CN115838388A - Preparation method of 3-N-acetyl etimicin - Google Patents
Preparation method of 3-N-acetyl etimicin Download PDFInfo
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- 229950009953 etimicin Drugs 0.000 title claims abstract description 42
- 238000002360 preparation method Methods 0.000 title claims abstract description 16
- 239000000243 solution Substances 0.000 claims abstract description 71
- 238000006243 chemical reaction Methods 0.000 claims abstract description 60
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 45
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims abstract description 27
- DNYGXMICFMACRA-UHFFFAOYSA-N gentamicin C1A Natural products O1C(CNC)CCC(N)C1OC1C(O)C(OC2C(C(NC)C(C)(O)CO2)O)C(N)CC1N DNYGXMICFMACRA-UHFFFAOYSA-N 0.000 claims abstract description 23
- VEGXETMJINRLTH-BOZYPMBZSA-N gentamycin C1a Chemical compound O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H](CC[C@@H](CN)O2)N)[C@@H](N)C[C@H]1N VEGXETMJINRLTH-BOZYPMBZSA-N 0.000 claims abstract description 22
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims abstract description 18
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims abstract description 18
- 239000000047 product Substances 0.000 claims abstract description 14
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- 229940039790 sodium oxalate Drugs 0.000 claims abstract description 6
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- 239000000413 hydrolysate Substances 0.000 claims abstract description 5
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 3
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims abstract description 3
- ZOIORXHNWRGPMV-UHFFFAOYSA-N acetic acid;zinc Chemical compound [Zn].CC(O)=O.CC(O)=O ZOIORXHNWRGPMV-UHFFFAOYSA-N 0.000 claims abstract description 3
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- 239000011230 binding agent Substances 0.000 claims abstract description 3
- 238000010668 complexation reaction Methods 0.000 claims abstract description 3
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- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims abstract description 3
- 238000002444 silanisation Methods 0.000 claims abstract description 3
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 239000012043 crude product Substances 0.000 claims description 9
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- 238000000926 separation method Methods 0.000 claims description 4
- 229940057499 anhydrous zinc acetate Drugs 0.000 claims description 3
- 238000000105 evaporative light scattering detection Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
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- 150000003839 salts Chemical class 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- DJWUNCQRNNEAKC-UHFFFAOYSA-L zinc acetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O DJWUNCQRNNEAKC-UHFFFAOYSA-L 0.000 claims description 3
- 239000008367 deionised water Substances 0.000 claims description 2
- 229910021641 deionized water Inorganic materials 0.000 claims description 2
- 238000011033 desalting Methods 0.000 claims description 2
- 239000012071 phase Substances 0.000 claims description 2
- 230000001376 precipitating effect Effects 0.000 claims description 2
- 229960000314 zinc acetate Drugs 0.000 claims description 2
- 239000012295 chemical reaction liquid Substances 0.000 claims 4
- 238000010025 steaming Methods 0.000 claims 3
- IKHGUXGNUITLKF-XPULMUKRSA-N acetaldehyde Chemical compound [14CH]([14CH3])=O IKHGUXGNUITLKF-XPULMUKRSA-N 0.000 claims 1
- 238000001704 evaporation Methods 0.000 claims 1
- 238000001179 sorption measurement Methods 0.000 claims 1
- OEBISAUVQBGQKC-ZIZSAZPJSA-N (2r,3r,4r,5r)-2-[(1s,2s,3r,4s,6r)-4-amino-3-[(2r,3r,6s)-3-amino-6-(aminomethyl)oxan-2-yl]oxy-6-(ethylamino)-2-hydroxycyclohexyl]oxy-5-methyl-4-(methylamino)oxane-3,5-diol;sulfuric acid Chemical compound OS(O)(=O)=O.O([C@@H]1[C@@H](N)C[C@H]([C@@H]([C@H]1O)O[C@@H]1[C@@H]([C@@H](NC)[C@@](C)(O)CO1)O)NCC)[C@H]1O[C@H](CN)CC[C@H]1N OEBISAUVQBGQKC-ZIZSAZPJSA-N 0.000 description 6
- 229940079593 drug Drugs 0.000 description 4
