3.8Hz, 2H), 5.20 (t, J =9.0Hz, 2H), 5.16 (d, J = 3.8Hz, 2H), 5.09 (dd, J = 10.8, 3.8Hz, 2H), 5.08 (t,J = 9.0Hz, 2H), 4.90 (t, J = 9.0Hz, 2H), 4.66-4.56 (m, 4H), 4.62 (d, J =9.0Hz, 2H), 4.27 (dt, J = 12.5, 3.8 Hz, 4H), 4.21 (t, J = 6.8Hz, 2H), 4.07(d, J = 6.8Hz, 4H), 3.41-3.16 (m, 4H), 2.13 (s, 6H), 2.11 (s, 6H), 2.07 (s,6H), 2.03 (s, 6H), 2.02 (s, 6H),2.00 (s, 6H),1.81 (s, 6H), 1.28 -1.16 (m,28H), 0.87 (d, J = 7.2 Hz, 6H). ESI-MS(m / z):1699.73[M+H] +

[0324] Structural characterization of M-Maltose-CuAAc-C9C9 (n=7):

[0325]

[0326] White foamy solid, separation yield 94.1%. 1H NMR (400 MHz, Chloroform-d) δ7.73(s, 2H), 5.96 (d, J = 9.0 Hz, 2H), 5.50-5.36 (m, 8H), 5.09 (t, J = 9.9 Hz,2H), 4.90 (dd, J = 10.6, 3.9 Hz, 2H), 4.64-4.54 (m, 4H), 4.54-4.47 (m, 2H), 4.27 (dt, J = 12.5, 3.9 Hz, 4H), 4.20 (t, J = 9.1 Hz, 2H), 4.08 (dd, J =12.5, 2.5 Hz, 4H), 4.02-3.97 (m, 2H), 3.39-3.11 (m, 4H), 2.12 (s, 6H), 2.11(s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.81 (s,6H), 1.30-1.18 (m, 32H), 0.88 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1727.76[M+H] +

[0327] Structural characterization of M-Maltose-CuAAc-C10C10 (n=8):

[0328]

[0329] White, foamy solid, with a separation yield of 85.1%. 1H NMR (400 MHz, Chloroform-d) δ7.73(s, 2H), 5.96 (d, J = 9.0 Hz, 2H), 5.52-5.35 (m, 8H), 5.09 (t, J = 9.8 Hz,2H), 4.90 (dd, J = 10.5, 4.0 Hz, 2H), 4.67-4.54 (m, 4H), 4.52 (dd, J = 12.5, 2.5 Hz, 2H), 4.27 (dt, J = 12.6, 3.7 Hz, 4H), 4.20 (dd, J = 9.8, 8.4 Hz, 2H), 4.11-4.02 (m, 4H), 4.00 (dt, J = 13.1, 2.5 Hz, 2H), 3.38-3.16 (m, 4H), 2.12 (s, 6H), 2.11 (s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.81 (s, 6H), 1.28-1.14 (m, 36H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):1755.80[M+H] +

[0330] Structural characterization of M-Cellobiose-CuAAc-C10C10 (n=8):

[0331]

[0332] White foamy solid, with a separation yield of 88.4%. 1H NMR (400 MHz, Chloroform-d) δ7.70(s,2H), 5.80 (d, J = 9.1 Hz, 2H), 5.34-5.32 (m, 4H), 5.12 (s, 4H), 5.01 (t, J=10.5 Hz, 2H) 4.87 (t, J= 10.2 Hz, 2H), 4.64-4.54 (m, 4H), 4.53 (d, J= 8.2Hz, 2H), 4.44 (d, J= 11.5 Hz, 2H), 4.30 (dd, J= 3.3, 10.5 Hz, 2H), 4.07 (d, J= 10.5 Hz, 2H), 3.98 (d, J= 11.4 Hz, 2H), 3.92-3.89 (m, 4H), 3.38-3.16 (m, 4H), 2.01 (s, 6H), 2.00 (s, 6H), 1.96 (s, 6H), 1.95 (s, 6H), 1.92 (s, 6H), 190 (s, 6H), 1.81 (s, 6H), 1.28 -1.16 (m, 36H), 0.87 (d, J = 7.2 Hz, 6H). ESI-MS(m / z):1755.80[M+H] +

[0333] Structural characterization of M-Lactose-CuAAc-C10C10 (n=8):

[0334]

[0335] White foamy solid, separation yield 91.7%. 1H NMR (400 MHz, Chloroform-d) δ7.82(s, 2H), 5.90-5.74 (m, 2H), 5.49-5.28(m, 10H), 4.89 (dd, J = 10.5, 4.0 Hz,2H), 4.97 (dd, J = 10.4, 3.4 Hz, 2H),4.53-4.46 (m, 4H), 4.19-4.04 (m, 6H),4.00 (ddd, J = 12.6, 11.1, 2.2 Hz, 4H), 3.84 (dd, J = 10.3, 3.2 Hz, 2H),3.38-3.16 (m, 4H), 2.12 (s, 6H), 2.11 (s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.81 (s, 6H), 1.28 -1.16 (m, 36H), 0.87 (d, J =7.2 Hz, 6H). ESI-MS(m / z):1755.80[M+H] +

[0336] Structural characterization of M-Melibiose-CuAAc-C10C10 (n=8):

[0337]

[0338] White foamy solid, separation yield 91.7%. 1H NMR (400 MHz, Chloroform-d) δ7.73(s, 2H), 5.44 (d, J = 3.8Hz, 2H), 5.36(dd, J = 10.8, 3.8Hz, 2H), 5.20 (t, J =9.0Hz, 2H), 5.16 (d, J = 3.8Hz, 2H), 5.09 (dd, J = 10.8, 3.8Hz, 2H), 5.08 (t,J = 9.0Hz, 2H), 4.90 (t, J = 9.0Hz, 2H), 4.66-4.56 (m, 4H), 4.62 (d, J =9.0Hz, 2H), 4.27 (dt, J = 12.5, 3.8 Hz, 4H), 4.21 (t, J = 6.8Hz, 2H), 4.07 (d, J = 6.8Hz, 4H), 3.38-3.16 (m, 4H), 2.13 (s, 6H), 2.11 (s, 6H), 2.07 (s,6H), 2.03 (s, 6H), 2.02 (s, 6H),2.00 (s, 6H),1.81 (s, 6H), 1.28 -1.16 (m,36H), 0.87 (d, J = 7.2 Hz, 6H). ESI-MS(m / z):1755.80[M+H] +

