A method of obtaining a crystalline mono - or dihydrochloride dihydrate 7-[2-(2 - aminothiazol-4-yl)-2(z)- methoxykynuramine] -3-[(1 - methyl-1 - pyrrolidinium) methyl] -ceph - 3-em - 4-carboxylate, essentially free from antihisamine and2isomer

 

(57) Abstract:

The inventive free2-isomer of an acid additive salt of 7-amino-3-[(1-methyl-1-pyrrolidino)methyl]-ceph-3-em-4-carboxylate f-ly I, where NH HCl, HJ or sulfuric acid, is subjected to the interaction with the hydrochloride of 2-(2-aminothiazol-4-yl)-2-methoxykynuramine essentially free from antihisamine, in aqueous-organic solvent at pH 5.0 to 7.5, followed by separation of the resulting reaction mixture the desired product in the form of its crystalline mono - or dihydrochloride dihydrate. Connection structure of f-crystals I: 13 C.p. f-crystals, 4 Il. table 1.

The invention proposes a method of chemical acylation, in particular the method of acylation in aqueous medium to obtain such antibiotic, as a hydrate of the dihydrochloride of cefepime, also known under the name 7-/2-(2-aminothiazol-4-yl)-(Z)-methoxykynuramine/-3- /(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate. The invention also provides stable crystalline salt of SYN isomer thiazolidine intermediate compounds that can be used in the synthesis of applicable antibiotics of a wide profile, and method for producing such intermediate soedinenie-4-yl)-(Z)-2-methoxyethoxy acid, attached to the 7-amino group of zefalosporinovy acid is well known acylation methods. In many cases, the acylation process includes mandatory protection of the amino group and the activation of the carboxylic acid side chain. Therefore, the literature provides a large number aminosidine groups for 2-amino thiazole cycle and a large number of activating groups for carboxylic acids. Search the latest protective groups and activating groups to obtain the target of the antibiotic is still the object of numerous publications that due to the cost and toxicity of certain activating groups. Thus, there is still a need for antibiotics of a wide profile based on a simple, stable, crystalline, economical and non-toxic side-chain with target geometric (Z)-configuration, easily connecting with the 7-amino group of a cephalosporin Adara. Below is a list of literature related to individual representatives of the thiazole side chain.

In U.S. patent N 4203899 (Ochiai and others) are disclosed compounds of the formula:

R where R1represents amino, protected amino, hydroxyl or protected hydroxyl; R5represent the EN or OM, where M is alkali metal.

In the patent application great Britain GB-2144424, 1985, revealed a number of derivatives of pyridine-cephalosporin in a variety of ways, including the use of the compounds of formula

R4HN or its salts, where R1means a hydrogen or halogen atom, R2means a hydrogen atom or a C1-C6-alkyl and R4represents hydrogen or aminosidine group, or activated derivative of this compound.

In the application for the European patent EP-160546, published. 1985, discloses a number of cephalosporin derivatives in a variety of ways, including the use of substituted derivatives examinationresources.net acid formula

R8HN or its reactive derivative, where R8represents a hydrogen atom or a protective group for amino group. Acceptable examples of such reactive derivatives, which are given in the application, include mixed anhydrides of the acids, galodamadruga acids, active esters, active amides and azides acids.

In U.S. patent N 4385181 (Farge and others), 1983, reveals a complex thioethers of the formula

R1HN where R1represents hydrogen or a protective radical,-carboxyethyl, phenyl or a great number of various geterotsiklicheskikh radicals listed in columns 4-8, and their SYN - and anti-isomers and mixtures thereof.

In addition to the cited references, there is also a large number of links, which provide various protective groups for 2-aminosalicylates and more activating (outgoing groups fragment carboxylic acids that can be used in the acylation derivative 7-aminocephalosporanic.

However, the most relevant here, the patent Czechoslovakia N 238950, published. 1987, (Chemical Abstracts, T. 110, page 544 (1989)), revealing the connection of the present invention of the formula

NH HCl and assumes that the connection has a SYN-configuration. The only proof of receipt compounds in the patent is in the content of chlorine in 99-105% of theoretical value.

In connection with the creation of new synthetic methods to obtain antibiotics creators of inventions, as well as other researchers in this area have felt the need for simple, convenient, economical, crystalline, stable and non-toxic source products for use in the manufacture of antibiotics. Initial attempts is the shaft groups were unsuccessful. However, the creators of the invention is now found that the compound according to the invention can be obtained in the specific reaction conditions. The opening was further confirmed after the creators of the invention could not reproduce the results of the patent Czechoslovakia. Target SYN-isomer hydrochloride of the acid chloride required to obtain the target of the antibiotic, get again failed. Moreover, additional experiments confirmed that, following the instructions of the prototype, it is impossible to obtain hydrochloride of the target SYN-isomers of carboxylic acids, essentially not containing anti-isomer and having a range of proton nuclear magnetic resonance, which is shown here.

A wide range of cefepime antibiotics is Aburaki and others in the U.S. patent N 4406899, 1983, as well as methods for their preparation described two schemes reactions in which the reactants and products require the use of blocking and deblokiruyuschee groups. According to the reaction scheme illustrated by real examples, the resulting product requires phase chromatographic purification for separating a mixture of2and3-isomers, and the resulting cefepime is zwitterionic form. However, black is 301 (Murray A. Kaplan, and others), 1990, disclosed a heat-resistant crystalline salt of cefepime in dry powder form having an excellent stability at room temperature and higher stability at elevated temperatures compared to zwitterionic form described Aburaki and others in the U.S. patent N 4406899.

In U.S. patent N 4868294 (Brundidge, and others), 1989, describes how to obtain 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate salts, essentially not containing2isomer, and their use in the method of acylation in aqueous medium to obtain antibiotic of cefepime in the form of a sulfate salt.

In U.S. patent N 4754031 (Angerbauer, and others), 1988, describes a method for several cephalosporin antibiotics, including of cefepime in zwitterionic form. Although this method is not applicable protective groups, however, for the activation reaction of acylation in aqueous medium used anhydride, and for zwitterionic form of cefepime use of chromatographic purification stages.

In U.S. patent N 4943631 (Brian E. Looker), 1990, describes an improved method of producing the antibiotic of cefepime in the form of hydroiodide. The formation of undesirable2isomer is governed by the application as an intermediate connection sulfoxy which includes two additional stages in comparison with the method of the prototype and in the way continue to use a protective group, requiring procedures for blocking and unblocking. In addition, as a method of cleaning method requires column chromatography, which is impractical on an industrial scale.

Obtaining a crystalline sulfate salt and zwitterion described above occur using essentially the same method of acylation in aqueous medium with the use of different blocking deblokiruyuschee groups and active esters. In all cases featured crystalline form of a hydrate of the dihydrochloride of cefepime should be obtained through purified zwitterion form of cefepime. Thus, there is a need to create a simple and inexpensive technique of direct acylation, not including additional stages of the reaction and removal of protective groups, stages of regulation stereochemistry and stages of chromatography was carried out, and, more importantly, methods of acylation, in which the target antibiotic hydrate of the dihydrochloride of cefepime get essentially without admixture of antisolar and2isomer.

The invention provides a method of chemical acylation, in particular, the method of acylation in aqueous medium to obtain such antibiotic, as a hydrate of the dihydrochloride of cefepime, basically no content is(2-aminothiazol-4-yl)-2-methoxyaminomethyl-Reid, hydrochloride, essentially not containing anti-isomer, which is used in the method of acylation to obtain Severinovich antibiotics of a wide profile.

In Fig. 1 shows the spectrum of the proton nuclear magnetic resonance SYN-2-(2-aminothiazol-4-yl)-2-methoxyaminomethyl - chloride, hydrochloride of example 10 in acetic acid-d4(100 MHz), Fig. 2 spectrum of nuclear magnetic resonance (proton) of the product of example 12 in acetic acid-d4(100 MHz), Fig. 3 spectrum of the proton NMR of the product of example 13 in acetic acid-d4(100 MHz), Fig. 4 spectrum of the proton NMR of the product of example 14 in acetic acid-d4(100 MHz).

The invention provides a method of acylation in the aquatic environment for N-acylation of 7-amino-3-/(1-methyl-1-pyrrolidino)methyl /ceph-3-em-4-carboxylate SYN-isomer of 2-(2-aminothiazol-4-yl)-2-methoxykynuramine, essentially not containing anti-isomer, obtaining thermally stable crystalline hydrate of the dihydrochloride of cefepime, essentially not containing anti-isomer and2isomer and is represented by the formula Y, where n is 1 or 2.

