CN115124552B - Preparation method of deacetylated cefathiamidine - Google Patents
Preparation method of deacetylated cefathiamidine Download PDFInfo
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- CN115124552B CN115124552B CN202210680073.4A CN202210680073A CN115124552B CN 115124552 B CN115124552 B CN 115124552B CN 202210680073 A CN202210680073 A CN 202210680073A CN 115124552 B CN115124552 B CN 115124552B
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- desacetylcefathiamidine
- bromoacetyl
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- cefathiamidine
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- JYXACOFERDBGGQ-RHSMWYFYSA-N cefathiamidine Chemical compound S1CC(COC(C)=O)=C(C(O)=O)N2C(=O)[C@@H](NC(=O)CSC(NC(C)C)=NC(C)C)[C@H]21 JYXACOFERDBGGQ-RHSMWYFYSA-N 0.000 title claims abstract description 44
- 229950005040 cefathiamidine Drugs 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 37
- LSTRKXWIZZZYAS-UHFFFAOYSA-N 2-bromoacetyl bromide Chemical compound BrCC(Br)=O LSTRKXWIZZZYAS-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000012043 crude product Substances 0.000 claims abstract description 13
- KREOCUNMMFZOOS-UHFFFAOYSA-N 1,3-di(propan-2-yl)thiourea Chemical compound CC(C)NC(S)=NC(C)C KREOCUNMMFZOOS-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 38
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 36
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 36
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 36
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 30
- 239000007787 solid Substances 0.000 claims description 27
- 238000006243 chemical reaction Methods 0.000 claims description 26
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 22
- 238000001914 filtration Methods 0.000 claims description 22
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
- 239000003960 organic solvent Substances 0.000 claims description 21
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 20
- 239000012071 phase Substances 0.000 claims description 13
- 238000003756 stirring Methods 0.000 claims description 13
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- 239000002904 solvent Substances 0.000 claims description 12
- 238000005406 washing Methods 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 11
- 239000011259 mixed solution Substances 0.000 claims description 11
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 10
- 229910052799 carbon Inorganic materials 0.000 claims description 10
- 239000002585 base Substances 0.000 claims description 9
- 239000003480 eluent Substances 0.000 claims description 9
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 8
- 238000002425 crystallisation Methods 0.000 claims description 8
- 230000008025 crystallization Effects 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 6
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-diisopropylethylamine Substances CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 6
- 238000007670 refining Methods 0.000 claims description 6
- 230000001276 controlling effect Effects 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 5
- 230000014759 maintenance of location Effects 0.000 claims description 5
- 238000002156 mixing Methods 0.000 claims description 5
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 5
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 4
- 239000003513 alkali Substances 0.000 claims description 4
- 238000001514 detection method Methods 0.000 claims description 4
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 4
- 239000002253 acid Substances 0.000 claims description 3
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical group O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 3
- 238000012856 packing Methods 0.000 claims description 3
- 239000008346 aqueous phase Substances 0.000 claims description 2
- 239000012046 mixed solvent Substances 0.000 claims description 2
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 2
- 230000001105 regulatory effect Effects 0.000 claims description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 2
- 125000003944 tolyl group Chemical group 0.000 claims 1
- 239000002994 raw material Substances 0.000 abstract description 6
- 229940079593 drug Drugs 0.000 abstract description 4
- 239000003814 drug Substances 0.000 abstract description 4
- 239000012535 impurity Substances 0.000 abstract description 4
- 239000013558 reference substance Substances 0.000 abstract description 2
- 239000000463 material Substances 0.000 description 16
- 239000013078 crystal Substances 0.000 description 7
- 239000007788 liquid Substances 0.000 description 7
- 238000001228 spectrum Methods 0.000 description 7
- 238000005481 NMR spectroscopy Methods 0.000 description 6
- 238000001291 vacuum drying Methods 0.000 description 5
- 229930186147 Cephalosporin Natural products 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 4
- 229940124587 cephalosporin Drugs 0.000 description 4
- 150000001780 cephalosporins Chemical class 0.000 description 4
