CN110563698A

CN110563698A - Novel protein cross-linking agent and preparation method thereof

Info

Publication number: CN110563698A
Application number: CN201910673959.4A
Authority: CN
Inventors: 聂承斌; 李宏林; 陆雪根
Original assignee: Suzhou Hao Fan Biological Ltd By Share Ltd
Current assignee: Suzhou Hao Fan Biological Ltd By Share Ltd
Priority date: 2019-07-25
Filing date: 2019-07-25
Publication date: 2019-12-13

Abstract

the invention discloses a preparation method of a novel protein cross-linking agent, which comprises the following steps; (a) adding Sulfo-NHS shown in the formula (I) into a first reaction solvent, and carrying out condensation reaction with 3- (2-pyridine dithio) propionic acid shown in the formula (II) under the action of a condensing agent and a catalyst to obtain Sulfo-SPDP shown in the formula (III). The invention takes Sulfo-NHS and 3- (2-pyridine dithio) propionic acid as raw materials, and the compound Sulfo-SPDP is obtained through condensation reaction, the synthetic route is short, the yield can reach 60 percent, the synthetic method has the characteristics of simple and convenient operation, easily controlled conditions, wide material source, high production efficiency and obvious production advantages.

Description

Novel protein cross-linking agent and preparation method thereof

Technical Field

The invention relates to the technical field of compound preparation, in particular to a novel protein cross-linking agent and a preparation method thereof.

background

N-hydroxysuccinimide-3- (2-pyridyl dithio) -propionate (SPDP) (IV) is a heterotype bifunctional protein cross-linking agent, wherein a 2-pyridyl dithio group in the structure can react with fatty thiol and can also perform exchange reaction with a thiol group in a protein structure so as to realize cross-linking with protein, an activated ester component at the other end can perform chemical reaction with amino groups and the like in chemical drugs, and finally, the bridge coupling of the protein and the chemical drugs is obtained, so that the drugs have a targeting effect and are key reagents of antibody coupling (ADC) drugs.

Because the reaction of protein participation is generally carried out in water or polar solvent, the water-soluble cross-linking agent can participate in the reaction better, thereby realizing the bridging coupling of the protein and the chemical drug, and the development of the water-soluble protein cross-linking agent has wide application prospect. The invention provides a synthetic method of a water-soluble cross-linking agent Sulfo-SPDP, wherein Sulfo-SPDP (III) is a protein cross-linking agent with good water solubility, and the related structural formula is shown as follows.

Disclosure of Invention

The present invention has been made to solve the above-mentioned problems occurring in the prior art, and an object of the present invention is to provide a simple and efficient method for synthesizing a novel protein crosslinking agent.

in order to achieve the purpose, the invention provides the following technical scheme: a novel protein crosslinking agent having the structure of formula (III):

a preparation method of a novel protein cross-linking agent comprises the following steps;

(a) Adding Sulfo-NHS shown in the formula (I) into a first reaction solvent, and carrying out condensation reaction with 3- (2-pyridine dithio) propionic acid shown in the formula (II) under the action of a condensing agent and a catalyst to obtain Sulfo-SPDP shown in the formula (III):

Preferably, in the step (a), the molar weight ratio of the Sulfo-NHS, the 3- (2-pyridyldithio) propionic acid, the condensing agent and the catalyst is 1:1 to 1.5:1 to 2:0.1 to 1, and the molar weight ratio is preferably 1:1.1:1.2: 0.1.

Preferably, in the step (a), the condensing agent is any one or more of EDCI, DCC, and DIC, and the condensing agent is preferably DIC.

Preferably, in step (a), the catalyst is any one or more of tetrabutylammonium chloride, benzyltriethylammonium chloride and tetrabutylammonium bromide, and the catalyst is preferably benzyltriethylammonium chloride.

Preferably, in the step (a), the condensation reaction temperature is 0-25 ℃, the condensation reaction time is 10-24 hours, the reaction temperature is preferably 25 ℃, and the reaction time is 17 hours.

preferably, in step (a), the first reaction solvent is: tetrahydrofuran, N-dimethylformamide and N, N-dimethylacetamide, and the first reaction solvent is preferably N, N-dimethylformamide.

preferably, in the step (a), after the condensation reaction is completed, the method may further comprise the following steps: evaporating the solvent, pulping and purifying, wherein the solvent used in pulping comprises one or more of ethyl acetate, acetone and methanol.