H2N 2HClZH2O

The advantages of this method of acylation in of them together. Withdrawal conventional amino - and carboxylamide groups and, accordingly, removal of additional chemical stages required for blocking and release is an advantage from the point of view of the efficiency of the method in General and from the point of view of reducing the cost due to the consumption of reagents in comparison with how-prototypes. In this way also provides and ensures the control of the stereochemical configuration of methoxyindole and double3connection cephalosporin nucleus without having to split undesirable cephalosporin by-products by chromatography and without the need of determining the stereochemistry sulfoxide intermediate compounds, such as described in U.S. patent N 4043631. Another advantage of the invention consists in the obtaining and use of unprotected crystalline hydrochloride SYN-isomer of 2-(2-aminothiazol-4-yl)-2-methoxykynuramine formula III, which avoids the use of unusual and sometimes difficult outgoing groups described in the prototypes. The use of simple chloride ion as the departing group avoids the use of potentially toxic waste groups such as 2-mercapto is to obtain the desired heat-resistant crystalline hydrate of the dihydrochloride of cefepime directly from the reaction mixture of the process of acylation without the need to obtain and allocate sulfate or zwitterion of cefepime. The main advantage of the featured embodiment of the present method of acylation in the aquatic environment is to get the targeted antibiotic without needing to apply any similitude funds or any soluble siciliane derivatives. This method successfully carried out without the use of protecting groups, governing the stereochemistry of the groups, solubilizers similarbank groups or chromatography to obtain water-soluble crystalline hydrate of the dihydrochloride of cefepime, essentially not containing anti-isomer and2-isomer, with high yield directly from the aqueous-organic reaction mixture.

The present invention also provides stable crystalline SYN-isomer of 2-(2-aminothiazol-4-yl)-2-methoxyimino - illorin, hydrochloride, essentially not containing antihisamine and represented by formula III

NH HCl

The result is essentially the absence of the anti-isomer of the compound III can be converted into a wide range of cephalosporins, in turn essentially not containing anti-isomer, without the need for chromatographic separation of SYN - and anisomerous. Due to the high stability of the compound IIi can be isolated and Mor what is the advantage in attaining the target of antibiotics, essentially not containing2isomer. Another advantage of the intermediate compounds of formula III is that it does not require blocking (protection) amino prior to acylation or release (removal of the protective group) of an amino group after acylation, affecting, therefore, on the effectiveness of the method. Another advantage of the acid chloride of formula III is in its application in the method of acylation to obtain broad-spectrum cephalosporins. Unlike other methods, for example, described in U.S. patent N 4406899 (Aburaki and others) of the intermediate compound of formula III has a simple and non-toxic waste group (chloride ion), do not require special precautions when removing from the target of the antibiotic, as is the case with most other known waste groups. In addition, some known intermediate compounds containing other waste groups, are synthesized with labor and other intermediate compounds containing waste type group 2-mercaptobenzothiazoles, as found, are toxic (Chem. Abstracts, 1989, T. III(3), 19243 R.)

A reaction scheme I

O(CH SYN-Isomer hydrochloride of the acid chloride of formula IIl can be obtained from the SYN-isomer of the acid of formula I, which have been welcome formula II, then, if desired, is isolated in the form of anhydrous crystalline compounds of formula II. Education hydrochloride successfully carried out using at least one molar equivalent of gaseous hydrogen chloride in an inert organic solvent, such as toluene, acetonitrile, dichloromethane, acetone, benzene, xylene, cyclohexane, hexane, dioxane or diethyl ether in the temperature range from -10 to 50aboutC. the Reaction is recommended in toluene, dichloromethane or acetonitrile, and the resulting hydrochloride of formula ll can be isolated or used in Situ. When conducting the reaction in acetonitrile, the resulting hydrochloride of the formula ll tends to hold the solvent. It is therefore desirable using hydrochloride acid of the formula ll of acetonitrile as soon as possible at the next stage, to avoid substitution MES atmospheric moisture. Most preferably carrying out the reaction in toluene or dichloromethane at a temperature from 0aboutWith up to room temperature.

The salt of the acid of formula II then successfully treated gloriouse means, most preferably oxalylamino in combination with dimethylformamide with the participating funds may lead to isomerization with the formation of undesirable anti-isomer or a mixture of SYN - and anti-isomer. In addition, the use of such gloriously funds, as pentachloride phosphorus can lead to chlorination in the 5-position of the thiazole cycle with subsequent undesirable contamination of the antibiotic. The creators of the present invention discovered that in addition to obtain hydrochloride of the acid of formula II a crucial role in the way to obtain the SYN isomer of compound III, essentially not containing anti-isomer, plays the correct selection glorieuses means and the reaction conditions such as solvent and temperature.

Methods of chlorination, usually used to activate the acid, are well known. Pentachloride phosphorus, which is the most widely used gloriouse means for chlorination of compound (II) is not suitable, because it leads to isomerization of methoxyimino with the formation of undesirable anti-isomer of compound III. This is clearly shown in the following examples 12, 13, 14, and 16. Another known method of chlorination is the use of oxalicacid in combination with dimethylformamide. However, the creators of the present invention discovered that the way of using oxalicacid, in which the catalyst used dimethylformamide, does not lead to the formation of Sacele intensive research, the creators of the present invention have found, the use of dimethylformamide at less than equimolar amount relative to the amount of oxalicacid has an adverse effect on education target SYN-isomer hydrochloride of the acid chloride of the formula III. Most preferably, if the molar amount of dimethylformamide to exceed the molar amount of dimethylformamide. The inventors also found that the use of excess amounts of dimethylformamide is also unfavorable for the reaction and the stability of the target product. Thus, the inventors of the open method of adjusting the volatility of the reaction products or the excess of the chloride ion formed by oxalylamino, or an excess of dimethylformamide, which plays a crucial role in the formation of stable crystalline SYN-isomer of compound III, essentially not containing anti-isomer. If the conversion of the compounds of formula II to the compound of formula III is incomplete, identified as product compound III will contain small amounts of residual SYN-isomer of the acid of formula II. The presence in the received quality of the product compound III some amount of unreacted compound (II) and a small culicivora antibiotic, essentially not containing anti-isomer of the antibiotic.

The creators of the present invention has also been found that the reaction temperature and the solvent for it is essential. It is recommended to carry out the reaction in an inert organic solvent, such as dichloromethane, chloroform or acetonitrile at temperatures below -10aboutC. Most preferably carrying out the reaction in dichloromethane and the temperature range from -15 to -40aboutC.

The use of SYN-isomer hydrochloride of the acid chloride of the formula III to produce useful antibiotics of a wide profile by conventional acylation reaction is illustrated by reaction scheme 2. More specifically, a reaction scheme 2 illustrates the application of the acid chloride of formula III to obtain the antibiotic of a wide profile of cefepime, basically bore containing the anti-isomer and2isomer. In addition, the hydrochloride of the acid chloride of the formula III may be used for the synthesis of cephalosporin antibiotics, characterized by SYN-configuration 2-(2-aminothiazol-4-yl)-2-methoxyimino - sludge attached to the 7-amino group of a cephalosporin nucleus, such as: cefodizime, aminoxy, Cefotaxime, cefpirome, cefpodoxime, rethinam, cefteram the IDA prototype is anti-isomer, and not target SYN-isomer, the creators of the present invention was replaced in the acylation reaction, illustrated by reaction scheme 2, the SYN-isomer of compounds of formula III product prototype, obtained, for example, in example 14. The final cephalosporin product obtained according to the methods of examples 17 and 18 were compared with the antibiotic cefepime, obtained by the method of the present invention. As can be seen from the comparison (example 19) anti-cefepime obtained according to the instructions of the prototype, not identical useful Shin-cefepime wide profile, synthesized using the present invention.

As used here and in the claims the value of the expression "not substantially contain" means that the compound contains less than about 5% of the undesired isomer. Preferably, if the compound contains less than about 1% of the undesired isomer.

According to the method of the invention N-acylation of compounds of formula IV SYN-isomer hydrochloride of the acid chloride of the formula III receive an antibiotic of a wide profile hydrate of the dihydrochloride of cefepime, essentially not containing anti-isomer and2-isomer, as illustrated by reaction scheme 2

< / BR>
7-Amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-SUB>4can be obtained according to the General method described by S. P. Brundidge, etc. in U.S. patent N 4868294.

Acceptable solvents that may be used in the method of acylation in aqueous medium include water and organic solvents, such as water with water-soluble organic solvent, for example methanol, ethanol, isopropanol, butanol, acetone, tetrahydrofuran, acetonitrile, dioxane, dimethylacetamide, dimethylformamide, etc., the pH Value of the reaction mixture control titration priemlimom organic and inorganic base, such as sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium hydroxide, ammonium hydroxide, primary amine, secondary amine, tertiary amine, etc., by neutralization of the resulting hydrochloric acid. Featured organic bases that may be used include, for example: diethylamine, triethylamine, diisopropylethylamine, N-methylmorpholine, 2,6-lutidine, N,N-dimethylaniline, N,N-diethylaniline, etc. To the most recommended bases include ammonium hydroxide, triethylamine or N-methylmorpholine.

Acylation with success carried out at pH 5-7,5, preferably at pH of 6.2 to 6.8. The way netpreteen at a temperature of from -10 to -40aboutC. Upon completion of the acylation reaction mixture is acidified with an appropriate acid, preferably sulfuric acid to a pH of 1.8 to 2.6 with the formation of the sulfate salt of the target cefepime antibiotic, essentially not containing anti-isomer and2isomer. If desired, the sulfate salt of cefepime may be converted into other salts of cefepime, for example, a hydrate of the dihydrochloride by the method of Kaplan and others in the U.S. patent N 4910301.

