KR100429585B1 - Orally available new cephalosporin antibiotics and their preparation - Google Patents

Abstract

The present invention provides novel cephalosporin compounds of formula (1), pharmaceutically acceptable non-toxic salts thereof, physiologically hydrolysable esters, hydrates, solvates and isomers thereof, pharmaceutical compositions containing them, and their Relates to a method of manufacture:

In the above formula,

A, R ₁ , R ₂ , W, Q and n have the meanings defined in the specification.

Description

Orally available new cephalosporin antibiotics and preparation method thereof {Orally available new cephalosporin antibiotics and their preparation}

The present invention relates to novel cephalosporin compounds useful as antibiotics. More specifically, the present invention provides a novel cephalosporin compound of formula (1), which has strong antimicrobial activity and broad antibacterial spectrum and improved pharmacokinetic properties and is also effective when used for oral administration, Permissible non-toxic salts thereof, physiologically hydrolysable esters thereof, hydrates, solvates and isomers thereof.

Formula 1

In the above formula,

A is an amino protecting group comprising hydrogen or amino acid,

R ₁ is hydrogen or may be mono- to trisubstituted by C _1-6 alkoxy, cyano, halogen, C _3-6 cycloalkyl or C _5-6 heteroaryl containing one or two nitrogen or oxygen atoms A C _1-6 alkyl group, C _3-4 alkenyl group, a C _3-4 alkynyl group unsubstituted or substituted by amino, or a C _3-6 cycloalkyl group,

R ₂ represents hydrogen or a carboxy protecting group,

W represents N or C-Z,

From here,

Z represents hydrogen, a substituted or unsubstituted amine group, hydroxyl group or C _1-4 alkyl group,

Q represents CH, CX or N, where X represents halogen,

n is 0 or 1;

The present invention also includes a method for preparing the compound of the formula (1) and an antimicrobial composition containing these as an active ingredient. Therefore, the manufacturing method and composition are the object of this invention.

Cephalosporin antibiotics are widely used in the treatment of infectious diseases caused by pathogenic bacteria in humans and animals, especially in the treatment of diseases caused by bacteria that are resistant to other antibiotics, such as penicillin compounds, and in the treatment of penicillin-sensitive patients. useful. In the treatment of these infectious diseases, in most cases it is preferable to use antibiotics that exhibit antimicrobial activity against both Gram-positive and Gram-negative microorganisms, and the antimicrobial activity of these cephalosporin antibiotics is 3 or 7 positions of the cefe ring. It is well known that it is greatly influenced by the kind of substituents present in. Thus, numerous attempts have been made to develop cephalosporin antibiotics in which various substituents have been introduced at the 3- or 7-position in an attempt to develop antibiotics that exhibit potent antimicrobial activity against a wide range of Gram-positive and Gram-negative bacteria.

For example, British Patent No. 1,399,086 describes the cephalosporin derivatives represented by the following formula (2) in a broad and comprehensive manner.

In the above formula,

R ₃ is hydrogen or an organic group,

R ₄ is an etherified monovalent organic group that is linked to oxygen through carbon,

B is -S- or> S-> O,

P is an organic group.

Since the invention of these compounds, many attempts have been made to develop antibiotics with improved antimicrobial properties, particularly against Gram-negative bacteria. As part of these attempts, British Patent No. 1,522,140 discloses syn-isomers or at least 90% of syn-isomers. The cephalosporin antibiotic of formula (3), which is present as a mixture of containing syn- and anti-isomers, is described.

In the above formula,

R ₅ is a furyl or thienyl group,

R ₆ is a C _1-4 alkyl, C _3-4 cycloalkyl, furylmethyl or thienylmethyl group,

R ₇ is hydrogen, carbamoyl, carboxymethyl, sulfonyl or methyl group.

Since then, numerous attempts have been made to develop antibiotics with a broad antimicrobial spectrum with enhanced antimicrobial activity to gram-positive bacteria as well as to gram-negative bacteria. As a result, many cephalosporin antibiotics having structures similar to those of formula (3) have been developed. These developments have attempted several changes, including the introduction of acylamido groups and specific groups at the C-3 position at the 7-position of the cephem nucleus of formula (3).

For example, Belgian Kingdom Patent No. 852,427 discloses that R ¹ in Formula (2) is substituted with various other organic groups including 2-aminothiazol-4-yl and the oxygen atom of the oxyimino group is attached to an aliphatic hydrocarbon group. A cephalosporin antibiotic compound is described in which the aliphatic hydrocarbon group itself may be substituted with a carboxy group, in which the C-3 substituent is acyloxymethyl, hydroxymethyl, formyl or an optionally substituted heterocyclic thiomethyl group. And so on.

Recently, a compound that exhibits strong activity against a wide range of pathogens and can also be used for oral administration with high blood concentration is intended to introduce a thioalkyl chain (SCH ₂ Ar, where Ar is an aromatic ring) at the C-3 position. There was an attempt.

That is, World Patent WO 95-07283 describes the cephalosporin derivative of formula (4) broadly and collectively, and in this patent, activity against a wide range of pathogens is introduced by introducing a thioalkyl chain at the C-3 position. It is raising.

