Adipyl-7-ADCA acid crystal as well as preparation method and application thereof

Abstract

The invention relates to the technical field of medical biology, and discloses an adipyl-7-ADCA acid crystal as well as a preparation method and application thereof. The preparation process comprises the following steps: slowly adding a dilute sulphuric acid solution into an adipyl-7-ADCA potassium salt aqueous solution, stirring at 50-60r / s, immediately stopping adding acid when the pH value of the solution reaches 2.51-2.57 and the turbidity of the solution is changed, growing crystal, adjusting the stirring speed to 25-30r / s, obtaining an initial crystal solution after 20 minutes, adjusting the pH value, filtering, washing and drying, and finally obtaining the adipyl-7-ADCA acid crystal. According to the preparation method, the yield is as high as 91%, and the crystal purity is as high as 97%; the crystal form is needle-shaped, the process conditions are mild, and the method is suitable for large-scale production; (I).

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical biotechnology, and in particular to adipoyl-7-ADCA acid crystal, a preparation method and application thereof. Background Art

[0002] β-lactam antibiotics are the most widely used and best-regarded class of antibiotics worldwide. β-lactam antibiotics include penicillins and cephalosporins. Cephalosporins are a general term for cephalosporins, which are derived from natural cephalosporin C, obtained through cultivation of Cephalosporium rubrum, through semi-synthetic modification of its side chains. Cephalosporins have a more stable molecular structure, a broad antimicrobial spectrum, strong antimicrobial activity, and low adverse reactions and side effects. They currently represent the largest-selling class of antibiotics in the anti-infective market.

[0003] Among cephalosporin products, 7-adipoylaminodesacetoxycephalosporanic acid (abbreviated as adipyl-7-ADCA) is a key API and a crucial intermediate in the semi-synthesis of cephalosporin antibiotics. After cleavage, adipyl-7-ADCA can be combined with various side chains to synthesize highly marketed drugs such as cephalexin, cephradine, cefadroxil, and cefetamet pivoxil. Its crystal form significantly influences the purity and content of related substances in subsequent synthesized drugs. The pursuit of high-content and high-purity adipyl-7-ADCA has long been a key research priority, and the preparation of adipyl-7-ADCA acid crystals is therefore crucial for the synthesis of cephalosporins.

[0004] Currently, there is little research on the preparation process of adipoyl-7-ADCA acid crystals by researchers at home and abroad, and there are relatively few related articles. Therefore, how to efficiently prepare adipoyl-7-ADCA acid crystals and provide a simple, safe and reliable preparation method is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0005] The embodiments of the present invention provide adipoyl-7-ADCA acid crystals, a preparation method thereof, and applications thereof, to solve the above-mentioned technical problems in the prior art.

[0006] In order to achieve the above technical purpose of the present invention, the technical solution adopted by the present invention is as follows: In one aspect, an embodiment of the present invention provides adipoyl-7-ADCA acid crystal, the structure of which is shown in formula (I): (I).

[0007] The second aspect of the present invention provides a method for preparing the adipoyl-7-ADCA acid crystals, and the preparation process is as follows: A dilute sulfuric acid solution is added to an aqueous solution of adipoyl-7-ADCA potassium salt, followed by crystallization to obtain diacyl-7-ADCA acid crystals. The specific reaction formula is as follows: .

[0008] Preferably, the specific preparation method includes: Step 1, preparing an aqueous solution of adipyl-7-ADCA potassium salt and a dilute sulfuric acid solution; wherein the pH of the aqueous solution of adipyl-7-ADCA potassium salt is 6.5-7.5; Step 2: Slowly add dilute sulfuric acid solution to the prepared adipoyl-7-ADCA potassium salt aqueous solution, stir at 50-60 r / s, and when the pH of the solution reaches 2.51-2.57 and the turbidity of the solution changes, immediately stop adding acid and carry out crystal growth. The stirring speed is adjusted to 25-30 r / s, and the initial crystal solution is obtained after 20 minutes; Step 3: adjusting the pH value of the initial crystallization solution, filtering, and obtaining adipoyl-7-ADCA acid crystal cake; Step 4: Wash and dry the adipoyl-7-ADCA acid crystal cake to obtain adipoyl-7-ADCA acid crystals.