[0339] Structural characterization of M-Maltose-CuAAc-C12C12 (n=10):

[0340]

[0341] White foamy solid, separation yield 85.3%. 1H NMR (400 MHz, Chloroform-d) δ7.73(s, 2H), 5.96 (d, J = 9.0 Hz, 2H), 5.51-5.36 (m, 8H), 5.09 (t, J = 9.9 Hz,2H), 4.90 (dd, J = 10.5, 4.0 Hz, 2H), 4.66-4.54 (m, 4H), 4.52 (dd, J = 12.4,2.5 Hz, 2H), 4.32-4.24 (m, 4H), 4.20 (dd, J = 9.8, 8.4 Hz, 2H), 4.13-4.03 (m,4H), 4.00 (dt, J = 10.1, 3.2 Hz, 2H), 3.35-3.17 (m, 4H), 2.12 (s, 6H), 2.11(s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.81 (s,6H), 1.30-1.16 (m, 44H), 0.88 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):1811.86[M+H] +

[0342] Structural characterization of M-Maltose-CuAAc-C14C14 (n=12):

[0343]

[0344] White foamy solid, separation yield 88.6%. 1H NMR (400 MHz, Chloroform-d) δ7.72(s, 2H), 5.96 (d, J = 8.9 Hz, 2H), 5.51-5.36 (m, 8H), 5.09 (t, J = 9.9 Hz,2H), 4.90 (dd, J = 10.6, 4.0 Hz, 2H), 4.67-4.54 (m, 4H), 4.52 (dd, J = 12.1,1.9 Hz, 2H), 4.27 (dt, J = 12.5, 3.6 Hz, 4H), 4.20 (dd, J = 9.8, 8.4 Hz, 2H), 4.11-4.03 (m, 4H), 4.03-3.98 (m, 2H), 3.34-3.19 (m, 4H), 2.12 (s, 6H), 2.11(s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.81 (s,6H), 1.30-1.16 (m, 52H), 0.88 (t, J = 6.8 Hz, 7H). ESI-MS(m / z):1868.92[M+H] +

[0345] Structural characterization of M-Maltose-CuAAc-C16C16 (n=14):

[0346]

[0347] White foamy solid, separation yield 86.0%. 1H NMR (400 MHz, Chloroform-d) δ7.72(s, 2H), 5.96 (d, J = 9.0 Hz, 2H), 5.55-5.38 (m, 8H), 5.09 (t, J = 9.9 Hz,2H), 4.90 (dd, J = 10.6, 4.0 Hz, 2H), 4.68-4.54 (m, 4H), 4.54-4.49 (m, 2H), 4.27 (dt, J = 12.6, 3.6 Hz, 4H), 4.20 (dd, J = 9.7, 8.4 Hz, 2H), 4.11-4.03(m, 4H), 4.02-3.98 (m, 2H), 3.36-3.12 (m, 4H), 2.12 (s, 6H), 2.11 (s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.81 (s, 6H), 1.30-1.16 (m, 60H), 0.88 (t, J = 6.7 Hz, 7H). ESI-MS(m / z):1924.98[M+H] +

[0348] Structural characterization of M-Maltose-CuAAc-C20C20 (n=18):

[0349]

[0350] White foamy solid, separation yield 84.0%. 1H NMR (400 MHz, Chloroform-d) δ7.72(s, 2H), 5.96 (d, J = 9.0 Hz, 2H), 5.55-5.38 (m, 8H), 5.09 (t, J = 9.9 Hz,2H), 4.90 (dd, J = 10.6, 4.0 Hz, 2H), 4.68-4.54 (m, 4H), 4.54-4.49 (m, 2H), 4.27 (dt, J = 12.6, 3.6 Hz, 4H), 4.20 (dd, J = 9.7, 8.4 Hz, 2H), 4.11-4.03(m, 4H), 4.02-3.98 (m, 2H), 3.36-3.12 (m, 4H), 2.12 (s, 6H), 2.11 (s, 6H), 2.07 (s, 6H), 2.04 (s, 6H), 2.03 (s, 6H), 2.02 (s, 6H), 1.81 (s, 6H), 1.30-1.16 (m, 76H), 0.88 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):2038.10[M+H] +

[0351] 6. Preparation method of D-CuAAc

[0352] (Maltose, Cellobiose, Lactose, Melibiose, Glucose):

[0353]

[0354] 6.1 Preparation of D-Maltose-CuAAc-C10C10:

[0355] 3.50 g (2.00 mmol) of M-CuAAc-C10C10 was dissolved in 35 ml of MeOH, and 22 mg (0.40 mmol) of MeONa was added. The mixture was kept at 25-30 °C with stirring. TLC monitoring showed that the starting material was completely converted after 2 h of reaction. An appropriate amount of Amberlite® IRC120H resin was added to adjust the pH to neutral, and stirring was continued for 10 min. The mixture was filtered, and the mother liquor was concentrated to obtain a white, foamy colloid. The crude product was purified by flash reversed-phase C18 chromatography to obtain 2.07 g of powdered solid (molar yield 88.8%).

[0356] By following the above method and only changing the type of substrate, the following D-CuAAc series compounds can be obtained.