In the above formula,

R ₈ represents an amine group having an amine group or a protecting group,

R ₉ represents hydrogen, lower alkyl or hydroxy protecting group,

R ₁₀ represents hydrogen or a carboxy protecting group,

R ₁₁ represents 3-pyridyl, 4-pyridyl, or an optionally substituted heteromonocyclic ring comprising two nitrogen atoms, one oxygen atom or a sulfur atom,

n 'is 0, 1 or 2,

Provided that when R ₂ is lower alkyl n 'is 1 or 2;

This patent introduces several heteroaromatic rings in the thioalkyl chain of C-3, but differs from the heterocycle according to the invention. That is, the present invention is characterized in that a substituted or unsubstituted pyrimidinyl group is introduced at the C-3 position, but there is no mention of this in the patent.

Accordingly, the present inventors have concentrated on developing a cephalosporin compound which shows not only strong antimicrobial activity against a wide range of pathogens including Gram-positive bacteria but also has improved pharmacokinetic properties and is effective in oral administration. The present invention has been accomplished by discovering that cephalosporin compounds represented by pyrimidinyl groups optionally substituted at positions meet these goals.

Accordingly, the present invention relates to a compound of formula (1) and a process for preparing the same.

It is an object of the present invention to provide novel cephalosporin compounds of formula (1), pharmaceutically acceptable non-toxic salts thereof, physiologically hydrolysable esters, hydrates, solvates and isomers thereof:

Formula 1

In the above formula,

A is an amino protecting group comprising hydrogen or amino acid,

R ₁ is hydrogen or may be mono- to trisubstituted by C _1-6 alkoxy, cyano, halogen, C _3-6 cycloalkyl or C _5-6 heteroaryl containing one or two nitrogen or oxygen atoms A C _1-6 alkyl group, C _3-4 alkenyl group, a C _3-4 alkynyl group unsubstituted or substituted by amino, or a C _3-6 cycloalkyl group,

R ₂ represents hydrogen or a carboxy protecting group,

W represents N or C-Z,

From here,

Z represents hydrogen, a substituted or unsubstituted amine group, hydroxyl group or C _1-4 alkyl group,

Q represents CH, CX or N, where X represents halogen,

n is 0 or 1;

The compound of formula (1) according to the present invention is a geometric isomer, which is a neo-isomer, or a mixture of neo- and anti-isomers containing 90% or more of the neo-isomer, and also a hydrate of the compound of formula (1) And solvates are also included within the scope of this invention. In addition, the aminothiazole group of the compound of formula (1) may form a tautomer with an iminothiazoline group as follows,

In addition, when Q is N, the aminothiazole group may form a tautomer with an iminothiadiazoline group as follows.

Therefore, such tautomers are also included in the scope of the present invention.

Pharmaceutically acceptable non-toxic salts of the compounds of formula (1) are salts with inorganic acids such as hydrochloric acid, bromic acid, phosphoric acid, sulfuric acid, acetic acid, trifluoroacetic acid, citric acid, formic acid, maleic acid, hydroxyl, succinic acid, benzoic acid, Organic carboxylic acids or salts of methanesulfonic acid, para-toluenesulfonic acid, such as tartaric acid, fumaric acid, manderic acid, ascorbic acid, dried acid, and salts of penicillin with other acids known and used in the art of cephalosporin. These acid addition salts can be prepared by conventional techniques. The compounds of formula (1) may also form nontoxic salts with bases. Bases used here include alkali metal hydroxides (e.g. sodium hydroxide, potassium hydroxide), alkali metal bicarbonates (e.g. sodium bicarbonate, potassium bicarbonate), alkali metal or alkyllitometal carbonates (e.g. sodium carbonate, potassium carbonate, calcium carbonate) Inorganic bases such as and the like and organic bases such as amino acids.

Examples of physiologically hydrolyzable esters of the compounds of formula (1) include indanyl, phthalidyl, methoxymethyl, pivaloyloxymethyl, glycyloxymethyl, phenylglycyloxymethyl, 5-methyl-2 Oxo-1,3-dioxoren-4-yl methyl and other physiologically hydrolysable esters known and used in the art of penicillin and cephalosporin. Such esters can be produced by known methods.

Representative examples of the compound of formula (1) according to the present invention are as follows.

I-1: 7-[(Z) -2- (2-amino-1,3-thiazol-4-yl) -2-methoxyimino-acetylamino] -3- (2,4-diaminopyri Midin-5-ylmethylsulfanyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid;

I-2: 7-[(Z) -2- (2-amino-1,3-thiazol-4-yl) -2-methoxyimino-acetylamino] -3- (2,4-diaminopyri Midin-6-ylmethylsulfanyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid;

I-3: 7-[(Z) -2- (2-amino-5-chloro-1,3-thiazol-4-yl) -2-methoxyimino-acetylamino] -3- (2,4 -Diaminopyrimidin-5-ylmethylsulfanyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid;

I-4: 7-[(Z) -2- (2-amino-5-chloro-1,3-thiazol-4-yl) -2-hydroxyimino-acetylamino] -3- (2,4 -Diaminopyrimidin-6-ylmethylsulfanyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid;

