KR20020085180A - Novel cephalosporin antibiotics and process for preparing same - Google Patents

Abstract

The present invention is useful as an antimicrobial agent, especially the novel cephalosporin compound of Formula 1, a method for preparing the compound of Formula 1, and the like having strong antibacterial activity against strains such as methicillin resistant Staphylococcus aureus (MRSA) It relates to an antimicrobial composition comprising as an active ingredient:

In the above formula

R ¹ , R ² , R ³ , R ^{3 ′} , Q, n and Ar are each as defined in the specification.

Description

New cephalosporin antibiotics and preparation method thereof {Novel cephalosporin antibiotics and process for preparing same}

The present invention relates to novel cephalosporin compounds useful as antibiotics. More specifically, the present invention is a novel cephalosporin compound of Formula 1 below, which is useful as an antibacterial agent and has strong antibacterial activity against strains such as methicillin resistant Staphylococcus aureus (MRSA). To nontoxic salts thereof, physiologically hydrolysable esters thereof, hydrates, solvates, and isomers thereof.

[Formula 1]

In the above formula

R ¹ and R ² each independently represent hydrogen, halogen, C ₁ -C ₆ -alkyl, C ₁ -C ₆ -alkylthio, aryl or arylthio, or one or two heteros selected from nitrogen and oxygen atoms C ₅ -C ₆ -heteroaryl containing atoms,

R ³ and R ^{3 ′} each independently represent hydrogen or a carboxy protecting group,

Q represents O, S, NH, NCH ₃ or CH ₂ ,

n represents an integer of 0 or 1,

Ar represents heteroaryl of the following structural formula,

From here

X, Y, Z, A, B, D, E, G and I each independently represent N or C (or CH), except that the 6-membered ring forms a pyrimidine structure,

R ⁴ represents hydrogen or C ₁ -C ₄ -alkyl or is unsubstituted or substituted by a substituent selected from hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkyl, aryl and pyridyl Indicates,

R ⁵ and R ⁶ each independently represent hydrogen, hydroxy, C ₁ -C ₄ -alkyl or C ₁ -C ₆ -alkylthio, or hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -Amino substituted or unsubstituted by a substituent selected from alkyl, aryl and pyridyl,

R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ each independently represent hydrogen or C ₁ -C ₆ -alkyl, or hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkyl, Amino substituted or unsubstituted by a substituent selected from aryl and pyridyl,

R ¹² , R ¹³ , R ¹⁴ and R ¹⁵ each independently represent hydrogen, hydroxy or C ₁ -C ₄ -alkyl, or hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkyl, Amino substituted or unsubstituted by a substituent selected from aryl and pyridyl,

Olefins in the C-3 position may exist in cis or trans form Represents a double bond or a single bond.

The present invention also relates to a method for preparing the compound of Formula 1 and an antimicrobial composition comprising the compound as an active ingredient.

Cephalosporin antibiotics are widely used to treat infectious diseases caused by pathogenic bacteria in humans and animals, and in particular, to treat diseases caused by bacteria that are resistant to other antibiotics such as penicillin and to treat penicillin-sensitive patients. Do. In most cases of treating infectious diseases, it is desirable to use antibiotics that exhibit antimicrobial activity against both Gram-positive and Gram-negative microorganisms, and the antimicrobial activity of cephalosporin-based antibiotics is 3- or 7-position 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-positions 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 derivative of formula (2) broadly and collectively.

In the above formula

R ¹⁸ is hydrogen or an organic group,

R ¹⁹ is an etherified monovalent organic group linked to oxygen through carbon,

A is -S- or> S-> O,

B is an organic group.

Since the invention of Formula 2, numerous attempts have been made to develop antibiotics of a broad antimicrobial spectrum, and as a result, many cephalosporin antibiotics have been developed. During this development, various changes were induced, such as the introduction of an acylamido group at the 7-position of the Cempec nucleus and a specific group at the 3-position.

Recently, Gram-positive bacteria, particularly methicillin-resistant Pseudomonas aeruginosa (MRSA), have become a serious problem of in vitro infection, and an attempt has been made to introduce an arylthio group at the C-3 position as a cephalosporin-based compound showing strong activity against this. For example, Japanese Patent Application Laid-Open No. 98-36375 discloses a broad and comprehensive description of cephalosporin derivatives of the following general formula (3), in which an arylthio group is introduced at the C-3 position to provide activity against a wide range of pathogens. It is raising.

