CN107827762B

CN107827762B - (2, 3-dioleoyl-propyl) trimethyl ammonium chloride and preparation method and application thereof

Info

Publication number: CN107827762B
Application number: CN201711121867.2A
Authority: CN
Inventors: 吉民; 刘海东; 宗玺; 李锐; 万广朋; 王冬冬; 杨苏; 于文渊; 张影; 胡海燕
Original assignee: Southeast Pharmaceuticals Co ltd
Current assignee: Southeast Pharmaceuticals Co ltd
Priority date: 2017-11-14
Filing date: 2017-11-14
Publication date: 2020-10-02
Anticipated expiration: 2037-11-14
Also published as: CN107827762A

Abstract

The invention discloses a preparation method of (2, 3-dioleoyl-propyl) trimethyl ammonium chloride, which adopts the following method to prepare: firstly, 3-dimethylamino-1, 2-propanediol reacts with oleic acid or oleoyl chloride to obtain 1, 2-dioleoyl-3-dimethylamino-propane, and then reacts with methyl chloride under 0.1-10 MPa to obtain (2, 3-dioleoyl-propyl) trimethyl ammonium chloride. The method is suitable for pilot scale production, and the prepared (2, 3-dioleoyl-propyl) trimethyl ammonium chloride is a phospholipid for cationic liposome and can be used as a pharmaceutical adjuvant for various types of medicines such as injections, tablets, capsules and the like.

Description

(2, 3-dioleoyl-propyl) trimethyl ammonium chloride and preparation method and application thereof

Technical Field

The invention belongs to the field of medical auxiliary materials. In particular to (2, 3-dioleoyl-propyl) trimethyl ammonium chloride and a preparation method and application thereof.

Background

Phospholipids are important components of biological membranes, and their inherent amphiphilic properties, which are both hydrophilic and lipophilic, allow phospholipids to spontaneously form closed bilayers in aqueous media, which become the biological membrane skeleton. By utilizing the property, a novel pharmaceutical preparation technology, namely liposome, is developed. Liposomes were originally discovered by Bangham, a scholarian of the United kingdom, who used liposomes for drug carriers in 1971, when phospholipids were dispersed in water by electron microscopy. The hydrogenated soybean lecithin is prepared by taking soybean lecithin as a raw material, extracting high-purity lecithin and then carrying out catalytic hydrogenation process. The product is light yellow or milk white powder. Due to the adoption of catalysis and hydrogenation processes, unsaturated chains of unsaturated fatty acids in the molecular structure of the soybean lecithin disappear, so that the chemical stability, the dispersibility and the emulsibility of the lecithin are greatly improved. Has decolorizing and deodorizing effects, and is suitable for storage and preservation, and can be used in medicine, high-grade cosmetics, and light industry. In particular to an emulsifier and a nutrient for intravenous fat. It is used as blood fat emulsifier to prevent arteriosclerosis, and has the advantages of easy digestion, easy absorption and less residue in internal organs. The structure of (2, 3-dioleoyl-propyl) trimethylammonium chloride is shown below:

disclosure of Invention

The invention provides an industrial preparation method of (2, 3-dioleoyl-propyl) trimethyl ammonium chloride creatively through a large number of experiments, 3-dimethylamino-1, 2-propanediol is used as a reaction raw material and reacts with oleic acid or oleoyl chloride to obtain 1, 2-dioleoyl-3-dimethylamino-propane, then the 1, 2-dioleoyl-3-dimethylamino-propane reacts with chloromethane at a certain temperature and pressure to obtain (2, 3-dioleoyl-propyl) trimethyl ammonium chloride, and the (2, 3-dioleoyl-propyl) trimethyl ammonium chloride with the purity of more than 99 percent can be obtained through a crystallization purification method. The preparation method provided by the invention is simple to operate, simple in purification steps and suitable for industrial production.