One aspect of the present invention is a method of acylation to obtain antibiotic broad profile of cefepime, essentially not containing anti-isomer and2-isomer, and the method consists in the reaction of 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate salts, essentially not containing2-isomer, with the SYN-isomer of 2-(2-aminothiazol-4-yl)-2-methoxykynuramine, hydrochloride, essentially not containing anti-isomer in water or preferably mixed aqueous-organic solvent at a carefully controlled pH value in the range of 5-7,5.

Another aspect of the invention provides stable crystalline SYN-isomer hydrochloride 2-(2-aminothiazol-4-yl)-2-methoxykynuramine, essentially not containing anti-isomer of the formula

practical SYN-isomer hydrochloride 2-(2-aminothiazol-4-yl)-2-methoxykynuramine, essentially not containing anti-isomer, and the method consists in the reaction of anhydrous crystalline hydrochloride SYN-isomer of 2-(2-aminothiazol-4-yl)-2-methoxyimino-susei acid with a mixture consisting of at least one molar equivalent of oxalicacid and at least one molar equivalent or a slight excess of dimethylformamide regarding the number of oxalicacid, in an inert organic solvent at a temperature below -10aboutWith the formation of stable crystalline SYN-isomer hydrochloride 2-(2-aminothiazol-4-yl)-2-methoxykynuramine, essentially not containing anti-isomer.

The complete N-acylation of the compounds of formula IV is confirmed by the well-known and available methods of detection, such as thin-layer chromatography, liquid chromatography, high pressure and spectral methods. According to the recommended method of the invention upon completion of the reaction, add enough acid, such as hydrochloric acid, sulfuric acid, etc. to ensure crystallization of the target salt of cefepime, and then diluted with a water-soluble solvent such as methyl ethyl ketone, acetone, isopropanol, butanol, etc., on Iwate sufficient amount of sulfuric acid by crystallization of the sulfate salt of cefepime, essentially not containing anti-isomer and2isomer. Sulphate of cefepime can then be turned into featured dihydrochloride monohydrate according to the method described Kapla and others in the U.S. patent N 4910301. Obtained by this method sulfate of cefepime may be neutralized with a weak base, preferably a weakly alkaline ion exchange resins used for such purposes, preferably selling ion exchange resins, such as: Amberlite LA-2, Dowex WGR, Bio-Rad AG3-X4A, Amberlite 1RA 93, Amberlite IRA 35, etc., with the formation of aqueous or aqueous-organic solution containing zwitterion form of cefepime. The solution is then treated with a sufficient quantity of hydrochloric acid, possibly in the presence of water-soluble organic solvent to cause crystallization featured crystalline dihydrochloride monohydrate of cefepime. Most preferably, if the aqueous-organic reaction mixture obtained by acylation in the aquatic environment this way is treated with sufficient amounts of hydrochloric acid with the intention of inducing and to ensure crystallization of the specified antibiotic, namely, a hydrate of the dihydrochloride of cefepime adding such water bodies which should be sufficient to complete crystallization of the specified antibiotic, and usually 2-9 volume to the amount of water in the aqueous-organic reaction medium with the formation of the heat resistant crystalline hydrate of the dihydrochloride of cefepime, essentially not containing anti-isomer and2isomer.

If you wish to get only the dihydrochloride monohydrate of cefepime aqueous-organic reaction mixture from acylation in aqueous medium preferably is treated with a sufficient quantity of hydrochloric acid and diluted with appropriate amount of water-soluble organic solvent of the above method to guarantee the crystallization target monohydrate form. Or, if you want to get a stable dihydrate dihydrochloride of cefepime, aqueous-organic reaction mixture can be successfully treated with hydrochloric acid to a greater equivalent concentration and therefore the amount of water-soluble organic solvent, in order to maintain the crystallization point of turbidity before adding an additional amount of organic solvent to complete the crystallization. But for professionals it is obvious that if not carefully regulate the extraction of the aqueous-organic reaction mixture, vozmogla one of the desired hydrate can be made from any hydrate or of a mixture of hydrates by recrystallization, are described here.

Crystalline dihydrochloride monohydrate of cefepime obtained by this method can be used to obtain a stable crystalline dihydrochloride dihydrate of cefepime by recrystallization at regulated concentrations of solvent and hydrochloric acid and the duration of aging in the point cloud (primary crystallization) as described here. Or crystalline dihydrochloride dehydrate obtained by this method can also be used for the preparation of stable crystalline dihydrochloride monohydrate of cefepime in other regulated the conditions described here. Thus, the method of the present invention can be used to obtain target as monohydrate and dihydrate specified antibiotic.

In contrast to the labile dihydrochloride dihydrate of cefepime described in U.S. patent N 4910301, which easily loses the second mole of water, the crystalline dihydrochloride dihydrate of cefepime, which can be obtained by this method, as found, has a well-defined crystal structure, which maintains the second mole of water. A new crystalline form of dihydrate (IG the conditions, for example, in air at 70aboutWith more than two months, in vacuum over R2O5at the 50aboutC for 48 h in a drying Cabinet at 70aboutC for 96 h and in terms of high and low relative humidity. Crystalline dihydrate has characteristic peaks in the infrared absorption spectrum at 3574 cm-1and 3432 cm-1as shown, the FT-IR diffusion reflective spectroscopy with CVG and in a ditch for a sample of 13 mm with a spectrometer Nicolet 20SX. Such is resistant to heat and moisture crystalline dihydrate of cefepime also characterized by a powder diffraction of x-rays, the results of which are shown in the table, where "d" refers to the interplanar distances and "T/To" refers to the relative percentage intensively. Data of x-ray analysis was obtained using x-ray diffractometer model Rigaku Geigerflex and when the radiation from the Nickel source with a copper filter (K ) with wavelength in 1,5425 .

Thus, one of the embodiments of the invention provides a method of producing antibiotic hydrate of the dihydrochloride of cefepime, essentially not containing anti-isomer and2-isomer, the method consists in the reaction of 7-and the mayor of the hydrochloride of 2-(2-aminothiazol-4-yl)-2-methoxykynuramine, essentially not containing anti-isomer, in mixed aqueous-organic solvent at pH 5-7,5.

Featured embodiment of the present invention additionally includes receiving the SYN-isomer hydrochloride 2-(2-amino-thiazol-4-or-2-methoxyimino - Teilhard, essentially not containing anti-isomer, the reaction of the anhydrous salt of a carboxylic SYN-isomer of 2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid with a mixture containing at least one molar equivalent of oxalicacid or at least one molar equivalent or a slight excess relative to oxalylamino of dimethylformamide, in an inert organic solvent at a temperature below -10aboutC.

The preferred embodiment of the present invention provides a method of producing antibiotic hydrate of the hydrochloride of cefepime, essentially not containing anti-isomer and2-isomer, the method consists in the reaction of 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate, essentially not containing2-isomer, with the SYN-isomer hydrochloride 2-(2-aminothiazol-4-yl)-2-methoxyaminomethyl - chloride, essentially not containing anti-isomer in aqueous-organic solvent, and optionally includes cobaltarthurite.

The most preferred embodiment of the invention provides a method of producing antibiotics, dihydrochloride monohydrate of cefepime and the dihydrochloride dihydrate of cefepime directly from the aqueous-organic reaction mixture of the present method.

The applicability of cefepime shown in U.S. patent N 4406899 (Aburaki and others). Sustainable digitata form of cefepime obtained by this method, has antibiotic properties above of cefepime for U.S. patent N 4406899, and finds use as an antibiotic in the same way.

It is obvious that the description and examples are for illustrative purposes and are not intended to limit the scope of the invention.

P R I m e R 1. SYN-2-(2-Amino-thiazol-4-yl)-2-methoxyimino acid, hydrochloride.

To a suspension of 25 g (124,25 mmole) of 2-(2-aminothiazol-4-yl)2-methoxyethoxy acid in toluene (250 ml) at 20-28aboutWith miss gaseous hcl, Hcl is injected under the surface of the reaction mixture in two portions 8.1 g (to 222.2 mmole) and 4.8 g (131,7 mmole) with stirring 30 minutes between adding portions. After incubation for 1 h at 20aboutThe product is filtered under nitrogen atmosphere, washed with toluene (50 ml) and hexane (250 ml) and drying in yl)-2-methoxykynuramine hydrochloride.

To a solution of 0.77 ml (10 mmol) of dimethylformamide in dichloromethane (40 ml) at 5aboutTo add to 0.89 ml (10 mmol) of 98-th oxalicacid in dichloromethane (4,1 ml). Added dropwise keep the temperature 4-5aboutC. To the resulting suspension cooled to -27aboutTo add 2.37 g (10 mmol) of the hydrochloride of 2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid of example 1. The suspension is stirred for 2.5 h at -25aboutC. Filtration in a nitrogen atmosphere and washing with dichloromethane (50 ml) and hexane (100 ml) to obtain 1.78 g (69,5%) of white crystalline title compound after drying in vacuum at 20aboutC.