In the above formula,

R ²⁰ represents substituted alkylthio, aryl, arylthio, aryloxy or heterocyclyl,

A represents protected amino, hydroxy or methylene,

R ²¹ represents protected carboxy or carboxylate,

R ²² is halo, cyano, amidino, guanidino, azido, nitro, substituted alkyl, alkenyl, dichloroalkyl, aryl, alkoxy, aryloxy, alkylthio, arylthio, alkylamino, acyl, carbamoyl , Carbamoyloxy, alkoxyimino, ureido, alkylsulfinyl, alkylsulfonyl or sulfamoyl, 2-substituted pyrimidinyl, quinazolinyl, purinyl, pyrazolo [3,4-d] pyrides Midinyl, pyrazolo [4,3-d] pyrimidinyl, [1,2,3] triazolo [4,5-d] pyrimidinyl or putridinyl,

m is 0 or 1;

In this patent, various heteroaromatic rings are bonded to the C-3 position via a thio group, while corresponding positions of the compound according to the present invention are bound to unsubstituted pyrimidinyl groups via methylene or propenyl. That is, the present invention is a structural feature that the pyrimidinyl group substituted or unsubstituted at the C-3 position is introduced through methylene or propenyl, but there is no mention of this in the patent.

Meanwhile, in order to develop a cephalosporin-based compound capable of exhibiting potent activity against severe inward infection by methicillin-resistant P. aeruginosa (MRSA) as described above, an acyl group is introduced at position 7- and a pyridine substituent is introduced at position C-3. Attempts have been made, a representative example of which is a compound of formula 4 as disclosed in European Patent EP 96-72742.

In the above formula,

Acyl substituent is Ar-S-CH ₂ -CO-, where Ar represents lipophilic substituted phenyl, pyridyl, benzthiazole,

R ²³ and R ²⁴ each represent hydrogen, alkyl or aminoalkylcarbonylamino,

R ²⁵ represents a group having a substituted aliphatic, aromatic, arylaliphatic or sugar moiety.

In this patent, various heteroaromatic rings are introduced into the thioaryl chain at the C-3 position, but this is a C-3 position substituent according to the present invention, that is, a substituted or unsubstituted pyrimidinyl group bonded through methylene or propenyl. Is different at all.

Another cephalosporin-based compound that exhibits potent activity against severe inward infection by methicillin-resistant P. aeruginosa (MRSA), which introduces an acyl group at position 7 and a quaternary ammonium substituent linked to propenyl at position C-3. There was an attempt to do so (WO 99-67255).

In the above formula,

R ²⁶ represents an organic substance not exceeding a molecular weight of 400,

R ²⁷ represents hydrogen, lower alkyl, or phenyl,

R ²⁸ represents an organic substance in which secondary, tertiary or quaternary nitrogen atoms are directly linked to propenyl groups and whose molecular weight does not exceed 400.

In the above patent, various organic substances connected through a nitrogen atom are introduced to the propenyl chain of the C-3 position, but the substituted or unsubstituted C-3 position substituent according to the present invention, that is, bonded through methylene or propenyl No mention is made of pyrimidinyl groups.

Accordingly, the present inventors have continuously tried to develop a cephalosporin compound exhibiting a wide range of antimicrobial activity against Gram-positive bacteria including MRSA. As a result, a pyrimidinyl group substituted or unsubstituted at the C-3 position is substituted with methylene or propenyl. The present invention was completed by discovering that the compound bound through the above-mentioned object is met.

Accordingly, 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.

The present invention also aims to provide a new process for the preparation of compounds of formula (I).

The present invention also aims to provide an antimicrobial composition comprising a compound of formula 1 as an active ingredient together with a pharmaceutically acceptable carrier.

The present invention relates to compounds of formula (1), pharmaceutically acceptable non-toxic salts thereof, physiologically hydrolyzable esters, hydrates, solvates and isomers thereof:

[Formula 1]

In the above formula

R ¹ and R ² each independently represent hydrogen, halogen, C ₁ -C ₆ -alkyl, C ₁ -C ₆ -alkylthio, aryl or arylthio, or one or two heteros selected from nitrogen and oxygen atoms C ₅ -C ₆ -heteroaryl containing atoms,

R ³ and R ^{3 ′} each independently represent hydrogen or a carboxy protecting group,

Q represents O, S, NH, NCH ₃ or CH ₂ ,

n represents an integer of 0 or 1,

Ar represents heteroaryl of the following structural formula,

From here

X, Y, Z, A, B, D, E, G and I each independently represent N or C (or CH), except that the 6-membered ring forms a pyrimidine structure,

R ⁴ represents hydrogen or C ₁ -C ₄ -alkyl or is unsubstituted or substituted by a substituent selected from hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkyl, aryl and pyridyl Indicates,