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- VEGXETMJINRLTH-ALRICIOSSA-N etimicin Chemical compound O1C[C@@](O)(C)[C@H](NC)[C@H](O)[C@H]1O[C@@H]1[C@H](O)[C@H](O[C@@H]2[C@@H](CC[C@@H](CN)O2)N)[C@@H](N)C[C@H]1N VEGXETMJINRLTH-ALRICIOSSA-N 0.000 description 4
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- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 2
- DYHSDKLCOJIUFX-UHFFFAOYSA-N tert-butoxycarbonyl anhydride Chemical compound CC(C)(C)OC(=O)OC(=O)OC(C)(C)C DYHSDKLCOJIUFX-UHFFFAOYSA-N 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 241000589291 Acinetobacter Species 0.000 description 1
- 125000006847 BOC protecting group Chemical group 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000588923 Citrobacter Species 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000606768 Haemophilus influenzae Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 241000588747 Klebsiella pneumoniae Species 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 206010057190 Respiratory tract infections Diseases 0.000 description 1
- 241000607720 Serratia Species 0.000 description 1
- 241000191940 Staphylococcus Species 0.000 description 1
- -1 acetyl etimicin Chemical compound 0.000 description 1
- 229940126575 aminoglycoside Drugs 0.000 description 1
- 229940126574 aminoglycoside antibiotic Drugs 0.000 description 1
- 239000002647 aminoglycoside antibiotic agent Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
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- 229910052739 hydrogen Inorganic materials 0.000 description 1
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- 210000003734 kidney Anatomy 0.000 description 1
- SYSQUGFVNFXIIT-UHFFFAOYSA-N n-[4-(1,3-benzoxazol-2-yl)phenyl]-4-nitrobenzenesulfonamide Chemical class C1=CC([N+](=O)[O-])=CC=C1S(=O)(=O)NC1=CC=C(C=2OC3=CC=CC=C3N=2)C=C1 SYSQUGFVNFXIIT-UHFFFAOYSA-N 0.000 description 1
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- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention relates to a preparation method of 3-N-acetyl etimicin, which comprises the following steps: step 1), reacting gentamicin C1a with zinc acetate to obtain a zinc complex; step 2), triethylamine is used as an acid-binding agent, and acetic anhydride is added for reaction to obtain an amino-acetylated product; step 3) reacting with sodium oxalate to release complexation, and separating and purifying to obtain a P1 intermediate; step 4), adding ethylene glycol dimethyl ether, hexamethyldisilazane and concentrated sulfuric acid for reaction to obtain a hydroxyl silanization product; step 5) adding dichloromethane and acetaldehyde for reaction, and then adding potassium borohydride and boric acid buffer solution for reaction to obtain an ethylate; and 6) hydrolyzing with a sodium hydroxide solution, passing the hydrolysate through a column, and separating and purifying a prepared liquid phase to obtain the 3-N-acetyl etimicin.
Description
The technical field is as follows:
the invention relates to the field of pharmaceutical chemistry, in particular to a preparation method of an aminoglycoside compound, and specifically relates to a preparation method of 3-N-ethyl gentamicin C1 a.
Background art:
etimicin sulfate (Etimicin sulfate) is a semi-synthetic aminoglycoside antibiotic, and is suitable for various infections caused by sensitive Escherichia coli, enterobacter, klebsiella pneumoniae, serratia, acinetobacter, citrobacter, proteus, pseudomonas aeruginosa, haemophilus influenzae, staphylococcus, etc. Clinical studies have shown that etimicin sulfate formulations have a superior therapeutic effect on infections such as respiratory tract infections, kidney and urogenital infections, soft tissue and other infections.
3-N-acetyl etimicin, having the formula:
the 3-N-acetyl etimicin is one of impurities in the production process of the etimicin sulfate bulk drug, and the existence of the 3-N-acetyl etimicin not only influences the yield of the etimicin sulfate, but also has potential risks on the quality of the etimicin sulfate bulk drug. Because of the similar structure and similar properties with etimicin, the etimicin is difficult to separate from the production. At present, no standard product of 3-N-acetyl etimicin is sold at home and abroad.
