CN115838363B - Purification method of mycophenolic acid - Google Patents
Purification method of mycophenolic acid Download PDFInfo
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- 229960000951 mycophenolic acid Drugs 0.000 title claims abstract description 111
- HPNSFSBZBAHARI-RUDMXATFSA-N mycophenolic acid Chemical compound OC1=C(C\C=C(/C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-RUDMXATFSA-N 0.000 title claims abstract description 111
- HPNSFSBZBAHARI-UHFFFAOYSA-N micophenolic acid Natural products OC1=C(CC=C(C)CCC(O)=O)C(OC)=C(C)C2=C1C(=O)OC2 HPNSFSBZBAHARI-UHFFFAOYSA-N 0.000 title claims abstract description 108
- 238000000034 method Methods 0.000 title claims abstract description 43
- 238000000746 purification Methods 0.000 title claims abstract description 37
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims abstract description 30
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims abstract description 26
- 238000001914 filtration Methods 0.000 claims abstract description 26
- 239000013078 crystal Substances 0.000 claims abstract description 25
- 238000000855 fermentation Methods 0.000 claims abstract description 25
- 230000004151 fermentation Effects 0.000 claims abstract description 25
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- 238000003756 stirring Methods 0.000 claims abstract description 19
- 238000001035 drying Methods 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims abstract description 13
- 235000019799 monosodium phosphate Nutrition 0.000 claims abstract description 13
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims abstract description 13
- 235000017557 sodium bicarbonate Nutrition 0.000 claims abstract description 13
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims abstract description 13
- 238000000605 extraction Methods 0.000 claims abstract description 12
- 238000005406 washing Methods 0.000 claims abstract description 11
- 238000001816 cooling Methods 0.000 claims abstract description 9
- 239000000706 filtrate Substances 0.000 claims description 30
- 239000000047 product Substances 0.000 claims description 24
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 12
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 9
- DKPFZGUDAPQIHT-UHFFFAOYSA-N butyl acetate Chemical compound CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 claims description 8
- 241000228145 Penicillium brevicompactum Species 0.000 claims description 7
- 238000002425 crystallisation Methods 0.000 claims description 7
- 230000008025 crystallization Effects 0.000 claims description 7
- 229940098377 penicillium brevicompactum Drugs 0.000 claims description 7
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 6
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 6
- GJRQTCIYDGXPES-UHFFFAOYSA-N iso-butyl acetate Natural products CC(C)COC(C)=O GJRQTCIYDGXPES-UHFFFAOYSA-N 0.000 claims description 3
- FGKJLKRYENPLQH-UHFFFAOYSA-M isocaproate Chemical compound CC(C)CCC([O-])=O FGKJLKRYENPLQH-UHFFFAOYSA-M 0.000 claims description 3
- OQAGVSWESNCJJT-UHFFFAOYSA-N isovaleric acid methyl ester Natural products COC(=O)CC(C)C OQAGVSWESNCJJT-UHFFFAOYSA-N 0.000 claims description 3
- 239000012141 concentrate Substances 0.000 claims 1
- 239000012535 impurity Substances 0.000 abstract description 11
- 238000002360 preparation method Methods 0.000 abstract description 8
- 239000003814 drug Substances 0.000 abstract description 2
- 239000000243 solution Substances 0.000 description 39
- 230000000052 comparative effect Effects 0.000 description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
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- 239000003153 chemical reaction reagent Substances 0.000 description 3
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- RTGDFNSFWBGLEC-SYZQJQIISA-N mycophenolate mofetil Chemical compound COC1=C(C)C=2COC(=O)C=2C(O)=C1C\C=C(/C)CCC(=O)OCCN1CCOCC1 RTGDFNSFWBGLEC-SYZQJQIISA-N 0.000 description 3
- 229960004866 mycophenolate mofetil Drugs 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical class O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical group CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- XUKUURHRXDUEBC-SXOMAYOGSA-N (3s,5r)-7-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-propan-2-ylpyrrol-1-yl]-3,5-dihydroxyheptanoic acid Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-SXOMAYOGSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- 101710130167 Mannitol-1-phosphate 5-dehydrogenase Proteins 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 230000000843 anti-fungal effect Effects 0.000 description 1
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- 230000001506 immunosuppresive effect Effects 0.000 description 1
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- 230000000670 limiting effect Effects 0.000 description 1
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- 239000000126 substance Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
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- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention belongs to the technical field of medicine preparation, and particularly relates to a purification method of mycophenolic acid. The invention provides a purification method of mycophenolic acid, which comprises the following steps: s1, filtering a fermentation culture solution containing mycophenolic acid, adding sodium dihydrogen phosphate, stirring, adjusting pH, adding an extractant for extraction, and separating a water layer to leave an extract for later use; s2, filtering the extract obtained in the step S1, washing with sodium bicarbonate solution, and separating a water layer; s3, concentrating the solution washed in the step S2 under reduced pressure, adding triethylamine, stirring, cooling, crystallizing and filtering; s4, drying the mycophenolic acid crystal obtained in the step S3 under reduced pressure. The purification method of the mycophenolic acid provided by the invention is simple, and can efficiently remove the Z isomer of the key impurity of the mycophenolic acid, so that the Z isomer content in the mycophenolic acid is below 0.10%.
