CN114480520A

CN114480520A - Synthesis method of aliphatic chain triazene in triazcins or analogs thereof

Info

Publication number: CN114480520A
Application number: CN202011267941.3A
Authority: CN
Inventors: 黄伟; 咸漠
Original assignee: Qingdao Institute of Bioenergy and Bioprocess Technology of CAS
Current assignee: Qingdao Institute of Bioenergy and Bioprocess Technology of CAS
Priority date: 2020-11-13
Filing date: 2020-11-13
Publication date: 2022-05-13

Abstract

A synthetic method of fatty chain triazene in triazcins or analogs thereof belongs to the technical field of chemical synthesis. In order to solve the problems of unstable reaction, high danger, higher requirements on equipment and reaction conditions, serious environmental pollution caused by process wastewater and the like of triazcins or analogues thereof in the prior art, the invention utilizes fatty chain olefine aldehyde and hydrazine hydrate to react in a room-temperature absolute ethanol system to generate undecane hydrazone, and long-chain fatty acid acetyl coenzyme A ligase Tri17 is used as a catalyst to convert the undecane hydrazone into the undecane triazene under a neutral condition in the presence of adenosine triphosphate, magnesium chloride, sodium chloride, nitrite and the like. The synthesis method is carried out under neutral conditions of normal temperature and normal pressure, the reaction condition is mild, and the operation is safe and pollution-free.

Description

Synthesis method of aliphatic chain triazene in triazcins or analogs thereof

Technical Field

The invention belongs to the technical field of chemical synthesis, and particularly relates to a synthetic method of fatty chain triazene in triazoin or analogues thereof.

Background

Triazenes are a unique class of polyazo compounds containing 3 consecutive nitrogen atoms. Due to the structural particularity, the novel composite material has wide application in various fields. The triazcins family compound is a natural product separated from streptomyces, and is characterized in that the tail end of a fatty chain is connected with a very rare hydroxytriazole alkene structure in nature, and biological activity tests prove that the triazcins family compound has the activity of inhibiting long-chain acyl coenzyme A synthetase so as to block the accumulation of intracellular liposome, and has good application prospect.

Nowadays, the triazacyclocin family compounds can only be obtained by fermentation, and the yield is low. At present, no report on chemical synthesis of triazacycline family compounds and analogues is found, the key structure of the compounds is a triazene part, the traditional chemical method generally needs amino diazotization and coupling processes in the process of synthesizing triazene, and the chemical diazotization reaction has certain problems, such as: 1. the reaction is generally carried out under the acidic condition (about pH2.0), the reaction is unstable, and the process has high pollution and explosive danger; 2. the requirements on equipment and reaction conditions are high, and the process wastewater seriously pollutes the environment. Therefore, it is necessary to research a safe, efficient and pollution-free triazene synthesis reaction of triazcins or triazenes analogues thereof under neutral conditions.

Disclosure of Invention

In order to solve the problems of unstable reaction, high risk, higher requirements on equipment and reaction conditions, serious environmental pollution caused by process wastewater and the like of triazcins or analogues thereof in the prior art, the invention provides a synthetic method of fatty chain triazene in triazcins or analogues thereof, wherein the synthetic method comprises two steps of chemical synthesis and enzymatic synthesis, and comprises the following specific steps:

firstly, dissolving hydrazine hydrate in absolute ethyl alcohol, dropwise adding undecylenic aldehyde while stirring at room temperature under the protection of nitrogen, continuously reacting for a period of time after dropwise adding, adding water, and sequentially extracting, drying, filtering and concentrating to obtain undecylenic hydrazone;

and step two, adding the undecane hydrazone obtained in the step one, long-chain fatty acid acetyl coenzyme A ligase Tri17, sodium nitrite, magnesium chloride, sodium chloride and adenosine triphosphate into Tris HCl buffer solution with the pH value of 7, and incubating at room temperature for at least 3 hours to obtain the undecane triazene.

In the embodiment of the invention, the concentration of hydrazine hydrate in the first step is 28 mmol/mL-32 mmol/mL, and the dosage is 1 mL-2 mL; the dosage of the absolute ethyl alcohol is 8 mL-10 mL.

Further limiting, the amount of hydrazine hydrate used in step one is 3 equivalents to the undecenal.

Further, the dropping rate of undecenal in the step one is 0.4mL/min to 0.6 mL/min.

Further limiting, the reaction time in the step one is 1.5-2.5 h.

Further, the amount of water added in step one is 8mL to 12 mL.

Further limiting, in the first step, 10-15 mL of dichloromethane is used for extraction for 2-3 times.