[0009] Preferably, the process of the adipoyl-7-ADCA potassium salt aqueous solution is: The adipoyl-7-ADCA fermentation broth prepared in advance is filtered, extracted and purified to obtain an adipoyl-7-ADCA potassium salt aqueous solution with a concentration of 50,000-70,000 ug / ml, and the temperature is controlled at 18-22°C.

[0010] Preferably, the process of filtering, extracting and purifying the adipoyl-7-ADCA fermentation broth comprises: filtering the adipoyl-7-ADCA fermentation broth through a pressurized plate frame, mixing the filtrate with butyl acetate and a surfactant under acidic conditions of pH=1.8 for extraction, and then washing and decolorizing, adding potassium carbonate and mixing and stirring for extraction to obtain an aqueous solution of adipoyl-7-ADCA potassium salt.

[0011] Preferably, the concentration of the dilute sulfuric acid is 25%-35%, and the temperature is controlled at 18-22°C.

[0012] Preferably, in step 3, the process of adjusting the pH of the initial crystallization solution comprises: The stirring speed was adjusted to 40-50 r / s, and then dilute sulfuric acid was slowly added to the initial crystallization solution. The pH change rate was controlled at 0.1-0.2 / min. When the pH reached 0.89-0.91, the addition of dilute sulfuric acid was stopped, and the crystallization solution was slowly cooled at a cooling rate of 0.2-0.3°C / min. After cooling to 5-7°C, stirring was continued for 30 minutes to obtain adipoyl-7-ADCA crystallization solution.

[0013] Preferably, in step three, the filtration is to pump the adipoyl-7-ADCA crystal solution into a plate and frame filter press using a feed pump, and perform pressure filtration at a pressure of 0.4 MPa to obtain adipoyl-7-ADCA acid crystal cake.

[0014] Preferably, in step 4, the washing and drying are as follows: first, purified water is adjusted to acidic water with a pH of 1, and then the adipoyl-7-ADCA acid crystal filter cake is washed twice with the acidic water, and the amount of acidic water is controlled to be 50% of the volume of the adipoyl-7-ADCA crystal liquid; then the filter cake is vacuum dried at 60° C., the vacuum degree is controlled at -0.09~-0.1MPa, and the moisture content of the drying end material is controlled to be below 0.5%, and finally adipoyl-7-ADCA acid crystals are obtained.

[0015] The third aspect of the embodiments of the present invention provides the use of the adipoyl-7-ADCA acid crystals or the adipoyl-7-ADCA acid crystals prepared by the method described in the preparation of cephalosporin drugs.

[0016] The technical solution provided by the present invention can have the following beneficial effects: The present invention can prepare adipoyl-7-ADCA acid crystals from an aqueous solution of adipoyl-7-ADCA potassium salt. This preparation method achieves a yield of up to 91% and a purity of up to 97%. The crystals are needle-shaped, and the process conditions are mild, making them suitable for large-scale production. The crystals produced by this process serve as semi-synthetic pharmaceutical intermediates for cephalosporin antibiotics and can be further reacted to synthesize commercially available drugs such as cephalexin, cephradine, cefadroxil, and cefetamet pivoxil. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.

[0018] Figure 1 The accompanying drawing is a scanning electron microscope image of the adipoyl-7-ADCA acid crystals prepared in Example 1; Figure 2 The accompanying drawing is a scanning electron microscope image of adipoyl-7-ADCA acid crystals prepared in Example 2; Figure 3 The accompanying drawing is a scanning electron microscope image of the adipoyl-7-ADCA acid crystals prepared in Example 3. DETAILED DESCRIPTION

[0019] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field are within the scope of protection of the present invention.

