阅读说明：本技术 地贝卡星中间体杂质的制备方法 (Preparation method of dibekacin intermediate impurity ) 是由 印靖 张寅� 陈宵鑫 徐乃林 王凯 石天晨 陈琴霞 于 2020-03-30 设计创作，主要内容包括：本发明公开了一种地贝卡星中间体杂质的制备方法,制备方法如下：在反应瓶中加入1,3,2’,6’,3’’-五-N-对甲苯磺酰基-3’,4’,2’’-三-氧-苄磺酰基-4’’,6’’-氧-环亚己基卡那霉素B、溶剂；再加入碘化钠、锌粉,在40℃～120℃下反应；反应液倒入水中分散、过滤、烘干得到粗品,纯化得到目标化合物。本发明对Tipson-Cohen反应进行了改进,最终获得了1,3,2’,6’-四-N-对甲苯磺酰基-2’’,3’’-(N-对甲苯磺酰基)吖丙啶-4’’,6’’-氧-环亚己基-3’,4’-二脱氧-3’,4’-二脱氢-卡那霉素B的专属合成方法,反应收率和纯度均较高。(The invention discloses a preparation method of an intermediate impurity of dibekacin, which comprises the following steps: adding 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-oxy-benzylsulfonyl-4 '',6'' -oxy-cyclohexylidene kanamycin B and a solvent into a reaction bottle; adding sodium iodide and zinc powder, and reacting at 40-120 deg.c; pouring the reaction solution into water for dispersion, filtering and drying to obtain a crude product, and purifying to obtain the target compound. The invention improves the Tipson-Cohen reaction, finally obtains the special synthetic method of 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B, and has higher reaction yield and purity.)

1. A preparation method of dibekacin intermediate impurities is characterized in that: the dibekacin intermediate impurity is 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -O-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B, and the preparation method is as follows:

adding 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-O-benzylsulfonyl-4 '',6'' -O-cyclohexylidene kanamycin B and a solvent into a reaction bottle, uniformly stirring, adding sodium iodide and zinc powder into the bottle, reacting at 40-120 ℃ for 3-24 hours, and obtaining a reaction solution after the reaction is finished; pouring the reaction solution into water for dispersion, filtering and drying to obtain a crude product, and purifying the crude product to obtain the target compound.

2. The method for preparing the dibekacin intermediate impurity according to claim 1, wherein: the amount of sodium iodide is 10 to 20 times the amount of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-oxy-benzylsulfonyl-4 '',6'' -oxy-cyclohexylidenecamycin B.

3. The method for preparing the dibekacin intermediate impurity according to claim 1, wherein: the amount of zinc powder is 1 to 3 times the amount of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-oxy-benzylsulfonyl-4 '',6'' -oxy-cyclohexylidene kanamycin B.

4. The method for preparing the dibekacin intermediate impurity according to claim 3, wherein: the amount of zinc powder is 1 to 1.2 times the amount of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-oxy-benzylsulfonyl-4 '',6'' -oxy-cyclohexylidenecamycin B.

5. The method for preparing the dibekacin intermediate impurity according to claim 1, wherein: the solvent is one or more of tetrahydrofuran, 1, 4-dioxane, ethanol, acetonitrile and N, N-dimethylformamide.

6. The method for preparing the dibekacin intermediate impurity according to claim 1, wherein: adding sodium iodide and zinc powder, and stirring at 55-65 deg.c for reaction.

7. The method for preparing the dibekacin intermediate impurity according to claim 1, wherein: separating and purifying by silica gel column, wherein the eluent is dichloromethane and ethyl acetate =4: 1.

Technical Field

The invention belongs to the field of medical chemistry, and particularly relates to a preparation method of an intermediate impurity of dibekacin.

Background

Dibekacin (3 ',4' -dideoxykanamycin B/Dibekacin, CAS: 34493-98-6) belongs to the semi-synthetic aminoglycoside antibiotics, with the chemical name O-3-amino-3-deoxy- α -D-glucopyranosyl- (1 → 6) -O- [2, 6-diamino-2, 3,4, 6-tetradeoxy- α -D-erythro-hexopyranosyl- (1 → 4) ] -2-deoxy-D-streptomycin. Dibekacin is a brand new aminoglycoside compound obtained by reforming kanamycin B of Mezechuf et al, and is mainly used for septicemia, respiratory tract infection, skin soft tissue infection, urinary tract infection, postoperative infection and the like caused by various sensitive enterobacteriaceae bacteria, pseudomonas aeruginosa, staphylococcus aureus, enterococcus and the like. As a novel broad-spectrum antibiotic, the antibiotic has a unique chemical structure and an antibacterial action mechanism, has small cross resistance with other antibiotics and antibacterial drugs, and has a synergistic effect in most cases.