[0357] 6.2 Structural characterization of D-CuAAc:

[0358] Structural characterization of D-Maltose-CuAAc-C3C3-Cy (abbreviation: D-Ma-C3C3-Cy):

[0359]

[0360] White powdery solid, with a separation yield of 87.6%. 1 H NMR (400 MHz, Methanol-d4) δ8.14(s, 2H), 5.68 (d, J = 9.1 Hz, 2H), 5.27 (d, J = 3.8 Hz, 2H), 4.53 (s, 4H),4.02 (t, J = 9.1 Hz, 2H), 3.96-3.82 (m, 8H), 3.79 (d, J = 8.8 Hz, 2H), 3.75-3.62 (m, 8H), 3.49 (dd, J = 9.7, 3.7 Hz, 2H), 3.30 (d, J = 1.9 Hz, 2H), 3.28(d, J = 2.0 Hz, 4H), 1.78-1.53 ​​(m, 10H), 1.30-1.03 (m, 20H), 0.95-0.78 (m,4H). ESI-MS(m / z):1135.59[M+H] +

[0361] Structural characterization of D-Glucose-CuAAc-C4C4 (abbreviation: D-Gl-C4C4, n=2):

[0362]

[0363] White powdery solid, with a separation yield of 87.0%. 1 H NMR (400 MHz, Methanol-d4) δ 8.11(s, 2H), 5.65 (d, J = 9.0 Hz, 2H), 4.63 (s, 4H), 3.97–3.87 (m, 4H), 3.75 (dd,J = 12.0, 4.5 Hz, 2H,), 3.66–3.50 (m, 6H), 3.28 (d, J = 2.0 Hz, 4H), 1.33-1.14(m, 12H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):675.36[M+H]+

[0364] Structural characterization of D-Glucose-CuAAc-C6C6 (abbreviation: D-Gl-C6C6, n=4):

[0365]

[0366] White powdery solid, with a separation yield of 88.5%. 1 H NMR (400 MHz, Methanol-d4) δ 8.11(s, 2H), 5.65 (d, J = 9.0 Hz, 2H), 4.63 (s, 4H), 3.97–3.87 (m, 4H), 3.75 (dd,J = 12.0, 4.5 Hz, 2H,), 3.66–3.50 (m, 6H), 3.28 (d, J = 2.0 Hz, 4H), 1.33-1.14(m, 12H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):731.42[M+H] +

[0367] Structural characterization of D-Maltose-CuAAc-C6C6 (abbreviation: D-Ma-C6C6, n=4):

[0368]

[0369] White powdery solid, with a separation yield of 87.5%. 1 H NMR (400 MHz, Methanol-d4) δ8.14(s, 2H), 5.68 (d, J = 9.1 Hz, 2H), 5.27 (d, J = 3.8 Hz, 2H), 4.53 (s, 4H),4.02 (t, J = 9.1 Hz, 2H), 3.96-3.82 (m, 8H), 3.79 (d, J = 8.8 Hz, 2H), 3.75-3.62 (m, 8H), 3.49 (dd, J = 9.7, 3.7 Hz, 2H), 3.30 (d, J = 1.9 Hz, 2H), 3.28(d, J = 2.0 Hz, 4H), 1.33-1.14 (m, 20H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1055.52[M+H] +

[0370] Structural characterization of D-Cellobiose-C6C6 (abbreviation: D-Ce-C6C6, n=4):

[0371]

[0372] White powdery solid, with a separation yield of 87.1%. 1 H NMR (400 MHz, Methanol-d4) δ8.18(s, 2H), 5.80 (d, J = 9.3 Hz, 2H),4.59(d, J = 7.8 Hz, 2H), 4.53 (s, 4H),3.91-413 (m, 8H),3.78 (dd, J = 12.0, 6.0 Hz, 2H), 3.35-3.58 (m,10H), 3.28 (d,J = 2.0 Hz, 4H), 1.33-1.14 (m, 20H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1055.52[M+H] +

[0373] Structural characterization of D-Lactose-C6C6 (abbreviation: D-La-C6C6, n=4):

[0374]

[0375] White powdery solid, with a separation yield of 88.0%. 1 H NMR (400 MHz, Methanol-d4) δ7.85(s, 2H), 5.50 (d, J = 7.7 Hz, 2H), 5.33-5.27 (m, 2H), 4.53 (s, 4H), 3.90-3.68(m, 8H), 3.68-3.48 (m, 12H), 3.47-3.38 (m, 2H), 3.34-3.27 (m, 2H), 3.28 (d, J= 2.0 Hz, 4H), 1.33-1.14 (m, 20H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1055.52[M+H] +

[0376] Structural characterization of D-Melibiose-C6C6 (abbreviation: D-Me-C6C6, n=4):

[0377]

[0378] White powdery solid, with a separation yield of 89.0%. 1 H NMR (400 MHz, Methanol-d4) δ8.14(s, 2H), 5.68 (d, J = 9.1 Hz, 2H), 5.27 (d, J = 3.8 Hz, 2H), 4.53 (s, 4H),4.02 (t, J = 9.1 Hz, 2H), 3.96-3.82 (m, 8H), 3.79 (d, J = 8.8 Hz, 2H), 3.75-3.62 (m, 8H), 3.49 (dd, J = 9.7, 3.7 Hz, 2H), 3.30 (d, J = 1.9 Hz, 2H), 3.28(d, J = 2.0 Hz, 4H), 1.33-1.14 (m, 20H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1055.52[M+H] +

[0379] Structural characterization of D-Maltose-CuAAc-C7C7 (abbreviation: D-Ma-C7C7, n=5):

[0380]

[0381] White powdery solid, with a separation yield of 89.0%. 1 H NMR (400 MHz, Methanol-d4) δ8.13(s, 2H), 5.68 (d, J = 9.1 Hz, 2H), 5.27 (d, J = 3.9 Hz, 2H), 4.53 (s, 4H),4.02 (t, J = 9.1 Hz, 2H), 3.94-3.82 (m, 8H), 3.79 (d, J = 8.8 Hz, 2H), 3.75-3.62 (m, 8H), 3.49 (dd, J = 9.7, 3.8 Hz, 2H), 3.30 (s, 2H), 3.29-3.24 (m,4H), 1.33-1.17 (m, 24H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1083.56[M+H] +