One object of the present invention is to provide a method for preparing (2, 3-dioleoyl-propyl) trimethylammonium chloride, comprising the steps of:

(1) 3-dimethylamino-1, 2-propanediol and oleic acid are reacted under the action of a condensation reagent to obtain 1, 2-dioleoyl-3-dimethylamino-propane, or the 3-dimethylamino-1, 2-propanediol is reacted with oleoyl chloride and alkali to prepare 1, 2-dioleoyl-3-dimethylamino-propane;

alternatively, the first and second electrodes may be,

the reaction solvent of the reaction is one or more selected from tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, trichloromethane, acetonitrile, 1, 4-dioxane, toluene, methyl tert-butyl ether, N-dimethylformamide and acetone N-methylpyrrolidone.

The condensation reagent used in the reaction is selected from DCC-DMAP system, HOBT-HATU system or N, N' -carbonyldiimidazole.

The alkali used in the reaction is selected from one or more of sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, barium hydroxide, sodium methoxide, sodium ethoxide, sodium tert-butoxide, potassium tert-butoxide, triethylamine, diisopropylethylamine, pyridine, N-dimethylpyridine, N-methylpiperidine, morpholine or N-methylmorpholine.

The reaction temperature of the reaction is-30 ℃ to 50 ℃, the reaction time is 1 to 10 hours, the reaction temperature is preferably-10 ℃ to 20 ℃, and the reaction time is 2 to 5 hours.

Reacting 1, 2-dioleoyl-3-dimethylamino-propane with methyl chloride to obtain (2, 3-dioleoyl-propyl) trimethyl ammonium chloride;

the reaction solvent of the reaction is selected from one or more of methanol, ethanol, isopropanol, water, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, trichloromethane, acetonitrile, 1, 4-dioxane, toluene, methyl tert-butyl ether, N-dimethylformamide and acetone N-methylpyrrolidone.

The reaction temperature of the above reaction is-20 ℃ to 200 ℃. The reaction pressure of the above reaction is 0.1 to 10 MPa. The reaction pressure is preferably 1.5MPa, the temperature is 60-90 ℃, and the reaction is carried out for 5-24 h.

At normal temperature and normal pressure, the methyl chloride is in a gas state, the methyl chloride can react with the 1, 2-dioleoyl-3-dimethylamino-propane in a liquid state by selectively controlling the pressure and the temperature, for example, the temperature is-30 ℃ to-25 ℃, the methyl chloride is in a liquid state, the normal pressure is selected, and when the temperature is higher than-23.7 ℃, the gaseous methyl chloride is converted into the liquid state to participate in the reaction by adjusting the pressure in the reaction kettle.

The preparation method of the invention can further comprise a purification step of performing crystallization purification on the (2, 3-dioleoyl-propyl) trimethyl ammonium chloride by using one or more of methanol, ethanol, isopropanol, water, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, trichloromethane, acetonitrile, 1, 4-dioxane, toluene, methyl tert-butyl ether, N-dimethylformamide, acetone and N-methylpyrrolidone.

Another object of the present invention is to provide a hydrogenated soybean lecithin, which is characterized by being prepared and purified by the method of the present invention, wherein the purity of (2, 3-dioleoyl-propyl) trimethyl ammonium chloride is not less than 99%.

The invention also aims to provide the application of the hydrogenated soybean lecithin as a medical auxiliary material for preparing medicines or health-care products.

The product obtained by the invention has stable quality, and meets the standards of medical injection and pharmaceutical excipients after degerming and vacuum drying.

Drawings

FIG. 1 is a HPLC chart of (2, 3-dioleoyl-propyl) trimethylammonium chloride prepared by the method of the present invention measured by external standard method.

Detailed Description

The following examples will aid the understanding of the present invention but should not be construed as limiting the invention.

Example 11 preparation of 2, 2-dioleoyl-3-dimethylamino-propane

The method comprises the following steps: adding 500g of 3-dimethylamino-1, 2-propanediol, 1600g of diisopropylethylamine and 15L of tetrahydrofuran into a reaction kettle, cooling to minus 5 ℃, replacing the gas in the kettle with nitrogen, slowly dropwise adding 2800g of tetrahydrofuran solution of oleoyl chloride, keeping the temperature in the reaction kettle not higher than 10 ℃, then slowly heating to room temperature, continuing to react for 2-3 hours, concentrating under reduced pressure, adding ethyl acetate and cold water, separating, washing an organic phase with saturated saline solution, separating, drying the organic phase with anhydrous magnesium sulfate, and filtering. The organic phase was concentrated and dried under reduced pressure to obtain 2580g of 1, 2-dioleoyl-3-dimethylamino-propane in a yield of 95%.