I-5: 7-[(Z) -2- (2-amino-1,3-thiazol-4-yl) -2-methoxyimino-acetylamino] -3- (2,4-diaminopyri Midin-5-ylmethylsulfanylmethyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid; And

I-6: 7-[(Z) -2- (2-amino-5-chloro-1,3-thiazol-4-yl) -2-hydroxyimino-acetylamino] -3- (2,4 -Diaminopyrimidin-5-ylmethylsulfanylmethyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid

The compound of formula (1) according to the present invention, a pharmaceutically acceptable non-toxic salt thereof, a physiologically hydrolysable ester, a hydrate, a solvate or an isomer thereof according to the present invention is a compound of formula (5a) in the presence of a base Reaction with a compound of formula (6a) gives a compound of formula (1a), or a compound of formula (5b) with a compound of formula (6b) in the presence of a base to give a compound of formula (1a) Or by reducing the compound of formula (1a) wherein m is 1 to obtain the compound of formula (1).

Formula 1

In the above formula,

A, R ₁ , R ₂ , W, Q and n are as described above,

X 'is a halogen atom,

P 'is a thiol, alkyl- or arylthioester,

L is a leaving group,

m is 0 or 1;

Wherein the aminoprotecting group A is acyl, substituted or unsubstituted ar (lower) alkyl (e.g. benzyl, diphenylmethyl, triphenylmethyl, 4-methoxybenzyl, etc.), halo (lower) alkyl (e.g. trichloro Chloromethyl, trichloroethyl, etc.), tetrahydropyranyl, substituted phenylthio, substituted alkylidene, substituted aralkylidene, substituted cycloidene and the like. Acyls suitable as amino protecting groups may be aliphatic and aromatic acyl groups or acyl groups having a heterocycle. Examples of such acyl groups include C _1-5 lower alkanoyl (eg formyl, acetyl, etc.), C _2-6 alkoxycarbonyl (eg methoxycarbonyl, ethoxycarbonyl, etc.), lower alkanesul Phenyl (for example, methylsulfonyl, ethylsulfonyl etc.) or ar (lower) alkoxycarbonyl (for example, benzyloxy carbonyl etc.) etc. are mentioned. The acyl described above may have suitable substituents, such as 1-3 halogens, hydroxy, cyano, nitro and the like. In addition, the reaction product of silane, boron, phosphorus compound and amino group may be an amino protecting group.

The carboxy protecting group R ² is usually suitable as long as it is easily removed under mild conditions, such as (lower) alkyl esters (eg methyl esters, t-butyl esters, etc.), (lower) alkenyl esters (eg vinyl esters, Allyl esters, etc.), (lower) alkylthio (lower) alkyl esters (e.g. methylthiomethyl esters, etc.), halo (lower) alkyl esters (e.g. 2,2,2-trichloroethyl esters, etc.), substituted or unsubstituted Ring aralkyl esters (eg, benzyl esters, p-nitrobenzyl esters, p-methoxybenzyl esters, etc.) or silyl esters.

The amino protecting group and the carboxy protecting group may be easily removed under mild reaction conditions such as hydrolysis and reduction to form a free amino group or a carboxy group. The amino protecting group and the carboxy protecting group may be appropriately selected and used according to the chemical properties of the compound of formula (1).

The leaving group L is, for example, halogen such as chlorine, fluorine and iodine, (lower) alkanoyloxy such as acetoxy, (lower) alkanesulfonyloxy such as methanesulfonyloxy, and arene such as pyrattoluenesulfonyloxy. Sulfonyloxy or alkoxycarbonyloxy.

The conversion of the halogen atom represented by X 'of formula (6a) to another halogen atom can be accomplished by conventional methods. For example, a compound of formula (6a) in which X 'is an iodine atom can be obtained by reacting a compound of general formula (6a) in which X' is a chlorine atom with an alkali metal iodide.

The compounds of the formulas (5a) and (5b) which are starting materials of the present invention can be prepared according to the following scheme (1). That is, the compound of formula (7) or a salt thereof can be prepared by activating with an acylating agent and reacting with the compound of formula (8a) or (8b).

In the above scheme,

A, R ¹ , R ² , Q, m and L are as described above.

The dotted lines in the structural formulas (8a) and (8b) represent 2-cefem or 3-cefem compounds, respectively, or mean mixtures thereof.

In the compounds of the formulas (8a) and (8b), the dotted lines indicate that the compounds of the formulas (8a) and (8b) alone are the following formulas (8a-1), (8a-2), (8b-1) and (8b-2). Or a mixture of a compound of formula (8a-1) with a compound of (8a-2) or a compound of formula (8b-1) with a compound of (8b-2).

Wherein R ² , m and L are as described above.