[0382] Structural characterization of D-Maltose-CuAAc-C8C8 (abbreviation: D-Ma-C8C8, n=6):

[0383]

[0384] White powdery solid, with a separation yield of 89.7%. 1 H NMR (400 MHz, Methanol-d4) δ8.13 (s,2H), 5.67 (d, J = 9.2 Hz, 2H), 5.26 (d, J = 3.8 Hz, 2H), 4.53 (s, 4H), 4.02(t, J = 9.1 Hz, 2H), 3.86 (ddt, J = 11.3, 8.0, 5.3 Hz, 8H), 3.79 (d, J = 8.8Hz, 2H), 3.75-3.62 (m, 8H), 3.49 (dd, J = 9.7, 3.8 Hz, 2H), 3.29 (d, J = 2.2Hz, 2H), 3.29-3.23 (m, 4H), 1.33-1.17 (m, 28H), 0.89 (t, J = 6.8 Hz, 6H).ESI-MS(m / z):1111.59[M+H] +

[0385] Structural characterization of D-Cellobiose-CuAAc-C8C8 (abbreviation: D-Ce-C8C8, n=6):

[0386]

[0387] White powdery solid, with a separation yield of 87.2%. 1 H NMR (400 MHz, Methanol-d4) δ8.18 (s,2H), 5.80 (d, J = 9.3 Hz, 2H),4.59(d, J = 7.8 Hz, 2H), 4.53 (s, 4H), 3.91-413(m, 8H),3.78 (dd, J = 12.0, 6.0 Hz, 2H), 3.35-3.58 (m,10H), 3.28 (d, J = 2.0Hz, 4H), 1.33-1.14 (m, 28H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1111.59[M+H] +

[0388] Structural characterization of D-Lactose-CuAAc-C8C8 (abbreviation: D-La-C8C8, n=6):

[0389]

[0390] White powdery solid, with a separation yield of 88.3%. 1 H NMR (400 MHz, Methanol-d4) δ7.85 (s,2H), 5.50 (d, J = 7.7 Hz, 2H), 5.33-5.27 (m, 2H), 4.53 (s, 4H), 3.90-3.68 (m,8H), 3.68-3.48 (m, 12H), 3.47-3.38 (m, 2H), 3.34-3.27 (m, 2H), 3.28 (d, J =2.0 Hz, 4H), 1.33-1.14 (m, 28H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1111.59[M+H] +

[0391] Structural characterization of D-Melibiose-CuAAc-C8C8 (abbreviation: D-Me-C8C8, n=6):

[0392]

[0393] White powdery solid, with a separation yield of 89.1%. 1 H NMR (400 MHz, Methanol-d4) δ8.14(s, 2H), 5.65 (d, J = 8.9 Hz, 2H), 5.40-5.35 (m, 2H), , 4.53 (s, 4H), 4.02 (m2H), 3.96-3.82 (m, 8H), 3.79-3.49 (m, 12H), 3.35-3.30 (m, 2H), 3.29 (d, J =2.0 Hz, 4H), 1.33-1.14 (m, 28H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1111.59[M+H] +

[0394] Structural characterization of D-Maltose-CuAAc-C9C9 (abbreviation: D-Ma-C9C9, n=7):

[0395]

[0396] White powdery solid, with a separation yield of 89.5%. 1H NMR (400 MHz, Methanol-d4) δ8.13 (s,2H), 5.67 (d, J = 9.1 Hz, 2H), 5.26 (d, J = 3.8 Hz, 2H), 4.53 (s, 4H), 4.02(t, J = 9.1 Hz, 2H), 3.87 (tdd, J = 13.2, 6.8, 2.1 Hz, 8H), 3.79 (d, J = 8.8Hz, 2H), 3.75-3.62 (m, 8H), 3.49 (dd, J = 9.7, 3.8 Hz, 2H), 3.29 (s, 2H), 3.27 (d, J = 2.0 Hz, 4H), 1.32-1.20 (m, 32H), 0.90 (t, J = 6.6 Hz, 6H). ESI-MS(m / z):1139.61[M+H] +

[0397] Structural characterization of D-Maltose-CuAAc-C10C10 (abbreviation: D-Ma-C10C10, n=8):

[0398]

[0399] White powdery solid, with a separation yield of 88.8%. 1 H NMR (400 MHz, Methanol-d4) δ8.14(s, 2H), 5.68 (d, J = 9.1 Hz, 2H), 5.26 (d, J = 3.8 Hz, 2H), 4.54 (s, 4H),4.02 (t, J = 9.1 Hz, 2H), 3.95-3.82 (m, 8H), 3.79 (d, J = 8.8 Hz, 2H), 3.75-3.62 (m, 8H), 3.49 (dd, J = 9.7, 3.8 Hz, 2H), 3.29 (s, 2H), 3.29-3.24 (m,4H), 1.30-1.20 (m, 36H), 0.92-0.87 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):1167.65[M+H] +

[0400] Structural characterization of D-Cellobiose-CuAAc-C10C10 (abbreviation: D-Ce-C10C10, n=8):

[0401]

[0402] White powdery solid, with a separation yield of 87.2%. 1 H NMR (400 MHz, Methanol-d4) δ8.18 (s,2H), 5.80 (d, J = 9.3 Hz, 2H),4.59(d, J = 7.8 Hz, 2H), 4.53 (s, 4H), 3.91-413(m, 8H),3.78 (dd, J = 12.0, 6.0 Hz, 2H), 3.35-3.58 (m,10H), 3.28 (d, J = 2.0Hz, 4H), 1.33-1.14 (m, 36H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1167.65[M+H] +

[0403] Structural characterization of D-Lactose-CuAAc-C10C10 (abbreviation: D-La-C10C10, n=8):