The method 2 comprises the following steps: adding oleic acid (751g, 251mmol, 3.0 equiv.), DCC (3.0 equiv.), DMAP (2.1 equiv.) and 1L chloroform into a reaction bottle, cooling to 5-10 ℃, replacing the gas in the reaction bottle with nitrogen, slowly dropping a chloroform solution of 3-dimethylamino-1, 2-propanediol (100g, 84mmol), keeping the temperature in the reaction kettle not higher than 10 ℃, then slowly raising the temperature to room temperature, continuing to react for 12-20 hours, concentrating under reduced pressure, purifying the crude product by silica gel chromatography to obtain 468g of 1, 2-dioleoyl-3-dimethylamino-propane, wherein the yield is 86%.

1H NMR(300MHz,d₆DMSO)＝5.53(m,4H),4.57(m,1H),2.11-2.76(m,14H),2.83(s,6H),1.21-1.36(m,40H),0.89(m,6H)；MS(m/z)[M+H]+calcd for C41H78NO4+648.6,found 648.5。

EXAMPLE 2 preparation of (2, 3-dioleoyl-propyl) trimethylammonium chloride

The influence of the reaction temperature, pressure and solvent on the product was examined with reference to Table 1. Adding 1, 2-dioleoyl-3-dimethylamino-propane and a reaction solvent into a high-pressure reaction kettle, replacing gas in the kettle with nitrogen, then replacing the nitrogen with methyl chloride, slowly increasing the pressure to 1.5MPa, keeping the pressure, slowly increasing the temperature to a set temperature, reacting at the temperature and the pressure for 5-24 hours, cooling to room temperature, emptying the pressure, transferring the reaction liquid into a normal-pressure reaction kettle, slowly adding a crystallization solvent, separating out white solid, continuously stirring at the room temperature for 2-5 hours, filtering, washing the solid with the crystallization solvent, drying the solid under reduced pressure to obtain (2, 3-dioleoyl-propyl) trimethyl ammonium chloride, and calculating the conversion rate of the raw materials.

TABLE 1 Effect of different temperatures and pressures on feedstock conversion

	1, 2-dioleoyl-3-dimethylamino-propane	Reaction solvent	Temperature of	Pressure of	Conversion rate
						1	50g	Tetrahydrofuran (THF)	50	1.5MPa	26.3％
2	50g	Tetrahydrofuran (THF)	60	1.5MPa	53.5％
						3	50g	Tetrahydrofuran (THF)	70	1.5MPa	87.2％
4	50g	Tetrahydrofuran (THF)	85	1.5MPa	95.3％
						5	50g	Tetrahydrofuran (THF)	90	1.5MPa	95.2％

The results show that the conversion of the starting material is highest at a pressure of 1.5MPa as the reaction temperature is increased to 85 ℃. The reaction conditions are selected for preparation. The influence of the reaction solvent and the purification solvent on the yield and purity of the product was examined with reference to the conditions of Table 3.

HPLC conditions:

using a C18 column, e.g. Phenomenex, Gemini, 5u C18110A: taking methanol-acetate buffer (pH 4.5) ═ 9:1 as mobile phase A, and chloroform-methanol-acetate buffer (pH 4.5) ═ 30:20:1 as mobile phase B; the column temperature is 40 ℃; gradient elution was performed as in table 2; an evaporative light scattering detector Agilent A380 at 40 ℃ under a nitrogen flow rate of 1.5L per minute.