The dibekacin can generate a plurality of impurities in the production process, and the research and control of the impurities in the production process are very necessary to ensure the safety and the effectiveness of the medicine. For example, Hamao Umezawa et al investigated 5-position dehydroxylation impurities and glycosidic bond hydrolysis impurities (Bull. chem. Soc. Jpn. 1979, 52(4): 1131-1134). As another example, TsutomuTsuchiya et al investigated the generation of impurities in the dideoxy step (Curbohydrate Research, 1976 (49): 141-.

As a common method for sugar ring ortho-dihydroxy dideoxy reaction, Tipson-Cohen reaction is a key step in the synthesis process of dibekacin, the existence of zinc powder easily causes nucleophilic substitution of 3 '-amino for 2' -sulfonyloxy, intramolecular ring synthesis of aziridine ring, so that aziridine kanamycin B derivative is one of important impurities in the reaction. Therefore, in order to ensure the quality of the dibekacin medicine finally, it is necessary to carry out reasonable and effective control on the impurity of the aziridine kanamycin B in the reaction in the step.

Chinese patent document CN 101575354B (application No. 200910084851.8) discloses a synthetic method of arbekacin and its intermediate dibekacin, in the invention, the synthetic route of dibekacin is as follows:

wherein the reaction c is Tipson-Cohen reaction, 8.0g of 1,3,2',6',3' -penta-N-benzylsulfonyl-3 ',4',2' -tri-O-benzylsulfonyl-4 ',6' -O-cyclohexylidenecarnycin B obtained in the step (2) is dissolved in a 250mL single-neck flask filled with 100mL of DMF, 80g of sodium iodide and 1.6g of zinc powder are weighed and put into the DMF reaction system for reaction at 100 ℃ for 2 hours, chloroform is added for dissolution, water washing and sodium thiosulfate washing are carried out, concentration is carried out, 1,3,2',6',3' -penta-N-benzylsulfonyl-2 ' -O-benzylsulfonyl-4 ',6' -O-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B6.0 g. In this step, an aziridine kanamycin B derivative impurity was produced.

With the advancement of the national work on drug consistency, the preparation method of each impurity compound is determined, qualified reference products are provided, and the quality control of the dibekacin drug substance can be more positively acted, but no relevant literature report of the exclusive synthesis method of 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B exists at home and abroad at present, so that a 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '', the 6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B synthesis method is used for contrasting with impurities in the actual production process and serving as a reference substance so as to control the impurities in the existing production process; higher purity is required as a control.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a preparation method of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-2 '' -oxy-p-toluenesulfonyl-4 '',6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B as an impurity in the preparation process of the dibekacin intermediate 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B.

The technical scheme for realizing the aim of the invention is a preparation method of the dibekacin intermediate impurity, the dibekacin intermediate impurity is 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B, and the preparation method is as follows:

adding 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-O-benzylsulfonyl-4 '',6'' -O-cyclohexylidene kanamycin B and a solvent into a reaction bottle, uniformly stirring, adding sodium iodide and zinc powder into the bottle, reacting at 40-120 ℃ for 3-24 hours, and obtaining a reaction solution after the reaction is finished; pouring the reaction solution into water for dispersion, filtering and drying to obtain a crude product, and purifying the crude product to obtain the target compound.

The amount of sodium iodide is 10 to 20 times the amount of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-oxy-benzylsulfonyl-4 '',6'' -oxy-cyclohexylidenecamycin B.

The amount of zinc powder is 1 to 3 times the amount of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-oxy-benzylsulfonyl-4 '',6'' -oxy-cyclohexylidene kanamycin B.

Further, the amount of zinc powder is 1 to 1.2 times the amount of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-oxy-benzylsulfonyl-4 '',6'' -oxy-cyclohexylidenecamycin B.

The solvent is one or more of tetrahydrofuran, 1, 4-dioxane, ethanol, acetonitrile and N, N-dimethylformamide.

Adding sodium iodide and zinc powder, and stirring at 55-65 deg.c for reaction.