[0404]

[0405] White powdery solid, with a separation yield of 88.3%. 1 H NMR (400 MHz, Methanol-d4) δ7.85 (s,2H), 5.50 (d, J = 7.7 Hz, 2H), 5.33-5.27 (m, 2H), 4.53 (s, 4H), 3.90-3.68 (m,8H), 3.68-3.48 (m, 12H), 3.47-3.38 (m, 2H), 3.34-3.27 (m, 2H), 3.28 (d, J =2.0 Hz, 4H), 1.33-1.14 (m, 36H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1167.65[M+H] +

[0406] Structural characterization of D-Melibiose-CuAAc-C10C10 (abbreviation: D-Me-C10C10, n=8):

[0407]

[0408] White powdery solid, with a separation yield of 89.1%. 1H NMR (400 MHz, Methanol-d4) δ8.14 (s,2H), 5.65 (d, J = 8.9 Hz, 2H), 5.40-5.35 (m, 2H), , 4.53 (s, 4H), 4.02 (m2H), 3.96-3.82 (m, 8H), 3.79-3.49 (m, 12H), 3.35-3.30 (m, 2H), 3.29 (d, J =2.0 Hz, 4H), 1.33-1.14 (m, 36H), 0.89 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1167.65[M+H] +

[0409] Structural characterization of D-Maltose-CuAAc-C12C12 (abbreviation: D-Ma-C12C12, n=10):

[0410]

[0411] White powdery solid, with a separation yield of 88.9%. 1 H NMR (400 MHz, Methanol-d4) δ8.14 (s,2H), 5.68 (d, J = 9.2 Hz, 2H), 5.26 (d, J = 3.8 Hz, 2H), 4.53 (s, 4H), 4.02(t, J = 9.1 Hz, 2H), 3.87 (tdd, J = 11.4, 7.3, 2.1 Hz, 8H), 3.79 (d, J = 8.8Hz, 2H), 3.76-3.62 (m, 8H), 3.49 (dd, J = 9.7, 3.7 Hz, 2H), 3.29 (s, 2H), 3.29-3.20 (m, 4H), 1.31-1.20 (m, 44H), 0.95-0.86 (t, J = 6.8 Hz, 6H). ESI-MS(m / z):1222.71[M+H] +

[0412] Structural characterization of D-Maltose-CuAAc-C14C14 (abbreviation: D-Ma-C14C14, n=12):

[0413]

[0414] White powdery solid, with a separation yield of 89.5%. 1H NMR (400 MHz, Methanol-d4) δ8.13(s, 2H), 5.67 (d, J = 9.0 Hz, 2H), 5.26 (d, J = 3.8 Hz, 2H), 4.53 (s, 4H),4.02 (t, J = 9.1 Hz, 2H), 3.95-3.82 (m, 8H), 3.79 (d, J = 8.8 Hz, 2H), 3.75-3.61 (m, 8H), 3.49 (dd, J = 9.7, 3.7 Hz, 2H), 3.29 (s, 2H), 3.28-3.24 (m,4H), 1.31-1.20 (m, 52H), 0.94-0.85 (m, 6H). ESI-MS(m / z):1279.77[M+H] +

[0415] Structural characterization of D-Maltose-CuAAc-C16C16 (abbreviation: D-Ma-C16C16, n=14):

[0416]

[0417] White powdery solid, with a separation yield of 89.8%. 1 H NMR (400 MHz, DMSO-d6) δ8.24 (s,2H), 5.48 (d, J = 6.0 Hz, 2H), 5.07 (d, J = 3.8 Hz, 2H), 4.47 (s, 4H), 3.86(td, J = 9.1, 5.3 Hz, 2H), 3.70 (q, J = 10.5, 9.5 Hz, 4H), 3.62 (dd, J =13.8, 9.0 Hz, 6H), 3.50 (td, J = 12.4, 10.7, 6.3 Hz, 6H), 3.40 (d, J = 9.2Hz, 2H), 3.31-3.28 (m, 2H), 3.24 (s, 4H), 3.09 (t, J = 9.2 Hz, 2H), 1.37-1.14(m, 60H), 0.85 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1335.83[M+H] +

[0418] Structural characterization of D-Maltose-CuAAc-C20C20 (abbreviation: D-Ma-C20C20, n=18):

[0419]

[0420] White powdery solid, with a separation yield of 85.1%. 1 H NMR (400 MHz, DMSO-d6) δ8.24 (s,2H), 5.48 (d, J = 6.0 Hz, 2H), 5.07 (d, J = 3.8 Hz, 2H), 4.47 (s, 4H), 3.86(td, J = 9.1, 5.3 Hz, 2H), 3.70 (q, J = 10.5, 9.5 Hz, 4H), 3.62 (dd, J =13.8, 9.0 Hz, 6H), 3.50 (td, J = 12.4, 10.7, 6.3 Hz, 6H), 3.40 (d, J = 9.2Hz, 2H), 3.31-3.28 (m, 2H), 3.24 (s, 4H), 3.09 (t, J = 9.2 Hz, 2H), 1.37-1.14(m, 76H), 0.85 (t, J = 6.7 Hz, 6H). ESI-MS(m / z):1448.95[M+H] +

[0421] Example 2: Evaluation of the micelle properties formed by the detergents (i.e., D-Ma, D-Gl, D-Ce, D-La, D-Me) in Example 1:

[0422] 1. CMC (Critical Micelle Concentration) Determination:

[0423] The small molecule detergent was dissolved in ultrapure water to prepare a 1.0 mM stock solution, which was then serially diluted with ultrapure water to different concentrations. 5 μL of acetonitrile stock solution (0.12 mM) containing pyrene was added to 1 ml of each detergent solution to ensure that each sample contained 0.6 μM pyrene. The samples were incubated overnight at 37°C with shaking. Subsequently, the above solutions were pipetted into 96-well plates, 100 μL per well. Using a microplate reader, the excitation wavelength was fixed at 335 nm, and the fluorescence values ​​at emission wavelengths of 371 and 382 nm were measured. Three sets of data were measured in parallel. The I371 / I382 versus lgC curve was fitted using Oringin software. The inflection point of the fluorescence value change with concentration corresponds to the concentration of the detergent. The CMC measurement results are shown in the table below (unit: μM).