[0020] The active ingredient in an aqueous solution of 7-ADCA is the potassium salt of 7-ADCA. 7-ADCA is a salt under neutral conditions and readily soluble in water. Under acidic conditions, it is an acid and readily soluble in organic solvents, with its solubility in organic solvents varying with pH. 7-ADCA is relatively stable under weakly acidic and neutral conditions but is easily destroyed under strongly acidic and alkaline conditions.

[0021] Based on the above situation, the present invention provides a method for preparing adipoyl-7-ADCA acid crystals, as shown in the following examples.

[0022] Example 1 A method for preparing adipoyl-7-ADCA acid crystals according to the present invention comprises the following steps: The pre-fermented adipyl-7-ADCA fermentation broth was filtered and purified by extraction to obtain an aqueous solution of adipyl-7-ADCA potassium salt (content 65086 μg / ml, purity 90.27% by HPLC, pH 7.13, temperature 19°C). The filtration and extraction purification process involved filtering the adipyl-7-ADCA fermentation broth, mixing the filtrate with butyl acetate and a surfactant (pentaethylenehexamine) under acidic conditions of pH 1.8, and stirring at a stirring speed of 53 rpm. The solution was then washed with water at a volume ratio of 10:1. The solution was then decolorized with activated carbon, and then extracted with potassium carbonate, stirring, to obtain an aqueous solution of adipyl-7-ADCA potassium salt.

[0023] 10L of adipoyl-7-ADCA potassium salt aqueous solution was injected into the reactor, and the reactor was stirred at 53 r / min. A 32.83% concentration of dilute sulfuric acid solution was slowly added. The pH change rate was controlled at about 0.16 / min. The pH value of the material was continuously tested until the material pH was 2.51. The acid addition was stopped, the stirring speed was adjusted to 28 r / min, and crystal growth began. After 20 minutes of crystal growth, the stirring speed was adjusted to 45 r / min, and dilute sulfuric acid was continued to be slowly added to the solution until the pH was 0.90. The acid addition was stopped, and the crystallization solution was slowly cooled at a cooling rate of about 0.26°C / min. After cooling to 6°C, stirring was continued for 30 minutes. The crystallization was completed to obtain adipoyl-7-ADCA crystal solution.

[0024] The adipyl-7-ADCA crystal solution was filtered to obtain an adipyl-7-ADCA acid crystal cake. The filter cake was washed with purified water to pH=1.0, filtered, and vacuum dried (temperature 60°C, vacuum degree -0.1MPa). The amount of acidic water used was 50% of the volume of the adipyl-7-ADCA crystal solution; 595.60g of adipyl-7-ADCA acid crystals (wet content 0.4%) were obtained. The product yield was 91.51%, and the purity of the crystals determined by high pressure liquid chromatography (HPLC) was 97.28%, which was 7.01% higher than that of the adipyl-7-ADCA potassium salt aqueous solution. The crystals are needle-shaped, and the scanning electron microscope photograph of the crystals is shown below. Figure 1 shown.

[0025] Example 2 The pre-fermented adipyl-7-ADCA fermentation broth was filtered, extracted, and purified to yield an aqueous solution of adipyl-7-ADCA potassium salt (content 60184 μg / ml, purity 90.84% ​​by HPLC, pH 6.81, temperature 20°C). The filtration and extraction purification process involved filtering the adipyl-7-ADCA fermentation broth, mixing the filtrate with butyl acetate and a surfactant (pentaethylenehexamine) under acidic conditions of pH 1.8, and stirring at 55 rpm. The solution was then washed with water at a water-to-material volume ratio of 10:1. The solution was then decolorized with activated carbon, and then extracted with potassium carbonate, stirring, to yield an aqueous solution of adipyl-7-ADCA potassium salt.