- 239000007795 chemical reaction product Substances 0.000 description 4
- 238000007599 discharging Methods 0.000 description 4
- 239000012065 filter cake Substances 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- HSHGZXNAXBPPDL-HZGVNTEJSA-N 7beta-aminocephalosporanic acid Chemical compound S1CC(COC(=O)C)=C(C([O-])=O)N2C(=O)[C@@H]([NH3+])[C@@H]12 HSHGZXNAXBPPDL-HZGVNTEJSA-N 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 229930182555 Penicillin Natural products 0.000 description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 239000006227 byproduct Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000011010 flushing procedure Methods 0.000 description 2
- 239000000543 intermediate Substances 0.000 description 2
- 238000010829 isocratic elution Methods 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- QAXZWHGWYSJAEI-UHFFFAOYSA-N n,n-dimethylformamide;ethanol Chemical compound CCO.CN(C)C=O QAXZWHGWYSJAEI-UHFFFAOYSA-N 0.000 description 2
- 229940049954 penicillin Drugs 0.000 description 2
- 238000002953 preparative HPLC Methods 0.000 description 2
- 238000004262 preparative liquid chromatography Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000004537 pulping Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- KWIUHFFTVRNATP-UHFFFAOYSA-N Betaine Natural products C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 1
- HZZVJAQRINQKSD-UHFFFAOYSA-N Clavulanic acid Natural products OC(=O)C1C(=CCO)OC2CC(=O)N21 HZZVJAQRINQKSD-UHFFFAOYSA-N 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- -1 N, N-diisopropylamidinothio Chemical group 0.000 description 1
- KWIUHFFTVRNATP-UHFFFAOYSA-O N,N,N-trimethylglycinium Chemical compound C[N+](C)(C)CC(O)=O KWIUHFFTVRNATP-UHFFFAOYSA-O 0.000 description 1
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 125000000738 acetamido group Chemical group [H]C([H])([H])C(=O)N([H])[*] 0.000 description 1
- 125000003668 acetyloxy group Chemical group [H]C([H])([H])C(=O)O[*] 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 229960003237 betaine Drugs 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 229960003324 clavulanic acid Drugs 0.000 description 1
- HZZVJAQRINQKSD-PBFISZAISA-N clavulanic acid Chemical compound OC(=O)[C@H]1C(=C/CO)/O[C@@H]2CC(=O)N21 HZZVJAQRINQKSD-PBFISZAISA-N 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000012527 feed solution Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 239000012450 pharmaceutical intermediate Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 210000001635 urinary tract Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
- C07D501/02—Preparation
- C07D501/04—Preparation from compounds already containing the ring or condensed ring systems, e.g. by dehydrogenation of the ring, by introduction, elimination or modification of substituents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
- C07D501/02—Preparation
- C07D501/12—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D501/00—Heterocyclic compounds containing 5-thia-1-azabicyclo [4.2.0] octane ring systems, i.e. compounds containing a ring system of the formula:, e.g. cephalosporins; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
- C07D501/14—Compounds having a nitrogen atom directly attached in position 7
- C07D501/16—Compounds having a nitrogen atom directly attached in position 7 with a double bond between positions 2 and 3
- C07D501/20—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids
- C07D501/24—7-Acylaminocephalosporanic or substituted 7-acylaminocephalosporanic acids in which the acyl radicals are derived from carboxylic acids with hydrocarbon radicals, substituted by hetero atoms or hetero rings, attached in position 3
- C07D501/26—Methylene radicals, substituted by oxygen atoms; Lactones thereof with the 2-carboxyl group
- C07D501/28—Methylene radicals, substituted by oxygen atoms; Lactones thereof with the 2-carboxyl group with the 7-amino radical acylated by an aliphatic carboxylic acid, which is substituted by hetero atoms
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Cephalosporin Compounds (AREA)
Abstract
The invention belongs to the technical field of pharmacy, and relates to a preparation method of deacetylated cefathiamidine. The preparation method comprises the following steps: reacting D-7-ACA with bromoacetyl bromide, and separating by high performance liquid chromatography to obtain N-bromoacetyl-D-7-ACA; reacting N-bromoacetyl-D-7-ACA with N, N' -diisopropylthiourea to obtain a crude product of the deacetylated cefathiamidine; the crude product of the deacetylated cefathiamidine is refined to obtain the purified deacetylated cefathiamidine. The purity of the deacetylated cefathiamidine obtained by the preparation method reaches more than 99.0%, and the deacetylated cefathiamidine can be used as a standard reference substance to accurately measure the content of deacetylated cefathiamidine impurities in cefathiamidine raw materials and preparations and ensure medication safety.