At present, the patent related to the preparation method of 3-N-acetyl etimicin (Chinese patent application No. 202011536901.4) mainly comprises the following steps: adding etimicin, ethanol and water into a round-bottom flask, uniformly stirring, cooling to 0 ℃, dropwise adding di-tert-butyl dicarbonate, continuously stirring, and concentrating a reaction system after the reaction is finished; adding formic acid into the reaction system, heating to 120 ℃, continuously reacting for 4 hours, and cooling to room temperature after the reaction is finished; respectively adding 10% sodium hydroxide, stirring, heating and refluxing, and cooling to room temperature after the reaction is finished; desalting, and separating to obtain the target product 3-N-acetyl etimicin. The BOC protecting group of the preparation method has no selectivity, and cannot distinguish 2 '-C-amino, 6' -C-amino and 3-C-amino, so that side reaction is generated in the preparation process without fail, the yield is low, and when 10% sodium hydroxide solution is used for deprotection, a large amount of 3-N-acetyl can be damaged due to too long reaction time, so that the yield is further reduced, and meanwhile, the problems of low yield or low purity and the like are caused due to the existence of isomers and higher separation difficulty.
The invention content is as follows:
the invention aims to provide a preparation method of 3-N-acetyl etimicin. The method has the advantages of good specificity, less side reaction and easy purification and separation.
The preparation method comprises the following steps:
step 1), reacting gentamicin C1a with zinc acetate to obtain a zinc complex;
step 2), triethylamine is used as an acid-binding agent, and acetic anhydride is added for reaction to obtain an amino-acetylated product;
step 3) reacting with sodium oxalate to release complexation, and separating and purifying to obtain a P1 intermediate;
step 4), adding ethylene glycol dimethyl ether, hexamethyldisilazane and concentrated sulfuric acid for reaction to obtain a hydroxyl silanization product; step 5) concentrating, removing the solvent, adding dichloromethane and acetaldehyde for reaction, and then adding potassium borohydride and boric acid buffer solution for reaction to obtain an ethylate;
and 6) hydrolyzing with a sodium hydroxide solution, passing the hydrolysate through a column, and separating and purifying a prepared liquid phase to obtain the 3-N-acetyl etimicin.
The preparation route of the method is as follows:
the method of the invention has the core of the hydrolysis in the step 6), the column passing of the hydrolysate and the purification step.
Wherein the hydrolysis step is as follows:
and adding 100mL of 10% sodium hydroxide solution into the reaction solution 6, concentrating and distilling to remove dichloromethane, adding 240mL of 20% sodium hydroxide solution, heating to 110 ℃, refluxing, and stirring for 3 hours to obtain a reaction solution 7.
Wherein, the step of passing the hydrolysate through the column is as follows:
absorbing the reaction solution 7 by macroporous resin, washing with brine-free water to remove salt, performing gradient elution by 5-25% ethanol, collecting eluent with purity of more than 80%, and performing vacuum concentration to obtain a crude product.
Wherein, the purification adopts preparation liquid phase separation and purification, and comprises the following steps:
2g of the crude product was placed in a centrifuge tube and dissolved with mobile phase A according to the following preparative liquid phase conditions:
the pure product of the 3-N-acetyl etimicin is obtained, and the detection shows that the area normalized content of the 3-N-acetyl etimicin is 97.18 percent (ELSD).
Compared with the prior art, the method has the following beneficial effects:
the 3-N-acetyl etimicin prepared by the method has the characteristics of good specificity, less side reaction, easy separation and purification of products, high yield and the like. Meanwhile, the impurities are deeply researched, the generation of the impurities is reduced by optimizing a hydrolysis process, and the method has great significance for improving the quality of medicines and improving the safety of clinical medication.
The beneficial effects of the present invention are further illustrated by the following comparison:
drawings
FIG. 1: 3-N-acetyl etimicin mass spectrum
FIG. 2: 3-N-acetyl etimicin hydrogen spectrum
FIG. 3: carbon spectrum of 3-N-acetyl etimicin
FIG. 4: HSQC spectrum of acetyl etimicin
FIG. 5: 3N-acetyl etimicin COSY spectra
FIG. 6: 3-N-acetyl etimicin thin layer chromatogram
FIG. 7: preparation diagram of 3-N-acetyl etimicin crude product
FIG. 8: 3-N-acetyl etimicin detection map
The specific implementation mode is as follows:
the invention is further illustrated by the following examples, which are not to be construed as limiting the invention thereto.