[0026] Pour 10L of adipoyl-7-ADCA potassium salt aqueous solution into the reactor, start stirring the reactor at 50r / min, slowly add 25% dilute sulfuric acid solution, control the pH change rate at 0.1 / min, continuously monitor the pH value of the material until the pH is 2.53, stop adding acid, adjust the stirring speed to 25r / min, and start crystal growth. After 20 minutes of crystal growth, adjust the stirring speed to 40r / min, continue to slowly add dilute sulfuric acid to the solution until the pH is 0.91, stop adding acid, and slowly cool the crystallization solution at a cooling rate of 0.3℃ / min. After cooling to 5℃, continue stirring for 30 minutes. Crystallization is completed to obtain adipoyl-7-ADCA crystal solution.

[0027] The adipyl-7-ADCA crystal solution was filtered to obtain an adipyl-7-ADCA acid crystal cake. The filter cake was washed with purified water to acidic water with a pH of 1.0, filtered, and vacuum dried (temperature 60°C, vacuum degree -0.095MPa). The amount of acidic water used was 50% of the volume of the adipyl-7-ADCA crystal solution to obtain 548.64g of adipyl-7-ADCA acid crystals (wet content 0.3%). The product yield was 91.16%, and the purity of the crystals as determined by high pressure liquid chromatography (HPLC) was 97.78%, which was 6.94% higher than that of the adipyl-7-ADCA potassium salt aqueous solution. The crystals are needle-shaped, and the scanning electron microscope photograph of the crystals is shown below. Figure 2 shown.

[0028] Example 3 The pre-fermented adipyl-7-ADCA fermentation broth was filtered, extracted, and purified to obtain an aqueous solution of adipyl-7-ADCA potassium salt (content 63,342 μg / ml, purity 90.45% by HPLC, pH 6.78, temperature 21°C). The filtration and extraction purification process involved filtering the adipyl-7-ADCA fermentation broth, mixing the filtrate with butyl acetate and a surfactant (pentaethylenehexamine) under acidic conditions of pH 1.8, and stirring at a stirring speed of 56 rpm. The solution was then washed with water at a water-to-material volume ratio of 10:1, decolorized with activated carbon, and then extracted with potassium carbonate to obtain an aqueous solution of adipyl-7-ADCA potassium salt.

[0029] Pour 10L of adipoyl-7-ADCA potassium salt aqueous solution into the reactor, start stirring the reactor at 60r / min, slowly add 35% dilute sulfuric acid solution, control the pH change rate at 0.2 / min, continuously monitor the pH value of the material until the pH is 2.57, stop adding acid, adjust the stirring speed to 30r / min, and start growing crystals. After growing the crystals for 20 minutes, adjust the stirring speed to 50r / min, continue to slowly add dilute sulfuric acid to the solution until the pH is 0.89, stop adding acid, and slowly cool the crystallization solution at a cooling rate of 0.2℃ / min. After cooling to 7℃, continue stirring for 30 minutes. The crystallization is completed to obtain adipoyl-7-ADCA crystal solution.

[0030] The adipyl-7-ADCA crystal solution was filtered under reduced pressure to obtain an adipyl-7-ADCA acid crystal cake. The filter cake was washed with purified water to acidic water with a pH of 1.0, filtered, and vacuum dried (temperature 60°C, vacuum degree -0.09MPa). The amount of acidic water used was 50% of the volume of the adipyl-7-ADCA crystal solution, and 585.91g of adipyl-7-ADCA acid crystals (wet content 0.2%) were obtained. The product yield was 92.50%, and the purity of the crystals was 97.00% as determined by high pressure liquid chromatography (HPLC), which was 6.55% higher than that of the adipyl-7-ADCA potassium salt aqueous solution. The crystals are needle-shaped, and the scanning electron microscope photograph of the crystals is shown below. Figure 3 shown.

[0031] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0032] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein, but is to be construed in the widest possible manner consistent with the principles and novel features disclosed herein.