Description
Technical Field
The invention belongs to the technical field of pharmacy, and relates to a preparation method of deacetylated cefathiamidine.
Background
Cefathiamidine, also known as cephalosporin 18, is a first-generation cephalosporin antibacterial agent which is self-developed in China, plays an antibacterial role by inhibiting the synthesis of bacterial cell walls, has a strong effect on staphylococcus aureus, streptococcus viridae and pneumococcus, has unique antibacterial activity on enterococcus, and is mainly used for respiratory systems, hepatobiliary systems, five sense organs and urinary tracts caused by sensitive bacteria. The chemical name is as follows: (6R, 7R) -3- [ (acetoxy) methyl ] -7- [ alpha- (N, N-diisopropylamidinothio) acetamido ] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid betaine of formula C 19H28N4O6S2.
At present, 7-ACA is commonly used as a raw material for producing cefathiamidine, the raw material reacts with bromoacetyl bromide to generate N-bromoacetyl-7-ACA, the N-bromoacetyl-7-ACA reacts with N, N' -diisopropylthiourea to generate a cefathiamidine crude product, and the crude product is further refined to obtain the cefathiamidine meeting pharmacopoeia requirements, wherein the reaction formula of the production process is as follows:
In the preparation of cefathiamidine, 7-ACA raw material is mixed with desacetyl 7-ACA (hereinafter referred to as D-7-ACA), and thus desacetyl cefathiamidine is an impurity which may be present in cefathiamidine. In addition, cefathiamidine may be degraded during storage to give desacetylcefathiamidine. Therefore, it is necessary to synthesize the desacetylcefathiamidine as a standard reference to accurately measure the content of the impurity in the cefathiamidine.
It is also contemplated that desacetylcefathiamidine is synthesized by the same chemical synthesis route as cefathiamidine, i.e., that desacetyl7-ACA is used as a raw material to synthesize desacetylcefathiamidine. However, as is known from the structural formula of D-7-ACA, D-7-ACA has-OH at the 3-position and-NH 2 at the 7-position, i.e., a plurality of reaction sites, and in the reaction of D-7-ACA with bromoacetyl bromide, the following products are produced:
I.e., bromoacetyl bromide will react with the-OH at the 3-position or the-NH 2 at the 7-position of D-7-ACA or with them simultaneously, and thus N-bromoacetyl-D-7-ACA cannot be obtained exclusively.
Disclosure of Invention
The Wikida pharmaceutical industry limited company of the national pharmaceutical group is a comprehensive pharmaceutical enterprise controlled by the Chinese pharmaceutical group, is one of the large antibiotic industry bases of the national pharmaceutical group, and mainly comprises cephalosporin, penicillin pharmaceutical intermediates, bulk drugs, powder injection and clavulanic acid pharmaceutical bulk drugs, thus being a leading-edge enterprise in China in terms of production and development of cephalosporin and penicillin antibiotics. Aiming at the problems, research and development personnel grope and separate the N-bromoacetyl-D-7-ACA, and then the N-bromoacetyl-D-7-ACA reacts with N, N' -diisopropylthiourea to obtain the deacetylated cefathiamidine.
Therefore, the invention aims to provide a preparation method of deacetylated cefathiamidine.