Separating and purifying by silica gel column, wherein the eluent is dichloromethane and ethyl acetate =4: 1.

The invention has the positive effects that:

(1) the preparation method is simple and feasible, has ideal yield and purity and correct structural formula, and can prepare a large amount of the intermediate impurities. The impurities are used as impurity reference substances in the dibekacin synthesis process, the Tipson-Cohen reaction impurity condition in the dibekacin synthesis process is reasonably and effectively controlled, and the safety and the effectiveness of the dibekacin raw material medicines are finally ensured.

(2) The invention improves the Tipson-Cohen reaction, screens a series of influence factors such as reaction raw materials, solvent types, reaction temperature, equivalent numbers, system concentration and the like, finally obtains a special synthesis method of 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B, and the prepared product has the purity of more than 96 percent and can be used as a reference substance.

Drawings

FIG. 1 is a HPLC chart of the compound prepared in example 1.

FIG. 2 shows the NMR spectrum of the compound prepared in example 1.

FIG. 3 is a NMR carbon spectrum of the compound prepared in example 1.

FIG. 4 is an infrared spectrum of the compound prepared in example 1.

Fig. 5 is a mass spectrum of the compound prepared in example 1.

Detailed Description

(example 1)

This example prepares 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B according to the following equation:

wherein Ts is p-toluenesulfonyl and Bs is benzylsulfonyl.

The preparation method of this example includes the following steps:

adding 50.0g (0.028mol) of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-O-benzylsulfonyl-4 '',6'' -O-cyclohexylidenecamycin B and 250mL of tetrahydrofuran into a 500mL four-neck flask, and stirring and heating to 60-65 ℃; then, 83.8g (0.56mol) of sodium iodide and 2.2g (0.033mol) of zinc powder were added to the flask, and the mixture was stirred at 60 ℃ to 65 ℃ to react for 8. + -. 0.5 hours, and the reaction was stopped to obtain a reaction solution.

Other solvents which do not adversely affect the reaction, such as 1, 4-dioxane, N-dimethylformamide, ethanol, acetonitrile, etc., may be used in addition to tetrahydrofuran; therefore, the reaction solvent can be one or a mixture of more than one of tetrahydrofuran, 1, 4-dioxane, ethanol, acetonitrile and N, N-dimethylformamide.

The reaction raw material 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-oxo-benzylsulfonyl-4 '',6'' -oxo-cyclohexadienylidenecamycin B can be prepared according to the preparation method of compound B (4) in the research on intermediate synthesis technology of dibekacin of Master thesis (Wufang Yuan, 2010, Zheng Zhou university). In this document the compound B (4) is named wrongly and should be 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-oxy-benzylsulfonyl-4 '',6'' -oxy-cyclohexylidenecarnycin B.

Pouring the reaction solution into 1250mL of water for dispersion, filtering and drying to obtain a crude product,

the water washing can remove the excessive sodium iodide.

The crude product obtained after drying was purified by silica gel column separation (eluent dichloromethane: ethyl acetate =4: 1) to obtain 28.0g (0.022 mol) of 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxo-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B, with a molar yield of 78.57% and a purity of 96.78% by HPLC (FIG. 1).

And (3) confirming the structure of the compound obtained by purifying the silica gel column, and detecting a sample of the compound by using a nuclear magnetic resonance hydrogen spectrum, a nuclear magnetic resonance carbon spectrum, an infrared spectrum and a mass spectrum, wherein the nuclear magnetic resonance hydrogen spectrum is shown in figure 2, the nuclear magnetic resonance carbon spectrum is shown in figure 3, the infrared spectrum is shown in figure 4, and the mass spectrum is shown in figure 5.

The specific test results are as follows:

¹H NMR (600MHz,DMSO-d6) : 7.83-7.81(d, 2H), 7.72-7.71(d, 2H), 7.63-7.57(m, 5H), 7.47-7.45(m, 4H), 7.36-7.32(m, 7H), 7.28-7.24(m, 4H), 5.59-5.58(d,1H), 5.50-5.48(d, 1H), 5.41-5.39(d, 1H), 5.12(s, 1H), 4.65-4.64(d, 1H), 4.43(s, 1H), 3.87-3.86(m, 1H), 3.77(s, 1H), 3.71-3.70(m, 1H), 3.57-3.55(m, 2H),3.11-3.09(m, 1H), 2.96-2.92(m, 3H), 2.89-2.82(m, 5H), 2.41(s, 3H), 2.40(s,3H), 2.37(s, 3H), 2.36(s, 3H), 2.34(s, 3H), 1.86-1.78(d, 2H), 1.54-1.46(m,4H), 1.33-1.29(m, 5H), 0.96-0.94(m, 1H)。