Example 1:
20.0g of gentamicin C1a is dissolved in 200mL of methanol to obtain solution 1. 20.6g of anhydrous zinc acetate is added into the solution 1, and the solution is stirred, dissolved and stirred for 2 hours to obtain a reaction solution 1.
And cooling the reaction solution 1 to 0-10 ℃, adding 20mL of triethylamine and 18.0mL of acetic anhydride, and stirring for 2 hours to obtain a reaction solution 2.
And (3) concentrating the reaction solution 2 to remove the solvent, adding 300mL of water to dissolve the reaction solution, adding 16.6g of sodium oxalate, stirring for 3 hours, precipitating and filtering to obtain a filtrate 2. The filtrate is concentrated to remove the solvent, and is separated by macroporous resin to obtain an intermediate P1.
100mL of ethylene glycol dimethyl ether, 120mL of hexamethyldisilazane and 0.2mL of concentrated sulfuric acid are added into the intermediate P1, and heating reflux is carried out for 4 hours at 90 ℃ to obtain a reaction solution 3.
The reaction solution 3 was concentrated to remove most of the solvent, and dissolved in 5.4mL of methylene chloride to obtain a solution 3. The solution 3 is cooled to 0-10 ℃, 4.8mL of 40% acetaldehyde is added, and the mixture is stirred for 1 hour to obtain a reaction solution 4.
7.2g of potassium borohydride was added to the reaction solution 4, and the mixture was stirred for 1 hour to obtain a reaction solution 5.
To the reaction solution 5, 30mL of a boric acid buffer solution having a pH of 10 (prepared by dissolving 10.0g of boric acid in 30mL of deionized water with stirring and adjusting pH =10 with sodium hydroxide) was added, and the mixture was stirred for 5 hours to obtain a reaction solution 6.
And adding 100mL of 10% sodium hydroxide solution into the reaction solution 6, concentrating and distilling to remove dichloromethane, adding 240mL of 20% sodium hydroxide solution, heating to 110 ℃, refluxing, and stirring for 3 hours to obtain a reaction solution 7.
Adsorbing the reaction solution 7 by macroporous resin, washing with brine-free water to remove salt, performing gradient elution by 5-25% ethanol, collecting eluent with purity of more than 80%, vacuum concentrating to obtain crude product,
the crude product was isolated and purified according to the following preparative liquid phase conditions:
the pure product of the 3-N-acetyl etimicin is obtained by liquid phase separation, and 1.2g is collected.
Comprehensive analysis:
the Exact Mass of the 3-N-acetyl etimicin is 519.3268, and the molecular formula is C 23 H 45 N 5 O 8 (ii) a Under the positive ion mode of the high resolution mass spectrum, the ion peak of the quasi-molecule is displayed to be 520.33417, namely ([ C) 23 H 45 N 5 O 8 ]+H) + In the negative ion mode, the excimer ion peak is 518.31812, i.e., ([ C) 23 H 45 N 5 O 8 ]-H) - All of which are consistent with the molecular formula of 3N-acetyl etimicin.
In NMR spectrum, the product is 1 H NMR has 20 sets of peaks (except solvent peaks) integrated from low-to high-field 1; 13 there are 23C signals (except for the solvent peak) in the C NMR spectrum from low to high field, thus containing a total of 23 carbon atoms; meanwhile, through HSQC and COSY analysis, the structural formula of the sample is consistent with that of 3N-acetyl etimicin and can be reasonably explained.
In conclusion, the product confirms that the structural formula of the sample is consistent with that of the 3N-acetyl etimicin by HRMS and NMR methods.
Claims (10)
1. A preparation method of 3-N-acetyl etimicin comprises the following steps:
step 1), reacting gentamicin C1a with zinc acetate to obtain a zinc complex;
step 2), triethylamine is used as an acid-binding agent, and acetic anhydride is added for reaction to obtain an amino-acetylated product;
step 3) reacting with sodium oxalate to release complexation, and separating and purifying to obtain a P1 intermediate;
step 4), adding ethylene glycol dimethyl ether, hexamethyldisilazane and concentrated sulfuric acid for reaction to obtain a hydroxyl silanization product;
step 5) adding dichloromethane and acetaldehyde for reaction, and then adding potassium borohydride and boric acid buffer solution for reaction to obtain an ethylate;
and 6) hydrolyzing with a sodium hydroxide solution, passing the hydrolysate through a column, and separating and purifying a prepared liquid phase to obtain the 3-N-acetyl etimicin.