According to the invention, the preparation method of the deacetylated cefathiamidine provided by the invention comprises the following steps:
(1) Preparation of N-bromoacetyl-D-7-ACA
Dissolving D-7-ACA in water at 0-40 ℃ in the presence of alkali, then adding a first organic solvent, dropwise adding bromoacetyl bromide, and stirring for reaction for 1-4 hours; after the reaction is finished, standing and layering to separate out a water phase or distilling under reduced pressure to remove an organic solvent, thereby obtaining the water phase; at room temperature, regulating the pH of the water phase to 0.5-2.5 by using acid, crystallizing and separating out solid, and filtering to obtain a solid;
Dissolving the solid in a second organic solvent, and separating by adopting a preparation high performance liquid chromatography to obtain N-bromoacetyl-D-7-ACA, wherein the preparation high performance liquid chromatography is as follows:
chromatographic column: c18 packing column;
eluent: water and acetonitrile mixed solution;
Collecting the elution fraction with retention time of 10-11min, mixing the eluates of the fraction with purity of more than or equal to 98%, concentrating, crystallizing and separating out solid, filtering, washing with water, and drying to obtain N-bromoacetyl-D-7-ACA solid;
(2) Preparation of desacetylcefathiamidine
Dissolving N-bromoacetyl-D-7-ACA in a third organic solvent in the presence of alkali at 0-40 ℃, and then adding N, N' -diisopropylthiourea; controlling the reaction temperature to be between 30 and 40 ℃ and stirring the mixture for reaction for 1 to 4 hours; after the reaction is finished, cooling to 0-10 ℃, crystallizing and separating out solid, and filtering to obtain a crude product of the deacetylated cefathiamidine;
(3) Refining of desacetylcefathiamidine
Dissolving the crude product of the deacetylated cefathiamidine in a fourth solvent at the temperature of 0-20 ℃, decolorizing with active carbon, adding a crystallization solvent into the filtrate after filtering, crystallizing and separating out the deacetylated cefathiamidine, and filtering, washing and drying to obtain the purified deacetylated cefathiamidine.
The preparation method of the desacetylcefathiamidine of the present invention is described in more detail below.
In the preparation of the N-bromoacetyl-D-7-ACA in the step (1), D-7-ACA is dissolved in water at 0 to 40 ℃, preferably 0 to 5 ℃ in the presence of a base, wherein the base is one or two selected from diethylamine, triethylamine, N-diisopropylethylamine, potassium carbonate, sodium carbonate and sodium bicarbonate, and preferably triethylamine and/or sodium bicarbonate. Then adding a first organic solvent which is one or two selected from toluene, dichloromethane, acetone, tetrahydrofuran and N, N-dimethylformamide, and preferably toluene or acetone; the ratio of the first organic solvent to water is 0.4-2.5:1 by volume.
Bromoacetyl bromide is added in a dropwise manner, and the molar ratio of D-7-ACA to bromoacetyl bromide is 1:1 to 3, and preferably 1:1.2 to 3.0. Under stirring, D-7-ACA reacts with bromoacetyl bromide for 1 to 4 hours, D-7-ACA has-OH at the 3-position and-NH 2 at the 7-position, namely a plurality of reaction sites, and in the reaction of D-7-ACA and bromoacetyl bromide, the following 3 products are produced:
Thus, N-bromoacetyl-D-7-ACA needs to be separated from the reaction product. After the reaction of D-7-ACA and bromoacetyl bromide is completed, standing and layering to separate out water phase, or distilling under reduced pressure to remove organic solvent, thereby obtaining water phase. The aqueous phase is adjusted to a pH of 0.5 to 2.5, preferably 1.0 to 2.0, at room temperature with an acid, which may be hydrochloric acid, for example 6M hydrochloric acid, to crystallize out a solid which is filtered off.
The solid is dissolved in a second organic solvent, which is one or two selected from N, N-dimethylformamide and dimethyl sulfoxide, preferably N, N-dimethylformamide. Separating by high performance liquid chromatography to obtain high purity N-bromoacetyl-D-7-ACA, wherein the conditions of the high performance liquid chromatography are as follows:
chromatographic column: c18 packing column;
Eluent: water and acetonitrile.
More specifically, the process is carried out,
Chromatographic column: c18 packed column (10 μm, 50mm x 250 mm);
eluent: water: acetonitrile=87:13 (V/V);
Detection wavelength: UV254nm;
Flow rate: 16-21L/min;
Collecting the elution fraction with retention time of 10-11min, mixing the eluates of the fraction with purity of more than or equal to 98%, concentrating, crystallizing and separating out solid, filtering, washing with water, and drying to obtain high-purity N-bromoacetyl-D-7-ACA solid.