Description
Technical Field
The invention belongs to the technical field of medicine preparation, and particularly relates to a purification method of mycophenolic acid.
Background
Mycophenolic acid (MycophenolicAcid, MPA, also known as mycophenolic acid) is an antifungal, antitumor and immunosuppressive antibiotic produced by penicillium brevicompactum, is a highly efficient, selective, non-competitive, reversible hypoxanthine mononucleotide dehydrogenase (MPDH) inhibitor, can inhibit the initial synthesis pathway of guanine nucleotides, deplete guanine nucleotides, and further block DNA synthesis. Clinically, it is often used in the form of Mycophenolate Mofetil (MMF) and mycophenolate sodium (MPS). The method for producing the mycophenolic acid comprises two ways of chemical synthesis and biological synthesis, wherein the chemical synthesis method is difficult to industrialize due to the large number of steps and low yield, and the international large-scale production adopts the fermentation liquor containing the mycophenolic acid prepared by the microbial fermentation production method of penicillium, and the mycophenolic acid is obtained through extraction and purification.
In the process of fermentation, extraction and purification of mycophenolic acid, an impurity Z isomer is generated simultaneously, is a key impurity in mycophenolic acid, is difficult to remove because of the high similarity of the structure of the mycophenolic acid and the mycophenolic acid, and finally is brought into mycophenolate mofetil or mycophenolate sodium, so that the quality I of a finished product is influenced. The Z isomer (sodium mycophenolate European Pharmacopeia 10.3 impurity B) has the following structure:
Chinese patent CN109020933B discloses a method for purifying mycophenolic acid, comprising the steps of: 1) Adding mycophenolic acid into a mixed solution of acetone and hydrochloric acid, heating, stirring and dissolving; 2) Adding active carbon, and stirring for decoloring; 3) Filtering; 4) Dropwise adding a solvent B into the filtrate; 5) Preserving heat and crystallizing; 6) Filtering, washing and drying to obtain the pure mycophenolic acid product. Chinese patent application CN103880798a discloses a method for purifying mycophenolic acid. The method comprises the following steps: 1) Providing an ethanol solution of mycophenolic acid having a temperature of 50 ℃ to 70 ℃; 2) Cooling the ethanol solution obtained in the step 1) to 10-30 ℃ to crystallize and separate out mycophenolic acid, and separating out mycophenolic acid crystals; 3) Dissolving the mycophenolic acid crystals obtained in the step 2) by using ethanol, mixing the obtained solution with water with the temperature of 5-12 ℃ to recrystallize and separate out the mycophenolic acid, and separating to obtain the mycophenolic acid recrystallized crystals.
In the above disclosed extraction and purification technology of mycophenolic acid, although the mycophenolic acid has a certain purification effect, the method has the defects of complex purification process, poor purification effect and the like, and only the removal process for impurity A is provided, so that a method for efficiently removing Z isomer which is a key impurity of mycophenolic acid is not disclosed.