[0424] 2. Dynamic light scattering experiment:

[0425] The detergent solid was dissolved in ultrapure water to prepare a solution, with a concentration of 1.0 mM for detergent solutions with a CMC ≥ 1 μM and a concentration of 0.1 mM for detergent solutions with a CMC < 1 μM. The micelle diameter was measured using a dynamic light scattering instrument (Malvin Zetasizer Pro). The results of the micelle diameter measurements are shown in the table below (D...). h (Unit: nm)

[0426] 3. Experimental Results (-Ma: maltose; -Ce: cellobiose; -La: lactose; -Me: melibiose; -Gl: glucose)

[0427] Positive controls: DDM (dodecyl-β-D-maltodextrin), LMNG (neopentyl lauryl maltose).

[0428] HLB refers to the hydrophilic-lipophilic balance value, and Water Sol refers to the solubility in water.

[0429]

[0430] Example 3: Membrane protein thermal stability experiments were conducted on some of the detergents characterized in Example 2:

[0431] Initial melting temperature (T) onest (): This refers to the temperature at which proteins begin to unfold;

[0432] Half-chain melting temperature (T) m This refers to the temperature at which 50% of the protein unfolds.

[0433] Protein aggregation initiation temperature (T) agg This refers to the initial temperature at which proteins begin to exhibit a tendency to aggregate.

[0434] T onest and T m The higher the value, the stronger the conformational stability of the detergent for membrane proteins; T agg The higher the value, the stronger the colloidal stability of the detergent on membrane proteins.

[0435] 1. Thermostability experiment of A2a receptor:

[0436] Positive control 1 was DDM (dodecyl-β-D-maltodextrin), and positive control 2 was LMNG (neopentyl lauryl maltose glycol).

[0437] Reagents: Membrane protein (A2a receptor, supplier Acro), membrane protein ligand (ZM241385, theophylline), detergent buffer (20mM HEPES, pH 7.4, 500mM NaCl, 2% glycerol)

[0438] Methods: Weigh an appropriate amount of detergent and prepare a 2% (w / v) detergent solution (50 μL) using detergent buffer; add membrane protein ligands (ligand concentration: ZM241385: 40 μM, theophylline: 1 mM), and incubate on ice for 10 min; add an appropriate amount of purified A2a receptor to a final mass of 1 μg, and incubate on ice for 10 min; transfer the sample to a multifunctional protein stability analyzer (NanoTemper PR Panta) and determine its thermal stability parameters (Tonest value, Tm value, and Tagg value). Each sample is measured in triplicate, and the average value is taken.

[0439] The results of the thermal stability parameters of the A2a receptor in various detergents are shown in the table below (unit: °C):

[0440]

[0441]

[0442] 2. Thermostability experiment of GLP-1 receptor membrane protein:

[0443] Positive control 1 was DDM (dodecyl-β-D-maltodextrin), and positive control 2 was LMNG (neopentyl lauryl maltose glycol).

[0444] Reagents: Membrane protein (GLP-1 receptor, supplier Acro), detergent buffer (20mM HEPES, pH 7.4, 500mM NaCl, 2% glycerol)

[0445] Methods: Weigh an appropriate amount of detergent and prepare a 2% (w / v) detergent solution (50 μL) using detergent buffer; incubate on ice for 10 min; add an appropriate amount of purified GLP-1 receptor to a final mass of 1 μg, and incubate on ice for 10 min; transfer the sample to a multifunctional protein stability analyzer (NanoTemper PR Panta), and determine its thermal stability parameters (Tonest value, Tm value, and Tagg value) using Thermo unfolding mode. Each sample was measured in triplicate, and the average value was taken.

[0446] The results of the thermal stability parameters of GLP-1 receptors in various detergents are shown in the table below (unit: °C):

[0447]

[0448] In the above embodiments, most of the detergents showed better results than the control detergent DDM in the thermal stability experiments of A2a receptor and GLP-1 receptor, and some detergents even outperformed LMNG, which is considered to have excellent performance in the stability of a variety of membrane proteins. This phenomenon was particularly significant in DTG-0102, DTG-0103, DTG-0104, DTG-0302, and DTG-0402.

[0449] Example 4: The particle size distribution of membrane protein-detergent complex (PDC) was tested on some of the detergents characterized in Example 2:

[0450] The particle size variation of PDCs can be tested using DLS technology to characterize the aggregation trend of PDCs; the smaller the aggregation trend of PDCs (particle size distribution shifts to the right, and the particle size increases), the stronger the stabilizing effect of the detergent on membrane proteins.

[0451] Positive control 1 was DDM (dodecyl-β-D-maltodextrin), and positive control 2 was LMNG (neopentyl lauryl maltose glycol).

[0452] Reagents: Membrane protein (A2a receptor, supplier Acro), membrane protein ligand (ZM241385), detergent buffer (20mM HEPES, pH 7.4, 500mM NaCl, 2% glycerol)

[0453] Methods: Weigh an appropriate amount of detergent and prepare a 2% (w / v) detergent solution (50 μL) using detergent buffer; add membrane protein ligand ZM241385 (ligand concentration: 40 μm) and incubate on ice for 10 min; add an appropriate amount of purified A2a receptor to a final mass of 1 μg and incubate on ice for 10 min; transfer the sample to a multifunctional protein stability analyzer (NanoTemper PR Panta) and use DLS technology in Isothermo Scan mode, maintaining a temperature of 45℃ and a holding time of 120 min. The particle size distribution of the membrane protein-detergent complex (PDC) was tested at holding times of 1 min and 120 min, respectively. Particle size distribution curves were plotted using Origin software. Results are shown in [Figure number missing]. Figure 1-6 .