In the process of separating N-bromoacetyl-D-7-ACA by adopting a preparation high performance liquid chromatography, the inventor has difficulty in separating because byproducts 1 and 2 are similar to the N-bromoacetyl-D-7-ACA in structure. Through repeated experiments, the chromatographic conditions are screened out, and the eluent with the purity of more than or equal to 98 percent is successfully separated.
For example, when the eluent is selected, when a mixed solution of methanol and water is used as a mobile phase, the liquid chromatogram thereof is shown in FIG. 1, and it can be seen that the peaks are irregular, the degree of separation is poor, and N-bromoacetyl-D-7-ACA cannot be separated from byproducts 1 and 2. When the mixed solution of acetonitrile and water is taken as a mobile phase, the liquid chromatogram of the mixed solution is shown as figure 2 at the flow rate of 35L/min, and the separation degree is only 0.2; however, when the flow rate was adjusted to a relatively low flow rate of 16-21L/min, the liquid chromatogram thereof was as shown in FIG. 3, and it was found that a good separation effect was obtained.
In the preparation of the deacetylated cefathiamidine in the step (2), N-bromoacetyl-D-7-ACA is dissolved in a third organic solvent at 0 ℃ to 40 ℃, preferably 0 ℃ to 5 ℃ in the presence of a base, wherein the base is one or two selected from diethylamine, triethylamine and N, N-diisopropylethylamine, and is preferably triethylamine; the third organic solvent is one or two selected from dichloromethane, chloroform, toluene, methanol, ethanol and isopropanol, and is preferably dichloromethane.
In a third organic solvent, N-bromoacetyl-D-7-ACA is dissolved in the presence of a base, followed by the addition of N, N' -diisopropylthiourea. Controlling the reaction temperature to be between 30 and 40 ℃, preferably between 35 and 40 ℃, and stirring and reacting for 1 to 4 hours; after the reaction is finished, the temperature is reduced to 0-10 ℃, preferably 0-5 ℃, solid crystals are separated out, and the crude product of the deacetylated cefathiamidine is obtained through filtration. Wherein, the mol ratio of N-bromoacetyl-D-7-ACA to N, N' -diisopropylthiourea is 1:1 to 3, and preferably 1:1 to 2.
In the refining of the desacetyl cefathiamidine in the step (3), the obtained crude desacetyl cefathiamidine is dissolved in a fourth solvent at the temperature of 0-20 ℃, preferably 0-5 ℃, then activated carbon is decolorized, a crystallization solvent is added into filtrate after filtration, the desacetyl cefathiamidine is crystallized and separated out, and the purified desacetyl cefathiamidine is obtained after filtration, washing and drying.
Wherein the fourth solvent is one or two selected from water, N-dimethylformamide, methanol, ethanol and isopropanol, and is preferably a water-ethanol mixed solvent; the crystallization solvent is selected from one or two of acetone, dichloromethane and ethyl acetate, and is preferably acetone.
The purity of the deacetylated cefathiamidine obtained by the preparation method reaches more than 99.0%, and the deacetylated cefathiamidine can be used as a standard reference substance to accurately measure the content of deacetylated cefathiamidine impurities in cefathiamidine raw materials and preparations and ensure medication safety.
Drawings
FIG. 1 is a high performance liquid chromatogram of the preparation of a reaction product of D-7-ACA and bromoacetyl bromide, wherein a mixed solution of methanol and water is a mobile phase;
FIG. 2 is a high performance liquid chromatogram of the preparation of a reaction product of D-7-ACA and bromoacetyl bromide, wherein a mixed solution of acetonitrile and water is a mobile phase, and the flow rate is 35L/min;
FIG. 3 is a high performance liquid chromatogram of the preparation of a reaction product of D-7-ACA and bromoacetyl bromide, wherein a mixed solution of acetonitrile and water is a mobile phase, and the flow rate is 16-21L/min;
FIG. 4 is a nuclear magnetic resonance hydrogen spectrum of N-bromoacetyl-D-7-ACA prepared in example 1 of the present invention;
FIG. 5 is a nuclear magnetic resonance carbon spectrum of N-bromoacetyl-D-7-ACA prepared in example 1 of the present invention;
FIG. 6 is a nuclear magnetic resonance hydrogen spectrum of desacetylcefathiamidine prepared in example 1 of the present invention;
FIG. 7 is a nuclear magnetic resonance carbon spectrum of desacetylcefathiamidine prepared in example 1 of the present invention.