In preparing compounds of formula (5), the active acylated derivatives of formula (7) are acid chlorides, acid anhydrides, mixed acid anhydrides (preferably methylchloroformate, mesitylenesulfonyl chloride, p-toluenesulfonyl Acid anhydrides formed with chlorides or chlorophosphates) or activated esters (preferably esters formed in the reaction with N-hydroxy benzotriazole in the presence of a condensing agent such as dicyclo hexyl carbodiimide). The acylation reaction may also proceed with the free acid of formula (7) in the presence of a condensing agent such as dicyclohexyl carbodiimide, carbonyl diimidazole. On the other hand, the acylation reaction usually proceeds well in the presence of organic bases such as tertiary amines, preferably triethylamine, dimethylaniline, pyridine or inorganic bases such as sodium bicarbonate and sodium carbonate, and the solvent used is methylene chloride and chloroform. Solvents such as halogenated carbon, tetrahydrofuran, acetonitrile, dimethylformamide or dimethyl acetamide and the like. Also mixed solvents of the above solvents may be used and may be used in the form of an aqueous solution.

The reaction temperature of this acylation step is -50 ° C to 50 ° C, preferably -30 ° C to 20 ° C, and the acylating agent of the compound of formula (7) is equimolar or slightly to one equivalent of the compound of formula (8). An excess of ie, 1.05 to 1.2 equivalents can be used.

In preparing the compound of formula (1), the amino protecting group or acid protecting group of the compound of formula (5) can be removed by conventional methods well known in the art of cephalosporin. That is, the protecting group can be removed by a hydrolysis or reduction method, and in the case where the protecting group contains an amido group, it is preferable to hydrolyze through aminohalogenation and amino etherification. Acid hydrolysis is useful for the removal of tri (di) phenylmethyl groups or alkoxy carbonyl groups, and is carried out using inorganic acids such as formic acid, organic acids such as trifluoroacetic acid, P-toluenesulfonic acid or hydrochloric acid.

From the reaction product of the reaction, the desired compound of formula (1) can be separated or purified by various methods such as recrystallization, iontophoresis, silica gel column chromatography or ion exchange resin chromatography.

Another object of the present invention is to provide a composition containing a compound of formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient together with a pharmaceutically acceptable carrier.

The compounds of the present invention can be administered in injectable and oral formulations as desired.

The compounds of formula (1) of the present invention can be formulated in a known manner using known pharmaceutical carriers and excipients and incorporated into unit dose forms or multidose containers. The formulation form may be in the form of a solution, suspension or emulsion in an oil or aqueous medium and may contain conventional dispersants, suspensions or stabilizers. In addition, for example, it may be in the form of dry powder which is dissolved before use with sterile, pyrogen-free water. The compounds of formula (1) may also be formulated into suppositories using conventional suppository bases such as cocoa butter or other glycerides. Solid dosage forms for oral administration may be capsules, tablets, pills, powders, and granules, and capsules and tablets are particularly useful. Tablets and pills are preferably prepared with enteric coatings. Solid dosage forms can be prepared by mixing the active compounds of formula (1) according to the invention with one or more inert diluents such as sucrose, lactose, starch and the like and carriers such as lubricants such as magnesium stearate, disintegrants, binders and the like. Can be.

One of the great features of the formula (1) according to the present invention is that the pharmaceutical composition containing the same is orally administered even when formulated in oral preparations. Pharmacokinetic experiments have demonstrated that oral administration of the pharmaceutical compositions of the present invention has confirmed that the concentration of the drug has a property that is maintained in the blood for a very long time.

If desired, the compounds of the present invention may be administered in combination with other antibacterial agents such as penicillin or cephalosporin.

When formulating a compound of the present invention in unit dose form, the unit dose form preferably contains about 50 to 1,500 mg of the active ingredient of the compound of formula (1). The dosage of the compound of formula (1) depends on the doctor's prescription depending on factors such as the patient's weight, age and the particular nature and severity of the disease. However, the dosage required for adult treatment typically ranges from about 50 to 5,000 mg per day, depending on the frequency and intensity of administration. A total dosage of about 150-3,000 mg per day, usually separated by a single dose for intramuscular or intravenous administration to adults, will be sufficient, but for some strain infections higher daily dosages may be desirable.

Compounds of formula (1) according to the invention and their nontoxic salts (preferably salts with alkali metal salts, alkaline earth metal salts, inorganic acid salts, organic acid salts and amino acids) are resistant to a wide range of pathogenic bacteria, including various Gram-positive and Gram-negative bacteria. Its strong antimicrobial activity and broad antimicrobial spectrum make it very useful for the prevention and treatment of diseases caused by bacterial infection in animals including humans.

The following Preparation Examples and Examples are merely to aid the understanding of the present invention, but they do not limit the present invention.

Example 1

7-[(Z) -2- (2-amino-1, 3-thiazol-4-yl) -2-methoxyimino-acetylamino] -3- (2,4-diaminopyrimidine-5- Synthesis of monomethylsulfanyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid.