Therefore, a purification method of mycophenolic acid needs to be developed, the impurity Z isomer generated in the fermentation and extraction purification processes of mycophenolic acid can be effectively removed, the preparation method is simple and controllable, and the purification effect is good.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a purification method of mycophenolic acid. The purification method of the mycophenolic acid provided by the invention is simple, the steps of the purification method are synergistic, and the Z isomer of key impurities of the mycophenolic acid can be removed efficiently, so that the content of the Z isomer in the mycophenolic acid is below 0.10%.
The technical scheme of the invention is as follows:
A method for purifying mycophenolic acid, comprising the steps of:
S1, filtering a fermentation culture solution containing mycophenolic acid, measuring the product quantity in the filtrate, adding sodium dihydrogen phosphate into the filtrate, stirring, adjusting the pH value, adding an extractant for extraction, separating a water layer, and leaving an extract for later use; the product quantity is the mass of the mycophenolic acid in the filtrate;
S2, filtering the extract obtained in the step S1 by using diatomite, washing by using sodium bicarbonate solution, and separating a water layer;
s3, concentrating the solution washed in the step S2 under reduced pressure to obtain a concentrated solution, adding triethylamine, stirring, cooling, crystallizing, and filtering to obtain a mycophenolic acid crystal;
s4, drying the mycophenolic acid crystal obtained in the step S3 under reduced pressure to obtain the purified mycophenolic acid crystal.
Further, the fermentation broth containing mycophenolic acid in the step S1 is a conventional fermentation broth obtained by fermenting penicillium brevicompactum (Penicillium Brevicompactum). The specific preparation method is as disclosed in example 2 in the specification of a fermentation process of mycophenolic acid in Chinese patent CN 109929890B.
Further, in the step S1, the filtering mode of the fermentation broth containing mycophenolic acid is plate-frame filtration.
Further, the addition amount of the sodium dihydrogen phosphate in the step S1 is 0.8-1.5% of the product amount in the filtrate.
Further, the addition amount of the sodium dihydrogen phosphate in the step S1 is 1% of the product amount in the filtrate.
Further, in the step S1, the stirring time after adding sodium dihydrogen phosphate is 1-2 hours; and (3) stirring, and then adjusting the pH to 4.0-5.0 by adopting a phosphoric acid solution with the mass concentration of 85%.
Further, the extractant in the step S1 is one of ethyl acetate, toluene, n-butyl acetate and isobutyl acetate.
Furthermore, the addition amount of the extractant in the step S1 is 1.5-2.5 times of the volume of the filtrate, and the extraction time is 2-3 hours.
Further, the mass concentration of the sodium bicarbonate solution in the step S2 is 0.3-1.0%, the addition amount is 25-35% of the volume of the extract liquid obtained in the step S1, and the washing time by adopting the sodium bicarbonate solution is 0.5-2 h.
Further, the mass concentration of the sodium bicarbonate solution in the step S2 is 0.5%, and the addition amount is 30% of the volume of the extract obtained in the step S1.
Further, in the step S3, the solution washed in the step S2 is decompressed and concentrated to 25 times of the product amount; the amount of the product is by mass, and the concentrated solution in step S2 is by volume.
Further, the addition amount of the triethylamine in the step S3 is 6-10% of the volume of the concentrated solution, and the stirring time is 30-60 min.
Further, in the step S3, the temperature at the time of decompression concentration is 40-55 ℃, and the temperature at the time of cooling crystallization is 5-15 ℃.
Further, the temperature in the step S4 is 40-55 ℃ during the drying under reduced pressure, and the mycophenolic acid crystals are dried under reduced pressure until the Loss On Drying (LOD) is less than or equal to 1%.
The invention provides a purification method of mycophenolic acid, which has simple overall purification process and excellent effect of removing Z isomer of impurities, so that the content of Z isomer in the mycophenolic acid is below 0.10%. In the purification process, bacterial residues and fermentation residues in the mycophenolic acid fermentation culture solution are removed better by adopting plate and frame filtration and the like, so that filtrate containing products is collected, and then Z isomer impurities in the mycophenolic acid can be removed effectively by adding sodium dihydrogen phosphate before extraction, washing the extract with sodium bicarbonate solution after extraction and adding a triethylamine solution with a specific addition amount before crystallization.