Detailed Description
The following examples are given to illustrate the process for the preparation of desacetylcefathiamidine according to the invention in further detail, and the scope of the invention is not limited to the following examples, which are given for illustrative purposes only and are not intended to limit the invention in any way.
Example 1
(1) Preparation of N-bromoacetyl-D-7-ACA
375ML of water was added to the reaction flask, cooled to 3 ℃, 15g (0.0651 mol) of D-7-ACA was added under stirring, 16.4g (0.1621 mol) of triethylamine was started to be added dropwise, D-7-ACA was dissolved and then 170mL of toluene was added; 16.6g (0.0822 mol) of bromoacetyl bromide is added dropwise, and the mixture is stirred and reacted for 4 hours at the temperature of 0 ℃ to 5 ℃; then stirring, standing for layering, separating out a water layer, dropwise adding 6M hydrochloric acid at room temperature, adjusting the pH of the feed liquid to 1.0, crystallizing and separating out solid, filtering and washing with water to obtain a solid wet material (34 g).
The solid wet material 34g was dissolved in 60mL of DMF to perform preparative high performance liquid chromatography to isolate N-bromoacetyl-D-7-ACA using the equipment DAC-50 preparative liquid chromatography instrument under the following chromatographic conditions:
chromatographic column: c18 packed column (10 μm, 50mm x 250 mm);
eluent: water: acetonitrile=87:13 (V/V);
Detection wavelength: UV254nm;
isocratic elution flow rate: 18L/min;
sample loading amount: 20-25 mL/time.
Collecting the elution fraction with retention time of 10-11min, mixing the eluates of the fraction with purity of more than or equal to 98%, concentrating under reduced pressure to remove acetonitrile and part of DMF, separating out solid during concentration, stirring for crystal growth for 30min, filtering, pulping and washing the filter cake with 400mL of water for 3-5 times. And (3) discharging wet materials, and putting the wet materials into a vacuum drying oven for drying to obtain 10.7g of intermediate N-bromoacetyl-D-7-ACA finished product, wherein the yield is 47% and the purity is 98.4%.
FIGS. 4 and 5 are a hydrogen spectrum and a carbon spectrum of N-bromoacetyl-D-7-ACA nuclear magnetic resonance, respectively.
(2) Preparation of desacetylcefathiamidine
To the dried reaction flask were added 100mL of methylene chloride and 10g (0.0285 mol) of N-bromoacetyl-D-7-ACA, cooled to 3℃and 3.2g (0.0316 mol) of triethylamine were added dropwise to the system solution; then 6.9g (0.0430 mol) of N, N' -diisopropylthiourea is added, the mixture is heated to 40 ℃ and the reaction is carried out for 4 hours with heat preservation; after the reaction is finished, cooling to 1 ℃ for crystallization; filtering, soaking and washing the filter cake with 40mL of dichloromethane for 3 times, and discharging to obtain 9.5g of deacetylated cefathiamidine crude product wet material.
(3) Refining of desacetylcefathiamidine
Dissolving 9.5g of the crude product of the deacetylated cefathiamidine in 50mL of ethanol-water (95% -5%), controlling the temperature to 3 ℃, adding 2g of active carbon, stirring and decoloring for 30min, filtering after decoloring, flushing the active carbon with 10mL of water, and combining the obtained filtrate; adding 300mL of acetone into the filtrate to precipitate crystals, filtering and washing to obtain a wet material of the desacetylcefathiamidine, placing the wet material into a vacuum drying oven, vacuumizing, cold pumping for 0.5 hour, and vacuum drying at 26-28 ℃ to obtain 4.5g of the desacetylcefathiamidine, wherein the yield is 36%, and the purity is 99.3%.
Fig. 6 and 7 are hydrogen and carbon spectra, respectively, of the nuclear magnetic resonance of desacetylcefathiamidine.