3-acetylsulfanyl-7- {2-methoxyimino-2- [2- (tritylamino) -1,3-thiazol-4-yl] acetylamino} -8-oxo-5-thia-1 1.0 mg (1.12 mmol) of aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid benzhydryl ester was dissolved in 9 ml of DMF and cooled to 0 ° C., followed by 0.467 ml of triethylamine. In a solution of 0.293 mL of morpholine dissolved in 10 mL of benzene, 23.3 mL was added and stirred for 30 minutes. 490 mg (1.2 equiv) of 2,4-diamino 5-bromomethyl pyrimidine hydrobromide was added followed by the slow dropwise addition of triethylamine (1.0 equiv). After stirring at 0 ° C. for 30 minutes, the mixture was diluted with ethyl acetate, washed with water, dried over magnesium sulfate, and the solvent was removed under reduced pressure. Ethyl ester was added and the solid obtained by filtration was dissolved in 1.5 ml of anisole, 3 ml of trifluoroacetic acid was added thereto, stirred for 60 minutes, cooled to 0 ° C., 20 ml of diethyl ether was filtered off, and the mixture was filtered. Purification by chromatography gave 150 mg (25% yield) of the title compound.

¹ H NMR (D ₂ O) δ 3.37 (d, 1H, 18 Hz), 3.64 (d, 1H, 18 Hz), 3.55 (d, 1H, 14 Hz), 3.89 (d, 1H, 14 Hz), 4.02 (s, 3H ), 5.17 (d, 1H, 4.5 Hz), 5.80 (d, 1H, 4.5 Hz), 7.03 (s, 1H), 7.0 (s, 1H)

Mass (FAB, m / e): 538

Example 2

7-[(Z) -2- (2-amino-1, 3-thiazol-4-yl) -2-methoxyimino-acetylamino] -3- (2,4-diaminopyrimidine-6- Synthesis of monomethylsulfanyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] ox-2-ene-2-carboxylic acid.

3-acetylsulfanyl-7- {2-methoxyimino-2- [2- (tritylamino) -1,3-thiazol-4-yl] acetylamino} -8-oxo-5-thia-1 623 mg (0.70 mmol) of aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid benzhydryl ester was dissolved in 6 ml of DMF and cooled to 0 ° C., followed by 0.467 ml of triethylamine and mor. In a solution of 0.293 ml of polyline dissolved in 10 ml of benzene, 2.1 ml was added and stirred for 30 minutes. 281 mg (1.1 equiv) of 2,4-diamino 6-bromomethyl pyrimidine hydrobromide was added followed by the slow dropwise addition of triethylamine (1.0 equiv). After stirring for 30 minutes at 0 ° C., the mixture was diluted with ethyl acetate, washed with water, dried over magnesium sulfate, and the solvent was removed under reduced pressure. The solid obtained by adding ethyl ester was filtered and dissolved in 1.5 ml of anisole, 1 ml of trifluoroacetic acid was added thereto, stirred for 60 minutes, cooled to 0 ° C., and the obtained solid was filtered through addition of 20 ml of diethyl ether. Purification by chromatography gave 150 mg (40% yield) of the title compound.

¹ H NMR (D ₂ O) δ 3.35 (d, 1H, 14.5 Hz), 3.60 (d, 1H, 14.5 Hz), 3.70 (d, 1H, 14.5 Hz), 3.73 (d, 1H, 14.5 Hz), 4.00 (s, 3H), 5.17 (d, 1H, 4.5 Hz), 5.80 (d, 1H, 4.5 Hz), 7.03 (s, 1H), 6.71 (s, 1H)

Mass (FAB, m / e): 538

Example 3

7-[(Z) -2- (2-amino-5-chloro-1,3-thiazol-4-yl) -2-methoxyimino-acetylamino] -3- (2,4-diaminopyri Synthesis of midin-5-ylmethylsulfanyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid.

3- (acetylsulfanyl) -7- {2- (5-chloro- [2- (tritylamino) -1,3-thiazol-4-yl] -2- (methoxyimino) acetyl) amino} 820 mg (0.911 mmol) of -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid benzhydryl ester was dissolved in 9 ml of DMF, After cooling, 1.0 ml of a solution of 0.467 ml of triethylamine and 0.293 ml of morpholine dissolved in 10 ml of benzene was added and stirred, followed by stirring for 30 minutes. 400 mg (1.2 equiv) of 2,4-diamino 5-bromomethyl pyrimidine hydrobromide was added followed by the slow dropwise addition of triethylamine (1.0 equiv). After stirring at 0 ° C. for 30 minutes, the mixture was diluted with ethyl acetate, washed with water, dried over magnesium sulfate, and the solvent was removed under reduced pressure. Ethyl ester was added, the solid obtained by filtration was dissolved in 1.5 ml of anisole, 3 ml of trifluoroacetic acid was added, stirred for 60 minutes, the temperature was cooled to 0 ° C. again, 20 ml of diethyl ether was filtered, and the obtained solid was filtered. Purification by high pressure liquid phase chromatography gave 150 mg (25%) of the title compound.

¹ H NMR (D ₂ O) δ 3.35 (d, 1H, 18 Hz), 3.64 (d, 1H, 18 Hz), 3.55 (d, 1H, 14 Hz), 3.90 (d, 1H, 14 Hz), 4.02 (s, 3H ), 5.17 (d, 1H, 4.5 Hz), 5.80 (d, 1H, 4.5 Hz), 7.70 (s, 1H).