TABLE 2

Time (minutes)	Mobile phase A (%)	Mobile phase B (%)
			0	85	15
30	0	100
			30.1	85	15
40	85	15

TABLE 3 Effect of different reaction and purification solvents on product yield and purity

	DOTAP	Reaction solvent	Crystallization solvent	Yield%	Purity of
						1	10.0g	Tetrahydrofuran (THF)	Methyl tert-butyl ether	89％	99％
2	10.0g	Tetrahydrofuran (THF)	Acetone (II)	91％	96.5％
						3	10.0g	Chloroform	Methyl tert-butyl ether	73％	99％
4	10.0g	Methylene dichloride	Methyl tert-butyl ether	87％	98.3％
						5	10.0g	Chloroform	Acetone (II)	83％	97.9％
6	10.0g	Methylene dichloride	Acetone (II)	92％	97.3％

The results show that the use of methyl tert-butyl ether as crystallization solvent enables a better product purity to be achieved. This condition was chosen to produce (2, 3-dioleoyl-propyl) trimethylammonium chloride.

Adding 2200g of 1, 2-dioleoyl-3-dimethylamino-propane and 13.2L of tetrahydrofuran into a high-pressure reaction kettle, replacing gas in the kettle with nitrogen, then replacing the nitrogen with methyl chloride, slowly increasing the pressure to 1.5MPa, keeping the pressure, slowly increasing the temperature to 85 ℃, reacting at the temperature and the pressure for 5-24 hours, cooling to room temperature, emptying the pressure, transferring the reaction liquid into a normal-pressure reaction kettle, slowly adding 10L of methyl tert-butyl ether, separating out white solid, continuously stirring at the room temperature for 2-5 hours, filtering, washing the solid with methyl tert-butyl ether, and drying the solid under reduced pressure to obtain 2107g of (2, 3-dioleoyl-propyl) trimethyl ammonium chloride, wherein the yield is 89%, and the HPLC purity of the product is 99.1% by adopting an external standard method, and the results are shown in figure 1.

1H NMR(300MHz,d₆DMSO)＝5.62(m,4H),4.63(m,1H),2.14-2.72(m,14H),3.62(s,9H),1.19-1.32(m,40H),0.91(m,6H)；MS(m/z)[M+H]+calcd for C41H78NO4+648.6,found 648.5。white solid。

Claims

A method for preparing (2, 3-dioleoyloxy-propyl) trimethylammonium chloride, comprising the steps of:

(1) 3-dimethylamino-1, 2-propanediol and oleic acid are reacted under the action of a condensation reagent to obtain 1, 2-dioleoyloxy-3-dimethylamino-propane, or the 3-dimethylamino-1, 2-propanediol is reacted with oleoyl chloride and alkali to prepare 1, 2-dioleoyloxy-3-dimethylamino-propane,

；

alternatively, the first and second electrodes may be,

；

(2) reacting 1, 2-dioleoyloxy-3-dimethylamino-propane with chloromethane to obtain (2, 3-dioleoyloxy-propyl) trimethyl ammonium chloride, wherein the reaction solvent is tetrahydrofuran, the reaction temperature is 60-90 ℃, the reaction pressure is 1.5MPa, the reaction time is 5-24h,

；

(3) and (3) crystallizing and purifying the product of the step (2) by using methyl tert-butyl ether.
2. The method according to claim 1, wherein the reaction solvent in step (1) is one or more selected from tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, chloroform, acetonitrile, 1, 4-dioxane, toluene, methyl tert-butyl ether, N-dimethylformamide, acetone, and N-methylpyrrolidone.
3. The process according to claim 1, wherein the condensation reagent used in step (1) is selected from the group consisting of DCC-DMAP system, HOBT-HATU system and N, N' -carbonyldiimidazole.
4. The method according to claim 1, wherein the base used in step (1) is one or more selected from the group consisting of sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, lithium hydroxide, sodium hydroxide, potassium hydroxide, calcium hydroxide, magnesium hydroxide, barium hydroxide, sodium methoxide, sodium ethoxide, sodium tert-butoxide, potassium tert-butoxide, triethylamine, diisopropylethylamine, pyridine, N-lutidine, N-methylpiperidine, morpholine and N-methylmorpholine.
5. The method according to claim 1, wherein the reaction temperature in the step (1) is from-30 ℃ to 80 ℃.