Compared with the prior art, the purification method of mycophenolic acid provided by the invention has the following advantages:
(1) The purification method of the mycophenolic acid provided by the invention is simple and easy to operate, the method is simple and easy to control, the extractant can be recycled, the production cost is low, the method is safe and environment-friendly, and the industrial popularization and application are facilitated.
(2) According to the invention, through the synergistic effect of a plurality of steps of the purification method, the Z isomer of the key impurity of the mycophenolic acid can be removed efficiently, so that the content of the Z isomer in the mycophenolic acid is below 0.10%.
Detailed Description
The invention is further illustrated by the following description of specific embodiments, which are not intended to be limiting, and various modifications or improvements can be made by those skilled in the art in light of the basic idea of the invention, but are within the scope of the invention as long as they do not depart from the basic idea of the invention.
In the following examples and comparative examples, the reagents not specifically described are conventional reagents, and can be purchased from conventional reagent manufacturing and selling companies.
Example 1 purification method of mycophenolic acid
The purification method of the mycophenolic acid comprises the following steps:
s1, filtering a fermentation culture solution containing mycophenolic acid through a plate frame, collecting a filtrate, measuring the product quantity in the filtrate, adding sodium dihydrogen phosphate with the product quantity of 0.8%, stirring for 1h, adjusting the pH value of the filtrate to 4.0-5.0 by using phosphoric acid with the mass concentration of 85%, extracting for 2h by using ethyl acetate with the volume of 1.5 times of the filtrate, separating a water layer, and leaving an extract for later use;
s2, filtering the extract obtained in the step S1 by using diatomite, adding sodium bicarbonate solution with the mass concentration of 0.4% and the volume of 30% of the extract, washing for 1 hour at room temperature, and separating a water layer;
S3, concentrating the solution washed in the step S2 to 25 times of the product amount (the product amount is calculated by mass, the concentrated solution in the step S2 is calculated by volume) under reduced pressure at 40 ℃, then adding triethylamine accounting for 6% of the volume of the concentrated solution, stirring for 30min, cooling to 5 ℃ for crystallization, and filtering to obtain mycophenolic acid crystals;
s4, drying the mycophenolic acid crystal obtained in the step S3 at 40 ℃ under reduced pressure until the Loss On Drying (LOD) is less than or equal to 1%.
The fermentation culture solution containing the mycophenolic acid in the step S1 is a conventional fermentation solution containing the mycophenolic acid obtained by fermenting penicillium brevicompactum (Penicillium Brevicompactum). The specific preparation method is as disclosed in example 2 in the specification of a fermentation process of mycophenolic acid in Chinese patent CN 109929890B.
Example 2 purification method of mycophenolic acid
The purification method of the mycophenolic acid comprises the following steps:
S1, filtering a fermentation culture solution containing mycophenolic acid through a plate frame, collecting a filtrate, measuring the product quantity in the filtrate, adding sodium dihydrogen phosphate with the product quantity of 1 percent into the filtrate, stirring for 1.5 hours, regulating the pH value of the filtrate to 4.0-5.0 by using a phosphoric acid solution with the mass concentration of 85 percent, extracting the filtrate with n-butyl acetate with the volume of 2 times of the filtrate for 3 hours, separating a water layer, and leaving an extract for later use;
s2, filtering the extract obtained in the step S1 by using diatomite, adding sodium bicarbonate solution with the mass concentration of 0.5% and the volume of 30% of the extract, washing for 1h at room temperature, and separating a water layer;
s3, concentrating the solution washed in the step S2 to 25 times of the product amount (the product amount is calculated by mass, the concentrated solution in the step S2 is calculated by volume) under reduced pressure at 45 ℃, then adding triethylamine accounting for 8% of the volume of the concentrated solution, stirring for 40min, cooling to 10 ℃ for crystallization, and filtering to obtain mycophenolic acid crystals;
S4, drying the mycophenolic acid crystal obtained in the step S3 at 45 ℃ under reduced pressure until the Loss On Drying (LOD) is less than or equal to 1%.