Example 2
(1) Preparation of N-bromoacetyl-D-7-ACA
Adding 100mL of water into a reaction bottle, cooling to 1 ℃, adding 15g (0.0651 mol) of D-7-ACA under stirring, starting to dropwise add 300g of sodium bicarbonate solution (16 g (0.1905 mol) of sodium bicarbonate contained therein) to dissolve D-7-ACA, and then adding 230mL of acetone; 20g (0.099 mol) of bromoacetyl bromide is added dropwise, and the mixture is stirred and reacted for 3 hours at the temperature of 0 ℃ to 5 ℃; then, acetone was distilled off by concentrating under reduced pressure, 8M hydrochloric acid was added dropwise at room temperature to adjust the pH of the feed solution to 1.3, and solid crystals were separated out, followed by filtration and washing with water to obtain a solid wet material (29 g).
The above solid wet material 29g was dissolved in 60mL of DMF to perform preparative high performance liquid chromatography to isolate N-bromoacetyl-D-7-ACA using equipment of DAC-50 preparative liquid chromatography instrument, the following chromatography:
chromatographic column: c18 packed column (10 μm, 50mm x 250 mm);
eluent: water: acetonitrile=87:13 (V/V);
Detection wavelength: UV254nm;
isocratic elution flow rate: 20L/min;
sample loading amount: 20-25 mL/time.
Collecting the elution fraction with retention time of 10-11min, mixing the eluates of the fraction with purity of more than or equal to 98%, concentrating under reduced pressure to remove acetonitrile and part of DMF, separating out solid during concentration, stirring for crystal growth for 30min, filtering, pulping and washing the filter cake with 400mL of water for 3-5 times. And (3) discharging wet materials, and putting the wet materials into a vacuum drying oven for drying to obtain 8g of intermediate N-bromoacetyl-D-7-ACA finished product, wherein the yield is 35%, and the purity is 98.7%.
(2) Preparation of desacetylcefathiamidine
To the dried reaction flask were added 60mL of methylene chloride, 10mL of methanol and 8g (0.0227 mol) of N-bromoacetyl-D-7-ACA, cooled to 5 ℃, and 2.4g (0.0239 mol) of triethylamine was added dropwise to the system solution; then 7.3g (0.0455 mol) of N, N' -diisopropylthiourea is added, the mixture is heated to 38 ℃ and the reaction is carried out for 4 hours with heat preservation; after the reaction is finished, cooling to 5 ℃ for crystallization; the mixture is filtered, the filter cake is soaked and washed 3 times with 40mL of dichloromethane, and 7.6g of deacetylated cefathiamidine crude product wet material is obtained after discharging.
(3) Refining of desacetylcefathiamidine
Dissolving 7.6g of the crude product of the deacetylated cefathiamidine in 50mL of mixed solution of ethanol-N, N-dimethylformamide (93% -7%), controlling the temperature to be 0-5 ℃, adding 2g of active carbon, stirring and decoloring for 30min, filtering after decoloring, flushing the active carbon with 10mL of mixed solution of ethanol-N, N-dimethylformamide (93% -7%), and combining the obtained filtrates; 400mL of acetone is added into the filtrate to separate out crystals, the crystals are filtered and washed to obtain a wet material of the desacetylcefathiamidine, the wet material is placed into a vacuum drying oven to be vacuumized, firstly, the wet material is cooled and vacuumized for 0.5 hour, and the wet material is dried in vacuum at 26-28 ℃ to obtain 3.2g of the desacetylcefathiamidine, the yield is 32%, and the purity is 99.2%.