¹³C NMR (600MHz,DMSO-d6) : 144.89, 142.89, 142.44, 142.14, 139.53,138.79, 138.18, 137.61, 133.17, 130.13, 129.59, 129.58, 129.47, 127.84,127.50, 126.76, 126.51, 126.24, 125.90, 125.80, 99.86, 97.52, 95.06, 83.09,80.47, 73.48, 67.30, 66.16, 62.52, 61.24, 52.29, 52.10, 49.63, 45.58, 40.25,37.32, 33.93, 27.38, 24.99, 22.39, 22.13, 21.13, 21.07, 21.02, 20.98, 20.92。

IR(KBr)cm^-1: 3441.61, 2928.14, 1629.29, 1598.84, 1449.68, 1329.68,1160.80, 1092.03, 1062.88, 814.92, 666.76, 552.04。

MS(ESI, m/z): 1299.40(M⁺+H₂O)。

the compound prepared in this example was confirmed to be 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -O-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B.

(example 2)

The preparation method of this example includes the following steps:

adding 50.0g (0.028mol) of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-O-benzylsulfonyl-4 '',6'' -O-cyclohexylidenecamycin B and 250mL of tetrahydrofuran into a 500mL four-neck flask, and stirring and heating to 60-65 ℃; then 41.9g (0.28mol) of sodium iodide and 2.2g (0.033mol) of zinc powder were added to the flask, and the mixture was stirred at 60 ℃ to 65 ℃ to react for 16. + -. 0.5 hours, and the reaction was stopped to obtain a reaction solution.

The reaction solution was poured into 1250mL of water to disperse, filtered and dried to obtain a crude product.

The crude product obtained after drying was purified by silica gel column separation (eluent dichloromethane: ethyl acetate =4: 1) to obtain 29.5g (0.023 mol) of 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxo-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B, with a molar yield of 82.22% and a purity of 96.08% by HPLC.

(example 3)

The preparation method of this example includes the following steps:

50.0g (0.028mol) of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-O-benzylsulfonyl-4 '',6'' -O-cyclohexylidenecamycin B and 250mL of tetrahydrofuran are added into a 500mL four-necked flask, and the temperature is raised to 40-45 ℃ by stirring; then, 83.8g (0.56mol) of sodium iodide and 2.2g (0.033mol) of zinc powder were added to the flask, and the mixture was stirred at 40 ℃ to 45 ℃ to react for 16. + -. 0.5 hours, and the reaction was stopped to obtain a reaction solution.

The reaction solution was poured into 1250mL of water to disperse, filtered and dried to obtain a crude product.

The crude product obtained after drying was purified by silica gel column separation (eluent dichloromethane: ethyl acetate =4: 1) to obtain 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B27.8 g g (0.022 mol), molar yield 77.48%, purity 96.54% by HPLC.

(example 4)

The preparation method of this example includes the following steps:

50.0g (0.028mol) of 1,3,2',6',3'' -penta-N-p-toluenesulfonyl-3 ',4',2'' -tri-O-benzylsulfonyl-4 '',6'' -O-cyclohexylidenecamycin B and 125mL of N, N-dimethylformamide are added into a 500mL four-necked flask, and the temperature is raised to 80-85 ℃ by stirring; then, 83.8g (0.56mol) of sodium iodide and 2.2g (0.033mol) of zinc powder were added to the flask, and the mixture was stirred at 100 ℃ to 120 ℃ for reaction for 4. + -. 0.5 hours, and the reaction was stopped to obtain a reaction solution.

The reaction solution was poured into 1250mL of water to disperse, filtered and dried to obtain a crude product.

The crude product obtained after drying was purified by silica gel column separation (eluent dichloromethane: ethyl acetate =4: 1) to obtain 29.3g (0.023 mol) of 1,3,2',6' -tetra-N-p-toluenesulfonyl-2 '',3'' - (N-p-toluenesulfonyl) aziridine-4 '',6'' -oxy-cyclohexylidene-3 ',4' -dideoxy-3 ',4' -didehydro-kanamycin B, with a molar yield of 81.67% and a purity of 96.63% by HPLC.