The obtained acid chloride acelerou hydrochloride diphenylmethylene ether 7-aminodesacetoxycephalosporanic acid in a solution of pyridine with the education which gives a single spot (TLC) product matching and inseparable with authentic sample target deacetoxycephalosporanic ether.

P R I m e R 3. SYN-2-(2-Aminothiazol-4-yl)-2-methoxykynuramine hydrochloride.

To a solution of 1.55 ml (20 mmol) of dimethylformamide in dichloromethane (80 ml) and 5aboutWith the type of 1.78 ml (20 mmol) of oxalicacid 98% purity in dichloromethane (8.2 ml). The time of addition at 5-8aboutWith extending t is klorid 2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid and 4.75 g, 20 mmol) of example 1. The suspension is stirred for 2.5 hours at a temperature of from -25 to -30aboutC. Filtration in a nitrogen atmosphere and washing with dichloromethane (75 ml) and hexane (100 ml) obtained after drying at 20aboutWith the vacuum 3.57 g (69,7%) of crystalline title compound.

A portion of the resulting hydrochloride of the acid chloride (dry) acelerou in a solution of pyridine hydrochloride diphenylmethylene ether 7-aminodesacetoxycephalosporanic acid and get giving a single spot (TLC) product matching and inseparable with authentic sample target datasetexception ether.

P R I m e R 4. Getting 7-/2-(2-aminothiazol-4-yl)-2-(Z)-methoxykynuramine /-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate (cefepime)

< / BR>
In 9 ml of a mixture of acetone-water (2:1) with triethylamine at pH 6.5 and 20aboutTo dissolve monohydrated 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carbox-Silat (0,85 g, 2 mmole) (obtained according to the method described in U.S. patent N 4714760 (S. P. Brundidge, etc.)/. Add hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxykynuramine (0.56 g, 2.2 mmole) (receipt see example 3) using triethylamine for keeping pH in the range of 5-7. The analysis of the obtained solution liquid hiloti to pH 2.2 to get to 0.63 g of the title of the antibiotic in the form of its sulfate salt (exit activity 51%), described in U.S. patent N 4406899 (Aburaki and others), 1983, and U.S. patent N 4910301 (Kaplan and others), 1990.

P R I m e R 5. SYN-2-(2-Aminothiazol-4-yl)-2-methoxybenzonitrile, hydrochloride.

To a solution of 9.75 ml (125,9 mmole) of dimethylformamide in dichloromethane (450 ml) and 5aboutWith added dropwise a solution 11,21 ml (125,9 mmole) of oxalicacid (98%) in dichloromethane (15 ml). The addition of at 5-7aboutWith ends 10 minutes To the resulting suspension was cooled to -25aboutTo one portion add 28.5 g (119,9 mmole) of the hydrochloride of SYN-2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid. The suspension is stirred for 3.5 h at -25 to -30aboutWith that filtered in a nitrogen atmosphere, washed with dichloromethane (100 ml) and hexane (400 ml) and dried at 20-25aboutWith the vacuum. The output of the crystal of the title compound 30,7 (72,5%).

The obtained acid chloride acelerou in a solution of pyridine hydrochloride diphenylmethylene ether 7-aminodesacetoxycephalosporanic acid and get essentially one spot (TLC) target deacetoxycephalosporanic ether when compared with an authentic sample.

The title acid chloride (200 mg, 0.8 mmole) hydrolyzing in water.1H-NMR spectrum is identical to the spectrum of the original Shin-acid.

P R and m is dimetilformamida in dichloromethane (350 ml) and 5aboutWith added dropwise 9,34 ml (105 mmol) of oxalicacid (98% purity) in dichloromethane (5 ml). The maximum temperature during the addition of 7aboutC. the Resulting suspension is stirred for 10 min at 5aboutC and then cooled to -27aboutC. One portion add the hydrochloride of 2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid (23,8 g, 100 mmol). The suspension is stirred for 2.5 h at -25 to -30aboutWith that filtered in a nitrogen atmosphere, washed with dichloromethane (25 ml) and hexane (125 ml) and after drying at 20aboutWith the vacuum get 21,39 g (83.5 per cent) of the crystalline hydrochloride of the acid chloride.

Calculated With 28,14; N Was 2.76; N 16,41; S To 12.52.

WITH6H7N3O2SCl2.

Found, 28,25; N. Of 2.93; N 16,32; S 12,67.

1H-NMR (DMSO-d6) 3-93 (CH3),? 7.04 baby mortality (N5).

P R I m e R 7. SYN-2-(2-Aminothiazol-4-yl)-2-methoxyimino acid, hydrochloride.

In the suspension 87 g (432,4 mmole) of SYN-2-(2-aminothiazol-4-yl)-2-methoxyimino Noi acid in toluene (870 ml) at 22aboutWith the two portions of the injected gaseous Hcl: 17,5 g (480 mmol) for 30 min and 15 g (410 mmol) over 20 min with stirring 20 min between the introduction portion. The suspension is stirred for 1.5 h at 25aboutWith the filter in the atmosphere astonomer connection.

Calculated C 30,32; N 3,39; N 17,68; S 13,49; Cl 14,92.

WITH6H8N3O3SCl.

Found, 30,51; N 3,39; N Comprised 17.54; S 13,37; CL Of 14.90.

P R I m e R 8. SYN-2-(2-Aminothiazol-4-yl)-2-methoxykynuramine hydrochloride.

The solution to 32.4 ml (419,7 mmol) of dimethylformamide in dichloromethane (400 ml) and 5aboutWith added dropwise with 37.4 ml (419,7 mmole) 98% oxalicacid. The resulting suspension is cooled to -25aboutWith and add at -25aboutTo a suspension of 95 g (399,7 mmole) of the hydrochloride of SYN-2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid of example 7. The suspension is stirred for 2.5 hours at a temperature of from -25 to -28aboutWith that filtered in a nitrogen atmosphere, washed with dichloromethane (100 ml) and hexane (500 ml) and after drying in vacuum at 20-25aboutWith the get of 84.3 g (82,3%) of crystalline title compound.

Calculated With 28,14; N Was 2.76; N 16,41; S To 12.52.

WITH6H7N3ABOUT2SCl2.

Found, With 27,90; N 3,10; N 16,14; S 12,27.

1H-NMR (DMSO-d6) 3,95 (CH3),? 7.04 baby mortality (N5).

P R I m e R 9. Obtain 7-(2-(2-aminothiazol-4-yl)-2-(Z)-methoxykynuramine /-3-(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate (cefepime).

To a solution of 240 ml of acetone and 80 ml of water add the so By using an automatic device for titration model Radiometer V80 configured at the end of the titration at pH 6.5 and filled with N-methylmorpholine, 4 portions with an interval of 5 minutes, maintaining the pH at 6.5 to add hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxykynuramine (20 g, 0,0785 mol) (receipt see example 5). Upon completion of addition, the fine suspension is stirred for further 20 minutes at room temperature. The addition of 21 ml of 6 n H2SO4the pH value of the reaction mixture is reduced to approximately 2.65. Is the precipitation of the title compound. Suspension tatrallyay and stirred for 20 minutes at room temperature. The addition of 16 ml of 6 n H2SO4set the pH of the suspension of 1.8 and stirring is continued for another 60 minutes the Suspension is filtered under vacuum and washed with 70 ml of a mixture of water-acetone (1:1) and then 70 ml of acetone to obtain 24,09 g (88.5% of stachion. the yield by weight) of the title compound, identical to the compound of example 4 and cefepime, described in U.S. patent N 4406899 (Aburaki and others), 1983, and in U.S. patent N 4910301 (plan and others), 1990.

P R I m e R 10. Receiving SYN-2-(2-aminothiazol-4-yl)-2-methoxyaminomethyl-reed hydrochloride.

To a solution of dimethylformamide (8,76 ml, 0,113 mol) in dichloromethane (375 ml) in 5aboutWith live 10 min and then cooled to -25aboutC. In a dry nitrogen atmosphere for 11 min servings add hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxime-noxubee acid (25 g) and the suspension stirred for 2.5 h at -25aboutC. the Product is filtered in a dry nitrogen atmosphere and the filter cake washed with dichloromethane (80 ml). Drying of the product under 20-25aboutC in vacuum over P2O5get 23,88 g (88,6%) of the title compound as a pale yellow crystalline substance.

Calculated With 28,14; N Was 2.76; N 16,41; S To 12.52; CL 27,68.

WITH6H7N3O2SCl2.

Found, With 28,06; N 2,71; N 16,26; S 12,30; Cl 27,23.

The product of the above experience is characterized by a spectrum of proton nuclear magnetic resonance (1H-NMR) in acetic acid-d4, shown in Fig. 1.

1H-NMR (CD4CO2D) : 4,14 (CH3), and 7.1 (N5).