Claims (9)
1. A method for preparing deacetylated cefathiamidine, comprising:
(1) Preparation of N-bromoacetyl-D-7-ACA
Dissolving D-7-ACA in water at 0-40 ℃ in the presence of alkali, then adding a first organic solvent, dropwise adding bromoacetyl bromide, and stirring for reaction for 1-4 hours; after the reaction is finished, standing and layering to separate out a water phase or distilling under reduced pressure to remove an organic solvent, thereby obtaining the water phase; at room temperature, regulating the pH of the water phase to 0.5-2.5 by using acid, crystallizing and separating out solid, and filtering to obtain a solid;
dissolving the solid in a second organic solvent, and separating by adopting a preparation high performance liquid chromatography to obtain N-bromoacetyl-D-7-ACA, wherein the preparation high performance liquid chromatography comprises the following conditions:
chromatographic column: c18 packing column;
eluent: a mixed solution of water and acetonitrile according to a volume ratio of 87:13;
Flow rate: 16-21L/min;
Collecting the elution fraction with retention time of 10-11min, mixing the eluates of the fraction with purity of more than or equal to 98%, concentrating, crystallizing and separating out solid, filtering, washing with water, and drying to obtain N-bromoacetyl-D-7-ACA solid;
The first organic solvent is one or two selected from toluene, dichloromethane, acetone and tetrahydrofuran, and the second organic solvent is one or two selected from N, N-dimethylformamide and dimethyl sulfoxide;
(2) Preparation of desacetylcefathiamidine
Dissolving N-bromoacetyl-D-7-ACA in a third organic solvent in the presence of alkali at 0-40 ℃, and then adding N, N' -diisopropylthiourea; controlling the reaction temperature to be between 30 and 40 ℃ and stirring the mixture for reaction for 1 to 4 hours; after the reaction is finished, cooling to 0-10 ℃, crystallizing and separating out solid, and filtering to obtain a crude product of the deacetylated cefathiamidine; wherein the third organic solvent is one or two selected from dichloromethane, chloroform, toluene, methanol, ethanol and isopropanol;
(3) Refining of desacetylcefathiamidine
Dissolving the obtained crude product of the desacetylcefathiamidine in a fourth solvent at the temperature of 0-20 ℃, decolorizing with active carbon, adding a crystallization solvent into filtrate after filtering, crystallizing and separating out the desacetylcefathiamidine, filtering, washing and drying to obtain the purified desacetylcefathiamidine, wherein the fourth solvent is one or two selected from water, N-dimethylformamide, methanol, ethanol and isopropanol, and the crystallization solvent is one or two selected from acetone, dichloromethane and ethyl acetate.
2. The method for preparing desacetylcefathiamidine according to claim 1, wherein in the preparation of N-bromoacetyl-D-7-ACA in the step (1), the conditions of the high performance liquid chromatography are as follows:
Chromatographic column: c18 packed column, 10 μm, 50mm x 250mm;
eluent: a mixed solution of water and acetonitrile according to a volume ratio of 87:13;
Detection wavelength: UV254nm;
flow rate: 16-21L/min.
3. The process for producing desacetylcefathiamidine according to claim 1, wherein in the production of N-bromoacetyl-D-7-ACA in the step (1), the base is one or two selected from the group consisting of diethylamine, triethylamine, N-diisopropylethylamine, potassium carbonate, sodium carbonate and sodium hydrogencarbonate.
4. The process for the preparation of desacetylcefathiamidine according to claim 1, wherein in the preparation of N-bromoacetyl-D-7-ACA in the step (1), the first organic solvent is toluene or acetone; the ratio of the first organic solvent to water is 0.4-2.5:1 by volume; the base is triethylamine and/or sodium bicarbonate.
5. The method for preparing desacetylcefathiamidine according to claim 1, wherein in the preparation of N-bromoacetyl-D-7-ACA in the step (1), the molar ratio of D-7-ACA to bromoacetyl bromide is 1:1 to 3.
6. The process for preparing desacetylcefathiamidine according to claim 1, wherein in the preparation of N-bromoacetyl-D-7-ACA in the step (1), the pH of the aqueous phase is adjusted to 1.0-2.0 with hydrochloric acid at room temperature, and the solid is crystallized and separated out and filtered to obtain a solid.
7. The process for producing desacetylcefathiamidine according to claim 1, wherein in the step (2) of producing desacetylcefathiamidine, the base is one or two selected from the group consisting of diethylamine, triethylamine and N, N-diisopropylethylamine, and the third organic solvent is dichloromethane.
8. The method for preparing desacetylcefathiamidine according to claim 1, wherein in the step (2) of preparing desacetylcefathiamidine, the molar ratio of N-bromoacetyl-D-7-ACA to N, N' -diisopropylthiourea is 1:1 to 3.
9. The process for producing desacetylcefathiamidine according to claim 1, wherein in the refining of desacetylcefathiamidine in the step (3), the fourth solvent is a water-ethanol mixed solvent; the crystallization solvent is acetone.
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