The preparation method of the mycophenolic acid-containing fermentation broth in the step S1 is the same as that of example 1.
Example 3 purification method of mycophenolic acid
The purification method of the mycophenolic acid comprises the following steps:
S1, filtering a fermentation culture solution containing mycophenolic acid through a plate frame, collecting a filtrate, measuring the product quantity in the filtrate, adding sodium dihydrogen phosphate with the product quantity of 1.5%, stirring for 2 hours, regulating the pH value of the filtrate to 4.0-5.0 by using a phosphoric acid solution with the mass concentration of 85%, extracting with isobutyl acetate with the volume of 2.5 times of the filtrate for 2 hours, separating a water layer, and leaving an extract for later use;
s2, filtering the extract obtained in the step S1 by using diatomite, adding sodium bicarbonate solution with the mass concentration of 0.8% and the volume of 30% of the extract, washing for 2 hours at room temperature, and separating a water layer;
S3, concentrating the solution washed in the step S2 to 25 times of the product amount (the product amount is calculated by mass, the concentrated solution in the step S2 is calculated by volume) under reduced pressure at 50 ℃, then stirring for 60min by 10% of triethylamine in the concentrated solution volume, cooling to 15 ℃ for crystallization, and filtering to obtain the mycophenolic acid crystals.
S4, drying the mycophenolic acid crystal obtained in the step S3 at 55 ℃ under reduced pressure until the Loss On Drying (LOD) is less than or equal to 1%.
The preparation method of the mycophenolic acid-containing fermentation broth in the step S1 is the same as that of example 1. Comparative example 1A conventional purification method of mycophenolic acid
The purification method of the mycophenolic acid comprises the following steps:
s1, filtering a fermentation culture solution containing mycophenolic acid through a plate frame, collecting a filtrate, measuring the product quantity in the filtrate, regulating the pH value of the filtrate to 4.0-5.0 by using a phosphoric acid solution with the mass concentration of 85%, extracting for 3 hours by using n-butyl acetate with the volume of 2 times of the filtrate, separating a water layer, and leaving an extract for later use;
s2, filtering the extract obtained in the step S1 by using diatomite;
S3, concentrating the extraction filtrate obtained in the step S2 under reduced pressure at 45 ℃ to 25 times (V/W) volume of the product amount, cooling to 10 ℃ for crystallization, and filtering to obtain mycophenolic acid crystals;
S4, drying the mycophenolic acid crystal obtained in the step S3 at 45 ℃ under reduced pressure until the Loss On Drying (LOD) is less than or equal to 1%.
The preparation method of the mycophenolic acid-containing fermentation broth in the step S1 is the same as that of example 1. Comparative example 2 purification method of mycophenolic acid
Compared with example 2, the difference of comparative example 2 is that triethylamine in the step S3 is replaced with n-hexane, and other parameters and operations are the same as those of example 2.
Comparative example 3 purification method of mycophenolic acid
Comparative example 2 differs from example 2 in that sodium dihydrogen phosphate was not added in step S1, and other parameters and operations were the same as in example 2.
Comparative example 4 purification method of mycophenolic acid
Comparative example 2 differs from example 2 in that no sodium bicarbonate solution washing was used in step S2, and other parameters and operations are the same as example 2.
Test example one, mycophenolic acid Performance detection
1. Test materials: examples 1 to 3, comparative examples 1 to 4 produced mycophenolic acid crystals.
2. The test method comprises the following steps: the content of Z isomer in the mycophenolic acid crystals prepared in examples 1 to 3, comparative examples 1 to 4 was examined by the method of examination of substances related to European pharmacopoeia 10.3 2813 (the content of impurity Z isomer in mycophenolic acid should not exceed 0.10%).
3. Test results
The test results are shown in Table 1.