2. The method of claim 1, wherein
Step 1) the method is as follows: dissolving gentamicin C1a in methanol with the volume 5-10 times of the weight of gentamicin C1a to obtain solution 1, adding anhydrous zinc acetate with the weight 1.5-3 times of that of gentamicin C1a into the solution 1, stirring, dissolving and stirring for 0.5-2 hours to obtain reaction solution 1.
3. The method of claim 1, wherein
Step 2) the method is as follows: cooling the reaction solution 1 to 0-10 ℃, adding triethylamine in a proportion of 1.5-3 times of the weight of the gentamicin C1a and acetic anhydride in a proportion of 3.5-4.5 times of the mass of the gentamicin C1a, and stirring for 1-3 hours to obtain a reaction solution 2.
4. The method of claim 1, wherein
Step 3) the method is as follows: and (3) concentrating the reaction liquid 2 to remove the solvent, adding water with the volume being 10-15 times of the weight of gentamicin C1a for dissolving, adding sodium oxalate with the weight being 2-4 times of the weight of gentamicin C1a for stirring for 3-5 hours, filtering to obtain filtrate 2, concentrating to remove the solvent, and performing gradient elution by macroporous resin ethanol to obtain an intermediate P1.
5. The method of claim 1, wherein
Step 4) the method is as follows: 3, 2', 6' -N, N, N-triacetyl gentamicin C1a was added as described in 3, 2', 6' -N, N, ethylene glycol dimethyl ether with the volume being 10-12 times of the weight of N-triacetyl gentamicin C1a, 3, 2', 6' -N, hexamethyldisilazane in a volume of 4-6 times the weight of N-triacetyl gentamicin C1a and concentrated sulfuric acid in a proportion of 0.05-0.5 times the amount of 3, 2', 6' -N, N, N-triacetyl gentamicin C1a are heated to 90-100 ℃ to be dissolved and refluxed for 3-5 hours to obtain reaction liquid 3.
6. The method of claim 1, wherein
Step 5) the method is as follows: concentrating the reaction solution 3, evaporating most of the solvent, and dissolving with dichloromethane which is 10-12 times of the weight of 3, 2', 6' -N, N, N-triacetyl gentamicin C1a in volume to obtain a solution 3; cooling the solution 3 to 0-10 ℃, adding 40% acetaldehyde solution with the volume 0.3-0.5 times of the weight of the 3, 2', 6' -N, N, N-triacetyl gentamicin C1a, and stirring for 1-2 hours to obtain a reaction solution 4; adding potassium borohydride which is 0.6 to 0.8 times of the weight of 3, 2', 6' -N, N, N-triacetyl gentamicin C1a into the reaction solution 4, and stirring for 0.5 to 3 hours to obtain a reaction solution 5; adding boric acid buffer solution with pH of 9-11 and the volume being 2-3 times of the weight of the 3, 2', 6' -N, N, N-triacetyl gentamicin C1a into the reaction solution 5, and stirring for 2-12 hours to obtain reaction solution 6.
7. The method according to claim 1, wherein the hydrolysis method in step 6) is as follows: adding 10 percent sodium hydroxide solution with the volume 5-8 times of the weight of 3, 2', 6' -N, N, N-triacetyl gentamicin C1a into the reaction liquid 6, concentrating and steaming to obtain a solution 4, adding 20 percent sodium hydroxide solution with the volume 10-15 times of the weight of 3, 2', 6' -N, N, N-triacetyl gentamicin C1a, heating to 100-120 ℃, refluxing, and reacting for 3-6 hours to obtain a reaction liquid 7.
8. The method according to claim 1, wherein the method of passing the hydrolysis solution through the column in step 6) is as follows: and (3) putting the reaction solution 7 on a macroporous resin adsorption column, desalting by using brine-free water, performing gradient elution by using 5-25% ethanol, collecting eluent with the purity of more than 80%, and performing vacuum concentration to obtain a crude product of the 3-N-acetyl etimicin.
9. The method according to claim 1, wherein the separation and purification of the liquid phase in step 6) is performed as follows:
2g of the crude product was placed in a centrifuge tube and dissolved with mobile phase A according to the following preparative liquid phase conditions:
the detector is an ELSD and the detector is,
gradient elution is carried out to obtain the pure product of the 3-N-acetyl etimicin.