Mass (FAB, m / e): 572

Example 4

7-[(Z) -2- (2-amino-5-chloro-1,3-thiazol-4-yl) -2-hydroxyimino-acetylamino] -3- (2,4-diaminopyri Synthesis of midin-6-ylmethylsulfanyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid.

3- (acetylsulfanyl) -7-({{2- {2-[(t-butoxycarbonyl) amino] -5-chloro-1,3-thiazol-4-yl} -2- (tri Tyloxy) imino} acetyl} amino) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid benzhydryl ester 1.0 g (1.063 mmol) ) Was dissolved in 10 ml of DMF, cooled to 0 ° C., 1.1 ml of a solution of 0.467 ml of triethylamine and 0.293 ml of morpholine in 10 ml of benzene was added and stirred for 30 minutes. 467 mg (1.2 equiv) of 2,4-diamino 6-bromomethyl pyrimidine hydrobromide was added. After stirring at 0 ° C. for 30 minutes, the mixture was diluted with ethyl acetate, washed with water, dried over magnesium sulfate, and the solvent was removed under reduced pressure. Ethyl ester was added, the solid obtained by filtration was dissolved in 1.5 ml of anisole, 1 ml of trifluoroacetic acid was added thereto, stirred for 60 minutes, the temperature was cooled to 0 ° C., and 20 ml of diethyl ether was filtered. Purification by high pressure liquid chromatography for extraction afforded 150 mg (40% yield) of the title compound.

¹ H NMR (D ₂ O) δ 3.35 (d, 1H, 14.5 Hz), 3.60 (d, 1H, 14.5 Hz), 3.70 (d, 1H, 14.5 Hz), 3.73 (d, 1H, 14.5 Hz), 5.15 (d, 1H, 4.5 Hz), 5.82 (d, 1H, 4.5 Hz), 7.71 (s, 1H).

Mass (FAB, m / e): 558

Example 5

7-[(Z) -2- (2-amino-1, 3-thiazol-4-yl) -2-methoxyimino-acetylamino] -3- (2,4-diaminopyrimidine-5- Synthesis of monomethylsulfanylmethyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid.

503 mg (1.5 mmol) of 2,4-diamino 5-bromomethyl dihydrobromide was added to 10 ml of a 4: 1 THF: DMF solvent and cooled to 0 ° C. Lithium thioacetate (3 equivalents) was dissolved in methanol and slowly added dropwise thereto. After confirming that the reaction was completed by TLC, NaOMe (1.0 equivalent) dissolved in methanol at 0 ° C. was added dropwise. After 10 minutes, it was cooled to -78 ° C and 4-methoxybenzyl 3- (iodomethyl) -7-({2- (methoxyimino) -2- [2- (tritylamino) -1,3- 927 mg (0.7 equiv) of thiazol-4-yl] acetyl} amino) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylate It was dissolved in THF and slowly added dropwise. After reacting for 30 minutes, the reaction was terminated with 1N hydrochloric acid solution, diluted with ethyl acetate and washed with brine. The organic layer was dried over magnesium sulfate, filtered, distilled under reduced pressure, and triturated with dichloromethane and diethyl ether to obtain a compound. The obtained compound was dissolved in 1 ml of anisole, cooled to 0 ° C., and slowly added dropwise trifluoroacetic acid, and then the temperature was raised to room temperature. After confirming that the reaction was completed by HPLC, the mixture was cooled to 0 ° C., 20 ml of diethyl ether was added thereto, and the solid obtained by filtration was purified by preparative high pressure liquid chromatography to obtain 20 mg of the title compound.

¹ H NMR (D ₂ O) δ 3.45 (m, 1H), 3.58-3.65 (m, 5H), 4.05 (s, 3H), 5.15 (m, 1H), 7.06 (s, 1H), 7.79-7.78 ( d, 1H, J = 4.6), 7.73 (s, 1H).

Mass (FAB, m / e): 552

Example 6

7-[(Z) -2- (2-amino-5-chloro-1,3-thiazol-4-yl) -2-hydroxyimino-acetylamino] -3- (2,4-diaminopyri Synthesis of midin-5-ylmethylsulfanylmethyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid.

839 mg (2.5 mmol) of 2,4-diamino 5-bromomethyl dihydrobromide was added to 10 ml of a 4: 1 THF: DMF solvent and cooled to 0 ° C. Lithium thioacetate (3 equivalents) was dissolved in methanol and slowly added dropwise thereto. After confirming that the reaction was completed by TLC, NaOMe (1.0 equivalent) dissolved in methanol at 0 ° C. was added dropwise. After 10 minutes, cooled to -78 ° C and 4-methoxybenzyl 7-({2- {2-[(t-butoxycarbonyl) amino] -5-chloro-1,3-thiazol-4-yl } -2-[(trityloxy) imino] acetyl} amino) -3- (iodomethyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2 1.94 g (0.8 equiv) of en-2-carboxylate was dissolved in THF and slowly added dropwise. After reacting for 30 minutes, the reaction was terminated with 1N hydrochloric acid solution, diluted with ethyl acetate and washed with brine. The organic layer was dried over magnesium sulfate, filtered, and distilled under reduced pressure, followed by polishing with dichloromethane and diethyl ether to obtain a compound. The obtained compound was dissolved in 1 ml of anisole, cooled to 0 ° C., and slowly added dropwise trifluoroacetic acid, and then the temperature was raised to room temperature. After confirming that the reaction was completed by HPLC, the mixture was cooled to 0 ° C., 20 ml of diethyl ether was added thereto, and the obtained solid was filtered and purified by preparative high pressure liquid chromatography to obtain 30 mg of the title compound.