TABLE 1Z isomer content in mycophenolic acid
| Group of | Z isomer content (%) |
| Example 1 | 0.04 |
| Example 2 | 0.02 |
| Example 3 | 0.05 |
| Comparative example 1 | 0.43 |
| Comparative example 2 | 0.18 |
| Comparative example 3 | 0.15 |
| Comparative example 4 | 0.14 |
As is clear from Table 1, the content of Z isomer in the purified mycophenolic acid crystals obtained in examples 1 to 3 is 0.10% or less, and example 2 is the most effective, but it is only 0.02%, which is the most effective example of the present invention. The Z isomer content of the purified mycophenolic acid crystals in comparative example 1 was 0.43% far exceeding the pharmacopoeia quality standard. In comparative examples 2 to 4, the Z isomer content in the purified mycophenolic acid crystals obtained after the purification process was changed to 0.10% or more.
The above embodiments are merely illustrative of the principles of the present invention and its effectiveness, and are not intended to limit the invention. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is intended that all equivalent modifications and variations of the invention be covered by the claims, which are within the ordinary skill of the art, be within the spirit and scope of the present disclosure.
Claims (6)
1. The purification method of the mycophenolic acid is characterized by comprising the following steps of:
S1, filtering a fermentation culture solution containing mycophenolic acid, adding sodium dihydrogen phosphate into the filtrate, stirring, adjusting the pH, adding an extractant for extraction, separating a water layer, and leaving an extract for later use;
S2, filtering the extract obtained in the step S1, washing with sodium bicarbonate solution, and separating a water layer;
s3, concentrating the solution washed in the step S2 under reduced pressure to obtain a concentrated solution, adding triethylamine, stirring, cooling, crystallizing, and filtering to obtain a mycophenolic acid crystal;
S4, drying the mycophenolic acid crystal obtained in the step S3 under reduced pressure to obtain a purified mycophenolic acid crystal; the addition amount of the sodium dihydrogen phosphate in the step S1 is 0.8-1.5% of the product amount in the filtrate; the stirring time after the sodium dihydrogen phosphate solution is added in the step S1 is 1-2 h; stirring, and then adjusting the pH to 4.0-5.0 by adopting a phosphoric acid solution with the mass concentration of 85%; the fermentation culture solution containing the mycophenolic acid in the step S1 is a fermentation solution containing the mycophenolic acid obtained by fermenting penicillium brevicompactum (Penicillium Brevicompactum).
2. The method for purifying mycophenolic acid according to claim 1, wherein the extractant in the step S1 is one of ethyl acetate, toluene, n-butyl acetate and isobutyl acetate; the addition amount of the extractant is 1.5-2.5 times of the volume of the filtrate, and the extraction time is 2-3 hours.
3. The method for purifying mycophenolic acid according to claim 1, wherein the mass concentration of the sodium bicarbonate solution in the step S2 is 0.3-1.0%, and the addition amount is 25% -35% of the volume of the extract obtained in the step S1; the washing time by adopting sodium bicarbonate solution is 0.5-2 h.
4. The method for purifying mycophenolic acid according to claim 1, wherein the amount of triethylamine in the step S3 is 6% to 10% of the volume of the concentrate.
5. The method for purifying mycophenolic acid according to claim 1, wherein the concentration under reduced pressure in the step S3 is performed at a temperature of 40 to 55℃and the crystallization under reduced temperature is performed at a temperature of 5 to 15 ℃.
6. The method for purifying mycophenolic acid according to claim 1, wherein the temperature at the time of drying under reduced pressure in the step S4 is 40-55 ℃, and the mycophenolic acid crystals are dried under reduced pressure until the loss on drying is less than or equal to 1%.
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| CN109020933A (en) * | 2017-06-09 | 2018-12-18 | 鲁南制药集团股份有限公司 | A kind of purification process of Mycophenolic Acid |
| CN110922371A (en) * | 2019-12-27 | 2020-03-27 | 广东蓝宝制药有限公司 | Preparation method of M2 crystal form meclofenol sodium |
| CN112645912A (en) * | 2020-12-27 | 2021-04-13 | 广东蓝宝制药有限公司 | Preparation method of high-purity M2 crystal form meclofenol sodium |
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