Method for preparing 1-oxo-4, 5-diazepane by enzyme catalytic hydrolysis
阅读说明：本技术 酶催化水解制备1-氧-4,5-二氮杂环庚烷的方法 (Method for preparing 1-oxo-4, 5-diazepane by enzyme catalytic hydrolysis ) 是由 范谦 任杰 晏三军 程柯 于 2019-09-19 设计创作，主要内容包括：本发明公开了一种酶催化水解制备1-氧-4,5-二氮杂环庚烷的方法,属于有机合成技术领域。本发明为解决现有技术中产率低,需要使用强酸或强碱等问题,提供了一种酶催化水解制备1-氧-4,5-二氮杂环庚烷的方法：使式I化合物在水解酶的催化下进行水解反应,分离纯化,得到1-氧-4,5-二氮杂环庚烷。本发明采用水解酶催化,反应选择性好、副反应少,产品收率可达90％以上,萃取液可直接作为后续反应的原料合成唑啉草酯；反应条件温和、易于控制,整个工艺操作简单,易于工业化。(The invention discloses a method for preparing 1-oxo-4, 5-diazepane by enzymatic hydrolysis, belonging to the technical field of organic synthesis. The invention provides a method for preparing 1-oxo-4, 5-diazepane by enzymatic hydrolysis, aiming at solving the problems of low yield, need of using strong acid or strong alkali and the like in the prior art: carrying out hydrolysis reaction on the compound of the formula I under the catalysis of hydrolase, and separating and purifying to obtain the 1-oxo-4, 5-diazepane. The invention adopts hydrolase catalysis, has good reaction selectivity and less side reaction, the product yield can reach more than 90 percent, and the extract can be directly used as the raw material for the subsequent reaction to synthesize pinoxaden; the reaction condition is mild and easy to control, and the whole process is simple to operate and easy to industrialize.)
1. The method for preparing 1-oxo-4, 5-diazepane by enzyme catalytic hydrolysis is characterized by comprising the following steps: the method comprises the following steps: carrying out hydrolysis reaction on the compound of the formula I under the catalysis of hydrolase, and separating and purifying to obtain 1-oxo-4, 5-diazepane;
wherein R is1And R2Each independently represents C1-C5Alkyl of (C)1-C5Haloalkyl, C2-C5Alkenyl radical, C2-C5Alkynyl, phenyl, C1-C5Alkylphenyl, halophenyl, C1-C5Alkoxyphenyl, benzyl, C1-C5Alkylbenzyl, halobenzyl, C1-C5Alkoxybenzyl radical, C1-C5alkoxy-C1-C5Alkyl or C3-C6Cycloalkyl, or R1And R2Together are C1-C4Alkylene, 1, 2-phenylene or 1, 8-naphthylene.
2. The process for the enzymatic hydrolysis of 1-oxo-4, 5-diazepane according to claim 1, wherein: r1And R2Each independently selected from methyl or ethyl.
3. The process for the enzymatic hydrolysis of 1-oxo-4, 5-diazepane according to claim 1, wherein: the hydrolase is at least one selected from alpha-chymotrypsin, papain, novacin 435, porcine pancreatic lipase, aspergillus oryzae lipase, neutral protease, porcine pepsin, bovine trypsin, urease, alkaline protease, Lipozyme TL 100 or bovine pancreatic lipase; preferably, it is novacin 435, porcine pancreatic lipase or urease.
4. The process for the enzymatic hydrolysis of 1-oxo-4, 5-diazepane according to claim 3, wherein: the hydrolase belongs to the enzyme classification e.c.3.5; preferably belonging to the enzyme class e.c.3.5.1.
5. The method for producing 1-oxo-4, 5-diazepane by enzymatic hydrolysis according to any one of claims 1 to 4, wherein: the addition amount of the hydrolase is 0.5 to 10 percent of the mass of the compound of the formula I; preferably 1 to 3 percent; more preferably 1% to 2%.
6. The process for the enzymatic hydrolysis of 1-oxo-4, 5-diazepane according to claim 1, wherein: the solvent of the hydrolysis reaction is water or a water-containing organic solvent; preferably, the solvent for the hydrolysis reaction is water.
7. The process for the enzymatic hydrolysis of 1-oxo-4, 5-diazepane according to claim 6, wherein: the water-containing organic solvent is one or more than one of tetrahydrofuran, dimethyl sulfoxide, N-dimethylformamide, 1, 3-dimethyl-2-imidazolidinone or N, N-dimethylacetamide.
8. The method for preparing 1-oxo-4, 5-diazepane by enzymatic hydrolysis according to any one of claims 1 to 7, wherein: the temperature of the hydrolysis reaction is 5-100 ℃; preferably 25-70 ℃; more preferably 40-60 ℃; most preferably 50 to 55 ℃.