¹ H NMR (D ₂ O) δ 3.31 (m, 1H), 3.58-3.54 (m, 5H), 5.15-5.16 (d, 1H, J = 4.6), 5.81 (d, 1H, J = 4.55), 7.65 (s, 1 H).

Mass (FAB, m / e): 572

Preparation Example 1

Synthesis of 2,4-diamino 5-carbetoxy pyrimidine

5.88 g (27.6 mmol) of 4-amino 5-carbetoxy 2-thiomethylpyrimidine was dissolved in 60 mL of dichloromethane and cooled to 0 ° C. 11.4 g (1.2 equivalents, ˜50%) of metachloroperbenzoic acid was dissolved in 40 ml of dichloromethane and slowly added dropwise thereto. After reacting at 0 ° C. for 2 hours, the reaction was terminated with 5% sodium thiosulfate solution, washed with sodium bicarbonate solution, and concentrated under reduced pressure. 28% ammonia water (100 ml) was added thereto, and the mixture was stirred at room temperature. After stirring for 2 hours, neutralization with acetic acid gave a solid. This solid was filtered off to give 2 g (yield: 42%) of the title compound.

¹ H NMR (DMSO) δ 1.25 (t, 3H), 4.19 (q, 2H), 7.23 (s, 1H), 7.38 (s, 1H), 7.45 (s, 1H)

Mass (FAB, m / e): 183

Preparation Example 2

Synthesis of 2,4-diamino 5-hydroxymethyl pyrimidine

1.2 g (6.6 mmol) of 2,4-diamino 5-hydroxymethyl pyrimidine were suspended in 1,1,1,3,3,3-hexamethyldisilazane. A catalytic amount of ammonium sulfate was added thereto, followed by reflux. After 24 hours hexamethyldisilazane was removed under reduced pressure and the product was dissolved in anhydrous tetrahydrofuran and cooled to 0 ° C. 250 mg (1.1 equiv) of lithium aluminum hydride was added dropwise, and the temperature was raised to room temperature. After reacting for 30 minutes, the reaction was terminated with water and acetone, the insoluble material was filtered off and the solvent was removed under reduced pressure to give 830 mg of the title compound.

¹ H NMR (DMSO) δ 4.19 (s, 2H), 5.79 (s, 2H), 6.07 (s, 2H), 7.57 (s, 1H)

Mass (FAB, m / e): 140

Preparation Example 3

Synthesis of 2,4-diamino 5-bromomethyl pyrimidine hydrobromide

165 mg (1.178 mmol) of 2,4-diamino 5-hydroxymethyl pyrimidine was dissolved in acetic acid (9 mL) and cooled to 5 ° C. 2.5 ml of bromic acid (30% acetic acid solution) was added dropwise. After reacting for 30 minutes at room temperature, the temperature was raised to 60 ° C. After reacting for 2 hours, the solvent was removed under reduced pressure, the compound was dissolved in a small amount of ethanol, and ether was added to give a solid. This solid was filtered off to give 408 mg of the title compound.

¹ H NMR (D ₂ O) δ 4.48 (s, 2H), 7.68 (s, 1H)

Preparation Example 4

Synthesis of 2,4-diphenylmethylamino 6-carboxy pyrimidine diphenylmethylamine salt

450 mg (2.33 mmol) of 2,4-dichloro 6-carboxy pyrimidine and diphenylmethylamine (5 equivalents) were dissolved in 30 ml of ethanol and refluxed. After reacting for 24 hours, the mixture was cooled to room temperature and the resulting solid was filtered to yield 710 mg (yield: 45%) of the title compound.

¹ H NMR (DMSO) δ 5.63 (s, 1H), 6.20 (d, 1H), 6.38 (d, 1H), 6.60 (s, 1H), 7.25-7.35 (m, 30H), 8.83 (d, 1H) , 9.58 (d, 1 H)

Preparation Example 5

Synthesis of 2,4-diphenylmethylamino 6-hydroxymethyl pyrimidine

710 mg (1.06 mmol) of 2,4-diphenylmethylamino 6-carboxy pyrimidine diphenylmethylamine salt was dissolved in anhydrous tetrahydrofuran, and lithium aluminum hydride (1.1 equiv) was added dropwise. After reacting for 3 hours, the reaction was terminated with water and acetone, the insoluble substance was filtered off, and the solvent was removed under reduced pressure to give 420 mg of the title compound.