[0454] In the above embodiments, most of the detergents outperformed the control detergent DDM in particle size distribution variation experiments, and some even outperformed LMNG, which is considered to have excellent performance in maintaining the stability of various membrane proteins. This phenomenon was particularly pronounced in DTG-0101, DTG-0102, and DTG-0103.

Claims

1. A compound I, ； in, R 3 for Or H; A 1 and A 2 The monosaccharide is independently a monosaccharide glycosyl or oligosaccharide glycosyl; the monosaccharide is a furanose or pyranose of a pentose, or a furanose or pyranose of a hexose; the oligosaccharide is obtained by dehydration polymerization of 2-10 of the monosaccharides; L 1 L 2 L 3 and L 4 Independently for C 1-6 Alkylene; R 1 C 1-25 Hydrophobic aliphatic groups or H; R 2 C 1-25 Hydrophobic aliphatic groups.

2. Compound I as claimed in claim 1, characterized in that, It meets one or more of the following conditions: (1) A 1 and A 2 In this context, the pentose is an aldose; (2) A 1 and A 2 In this context, the hexose is a ketose or an aldose, preferably an aldose; (3) A 1 and A 2 In this context, the monosaccharide is a furanose or pyranose of a hexose, preferably a pyranose of a hexose; (4) A 1 and A 2 In this context, the furanose of the hexose is the furanose of D-glucose; (5) A 1 and A 2 In this context, the pyranose of the hexose is either the pyranose of glucose or the pyranose of galactose; (6) A 1 and A 2 In this process, the oligosaccharide is obtained by dehydration polymerization of 2-5 of the monosaccharides, preferably by dehydration polymerization of 2 of the monosaccharides; (7) A 1 and A 2 In the oligosaccharide, the "monosaccharide dehydration polymerization" is the "dehydration polymerization" of a hemiacetal or hemiketal hydroxyl group of a monosaccharide with any hydroxyl group of another monosaccharide. (8) R 1 and R 2 In, the C 1-25 The hydrophobic aliphatic group is independently C 1-25 Alkyl, C 1-25 Heteroalkyl or -C 1-20 Alkylene-C 3-8 cycloalkyl; The C 1-25 The heteroalkyl group contains 1-5 heteroatoms, wherein each heteroatom is independently O or S; The C 3-8 Cycloalkyl groups are preferably C 4-6 Cycloalkyl groups, such as cyclohexyl; (9) R 1 C 1-25 Hydrophobic aliphatic groups; (10) R 3 for .

3. Compound I as described in claim 2, characterized in that, It meets one or more of the following conditions: (1) A 1 and A 2 In the context of the hexose, when the hexose is a ketose, the ketose is allulose, fructose, sorbitol, or tagatose; (2) A 1 and A 2 In the context of the hexose being aldose, the aldose is allose, acetose, glucose, mannose, gulose, idose, galactose, or tarose. (3) A 1 and A 2 In the case where the furanose of the hexose is the furanose of D-glucose, the furanose of the D-glucose is α-D-furanose or β-D-furanose; (4) A 1 and A 2 In the case where the pyranose of the hexose is the pyranose of glucose, the pyranose of glucose is the pyranose of D-glucose, such as α-D-glucose pyranose or β-D-glucose pyranose; (5) A 1 and A 2 In the case where the pyranose of the hexose is a pyranose of galactose, the pyranose of the galactose is a pyranose of D-galactose, such as α-D-galactopyranose or β-D-galactopyranose; (6) A 1 and A 2 In the oligosaccharide, the "monosaccharide dehydration polymerization" is the "dehydration polymerization" of a hemiacetal or hemiketal hydroxyl group of one monosaccharide with a hydroxyl group at the 4 or 6 position of another monosaccharide. (7) R 1 and R 2 In, the C 1-25 Heteroalkyl , or ; (8) R 1 and R 2 In the context, the -C 1-20 Alkylene-C 3-8 Cycloalkyl group is -C 1-10 Alkylene-C 3-8 cycloalkyl, preferably -C 1-6 Alkylene-C 3-8 cycloalkyl, for example ; (9) R 1 and R 2 In, the C 1-25 Alkyl group is C 4-20 Alkyl, for example , , , , , , , , or ; The C 1-25 Alkyl groups are preferably C 5-10 Alkyl, for example , , or ; (10) L 1 It is methylene; (11) L 2 It is methylene; (12) L 3 It is methylene; (13) L 4 It is methylene; (14) R 1 and R 2 Independently for 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. Compound I as described in claim 3, characterized in that, It meets one or more of the following conditions: (1) A 1 and A 2 In this context, when the hexose is an aldose, the aldose is glucose or galactose; the glucose is preferably D-glucose; the galactose is preferably D-galactose. (2) A 1 and A 2 In the oligosaccharide, the "monosaccharide dehydration polymerization" is the "dehydration polymerization" of the α-hydroxyl or β-hydroxyl group of the hemiacetal of one monosaccharide with the 4- or 6-hydroxyl group of another monosaccharide.

5. Compound I as claimed in claim 1, characterized in that, It meets one or more of the following conditions: (1) A 1 and A 2 In the text, the monosaccharide is... ; (2) A 1 and A 2 In this context, the monosaccharide glycosyl group is... ; (3) A 1 and A 2 In this context, the oligosaccharide is cellobiose, maltose, lactose, melibiose, or sucrose, preferably maltose, cellobiose, or lactose, and more preferably maltose; The maltose is preferably α-maltose or β-maltose; the β-maltose is preferably β-D-maltose. The cellobiose is preferably α-cellobiose or β-cellobiose; the β-cellobiose is preferably β-D-cellobiose. The lactose is preferably α-lactose or β-lactose; the β-lactose is preferably β-D-lactose. The melibiose is preferably α-melibiose or β-melibiose; β-melibiose is preferably β-D-melibiose. (4) A 1 and A 2 In this context, the oligosaccharide glycosyl group is... , , or ; (5) A 1 and A 2 Independently , , , , or ; (6) R 1 and R 2 Independently , , , , , , , , , or .