9. The enzymatic method of claim 8, wherein: the hydrolysis reaction time is 12-96 hours; preferably 24 to 48 hours.
10. The method for producing 1-oxo-4, 5-diazepane by enzymatic hydrolysis according to any one of claims 1 to 9, wherein: the separation and purification comprises the step of adding alkali to adjust the pH value to 9-10, wherein the alkali is at least one of sodium carbonate, potassium carbonate, sodium hydroxide or potassium hydroxide.
Technical Field
The invention belongs to the technical field of organic synthesis, and particularly relates to a method for preparing 1-oxo-4, 5-diazepane through enzymatic hydrolysis.
Background
1-oxo-4, 5-diazepanes are intermediates in the preparation of tetrahydropyrazolidinedione herbicides, such as pinoxaden (WO 99047525).
The current methods for synthesizing 1-oxo-4, 5-diazepane mainly comprise:
CN1604896A reports the method of the above reaction formula, which uses formula I as raw material, and hydrolyzes with halogen acid in polar solvent to obtain the corresponding salt formula III of formula II, the method needs to use more than 2 equivalents of acid, the product is salt, it is easy to absorb moisture, and it is not easy to store; and when the intermediate is used, neutralization and dissociation are needed, the operation is complicated, the market competitiveness is lost, a large amount of three wastes are generated, the operation environment is severe, and the intermediate belongs to a obsolete process.
CN101039926B reports the method of the above reaction formula, which uses formula I as raw material, and hydrolyzes with more than 2 equivalents of sodium hydroxide or potassium hydroxide in water to obtain formula II, although the process is superior to CN1604896A, the yield is not high (49-69%) due to too strong sodium hydroxide or potassium hydroxide, and the cost of raw material is increased, so it cannot be applied to scale-up production.
Therefore, a method for preparing 1-oxo-4, 5-diazepane with mild reaction conditions, high yield and easy post-treatment is needed.
Disclosure of Invention
Aiming at the problems in the prior art, the inventor surprisingly discovers that the compound in the formula I can be efficiently hydrolyzed under the catalysis of hydrolase and mild reaction conditions to obtain the compound in the formula II, so that the problems of easy moisture absorption of 1-oxo-4, 5-diazepane hydrohalide salt, low yield and serious equipment corrosion can be solved.
Based on this, the present invention provides a method for preparing 1-oxo-4, 5-diazepane by enzymatic hydrolysis, which comprises the following steps: carrying out hydrolysis reaction on the compound of the formula I under the catalysis of hydrolase, and separating and purifying to obtain 1-oxo-4, 5-diazepane;
wherein R is1And R2Each independently represents C1-C5Alkyl of (C)1-C5Haloalkyl, C2-C5Alkenyl radical, C2-C5Alkynyl, phenyl, C1-C5Alkylphenyl, halophenyl, C1-C5Alkoxyphenyl, benzyl, C1-C5Alkylbenzyl, halobenzyl, C1-C5Alkoxybenzyl radical, C1-C5alkoxy-C1-C5Alkyl or C3-C6Cycloalkyl, or R1And R2Together are C1-C4Alkylene, 1, 2-phenylene or 1, 8-naphthylene.
Preferably, in the above process, R1And R2Each independently selected from methyl or ethyl, especially methyl, to facilitate isolation and purification of the compound of formula II.
In the method, a plurality of hydrolytic enzymes are tested, and the hydrolytic enzymes are at least one selected from alpha-chymotrypsin, papain, novacin 435, porcine pancreatic lipase, aspergillus oryzae lipase, neutral protease, porcine pepsin, bovine trypsin, urease, alkaline protease, Lipozyme TL 100 or bovine pancreatic lipase.
The catalytic effect of different hydrolases is different, and in order to improve the product yield, it is preferable that in the above method, the hydrolase is selected from the group consisting of novacin 435, porcine pancreatic lipase or urease.
"hydrolase" refers to an enzyme that catalyzes a type of hydrolysis reaction, i.e., an enzyme belonging to EC 3, under the classification system of the International Enzyme Committee (EC).
Wherein, in the above method, the hydrolase belongs to the enzyme classification E.C.3.5.
Preferably, in the above process, the hydrolase belongs to the enzyme class e.c. 3.5.1.
In the method, the addition amount of the hydrolase is 0.5-10% of the mass of the compound shown in the formula I.
Preferably, in order to improve the product yield, in the method, the addition amount of the hydrolase is 1-3% of the mass of the compound shown in the formula I.
More preferably, in order to further improve the product yield, in the method, the addition amount of the hydrolase is 1 to 2 percent of the mass of the compound shown in the formula I.
In the method, the solvent for the hydrolysis reaction is water or a water-containing organic solvent.
Preferably, in the above method, the solvent for the hydrolysis reaction is water.
In the method, the dosage of the solvent is 1-100 times of the mass of the compound shown in the formula I.
Preferably, in the method, the amount of the solvent is 1-10 times of the amount of the compound of formula I.
Wherein, in the method, the aqueous organic solvent is one or more selected from tetrahydrofuran, dimethyl sulfoxide, N-dimethylformamide, 1, 3-dimethyl-2-imidazolidinone or N, N-dimethylacetamide.