The residual level of hydrochloride acid in CH3(4,11) the integral is 5.1% Trace level isomeric H5 visible when to 7.67 h/million

P R I m e R 11. SYN-2-(2-Aminothiazol-4-yl)-2-methoxykynuramine hydrochloride.

To a solution of dimethylformamide (17,92 ml, 231,9 mmole) in dichloromethane (375 ml) in 5aboutWith add oxalicacid (19,76 ml, 220,8 SUP>C and then cooled to -25aboutC. Add the hydrochloride of 2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid (25 g, 105,2 mmole). The resulting solution was seeded with the title compound with a suspension of the product. The suspension is stirred for 2.5 h at -25aboutWith that filtered in a dry nitrogen atmosphere, washed with dichloromethane (150 ml) and after drying in vacuum at 20-25aboutTo get being 9.61 g (35,7%) of crystalline title compound.

A portion of the dry hydrochloride of the acid chloride acelerou in a solution of pyridine hydrochloride diphenylmethylene ether 7-aminodesacetoxycephalosporanic acid and get the product with essentially a single spot (TLC), matching and inseparable with authentic sample target deacetoxycephalosporanic ether.

P R I m e R 12. Obtaining 2-(2-aminothiazol-4-yl)-2-methoxykynuramine hydrochloride.

The experimental procedure of example 1 of the patent Czechoslovakia N 238950 reproduced as follows.

In 30 ml of benzene at the 21aboutTo form a suspension of SYN-2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid (4 g) with a value of KF is 0.06% Add one drop of dimethylformamide followed by the addition of one portion of 5 g of powdery pin 40aboutWith the complete dissolution of the reagents. The solution is allowed to cool and when the 36aboutWith a precipitate. After stirring 30 minutes the temperature reaches 22aboutS. Light yellow precipitate is filtered in a dry nitrogen atmosphere and washed with 30 ml of benzene and 20 ml of heptane. The output is 2,88 g after drying in vacuum for 18 h at 20-25aboutWith over R2ABOUT5.

The product of the above experience is characterized by a spectrum of proton nuclear magnetic resonance (1H-NMR) in acetic acid-d4, shown in Fig. 2, where N5 is manifested when 7,56 ppm and CH3when 4,34 h/million This range meets the capital connection configuration of the anti-isomer and not SYN-isomer, as indicated in the patent Czechoslovakia.

P R I m e p 13. Obtaining 2-(2-aminothiazol-4-yl)-2-methoxykynuramine hydrochloride.

The experimental procedure of example 2 of the patent Czechoslovakia N 238950 reproduced as follows.

In 20 ml of acetonitrile, which set the value KF of 0.22% obrazuyuschim-2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid (4 g), having a value of KF 0,06% At 20aboutTo add one drop of dimethylformamide and after adding 6 g of powdered pin 20aboutAnd when the 33aboutWith a precipitate. After stirring 30 min, the product is collected in a dry nitrogen atmosphere and washed with 30 ml of benzene and 20 ml of heptane. After drying at 20-25aboutC in vacuum over P2ABOUT5for 18 h obtain 1.86 g of product.

The product of the above experience is characterized by a spectrum of proton nuclear magnetic resonance (1H-NMR) in acetic acid-d4, shown in Fig. 3, where N5 is manifested when 7,56 ppm and CH3when 5,31 h/million1H-NMR spectrum corresponds capital connection configuration of the anti-isomer and not SYN-isomer, as indicated in the patent Czechoslovakia.

P R I m e R 14. Obtaining 2-(2-aminothiazol-4-yl)-2-methoxykynuramine hydrochloride.

The experimental procedure of example 3 of the patent Czechoslovakia N 238950 reproduced as follows.

To 30 ml of dichloromethane added concentrated hydrochloric acid (0.16 ml) and after cooling to -10aboutWith portions added 6.5 g of pentachloride phosphorus. After warming to 0aboutTo one portion add 4 g of SYN-2-(2-aminothiazol-4-yl)-2-methoxyimino acid with a value of KF 0,06% the temperature is increased to 2aboutC. Complete dissolution occurs through SUP>aboutWith that filtered in a dry nitrogen atmosphere, washed with 30 ml of benzene and 20 ml of heptane and after drying for 18 h in vacuum over P2ABOUT5and when 20-25aboutTo obtain 3.42 g of light yellow powder.

The product of the above experience is characterized by a spectrum of proton nuclear magnetic resonance (1H-NMR) in acetic acid-d4, shown in Fig. 4, where N5 is manifested when 7,56 ppm and CH3when or 4.31 h/million 1H-NMR spectrum corresponds capital connection configuration of the anti-isomer and not SYN-isomer, as indicated in the patent Czechoslovakia.

P R I m e R 15. Attempt to obtain 2-(2-aminothiazol-4-yl)-2-methoxyaminomethyl - chloride hydrochloride.

The General methodology described in example 7 of U.S. patent N 4203899 to prevent protected aminothiazolyl acid into the corresponding acid chloride used unprotected aminothiazoline acid as follows.

In 30,5 ml of benzene suspended hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid and cooled to 20aboutC. Add oxalicacid (2,09 ml, 0,024 mol) followed by addition of dimethylformamide (0.5 ml, 0,0065 mol). The temperature rises to 22aboutWith with vigorous evolution of gas. In centerour in vacuum to remove solvent and the resulting yellow product is dried 16 h in vacuo over P2ABOUT5and when 20-25aboutC. Output 2,59,

The product of the above experience is characterized by a spectrum of proton nuclear magnetic resonance in acetic acid-d4in which N5 is manifested when a 7.6 h/min, and CH3at 4.37 h/million Range product meets the capital connection configuration of the anti-isomer.

P R I m e R 16. Attempt to obtain 2-(2-aminothiazol-4-yl)-2-methoxyaminomethyl - chloride hydrochloride.

The General methodology described in example 59 U.S. patent N 4203899 to turn protected aminothiazolyl acid into the corresponding acid chloride applied to unprotected aminothiazoline acid as follows.

In 25 ml of dichloromethane suspended hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid (2.38 g of 0.01 mol) and after cooling to 4aboutWith the type of 2.08 g (0,01 mol) of pentachloride phosphorus. Under ice cooling, the temperature rises to 6aboutFrom and after cooling again to 4aboutWith the suspension stirred for 1 h the Precipitate is filtered in a dry nitrogen atmosphere, washed with dichloromethane (10 ml) and after drying in vacuum at 20-25aboutTo obtain 1.4 g of pale yellow solid.

The product of the above experience oharra H5 occurs when to 7.61 ppm, and CH3when 4,34 h/million Range product meets the capital connection configuration of the anti-isomer. Additionally, the product is contaminated neprevyshenie acid (1H-NMR has the N5 when 7,07 ppm and CH3when 4,06 ppm), which further confirmed by combining with the original acid.

P R I m e R 17. Acylation of 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate, HL-salt 2-(2-aminothiazol-4-yl)-2-methoxykynuramine hydrochloride (anti-form example 14).

To a pre-cooled solution of 9 ml of acetone and 3.4 ml of water at 10aboutWith add HL-salt of 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxy - LVL (1.13 g, and 2.26 mmole). At 0aboutWith five portions was added the hydrochloride of 2-(2-aminothiazol-4-yl)-2-methoxyaminomethyl - chloride (1,09 g, 4,21 mmole) (receipt see example 14) together with triethylamine (0,37 ml of 2.66 mmole) to maintain the pH at 6-7. The reaction mixture was stirred 15 min at room temperature. The analysis of the obtained solution liquid chromatography high pressure (18column with a gradient of 2-25% acetonitrile in 0.005 M NH4H2PO4) showed the presence of (ploddy peak) to 72.4% of anticipate with a retention time of 13,08 min and the lack of give in the Auteuil to pH 1.9 to obtain 1.48 g of anti-cefepime in the form of a sulfate salt. The product identification is confirmed1H-NMR spectroscopy (DMSO-d6), according to which the product contains of 0.58 mol of the salt of triethylamine.

P R I m e R 18. Acylation of 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate, HL-salt 2-(2-aminothiazol-4-yl)-2-methoxykynuramine hydrochloride (anti-isomer of example 14).

To a pre-cooled solution of 108 ml of acetone and 40.5 ml of water is added at 10aboutWith HL-salt of 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate (13.5 g, 0,0317 mol). The addition of 2.7 ml of 14% NH4OH install in suspended solids pH 7. At a temperature of 10aboutC for 60 minutes servings add the hydrochloride of 2-(2-aminothiazol-4-yl)-2-methoxyaminomethyl - chloride (of 13.05 g of 0.015 mole) obtained according to the method of example 14/ using 14% NH4OH (27 ml) to maintain a pH of 6.3-7 for the first half hour of addition and pH of 6.1 and 6.6 during the second half of the addition. The reaction mixture is stirred for 30 minutes at room temperature. The reaction mixture was filtered and washed with 6 ml of a mixture of acetone-water (2:1) and then 6 N. N2SO4(15 ml), which is slowly added to the filtrate with the establishment of the pH of 1.87 to 1.9. After stirring 1 h, the insoluble part hoteltravel the n add 1 l of acetone and the resulting mixture stirred for 40 min at 5-8aboutC. the Product is filtered, washed twice 24 ml of a mixture of acetone-water (4:1), 60 ml of acetone, and after drying in a vacuum get 20,64 g (116% from stachion. mass anti-cefepime in the form of sulfate salts (95,4% purity according ghvd).1H-NMR spectrum corresponds to the structure of anticipate containing 3 mol of ammonium salts.