Further defined, step one is dried using anhydrous sodium sulfate.

In the embodiment of the invention, in the second step, 0.45-0.55 mg of undecane hydrazone, 0.1-0.3 mg of sodium nitrite, 0.2-0.3 mg of magnesium chloride, 12-16 mg of sodium chloride, 1.3-1.5 mg of adenosine triphosphate and 150-250 μ L of long-chain fatty acid acetyl coenzyme A ligase Tri17 obtained in the first step are added into 15mL of Tris HCl buffer solution together.

Further, the concentration of the Tris HCl buffer solution in the second step is 45-50 mM.

Advantageous effects

The method for synthesizing fatty chain triazene in triazene or analogues thereof provided by the invention is carried out under neutral conditions, has the characteristics of stability, safety, high efficiency, no pollution and the like, and solves the problems that the traditional chemical method is carried out under acidic conditions generally, the reaction is unstable, the reaction process has high pollution and high explosive risk, the requirements on equipment and reaction conditions are high, and the process wastewater seriously pollutes the environment.

Drawings

FIG. 1: the invention relates to a schematic diagram of a synthetic reaction of fatty chain triazene in triazene or an analogue thereof, wherein a compound I is undecenal; the compound II is undecane hydrazone; the compound III is undec triazene; r has the structure

FIG. 2: the chemical synthesis route is characterized in that the compound I is undecenal; the compound II is undecane hydrazone;

FIG. 3: an enzyme catalysis synthesis route, wherein the compound II is undecane hydrazone; the compound III is undec triazene;

FIG. 4: and detecting the undecenetricene by mass spectrum.

Detailed Description

The synthesis method of the present invention is described in detail below by way of examples, and the reaction scheme of the method is shown in fig. 1, and the specific steps are as follows:

1) chemical synthesis: hydrazine hydrate (1.5mL,30mmol/mL) is dissolved in 10mL of absolute ethanol, 2mL of undecylenic aldehyde (10mmol/mL) is added dropwise under the protection of nitrogen, and the reaction solution is stirred at room temperature for 2 hours; after the reaction is finished, 10mL of water is added, 10mL of dichloromethane is added for extraction for three times, the organic phase is dried by anhydrous sodium sulfate, filtered and concentrated to obtain a white solid, namely the undecanone hydrazone, and LC analysis is carried out, wherein the conversion rate is about 90%.

2) Enzyme-catalyzed synthesis: 0.5mg of the undecane hydrazone obtained in the step 1) is taken, 0.2mg of sodium nitrite, 0.25mg of magnesium chloride, 15mg of sodium chloride, 1.4mg of Adenosine Triphosphate (ATP) and 200 muL of long-chain fatty acid acetyl coenzyme A ligase Tri17 are added and dissolved in 15mL of 50mM Tris HCl buffer solution, incubation is carried out for 3 hours at room temperature, and the undecane triazene is obtained and analyzed by MS, and the conversion rate is more than 80%.

The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes performed by the present specification and drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.

Claims

1. A synthetic method of fatty chain triazene in triazcins or analogues thereof is characterized in that the synthetic method comprises two steps of chemical synthesis and enzymatic synthesis, and the specific steps are as follows:

2. The synthesis method according to claim 1, wherein the concentration of hydrazine hydrate in the first step is 28mmol/mL to 32mmol/mL, and the amount is 1mL to 2 mL; the dosage of the absolute ethyl alcohol is 8 mL-10 mL.

3. The method of claim 1, wherein hydrazine hydrate is used in an amount of 3 equivalents to undecenal in step one.

4. The method of claim 1, wherein the undecenal is added at a rate of 0.4mL/min to 0.6mL/min in step one.

5. The synthesis method according to claim 1, wherein the reaction time in the first step is 1.5-2.5 h.

6. The method of claim 1, wherein the amount of water added in step one is 8mL to 12 mL.

7. The method of claim 1, wherein in step one, 10 to 15mL of dichloromethane are used for 2 to 3 times of extraction.

8. The method of claim 1, wherein step one is performed by drying with anhydrous sodium sulfate.

9. The method of claim 1, wherein 0.45-0.55 mg of undecanohydrazone, 0.1-0.3 mg of sodium nitrite, 0.2-0.3 mg of magnesium chloride, 12-16 mg of sodium chloride, 1.3-1.5 mg of adenosine triphosphate and 150-250 μ L of long chain fatty acid acetyl-CoA ligase Tri17 obtained in the first step are added together to 15ml Tris HCl buffer solution in the second step.

10. The method of claim 1, wherein the concentration of Tris HCl buffer in step two is 45-50 mM.