6. Compound I as claimed in claim 1, characterized in that, Compound I satisfies any of the following schemes: Option (1) A 1 and A 2 In this process, the oligosaccharide is obtained by dehydration polymerization of 2-10 monosaccharides; the monosaccharide is a furanose or pyranose of a hexose, preferably a pyranose of a hexose; the oligosaccharide is preferably obtained by dehydration polymerization of 2-5 monosaccharides. The pyranose of the hexose is preferably D-glucose pyranose or D-galactopyranose; The D-glucose pyranose is preferably α-D-glucose pyranose or β-D-glucose pyranose; The D-galactopyranose is preferably α-D-galactopyranose or β-D-galactopyranose; In the oligosaccharide, the linkage between each monosaccharide is preferably an α-1,4 glycosidic bond, a β-1,4 glycosidic bond, or an α-1,6 glycosidic bond. The structure of compound I in scheme (2) is as follows: ， Among them, A 1 A 2 L 1 L 2 L 3 L 4 R 1 and R 2 The definition is as described in any one of claims 1-5; The structure of compound I in scheme (3) is as follows: ； Among them, A 1 and A 2 The oligosaccharide is independently a monosaccharide glycosyl or an oligosaccharide glycosyl; the monosaccharide is a pyranose of a hexose; the oligosaccharide is obtained by dehydration polymerization of 2-10 of the monosaccharides; preferably, the oligosaccharide is obtained by dehydration polymerization of 2-5 of the monosaccharides. In the oligosaccharide, the linkage between each monosaccharide is preferably an α-1,4 glycosidic bond, a β-1,4 glycosidic bond, or an α-1,6 glycosidic bond; L 1 L 2 L 3 and L 4 Independently for C 1-3 Alkylene; R 1 C 4-20 alkyl; R 2 C 4-20 alkyl; The structure of compound I in scheme (4) is as follows: ； Among them, A 1 and A 2 Independently, it is an oligosaccharide glycosyl group; the oligosaccharide is obtained by dehydration polymerization of 2-5 monosaccharides; the monosaccharide is a pyranose of a hexose, such as α-D-glucopyranose, β-D-glucopyranose, α-D-galactopyranose or β-D-galactopyranose; In the oligosaccharide, the linkage between each monosaccharide is preferably an α-1,4 glycosidic bond or a β-1,4 glycosidic bond; L 1 L 2 L 3 and L 4 Independently for C 1-3 Alkyl groups, such as methylene groups; R 1 C 5-10 alkyl; R 2 C 5-10 alkyl; Preferably, the oligosaccharide is maltose, cellobiose, or lactose, such as β-D-maltose, β-D-cellobiose, or β-D-lactose.

7. Compound I as claimed in claim 1, characterized in that, Compound I is any of the following compounds: , , , , , , , , , , , , , , , , , , or .

8. Use of compound I as claimed in any one of claims 1-7 in the preparation of a detergent formulation, or as a detergent.

9. The use as described in claim 8, characterized in that, It meets one or more of the following conditions: (1) The detergent preparation or the detergent is used to stabilize membrane proteins, or to improve the stability of membrane proteins, or to improve the thermal stability of membrane proteins; the membrane protein is preferably a G protein-coupled receptor or a GLP-1 receptor; the G protein-coupled receptor is preferably an A2a receptor; (2) The detergent or the detergent works by the following steps: causing the biomembrane to disintegrate and release membrane proteins, and providing a hydrophobic environment for the membrane proteins in the demembrane state in the solution, thereby extracting the membrane proteins on the biomembrane into the solution; The solution is preferably an aqueous solution; the membrane protein is preferably a G protein-coupled receptor, such as the A2a receptor; (3) The detergent formulation or the detergent forms micelles in the solution, wherein the diameter of the micelles is preferably 4-50 nm, for example 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; the micelles are preferably amphiphilic micelles; the solution is preferably an aqueous solution.

10. A method for preparing compound I according to any one of claims 1-7, comprising the following steps: (1) In a solvent, dimethyl malonate, base and monohalogenated product with hydrophobic aliphatic group are subjected to alkylation reaction. After the reaction is completed, the first intermediate is separated. The first intermediate refers to the intermediate compound obtained by di-substitution of the active methylene group of dimethyl malonate with hydrophobic aliphatic group. (2) In a solvent, the first intermediate and the reducing agent are reduced to obtain the second intermediate; the substance obtained in this step is the intermediate compound obtained after both ester groups on the first intermediate are reduced to primary alcohols. (3) In a solvent, the second intermediate, the base and 3-bromopropyne are subjected to an etherification reaction. After the reaction is completed, a hydrophobic fragment is separated. The hydrophobic fragment refers to the third intermediate compound obtained after both primary hydroxyl groups of the second intermediate are etherified. ； (4) In a solvent, a monosaccharide or oligosaccharide with hydroxyl protected, an azide reagent and a Lewis acid are subjected to a terminal azide reaction. After the reaction is completed, a hydrophilic fragment is separated. The hydrophilic fragment refers to the fourth intermediate compound obtained after the terminal position of the monosaccharide or oligosaccharide with hydroxyl protected is replaced by an azide group. ； (5) In a solvent, the third intermediate, the fourth intermediate, sodium ascorbate and copper sulfate are subjected to a Click reaction. After the reaction is completed, the fifth intermediate is separated. The fifth intermediate obtained in this step is an intermediate compound obtained by linking the hydrophobic fragment and the hydrophilic fragment through a triazole group using a Click reaction. (6) The fifth intermediate and sodium methoxide are subjected to a methanololysis reaction in a solvent. After the reaction is completed, compound I is separated. 。 11. A compound II having any of the following structures: , , , , , , , , , , , , , , , , , , or .

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