P R I m e R 19. Comparison of the product of example 9 (SYN-isomer of cefepime) with the product of example 17 (anti-isomer of cefepime) showed the following differences in physical parameters.

Chromatography high pressure isomers of cefepime carried out on a column of waters M. Bondapak18(3.9 x 300 mm) using as solvent system 1000 ml of water containing 2,88 g (0,013 mol) of sodium salt heptanesulfonic acid, with a pH of 4 by addition of acetic acid and 100 ml of acetonitrile, at a flow rate of 2 ml/min. and the Products analyzed visually using the detector with variable wavelength forms a waters model 450 installed at 254 , and obtain the following results.

Retention time, min

the SYN-Isomer of cefepime

(example 9) 10,5

anti-Isomer of cefepime

(example 17) of 37.8

Spectra1H-NMR spectrum of SYN - and anti-isomers of cefepime on metasymbol group received glavmetalsnabsbyt. Specified chemical shifts correlate with DMSO at 2,49 h/million numbering System set forth in the formula and the table is given only for convenience.

SYN - and anti-cefepime

NH Wavy line represents the SYN - and anti-isomers on methoxyimino.

Comparison of the chemical shifts in the1H-NMR spectra (M. D.)

Designation Shin-Cefepime anti-Cefepime

C2-H2 4,04; 3,65 4,02,3,65

C6-H 5,33 5,31

C7-H 5,88 5,85

C11-N2 4,6; or 4.31 4,59; 4,3

C12-H3 2,93 2,93

C13-H4 3,7; 3,4 3,6; 3,3

C14-H4 2,10 2,1

C18-H4 6,88 EUR 7.57

C20-H3 3,92 4,05

NH 9,83 9,56

NH2 8,6 8,7

1H-NMR spectra of the two isomers of cefepime for methoxyimino differ significantly. SN(18) thiazole cycle SYN-(Z)-methoxymetopon isomer when 6,88 shifted to high field at EUR 7.57 ppm for anti-(E)-methoxymetopon isomer of CH(18).

P R I m e R 20. SYN-2-(2-Aminothiazol-4-yl)-2-methoxyimino acid, hydrochloride.

SYN-2-(2-Aminothiazol-4-yl)-2-metoxi - linoxyn acid (95,3 g, 424 mmole) in dichloromethane (570 ml) crushed in a mixer (15 min under nitrogen). The obtained fine suspension was diluted with dichloromethane (100 ml) and transferred under nitrogen into the reactor Bucha 1 l jacketed. In the reactor creates the nitrogen pressure in 5 psi (0.35 kg/cm2aboutC. the Mixture is stirred for further 30 min at 0aboutC, filtered and washed under nitrogen dichloromethane (350 ml). Drying the solid product 18 h in a vacuum get 110,9 g (111% uncorrected yield) of the title compound in the form of a snow-white powder.

Calculated With 30,32; N 3,39; N 17,68; S 13,49; Cl 14,91.

WITH6H8N3SCl.

Found, 29,37; N 3,17; N 16,34; S 12,70; CL 16,99.

1H-NMR (DMSO-d6) of 4.05 (s, 3H, CH3) and 5.9 (s, 15 mol. residual CH2CL2), and 7.1 (s, 1H, C-5 H). The signals are also visible when 4,18 (s, 3H, CH3) and 7.7 (s, 1H, C-5 H), which corresponds to approximately 2% of the anti-isomer.

P R I m e R 21. SYN-2-(2-Amino-thiazol-4-yl)-2-methoxyimino acid, hydrochloride.

SYN-2-(2-Aminothiazol-4-yl)-2-metoxi - linoxyn acid (25 g, 124 mmole) in acetonitrile (125 ml) titrated under nitrogen 1,39 m solution of Hcl in acetonitrile (89,2 ml, 123,9 mmole) and maintained at 10-15aboutC. the Mixture is stirred for another 30 min at 10-15aboutC, filtered and washed under nitrogen with acetonitrile (200 ml). Drying the solid product 3 h in vacuum at 45aboutWith the gain of 29.5 g (97,4%), uncorrected yield) of the title compound in the form of a snow-white powder.

3) and 7.8 (s, 1H, C-5H), corresponding to approx. 0.5% of the anti-isomer.

P R I m e R 22. SYN-2-(2-Aminothiazol-4-yl)-2-methoxykynuramine hydrochloride.

Hydrochloride of 2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid (56,24 g, 210 mmol), containing about 11 wt./wt. residual acetonitrile in dichloromethane (450 ml) crushed 3 min under nitrogen in the mixer, then cooled to -35aboutWith and transferred under nitrogen for 5 min in a well-mixed suspension of the reagent Vilsmaier, also cooled to -35aboutC. Suspension of the reagent Vilsmeier get added at 0aboutFrom oxalicacid (28,2 g, 221 mmol) in portions to a solution of dimethylformamide (16,89 g, 231 mmol) in dichloromethane and then cooled to -35aboutC. during the addition the reaction temperature is increased up to -28aboutC. After addition, the reaction mixture is seeded with the product. After stirring 2.5 h in the temperature range from -28 to -35aboutThe mixture is filtered and the filter cake washed under nitrogen with dichloromethane (200 ml). Through pie 30 min flow of nitrogen, then dried for 12 h at room temperature in vacuum. The title compound is obtained in the form of a snow-white powder (42.9 g, yield 72%).1H-NMR (CD3OD) 4-06 the ri of 7.8 (s, C-5H), corresponding to approx. 0.5% of the anti-isomer. After derivatization with diethylamine in acetonitrile Ehud showed the title compound (SYN-isomer in the form of its diethylamino derived) with a retention time of 9.6 min, hydrochloride acid with a retention time of 2.8 min and anti-isomer (in the form of diethylamino derived) with a retention time of 16.4 minutes. The ratio of the SYN-isomer: hydrochloride acid: anti-isomer is 90:5:<1.

In the mixer 20 min crushed under nitrogen SYN-2-(2-aminothiazol-4-yl)-2-methoxyimino acid (84,7 g, 421 mmol) in dichloromethane (570 ml). The obtained fine suspension was diluted with dichloromethane (100 ml) and under nitrogen is transferred into the reactor Bucha 1 l jacketed. In the reactor creates the nitrogen pressure in 5 psi (0.35 kg/cm2) and the mixture is stirred with a speed of 375 rpm./min and cooled to -2aboutC. In the head space of the reactor at a rate of 0.2 g per minute injected hydrogen chloride (15.3 g, 421 mmole). There is a temperature increase of 2aboutC. the Mixture is stirred for further 30 min at 0aboutWith crushed 3 minutes in the mixer, cooled to -35aboutC and under nitrogen is transferred for 5 min to mix well in the eating portions at 0aboutFrom oxalicacid (56,1 g, 439 mmole) to a solution of dimethylformamide (33,8 g, 462 mmole) in dichloromethane (880 ml) and then cooled to -35aboutC. during the addition the reaction temperature is increased up to -28aboutC. After addition, the reaction mixture is seeded with the product. After additional aging in the temperature range from -28 to -35aboutC for 2.5 h the mixture fillout and the filter cake washed under nitrogen dichloromethane (350 ml). Through the filter of Perak 30 min flow of nitrogen and then the solid product is dried for 12 h in vacuum at room temperature. The title compound is obtained in the form of a snow-white powder (95,2 g, unadjusted yield 89%).

Calculated C 28,14; N Was 2.76; N 16,41; S To 12.52; CL 27,68.

WITH6H7N3O2SCl2.

Found, 28,11; N 2,62; N 16,20; S 12,22; CL 26,74.

1H-NMR (DOD) : 4,06 (s, 3H, CH3), 7,12 (s, 1H,-5H). Signals are also visible when 7,18 (s, C-5H), corresponding to approx. 4% hydrochloride acid, and at 7.8 (s, C-5H), corresponding to approx. 2% anti-isomer. After derivatization with diethylamino analysis Ehud showed the title compound (in the form of its diethylamino derived) with a retention time of 9.6 min, initial acid over time abstain the-isomer of the original acid-anti-isomer is 90:4:2.

P R I m e R 24. Getting 7-/2-(2-aminothiazol-4-yl)-2-(Z)-methoxykynuramine /-3-/(1-methyl-1-pyrrolidinyl)methyl/ceph-3-em-4-carboxylate the dihydrochloride hydrate.