¹ H NMR (DMSO) δ 4.13 (s, 2H), 5.63 (s, 1H)

Preparation Example 6

Synthesis of 2,4-diamino 6-bromomethyl pyrimidine hydrobromide

840 mg (1.78 mmol) of 2,4-diamino 5-hydroxymethyl pyrimidine was dissolved in acetic acid (12 mL) and then cooled to 5 ° C. To this was added dropwise 3 ml of bromic acid (30% acetic acid solution). After reacting for 30 minutes at room temperature, the temperature was raised to 70 ° C. After reacting for 3 hours, the solvent was removed under reduced pressure, dissolved in a small amount of ethanol, and ether was added to give a solid. This solid was filtered off to give 500 mg of the title compound.

¹ H NMR (D ₂ O) δ 4.38 (s, 2H), 6.33 (s, 1H).

Reference Example 1) Minimum Inhibitory Concentration (MIC)

The usefulness of the compound according to the present invention is Ceftazidime, an injectable sephatic agent having an excellent effect on Gram-negative bacteria, and Cefdinir, an oral sepase having excellent activity on both Gram-positive and Gram-negative bacteria. ) As a control agent was evaluated by obtaining the minimum inhibitory concentration (Minimum Inhibitory Concentration) for the standard strains of the compounds (I-1 and I-2) prepared in Examples 1 and 2. In other words, the minimum inhibitory concentration was diluted by 2-fold dilution method, and then dispersed in Mueller-Hinton agar medium. Then, 2 μl of the test strain having 10 ⁷ cfu (colony forming unit) per ml was inoculated. It was obtained by incubating for 20 hours at 37 ℃ and the results are shown in Table 1. As a result of the minimum inhibitory concentration test for the strains, the compounds according to the present invention showed improved Gram-positive bacteria activity compared to ceftazidime, and had improved activity against Haemophilus influenzae bacteria, which are the main causative agents of respiratory infections, compared to ceftidirin. appear.

Susceptibility test results for the standard strain (㎍ / ㎖) Staphylococcus aureus Staphylococcus pneumoniae E. coli Proteus vulgaris H. influenzae 1-1 0.5 2 0.031 0.063 0.031 1-2 2 2 0.031 0.016 0.031 Ceftazidime 4 8 0.031 0.031 0.063 Cefdinir 0.031 4 0.016 0.031 0.25

Reference Example 2) Pharmacokinetics

Pharmacokinetic experiments were conducted in male rats weighing 230 ± 10 g. 1-1 compound was prepared as a 1% solution and then orally administered at a dose of 20 mg / kg. After administration, blood was collected according to a predetermined time to obtain blood concentration by HPLC quantitative analysis, and based on this, pharmacokinetic parameters were calculated and shown in Table 2 below. Compound 1-1 showed high oral absorption and long half-life in rats.

Pharmacokinetic Parameters in Rats Maximum concentration (㎍ / ㎖) Half-life (minutes) AUC (μgmin / ml) 1-1 3.2 69 379 Cefdinir 2.9 74 383

Claims (5)

A cephalosporin compound of formula (1), a pharmaceutically acceptable nontoxic salt thereof, a physiologically hydrolysable ester, hydrate, solvate or isomer thereof: Formula 1 In the above formula, A is an amino protecting group comprising hydrogen or amino acid, R ₁ is hydrogen or may be mono- to trisubstituted by C _1-6 alkoxy, cyano, halogen, C _3-6 cycloalkyl or C _5-6 heteroaryl containing one or two nitrogen or oxygen atoms A C _1-6 alkyl group, C _3-4 alkenyl group, a C _3-4 alkynyl group unsubstituted or substituted by amino, or a C _3-6 cycloalkyl group, R ₂ represents hydrogen or a carboxy protecting group, W represents N or C-Z, but together they form a substituted or unsubstituted pyrimidinyl group, From here, Z represents hydrogen, a substituted or unsubstituted amine group, hydroxyl group or C _1-4 alkyl group, Q represents CH, CX or N, where X represents halogen, n is 1. The compound of claim 1, wherein 7-[(Z) -2- (2-amino-1,3-thiazol-4-yl) -2-methoxyimino-acetylamino] -3- (2,4-dia Minopyrimidin-5-ylmethylsulfanylmethyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid or 7-[(Z) -2- (2-amino-5-chloro-1,3-thiazol-4-yl) -2-hydroxyimino-acetylamino] -3- (2,4-diaminopyrimidin-5-ylmethyl Sulfanylmethyl) -8-oxo-5-thia-1-aza-bicyclo [4,2,0] oct-2-ene-2-carboxylic acid. The compound of formula (5a) is reacted with a compound of formula (6a) in the presence of a base to give a compound of formula (1a), or the compound of formula (5b) in the presence of a base of formula (6b) Reacting with a compound to give a compound of formula (1a) or reducing a compound of formula (1a) with m equal to 1 to give a compound of formula (1) How to prepare the compound: Formula 5a Formula 5b Formula 6a Formula 6b Formula 1a In the above formula, A, R ₁ , R ₂ , W, Q and n are as described above, X 'is a halogen atom, P 'is a thiol, alkyl- or arylthioester, L is a leaving group, m is 0 or 1; An antibiotic composition comprising as an active ingredient a compound of formula (1) and a pharmaceutically acceptable carrier. The antibiotic composition according to claim 4, formulated into an oral preparation containing 50 to 1500 mg of the compound of formula (1).