10. The method of claim 1, comprising the steps of:
dissolving 20.0g of gentamicin C1a in 200mL of methanol to obtain a solution 1, adding 20.6g of anhydrous zinc acetate into the solution 1, stirring, dissolving and stirring for 2 hours to obtain a reaction solution 1, cooling the reaction solution 1 to 0-10 ℃, adding 20mL of triethylamine and 18.0mL of acetic anhydride, stirring for 2 hours to obtain a reaction solution 2, concentrating the reaction solution 2 to remove a solvent, adding 300mL of water to dissolve the reaction solution, adding 16.6g of sodium oxalate, stirring for 3 hours, precipitating and filtering to obtain a filtrate 2, concentrating the filtrate to remove the solvent, separating the filtrate by using macroporous resin to obtain an intermediate P1, adding 100mL of ethylene glycol dimethyl ether, 120mL of hexamethyldisilazane and 0.2mL of concentrated sulfuric acid into the intermediate P1, heating and refluxing at 90 ℃ for 4 hours to obtain a reaction solution 3, concentrating and steaming most of the reaction solution 3, dissolving by using 5.4mL of dichloromethane to obtain a solution 3, cooling the solution 3 to 0-10 ℃, adding 4.8mL of 40% acetaldehyde, stirring for 1 hour to obtain a reaction solution 4, adding 7.2g of potassium borohydride into the reaction solution 4, stirring for 1 hour to obtain a reaction solution 5, adding 30mL of a boric acid buffer solution with the pH value of 10 into the reaction solution 5 (prepared by adding 30mL of deionized water into 10.0g of boric acid, stirring and dissolving, and adjusting the pH value to be =10 by using sodium hydroxide), stirring for 5 hours to obtain a reaction solution 6, adding 100mL of a 10% sodium hydroxide solution into the reaction solution 6, concentrating and steaming dichloromethane, adding 240mL of 20% sodium hydroxide solution, heating to 110 ℃, refluxing, stirring for 3 hours to obtain a reaction solution 7, adsorbing the reaction solution 7 by using macroporous resin, washing with brine-free water to remove salt, carrying out gradient elution by using 5% -25% ethanol, collecting eluent with the purity of more than 80%, carrying out vacuum concentration to obtain a crude product, carrying out preparation liquid phase separation on the crude product to obtain a pure 3-N-acetyl etimicin, collecting 1.2g in total,
preparation of liquid phase conditions: the detector is an ELSD and the detector is,
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| WO2009049490A1 (en) * | 2007-10-09 | 2009-04-23 | Changzhou Fangyuan Pharmaceutical Co., Ltd. | Use of aminoglycoside antibiotic in the preparation of pharmaceutical composition for treating drug-resistant bacterial infection |
| CN110372760A (en) * | 2019-08-14 | 2019-10-25 | 无锡济民可信山禾药业股份有限公司 | A kind of synthetic method of 3-N- ethyl Gentamicin C1a |
| CN111440219A (en) * | 2020-04-20 | 2020-07-24 | 无锡济民可信山禾药业股份有限公司 | Method for separating and purifying high-purity 3,2 ', 6' -tri-N-acetyl etimicin |
| CN112608350A (en) * | 2020-12-23 | 2021-04-06 | 无锡济煜山禾药业股份有限公司 | Synthetic method of 3-N-acetyl etimicin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2009049490A1 (en) * | 2007-10-09 | 2009-04-23 | Changzhou Fangyuan Pharmaceutical Co., Ltd. | Use of aminoglycoside antibiotic in the preparation of pharmaceutical composition for treating drug-resistant bacterial infection |
| CN110372760A (en) * | 2019-08-14 | 2019-10-25 | 无锡济民可信山禾药业股份有限公司 | A kind of synthetic method of 3-N- ethyl Gentamicin C1a |
| CN111440219A (en) * | 2020-04-20 | 2020-07-24 | 无锡济民可信山禾药业股份有限公司 | Method for separating and purifying high-purity 3,2 ', 6' -tri-N-acetyl etimicin |
| CN112608350A (en) * | 2020-12-23 | 2021-04-06 | 无锡济煜山禾药业股份有限公司 | Synthetic method of 3-N-acetyl etimicin |
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