CN112608350B

CN112608350B - Synthetic method of 3-N-acetyl etimicin

Info

Publication number: CN112608350B
Application number: CN202011536901.4A
Authority: CN
Inventors: 王晓霞; 朱音; 於江华; 华梦丹; 毛菊红; 过晓磊
Original assignee: Wuxi Jiyu Shanhe Pharmaceutical Co ltd
Current assignee: Wuxi Jiyu Shanhe Pharmaceutical Co ltd
Priority date: 2020-12-23
Filing date: 2020-12-23
Publication date: 2022-12-02
Anticipated expiration: 2040-12-23
Also published as: CN112608350A

Abstract

The invention relates to a synthesis method of 3-N-acetyl etimicin, which comprises the following steps: putting etimicin, di-tert-butyl dicarbonate ethanol and water into a round-bottom flask for refluxing, and distilling part of the solvent after reactants are completed. Formic acid was added and left to stir at 0 ℃ for 4h. After the reaction is finished, adding 5 percent of sodium hydroxide, stirring and heating to 125 ℃ for reaction for 12 hours, heating and refluxing, cooling to room temperature, and concentrating the reaction system. The target compound 3-N-acetyl etimicin is obtained by separation.

Description

Synthetic method of 3-N-acetyl etimicin

Technical Field

The invention relates to an organic chemical synthesis method, in particular to a method for synthesizing 3-N-acetyl etimicin by etimicin.

Background

Aminoglycoside antibiotics have strong toxic and side effects on the eighth pair of cranial nerves, kidney and other tissues, and can cause serious damage to the hearing and the kidney after long-term and large-scale use, so that Chinese scientific research personnel develop a large amount of semi-synthesis modification work on the medicaments.

Etimicin sulfate, also known as epirubicin, 89-07 or E-402, is one of semisynthetic aminoglycoside antibiotics, and has good antibacterial effect on gram-positive bacteria and gram-negative bacteria. The chemical name of the compound is O-2-amino-2, 3,4, 6-tetradeoxy-6-amino-alpha-D-erythro-hexopyranosyl- (1 → 4) -O- [ 3-deoxy-4-C-methyl-3 (methylamino) -beta-L-arabinopyranosyl- (1 → 6) ] -2-deoxy-N-ethyl-L-streptomycin sulfate, and the structural formula is shown as follows

The etimicin sulfate parent nucleus belongs to 4, 6-disubstituted deoxystreptomycin amino glycoside antibiotics, and the synthetic principle is that gentamicin C _1a The deoxystreptomycin 1-N position in the molecule is introduced with ethyl, so that the deoxystreptomycin has better antibacterial effect on various drug-resistant bacteria, and meanwhile, the side effect of otonephrotoxicity is obviously reduced. It has high efficiency, low toxicity, high resistance to drug-resistant bacteria, high curative effect on respiratory tract infection, urinary system infection, skin and soft tissue infection, low adverse reaction, high cross drug resistance and far less ototoxicity than other similar products, and is a safe and effective new anti-infective drug suitable for various infections caused by sensitive gram-negative bacilli, such as bronchitis, lung infection, cystitis, pyelonephritis and the like. Etimicin sulfate was awarded by the State Council of the people's republic of China State Council in 2001 to the second-class prize of the national science and technology progress (certificate number: J-235-2-03-R09).

The traditional antibiotics with large toxic and side effects and drug resistance are partially replaced successfully in the market of the drug, and the drug requirements of the market are met; but also improves the development level of antibiotic medicines in China and initiates a demonstration and leading function for the independent research of the medicine innovation of the same pharmaceutical industry.

The adverse reaction generated in clinical use of the medicine is not only related to the pharmacological activity of the medicine, but also has a great relationship with impurities in the medicine.

3-N-acetyl etimicin with the structure as follows

Is a byproduct in the synthesis process of etimicin sulfate, is frequently used in the detection process, but the product has low content and is difficult to obtain, and no 3-N-acetyl etimicin standard product is sold at home and abroad and no relevant literature report exists. In the safety research process of the etimicin sulfate bulk drug and the preparation thereof, the reference substance of trace impurities is inevitably required to be used. Therefore, the development of the preparation process of the 3-N-acetyl etimicin has great significance for improving the quality of the medicine and the safety of clinical medication.

Disclosure of Invention

The invention aims to provide a preparation method of 3-N-acetyl etimicin.

The preparation method of the invention has the following synthetic route:

the method comprises the following operation steps:

(1) Adding etimicin, ethanol and water into a round-bottom flask, uniformly stirring, cooling to 0 ℃, dropwise adding di-tert-butyl dicarbonate, continuously stirring, and concentrating a reaction system after the reaction is finished;

(2) Adding formic acid into the reaction system, heating to 120 ℃, continuously reacting for 4 hours, and cooling to room temperature after the reaction is finished;

(3) Respectively adding 10% sodium hydroxide, stirring, heating and refluxing, and cooling to room temperature after the reaction is finished;

(4) Desalting, and separating to obtain the target product 3-N-acetyl etimicin.