The invention provides a novel protein cross-linking agent and a preparation method thereof, and the novel protein cross-linking agent has the beneficial effects that:

1. The invention takes Sulfo-NHS and 3- (2-pyridine dithio) propionic acid as raw materials, and the compound Sulfo-SPDP is obtained through condensation reaction, the synthetic route is simple and short, and the yield can reach 60%;

2. The synthesis method has the characteristics of simple and convenient operation, easily controlled conditions, wide material sources, high production efficiency and obvious production advantages.

Detailed Description

The present invention will be described in further detail with reference to the following specific examples. The procedures, conditions, experimental methods and the like for carrying out the present invention are general knowledge and common general knowledge in the art except for the contents specifically mentioned below, and the present invention is not particularly limited.

Example 1

Dissolving 0.46g of Sulfo-NHS and 0.5g of 3- (2-pyridyldithio) propionic acid in 7mLN, N-dimethylformamide in a single-neck flask, stirring for a while, adding 0.35g of DIC, reacting at room temperature for 24 hours, detecting that the raw materials are completely reacted by TLC, evaporating to dryness of N, N-dimethylformamide, pulping with ethyl acetate once, and pulping with acetone twice to obtain 0.42g of Sulfo-SPDP (III), wherein the yield is 48%;

1H NMR(500MHz,DMSO-d6)δ8.51-8.49(m,1H),7.68-7.64(m,2H),7.15-7.12(m,1H),4.30-4.27(m,1H),3.33-3.25(m,2H),3.18–3.14(m,2H),3.10–3.07(m,2H)。

Example 2

Dissolving 0.46g of Sulfo-NHS, 0.5g of 3- (2-pyridyldithio) propionic acid and 0.05g of benzyltriethylammonium chloride in 7mL of N, N-dimethylformamide in a single-mouth bottle, stirring for a moment, adding 0.35g of DIC, reacting at room temperature for 17 hours, detecting the reaction completion of raw materials by TLC, evaporating N, N-dimethylformamide to dryness, pulping with ethyl acetate once, and pulping with acetone twice to obtain 0.53g of Sulfo-SPDP (III), wherein the yield is 60%;

Example 3

Dissolving 0.46g of Sulfo-NHS, 0.5g of 3- (2-pyridyldithio) propionic acid and 0.05g of benzyltriethylammonium chloride in 7mL of N, N-dimethylformamide in a single-mouth bottle, stirring for a moment, adding 0.52g of DCC, reacting at room temperature for 17 hours, detecting the reaction completion of raw materials by TLC, evaporating N, N-dimethylformamide to dryness, pulping with ethyl acetate once, and beating with acetone twice to obtain 0.31g of Sulfo-SPDP (III), wherein the yield is 35%;

In conclusion, the method for preparing the novel protein cross-linking agent has the advantages of short synthetic route, simple and convenient operation, low cost and high yield.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims

1. A novel protein crosslinking agent characterized by: the structure is shown as the structural formula (III):

2. A method for preparing a novel protein cross-linking agent is characterized by comprising the following steps: comprises the following steps;

(a) Adding Sulfo-NHS shown in the formula (I) into a first reaction solvent, and carrying out condensation reaction with 3- (2-pyridine dithio) propionic acid shown in the formula (II) under the action of a condensing agent and a catalyst to obtain a compound shown in the formula (III)

Sulfo-SPDP, the reaction sequence is shown in scheme (1):

3. the method for preparing a novel protein crosslinking agent according to claim 2, wherein: in the step (a), the molar ratio of the Sulfo-NHS, the 3- (2-pyridyldithio) propionic acid, the condensing agent and the catalyst is 1: 1-1.5: 1-2: 0.1-1.

4. The method for preparing a novel protein crosslinking agent according to claim 2, wherein: in the step (a), the condensing agent is any one or more of EDCI, DCC and DIC.

5. The method for preparing a novel protein crosslinking agent according to claim 2, wherein: in the step (a), the catalyst is one or more of tetrabutylammonium chloride, benzyltriethylammonium chloride and tetrabutylammonium bromide.

6. the method for preparing a novel protein crosslinking agent according to claim 2, wherein: in the step (a), the condensation reaction temperature is 0-25 ℃, and the condensation reaction time is 10-24 hours.

7. the method for preparing a novel protein crosslinking agent according to claim 2, wherein: in step (a), the first reaction solvent is: any one or more of tetrahydrofuran, N-dimethylformamide and N, N-dimethylacetamide.

8. The method for preparing a novel protein crosslinking agent according to claim 2, wherein: in the step (a), after the condensation reaction is completed, the method may further include the steps of: evaporating the solvent, pulping and purifying, wherein the solvent used in pulping comprises one or more of ethyl acetate, acetone and methanol.