Wherein, in the above method, the molar ratio of water to the compound of formula I in the aqueous organic solvent is not less than 5: 1.
the enzyme catalysis reaction is generally carried out at a suitable temperature, and the temperature of the hydrolysis reaction in the method is 5-100 ℃ through tests.
Preferably, in order to improve the product yield, in the method, the temperature of the hydrolysis reaction is 25-70 ℃.
More preferably, in order to further improve the product yield, in the method, the temperature of the hydrolysis reaction is 40-60 ℃.
Most preferably, in the method, the temperature of the hydrolysis reaction is 50-55 ℃.
In the method, the hydrolysis reaction time is 12-96 hours.
Preferably, in the method, the hydrolysis reaction time is 24 to 48 hours.
In the method, the separation and purification comprises the step of adding alkali to adjust the pH value to 9-10, wherein the alkali is at least one of sodium carbonate, potassium carbonate, sodium hydroxide or potassium hydroxide, and then the extraction is carried out by adopting xylene, toluene, chlorobenzene, dichloromethane, 1, 2-dichloroethane and the like, and the extract can be directly used as a raw material for subsequent reaction to synthesize pinoxaden.
The invention has the beneficial effects that:
the raw materials are cheap and easy to obtain, the hydrolase is used for catalyzing, the reaction selectivity is good, the side reactions are few, and the product yield can reach more than 90% by optimizing the reaction conditions; the reaction condition is mild and easy to control, and the whole process is simple to operate and easy to industrialize.
Detailed Description
The method of the present invention for preparing 1-oxo-4, 5-diazepane by enzymatic hydrolysis is further illustrated by the following examples, but it should not be construed that the scope of the above subject matter is limited to the following examples, and that all techniques realized based on the above disclosure are within the scope of the present invention.
Sources of raw materials and reagents in the examples:
alpha-chymotrypsin, papain, bovine pancreatic lipase, porcine pancreatic lipase, aspergillus oryzae lipase, neutral protease, porcine pepsin, bovine trypsin and urease were purchased from alatin.
Novozyme 435, alkaline protease (Novozyme Alcalase 2.4L) and Lipozyme TL 100 were purchased from Novozyme.
Example 1: screening for hydrolytic enzymes
Adding 4, 5-diacetyl- [1,4,5 ] into a reaction bottle]-oxadihetero10g (0.054mol) of water and 25g of water, adding 0.1g of hydrolase, reacting for 24 hours at 50 ℃, controlling the temperature to be 30-35 ℃, adjusting the pH value to 9-10 by using 30% sodium hydroxide, and addingThe reaction solution was extracted with 10mL of toluene, and the product yield was determined by gas chromatography, and the results are shown in Table 1 below.
TABLE 1 screening of hydrolases
As can be seen from Table 1, the target products can be obtained by using different hydrolases, but the yield is greatly different; wherein, the effects of the novacin 435, the bovine trypsin, the porcine pancreatic lipase and the urease are better and reach more than 70 percent; the urease effect is optimal, and the yield reaches more than 90%.
Example 2: screening of reaction temperature
Adding 4, 5-diacetyl- [1,4,5 ] into a reaction bottle]-oxadihetero10g (0.054mol) of urease and 25g of water are added, 0.1g of urease is added, after the reaction is carried out for 24 hours at a certain temperature, the pH value is adjusted to 9-10 by 30 percent sodium hydroxide at the temperature of 30-35 ℃, 10mL of toluene is added to extract reaction liquid, and the product yield is detected by gas chromatography, and the results are shown in the following table 2.
TABLE 2 screening of reaction temperatures
Experimental group
Reaction temperature/. degree.C
Yield/%
13
25
63.3
14
40
82.4
15
45
86.2
9
50
92.8
16
55
94.7
17
60
77.8
18
70
54.1
As can be seen from Table 2, the yield of the product increases and then decreases with the temperature increase by using urease as a catalyst, and the reaction temperature is preferably 40-60 ℃; the effect is best between 50 ℃ and 55 ℃, and the yield reaches more than 90%.
Example 3: screening of enzyme dosage
Adding 4, 5-diacetyl- [1,4,5 ] into a reaction bottle]-oxadihetero10g (0.054mol) of water and 25g of water are added with a certain mass of urease, and after 24 hours of reaction at 55 ℃, the temperature is controlled at 30-35 DEG CThen, 30% sodium hydroxide is used for adjusting the pH value to 9-10, 10mL of toluene is added for extracting the reaction solution, and the product yield is detected by gas chromatography, and the results are shown in the following table 3.
TABLE 3 screening of enzyme dosage
As can be seen from Table 3, the amount of the hydrolase used had a large influence on the reaction results, and when the amount of the enzyme was less than 1 wt%, the yield was very low, and excessive amount of the enzyme decreased the reaction yield; the effect is better when the dosage of the hydrolase is 1 wt% -2 wt%.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention in any way, and all simple modifications and equivalent variations of the above embodiments according to the technical spirit of the present invention are included in the scope of the present invention.
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