In 60 ml of water is gradually dissolve 7-amino-3-/(1-methyl-1-pyrrolidino)methyl /ceph-3-em-4-carboxylate, hydroiodide (15,01 g, 35,29 mmole), maintaining the pH below 6.5 the use of triethylamine. Add acetone (120 ml) and the resulting aqueous-acetone solution is cooled in the temperature range from -15 to -20aboutC. Maintaining the pH below 7.5 and above 5 using autotitrator model Radiometer V80 filled with triethylamine and configured at the end of the titration at pH 6.5, add hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxykynuramine (14.6 g active, to 45.4 mmole), lasting 2 hours and 7 minutes, the Reaction mixture is left to gradually heated from 12aboutWith up to 5aboutWith up to completion of acylation (according ghvd), after which the reaction mixture is filtered.

The filtrate is acidified with 12 N. hydrochloric acid (about 17.7 ml, 0,212 mol), and then to initiate crystallization add acetone (210 ml). The suspension is stirred for 1.25 h and then diluted with additional kolichestvo acetone (195 ml). The suspension is cooled to 0-5aboutC, stirred for 0.75 h and filtered. Pollpri 45aboutC. Dihydrochloride hydrated salt form of the title compounds emit from 95.5% purity (12.8 g, stoichiometric yield mass 63,8%).

P R I m e R 25. Getting 7-/2-(2-aminothiazol-4-yl)-2-(Z)-methoxykynuramine /-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate the dihydrochloride hydrate.

To a cold (-30aboutC) a solution of acetone (120 ml) and water (40 ml) simultaneously and separately dry for 30 min add hydroiodic 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-car - barcelata (15 g, 35 mmol) and hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxime-nacetylmuramic (10,49 g active, 40.9 mmole). With the help of autotitrator model Radiometer V80 filled with triethylamine and configured at the end of the titration at pH 6.5, the pH value of the reaction mixture is maintained within the range of 5.5-7 with cooling from -20 to -40aboutC. Upon completion of addition of the reagents obtained small opaque suspension is heated to 0-5aboutC and stirred until dissolved solids (20 min), and by this time high-performance liquid chromatography indicates the completion of the acylation. Then the reaction mixture is filtered.

The filtrate is acidified with 12 N. hydrochloric acid (17.6 ml, 0,212 e 30 min, diluted with an additional amount of acetone (315 ml), stirred for 30 min at room temperature and cooled 1 h at 0-5aboutC. After vacuum filtration, the product is washed with acetone (250 ml) and dried in vacuum at 45aboutC. the Title compound isolated in the form of its dihydrochloride, hydrate purity (17,68 g, stoichiometric yield mass 87,7%). The water content by the Karl Fischer method of 4.45%

P R I m e R 26. Getting 7-/2-(2-aminothiazol-4-yl)-2-(Z)-methoxyimino-amido /-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate the dihydrochloride hydrate.

In (60 ml) slowly at room temperature dissolve hydroiodic 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/-ceph-3-em-4-carboxylate (14,75 g, to 34.7 mmole), maintaining the pH below 6.5 the use of triethylamine. Red-orange solution discolor at 0-5aboutWith activated charcoal (3 g). The charcoal is filtered off and the resulting light amber solution store at 0-5aboutC. Coal cake is washed with 22.5 ml of water. The washing solution is mixed with 68 ml of acetone and the resulting aqueous-acetone solution is cooled to -30aboutC. by Setting the temperature between -20aboutC to -30aboutWith and maintaining a pH in the range 5.5 to 6.5 using autotitrator model Radiometer V80 filled with triethylamine and configured at the end of t is)methyl/ceph-3-em-4-car - boxylic to chilled water-acetone solution from washing the coal cake while adding hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxykynuramine (9,06 g active 35,4 mmole) and 180 ml of acetone (steady stream). Upon completion of addition of the reagents, the reaction mixture is heated to 0-5aboutC and stirred until complete dissolution of the solids. After acylation (according to HPLC), the reaction mixture was filtered.

The filtrate is acidified with 12 N. hydrochloric acid (17.6 ml, 0,212 mol), then at room temperature for hours, diluted with acetone (690 ml) and incubated for one hour at room temperature. The product is filtered under vacuum, washed with acetone (250 ml) and dried for 15 h under vacuum at about 45aboutC. the Title compound selected purity 89,6% (15,44 g, stoichiometric yield mass 76,4%). Water content by Karl Fischer method 4,07%

P R I m e R 27. Getting 7-/2-(2-aminothiazol-4-yl)-2-(Z)-methoxykynuramine /-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate the dihydrochloride hydrate

To a cold (-30aboutC) a solution of acetone (120 ml) and water (40 ml) simultaneously or separately dry for 30 min was added the hydrochloride of 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-car - barcelata (14,34 g, 43 mmole) and hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxime-nacetylmuramic (br11.01 g active, 43 mmole). With the help of autotitrator model Radiometer V80, zapolnenie 5.5-7, and cooling the mixture in the range from -20 to -40aboutC. Upon completion of addition of the reagents obtained small nepoznato suspension is heated to 0-5aboutC and stirred until dissolved solids (20 min), and at this point, according to high performance liquid chromatography acylation is completed. Then the reaction mixture is filtered.

The filtrate is acidified with 12 N. hydrochloric acid (21,4 ml, 0,257 mole) and to initiate crystallization add acetone (75 ml). Crystallization slurry is stirred for 1 hour, and then for 30 min diluted with acetone (505 ml). After stirring 30 min at room temperature, the suspension is cooled for 1 h to 0-5aboutC and filtered. The product is washed with acetone (250 ml) and dried in vacuum at 45aboutC. the Title compound isolated in the form of its dihydrochloride, hydrate purity of 93.5% (18,81 g, stoichiometric yield mass of 85.1%). Water content by Karl Fischer method 4,2%

P R I m e R 28. The transformation of the dihydrochloride monohydrate of cefepime in dehydrate the dihydrochloride of cefepime.

In 1200 ml of deionized water dissolve monohydrate, the dihydrochloride of cefepime (300 g, purity of 99.9% according to HPLC, KF 3.8 percent). Add 6 N. hydrochloric acid (132 ml, 1.5 equivalents). The solution is filtered and washed with deionized water (300 additional amount of acetone (4000 ml). The solution is kept in the point cloud to the moment of intensive formation of dihydrate crystals (needles under a microscope, at the point of turbidity possible persecution). Within 25 min add an additional amount of acetone (8000 ml) and the thick suspension is stirred for one hour at 25aboutC.

The identity of the dihydrate crystals (needles) was confirmed by comparison of the crystals under a microscope with an authentic sample. The suspension is filtered and washed with acetone (2 × 1500 ml). The filter cake is dried for 15 h in vacuum at 40aboutC. the Yield of the dihydrochloride dihydrate of cefepime 305,10 g (98,6%), purity according to HPLC 99% KF 6,5%

P R I m e R 29. The transformation of the dihydrochloride dihydrate of cefepime in the monohydrate of the dihydrochloride of cefepime.

In 75 ml demonizirovannyj water dissolve dehydrate the dihydrochloride of cefepime (15 g, purity according to HPLC 99,2% KF 6,4%). Add 6 N. hydrochloric acid (0.9 ml, 0.2 equivalent) and the solution is filtered through a filter of 0.45 μm.

To the filtered solution dropwise for 20 minutes was added acetone (200 ml) with formation of a turbid solution (in this moment possible persecution). Without keeping in this state dropwise over 40 min add more acetone (400 ml) and the suspension is cooled for one hour VNIIM under a microscope with an authentic sample. The suspension is filtered and washed with acetone (2 x 60 ml). The filter cake is dried in vacuum for 15 h at 40aboutC. the Yield of the dihydrochloride monohydrate of cefepime 13,28 g (91,8%) and the structure of crystals, as confirmed, is the same as that described in U.S. patent N 4910301 (plan and others).

P R I m e R 30. Getting 7-/2-(2-aminothiazol-4-yl)-2-(Z)-methoxykynuramine /-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-carboxylate the dihydrochloride hydrate.

To a cold (-22aboutC) a solution of acetone (120 ml) and water (40 ml) for 25 min at the same time or separately in dry form is added hydroiodic 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-car - barcelata (14,67 g active 0,0345 mol) and hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxykynuramine Ltd (9.93 g of the active, 0,0388 mol). With the help of autotitrator model Radiometer V80 filled with triethylamine and configured at the end of the titration at pH 6.5, in the reaction mixture maintain pH in the range of 5-7 during cooling from -20 to -30aboutC. Upon completion of addition of the reagents obtained opaque suspension is heated to 0-5aboutWith up to complete dissolution of the solids, and at this point, according to high performance liquid chromatography acylation is completed. The reaction mixture was filter 12 N. hydrochloric acid (8,8 ml, 0,106 mol) and diluted with acetone (114 ml) until the turbidity of the filtrate. The filtrate tatrallyay crystals of the dihydrochloride dihydrate of cefepime (0.5 g) and the suspension is heated for about 3 hours at 40aboutC. the Mixture is cooled for 1 h at 0-5aboutC and filtered. The product is washed with acetone and dried in vacuum at room temperature. The dihydrochloride of the title compound crystallized with purity of 93.3% (7,34 g, stoichiometric yield mass 67,3%). Water content analysis according to Karl Fischer -4,1% and FT-IR (Diffusive reflection with CVG) analysis with absorption peaks at 3574 cm and 3432 cm-1indicate that the product is a mixture of the monohydrate (granulopenia crystals) and dihydrate (needle-like crystals) of the title compound.