Wherein the molar ratio of etimicin to di-tert-butyl dicarbonate in the step (1) is 1.5-1.

Preferably, the molar ratio of etimicin to di-tert-butyl dicarbonate in step (1) is 1.

Wherein, the temperature in the step (1) is between 5 ℃ below zero and 5 ℃ and the reflux is carried out for 4 to 12 hours.

Preferably, the temperature in step (1) is refluxed at 0 ℃ for 10h.

Wherein the molar ratio of etimicin to formic acid in the step (2) is 1.

Preferably, the molar ratio of etimicin to formic acid in step (2) is 1.

Wherein the temperature in the step (2) is between 80 and 140 ℃ for refluxing for 1.5 to 6 hours.

Preferably, the temperature in step (2) is refluxed at 120 ℃ for 4h.

Wherein, the concentration of the sodium hydroxide solution in the step (3) is 5 to 30 percent.

Preferably, the concentration of the sodium hydroxide solution in the step (3) is 10%.

Wherein the heating temperature in the step (3) is 110-140 ℃, and the reflux is carried out for 10-16 h.

Preferably, the heating temperature in step (3) is 125 ℃, and the reflux is carried out for 12h.

Wherein, the separation in the step (4) is to separate the 3-N-acetyl etimicin by a silica gel column and a preparative liquid phase.

Preferably, the separation in the step (4) is carried out by silica gel column separation to obtain 3-N-acetyl etimicin and obtain high-purity 3-N-ethyl gentamicin C _1a 。

Further preferably, the preparation method of the invention comprises the following steps:

a. adding 5g of etimicin, ethanol and water into a round-bottom flask, uniformly stirring, cooling to-5 ℃, dropwise adding 3.27-6.54 g of di-tert-butyl dicarbonate, continuously stirring for 5-7 hours, and concentrating a reaction system after the reaction is finished.

b. 10-26.5 ml of formic acid is added into the reaction system, the temperature is raised to 80-140 ℃, the reaction is continued for 3-5 h, and the temperature is reduced to room temperature after the reaction is finished.

c. Respectively adding 10ml of 5-25% sodium hydroxide, stirring, heating, refluxing, and cooling to room temperature after the reaction is finished.

d. Desalting, and separating by a silica gel column to obtain the target product 3-N-acetyl etimicin.

N-6-acetamido-3, 5-dihydroxy-5-methyl-4- (methylamino) tetrahydro-2H-pyran-2-yl) oxy) -4- (ethylamino) -2-hydroxychlorohexyl) oxy) -5-pivaloylamide tetrahydro-2H-pyran-2-yl) methyl) pivaloylamide.

The chinese names of compounds 1-5 appearing in the above synthetic route:

compound 1: etimicin

Compound 2:3',5' -bis (pivaloylamide) etimicin

Compound 3:3',5' -bis (pivaloylamide) -3-acetyletimicin

Compound 4: 3-N-acetyl etimicin

The above-mentioned compounds 1 to 4,1,4 are known compounds, and 2,3 are unknown compounds.

Compared with the prior art, the preparation method of the invention has the following beneficial effects:

at present, no report of a relevant synthesis process of 3-N-acetyl etimicin exists, the synthesis process of the only 3-N-acetyl etimicin has the advantages of simple synthesis route, convenient operation, short time, low cost and little environmental pollution, and the 3-N-acetyl etimicin prepared by the method has the characteristics of high purity, high yield and the like, and has great significance for improving the medicine quality and the safety of clinical medication.

Drawings

FIG. 1TLC chart of reaction system

FIG. 2 LC-MS detection of reaction system, total ion flow diagram

FIG. 3 first-order mass spectrum at 9.3min under positive ion mode under the detection condition of reaction system LC-MS

FIG. 4 shows the first-order mass spectrum at 11.2min (Etimicin) in the positive ion mode under the detection condition of the reaction system LC-MS

FIG. 5 is a primary mass spectrum of peak 3 at 12.4min in positive ion mode under LC-MS detection of the reaction system

FIG. 6 TLC chart of target Compound 1

FIG. 7 TLC chart for target Compound 2

FIG. 8 structural formula of N-acetyl etimicin

Detailed description of the preferred embodiments

The present invention is further illustrated by the following specific examples, which are not to be construed as limiting the invention thereto.