Claims

1. Adipoyl-7-ADCA acid crystal, the structure of which is shown in formula (I): （I）。 2. The method for preparing adipoyl-7-ADCA acid crystals according to claim 1, wherein The preparation process is as follows: A dilute sulfuric acid solution is added to an aqueous solution of adipoyl-7-ADCA potassium salt, followed by crystallization to obtain diacyl-7-ADCA acid crystals. The specific reaction formula is as follows: 。 3. The preparation method of adipoyl-7-ADCA acid crystals according to claim 2, wherein the specific preparation method comprises: Step 1, preparing an aqueous solution of adipyl-7-ADCA potassium salt and a dilute sulfuric acid solution; wherein the pH of the aqueous solution of adipyl-7-ADCA potassium salt is 6.5-7.5; Step 2: Slowly add dilute sulfuric acid solution to the prepared adipoyl-7-ADCA potassium salt aqueous solution, stir at 50-60 r / s, and when the pH of the solution reaches 2.51-2.57 and the turbidity of the solution changes, immediately stop adding acid and carry out crystal growth. The stirring speed is adjusted to 25-30 r / s, and the initial crystal solution is obtained after 20 minutes; Step 3: adjusting the pH value of the initial crystallization solution, filtering, and obtaining adipoyl-7-ADCA acid crystal cake; Step 4: Wash and dry the adipoyl-7-ADCA acid crystal cake to obtain adipoyl-7-ADCA acid crystals.

4. The method for preparing adipoyl-7-ADCA acid crystals according to claim 3, wherein The process of adipoyl-7-ADCA potassium salt aqueous solution is: The adipoyl-7-ADCA fermentation broth prepared in advance is filtered, extracted and purified to obtain an adipoyl-7-ADCA potassium salt aqueous solution with a concentration of 50,000-70,000 ug / ml, and the temperature is controlled at 18-22°C.

5. The method for preparing adipoyl-7-ADCA acid crystals according to claim 4, wherein The process of filtering, extracting and purifying the adipoyl-7-ADCA fermentation broth includes: filtering the adipoyl-7-ADCA fermentation broth, mixing the filtrate with butyl acetate and a surfactant under acidic conditions of pH=1.8 for extraction, and then washing and decolorizing the filtrate with potassium carbonate, mixing and stirring for extraction to obtain an adipoyl-7-ADCA potassium salt aqueous solution.

6. The method for preparing adipoyl-7-ADCA acid crystals according to claim 5, wherein The concentration of the dilute sulfuric acid is 25%-35%, and the temperature is controlled at 18-22°C.

7. The method for preparing adipoyl-7-ADCA acid crystals according to claim 6, wherein In step 3, the process of adjusting the pH of the initial crystallization solution includes: The stirring speed was adjusted to 40-50 r / s, and then dilute sulfuric acid was slowly added to the initial crystallization solution so that the pH change rate of the crystallization solution was controlled at 0.1-0.2 / min. When the pH reached 0.89-0.91, the addition of dilute sulfuric acid was stopped, and the crystallization solution was slowly cooled at a cooling rate of 0.2-0.3°C / min. After cooling to 5-7°C, stirring was continued for 30 minutes to obtain adipoyl-7-ADCA crystallization solution.

8. The method for preparing adipoyl-7-ADCA acid crystals according to claim 7, wherein In step 3, the filtration is performed under a pressure of 0.4 MPa to obtain a crystalline filter cake of adipoyl-7-ADCA acid.

9. The method for preparing adipoyl-7-ADCA acid crystals according to claim 7, wherein In step 4, the washing and drying are as follows: first, purified water is adjusted to acidic water with a pH of 1, and then the adipoyl-7-ADCA acid crystal filter cake is washed twice with the acidic water, and the amount of acidic water used is 50% of the volume of the adipoyl-7-ADCA crystal liquid; then the filter cake is vacuum dried at 60°C, the vacuum degree is controlled at -0.09~-0.1MPa, and the moisture content of the drying end material is controlled to be below 0.5%, and finally adipoyl-7-ADCA acid crystals are obtained.

10. Use of the adipoyl-7-ADCA acid crystals according to claim 1 or the adipoyl-7-ADCA acid crystals prepared by the method according to any one of claims 2 to 9 in the preparation of cephalosporins.

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