Method C.

The second portion of the filtrate acidified with 12 N. hydrochloric acid (8,8 ml, 0,106 mole) and for 1 h, diluted with acetone (206 ml). Suspension stand before crystallization, and then cooled to 0-5aboutWith and incubated for 1 h the Suspension was filtered, the product washed with acetone and dried in vacuum at 45aboutC. the Dihydrochloride of the title compound crystallized purity 95.3 per cent (8.6 g, stoichiometric yield mass 85,3%). Water content analysis according to Karl Fischer 4.6% and FT-IR is possible with the content dihydrate less than 0.4%

P R I m e R 31. Getting 7-/2-(2-aminothiazol-4-yl)-2-(Z)-methoxykynuramine /-3-/(1-methyl-1-pyrrolidinyl)methyl/ceph-3-em-4-carboxylate the dihydrochloride dihydrate.

To a cold (-22aboutC) a solution of acetone (120 ml) and water (40 ml) for 25 min at the same time or separately in dry form is added hydroiodic 7-amino-3-/(1-methyl-1-pyrrolidino)methyl/ceph-3-em-4-car - barcelata (14,61 g active 0,0344 mol) and hydrochloride SYN-2-(2-aminothiazol-4-yl)-2-methoxykynuramine (9,94 g active 0,0388 mol). With the help of autotitrator model Radiometer V 80 filled with triethylamine and configured at the end of the titration at pH 6.5, in the reaction mixture maintain pH in the range of 5-7 during cooling from -20 to -30aboutC. Upon completion of addition of the reagents obtained opaque suspension is heated to 0-5aboutWith up to complete dissolution of the solids, and at this point, according to high performance liquid chromatography acylation is completed. The reaction mixture is filtered and the filtrate is divided into two equal portions.

Method A.

One portion of the obtained filtrate is acidified with 12 N. hydrochloric acid (11.7 ml, 0,404 mol) and diluted with acetone until the turbidity of the filtrate. The filtrate tatrallyay crystals of the dihydrate digikey, dilute with acetone and stirred for 15 hours, the Slurry is heated under 40aboutC for 1 h and diluted with acetone. The total amount of acetone in 280 ml is used for crystallization of the product. After gradual cooling for 1 h to 0-5aboutThe mixture is filtered, washed with acetone (125 ml) and dried in vacuum at 45aboutC.

The main compound in the form of cefepime of dihydrochloride allocate the number of 97.8% (8,19 g, stoichiometric yield mass of 80.9%). Water content analysis according to Karl Fischer 6.5% and FT-IR (Diffusive reflection with CVG) analysis with absorption peaks at 3574 and 3432 cm-1indicate that the product is digitalnow shape (needle-like crystals) of the title compound.

Method C. a Second portion of the filtrate acidified with 12 N. hydrochloric acid (14.6 ml, 0,1752 mole) and add acetone under stirring until turbidity. The filtrate tatrallyay crystals of the dihydrochloride dihydrate of cefepime (0.3 g) and stirred at room temperature for 1.5 hours the Mixture is further diluted with acetone, stirred at room temperature for 15 h and then heat - live at the 40aboutC for 1 h further diluted with a mixture of acetone to the volume and dried in vacuum at 45aboutC. Dehydrate the dihydrochloride of cefepime distinguish with purity in 96.3% (8,68 g, stoichiometric yield mass 85,7%). Water content analysis according to Karl Fischer 6.7% and FT-IR (Diffusive reflection with CVG) analysis shows that the product is dihydrates form the title compound (needle-like crystals).

1. A method of obtaining a crystalline mono - or dihydrochloride dihydrate 7-[2-(2-aminothiazol-4-yl)-2-(Z) -methoxykynuramine] -3-[(1-methyl-1-pyrrolidinium)methyl] -ceph-3-em-4-carboxylate, essentially free from antihisamine and2-isomer of an acid additive salt of 7-amino-3-[(1-methyl-1-pyrrolidinium)methyl]-ceph-3-em-4-carboxylate of General formula

< / BR>
where HX denotes HCl, HJ or H2SO4,

subjected to interaction with the hydrochloride of 2-(2-aminothiazol-4-yl)-2 - methoxykynuramine essentially free from antihisamine, in aqueous-organic solvent at a pH of 5.0 to 7.5, followed by separation of the resulting reaction mixture the desired product in the form of its crystalline mono - or dihydrochloride dihydrate.

2. The method according to p. 1, characterized in that the hydrochloride of 2-(2-aminothiazol-4-yl)-2-methoxykynuramine essentially free from antisolar receive interaction besod is ina least 1 mol of oxalicacid and at least 1 mol or a slight excess relative to oxalylamino of dimethylformamide, in an inert organic solvent at a temperature below -10oC.

3. The method according to p. 2, characterized in that the number of oxalicacid is 1 to 2 mol relative to the anhydrous hydrochloride SYN-isomer of 2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid, and the number of dimethylformamide is small equimolar excess relative oxalicacid.

4. The method according to p. 3, characterized in that the number of oxalicacid is 1.05 mol and the number of dimethylformamide 1,075 mol relative to the anhydrous hydrochloride SYN-isomer of 2-(2-aminothiazol-4-yl)-2-methoxyethoxy acid.

5. The method according to p. 2, characterized in that the temperature is minus 15

minus 40oC.

6.The method according to p. 1, wherein the inert organic solvent is a dichloromethane or acetonitrile.

7. The method according to p. 1, characterized in that the reaction mixture is further treated with the necessary amount of acid and water-soluble organic solvent, followed by separation of the target product.

8. The method according to p. 7, wherein the acid is a sulfuric acid.

9. The method according to p. 8, of the cation followed by the addition of the necessary quantity of hydrochloric acid with the formation of the target product.

10. The method according to p. 1, wherein the aqueous organic solvent is a water-acetone.

12. The method according to p. 1, characterized in that the selected target product contains 2,7 7,0 wt. water.

13. The method according to p. 1, characterized in that the selected target product is a dihydrochloride monohydrate of 7-[2-(2-aminothiazol-4-yl)-2-(Z)- methoxykynuramine] -3-[(1 - methyl-1-pyrrolidino) -methyl] -ceph-3 - em-4-carboxylate.

14. The method according to p. 1, characterized in that the selected target product is a dihydrochloride dihydrate 7 [2-(2 - aminothiazol-4-yl) -2-(Z)- methoxykynuramine] -3-[(1 - methyl-1 - pyrrolidinium) methyl]-ceph-3 - em-4-carboxylate.

 

Same patents:

The invention relates to a method for producing derivatives of cafema General formula I(I) where R1is amino or protected amino group,

R2is lower alkyl or fluorinated methyl,

R3- COO-, carboxy or protected carboxy,

R4hydroxy(lower)alkyl or protected hydroxy(lower)alkyl,

R5is amino or protected amino group,

R6is hydrogen or methyl, or its salt additive triperoxonane acid, its additive salts of sulfuric acid, or its salts with trifurcation having antibacterial activity

FIELD: organic synthesis.

SUBSTANCE: invention provides substituted 7-acylaminocephalosporins of formula I:

(I), where W denotes CH or B; V denotes NO; R1 hydrogen or С14-alkyl; R3 hydrogen or ester residue; and R2 one of the following groups: , , , , in which X, R5, R6, R'6, R7, and Hal have meanings indicated in claims, in free state, in the form of salts and/or solvates, or, if such forms are stable, in the form of internal salt, quaternary salt, or their hydrates, possessing antimicrobial activity. Invention also discloses a method for preparing such compounds and a pharmaceutical connected containing them.

EFFECT: increased choice of antimicrobial preparations.

13 cl, 10 tbl, 225 ex

FIELD: chemistry.

SUBSTANCE: invention relates to a method of producing 7-[2-(2-aminothiazol-4-yl)-2(Z)-methoxyimino acetamido]-3-[(1-methyl-1-pyrrolidino)methyl]-cef-3-em-4-carboxylate dihydrochloride monohydrate (cefepime dihydrochloride monohydrate) of formula (I) The method is realised by crystallising 7-[2-(2-aminothiazol-4-yl)-2(z)-methoxyimino acetamido]-3-[(1-methyl-1-pyrrolidino)methyl]-cef-3-em-4-carboxylate dihydrochloride monohydrate from aqueous or aqueous-organic solutions thereof by adding sodium chloride to the solution in an amount which provides sodium chloride concentration in the formed solution ranging from 10.0 to 15.0%. The precipitated cefepime dihydrochloride is then filtered, washed and dried.

EFFECT: improved method of producing cefepime dihydrochloride monohydrate.

2 cl, 2 ex

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