Example 1, 3-N-Acetyletimicin

a. Adding 5g of etimicin, ethanol and water into a round-bottom flask, uniformly stirring, cooling to-5 ℃, dropwise adding 4.8g of di-tert-butyl dicarbonate, continuously stirring for 5 hours, and concentrating a reaction system after the reaction is finished.

b. Adding 15ml of formic acid into the reaction system, heating to 140 ℃, continuously reacting for 5h, and cooling to room temperature after the reaction is finished.

c. Respectively adding 10ml of 5 percent sodium hydroxide, stirring, heating and refluxing, and cooling to room temperature after the reaction is finished.

d. Desalting, and separating by silica gel column to obtain the target product 3-N-acetyl etimicin 50%.

Example 2, 3-N-Acetyletimicin

a. Adding 5g of etimicin, ethanol and water into a round-bottom flask, uniformly stirring, cooling to 5 ℃, dropwise adding 4.8g of di-tert-butyl dicarbonate, continuously stirring for 7 hours, and concentrating a reaction system after the reaction is finished.

b. Adding 26.5ml of formic acid into the reaction system, heating to 80 ℃, continuously reacting for 5 hours, and cooling to room temperature after the reaction is finished.

c. Respectively adding 10ml of 25% sodium hydroxide, stirring, heating, refluxing, and cooling to room temperature after the reaction is finished.

d. Desalting, and separating by a silica gel column to obtain 60 percent of the target product 3-N-acetyl etimicin.

Example 3, 3-N-Acetyletimicin

a. Adding 5g of etimicin, ethanol and water into a round-bottom flask, uniformly stirring, cooling to 0 ℃, dropwise adding 4.8g of di-tert-butyl dicarbonate, continuously stirring for 6 hours, and concentrating a reaction system after the reaction is finished.

b. Adding 22.7ml of formic acid into the reaction system, heating to 120 ℃, continuously reacting for 4 hours, and cooling to room temperature after the reaction is finished.

c. 10ml of 10 percent sodium hydroxide is respectively added, stirred, heated and refluxed, and cooled to room temperature after the reaction is finished.

d. Desalting, and separating by a silica gel column to obtain the target product 3-N-acetyl etimicin 99 percent.

Example 4, 3-N-acetyl etimicin isolation and detection method.

1. The reaction system is shown in FIG. 1 by thin layer chromatography.

2. LC-MS detection of reaction system

A detection instrument: shimadzu LC-MS 2020 single quadrupole mass spectrometer

Chromatographic conditions are as follows: methanol-Water (6.7 mM ammonium acetate, adjusted pH to 9.8 with Ammonia Water)

Mass spectrum conditions: SCAN Mode-SCAN Positive Mode, SCAN range 100-1200 Da, event time 0.5s, interface temperature: 350 ℃, DL temperature: the flow rate of the atomizer was 1.5L/min and the flow rate of the dryer was 15L/min at 250 ℃. See fig. 2-5.

3. With chloroform-methanol-ammonia =1 as a developing solvent, iodine vapor was developed, and two compounds were mainly separated in the control. A sample (150 ml) was taken from the supplier, mixed with 300g of silica gel (200-300 mesh) and chromatographed on 900g of normal phase silica gel column (. Phi. 7cm. Multidot.120cm). The target compound was followed by TLC, a control provided by the trusted party, using chloroform-methanol-ammonia =10 =1 as the elution phase, to give a fraction Fr1 (15 g) containing two target compounds. The Fr1 site was separated by normal phase silica gel column chromatography, and the target compound was followed by control TLC using chloroform-methanol-ammonia =8. Fr1.3 was stirred with 10 g of silica gel, loaded on the column 100g of silica gel, eluted with chloroform-methanol-ammonia = 8.5.

4. The nuclear magnetic detection shows that the target compound 1 is 3-N-acetyl etimicin, which is shown in figure 8.

Claims

1. A synthetic method of 3-N-acetyl etimicin comprises the following steps:

a. adding etimicin, ethanol and water into a round-bottom flask, uniformly stirring, cooling to 0 ℃, dropwise adding di-tert-butyl dicarbonate, continuously stirring, and concentrating a reaction system after the reaction is finished;

b. adding formic acid into the reaction system, heating to 120 ℃, continuously reacting for 4 hours, and cooling to room temperature after the reaction is finished;

c. respectively adding 10% sodium hydroxide, stirring, heating and refluxing, and cooling to room temperature after the reaction is finished;

d. desalting, namely separating to obtain a target product 3-N-acetyl etimicin;

wherein, the molar ratio of the etimicin to the di-tert-butyl dicarbonate in the step a is 1.5-1, the reaction temperature is-5 ℃, and the reflux is carried out for 6h; step b, the molar ratio of etimicin to formic acid is 1; in the step d, the separation is to obtain the 3-N-acetyl etimicin through silica gel column or preparative liquid phase separation.