R ⁵ and R ⁶ each independently represent hydrogen, hydroxy, C ₁ -C ₄ -alkyl or C ₁ -C ₆ -alkylthio, or hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -Amino substituted or unsubstituted by a substituent selected from alkyl, aryl and pyridyl,

R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ each independently represent hydrogen or C ₁ -C ₆ -alkyl, or hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkyl, Amino substituted or unsubstituted by a substituent selected from aryl and pyridyl,

R ¹² , R ¹³ , R ¹⁴ and R ¹⁵ each independently represent hydrogen, hydroxy or C ₁ -C ₄ -alkyl, or hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkyl, Amino substituted or unsubstituted by a substituent selected from aryl and pyridyl,

Olefins in the C-3 position may exist in cis or trans form Represents a double bond or a single bond.

Pharmaceutically acceptable non-toxic salts of the compound of formula 1 according to the present invention are salts with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, acetic acid, trifluoroacetic acid, citric acid, formic acid, maleic acid, hydroxyl acid, 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 with other acids known and used in the art of penicillin and cephalosporin do. These acid addition salts can be prepared according to a conventional conversion method. The compounds of formula 1 may also form non-toxic salts with bases. Bases that can be used include alkali metal hydroxides (e.g. sodium hydroxide, potassium hydroxide), alkali metal hydrogencarbonates (e.g. sodium bicarbonate, potassium bicarbonate), alkali metal carbonates (e.g. sodium carbonate, potassium carbonate), alkaline earth metal carbonates ( Examples include inorganic bases such as calcium carbonate, magnesium carbonate) and organic bases such as amino acids.

Examples of physiologically hydrolyzable esters of the compound of formula (I) include indanyl, phthalidyl, methoxymethyl, pivaloyloxymethyl, glycyloxymethyl, phenylglycyloxymethyl, 5-methyl-2-oxo- 1,3-dioxoren-4-ylmethyl esters and other physiologically hydrolysable esters known and used in the art of penicillin and cephalosporin. Such esters can also be prepared according to known methods.

The compounds according to the invention may have asymmetric carbon centers at the 6- and 7-positions of the cefe ring and thus may be present in R or S isomers, diastereomers, or mixtures thereof including racemates. In addition, when n = 1 may be present in the trans or cis form depending on the geometry of the double bond. Accordingly, the scope of the present invention includes each of these isomers and mixtures thereof.

When R ³ and R ^{3 ′} in the structure of Formula 1 are carboxy protecting groups, they are usually suitable as long as they can be easily removed under mild conditions, such as (lower) alkyl (eg methyl, t-butyl, etc.), (Lower) alkenyl (eg, vinyl, allyl, etc.), (lower) alkylthio (lower) alkyl (eg, methylthiomethyl, etc.), halo (lower) alkyl (eg, 2,2,2-trichloroethyl, etc.) ), Substituted or unsubstituted aralkyl (e.g. benzyl, p-nitrobenzyl, p-methoxybenzyl, etc.), (lower) alkoxycarbonyl (e.g. t -butoxycarbonyl, etc.), tri (di) phenyl (Lower) alkyl, silyl, and the like. The carboxy protecting group can be easily removed under mild reaction conditions such as hydrolysis and reduction to form a free carboxyl group. The carboxy protecting group is appropriately selected and used according to the chemical properties of the compound of Formula 1. In particular, tri (di) phenyl (lower) alkyl group or (lower) alkoxycarbonyl group is easily removed by acid hydrolysis, where organic acids such as formic acid, trifluoroacetic acid and p-toluenesulfonic acid or inorganic acids such as hydrochloric acid can be used. Can be.

Representative of the compound of formula 1 according to the invention may be mentioned:

( 6R, 7R ) -7-{[2-({4-[( E ) -carboxyethenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -8-oxo-3-[( E ) -3- ( 1H -pyrazolo [3,4- d ] -pyrimidin-4-ylsulfanyl) -1-propenyl] -5-thia-1-azabicyclo [4.2.0] oct-2- En-2-carboxylic acid;

( 6R, 7R ) -7-{[2-({4-[( E ) -2-carboxycytyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -3-{( E )- 3-[(2,6-diamino-4-pyrimidinyl) sulfanyl] -1-propenyl} -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene- 2-carboxylic acid;

( 6R, 7R ) -7-{[2-({4-[( E ) -carboxyethenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -8-oxo-3-[( E ) -3- (4,5,6-triamino-1λ ⁵ -pyrimidin-1-yl) -1-propenyl] -5-thia-1-azabicyclo [4.2.0] oct-2-ene- 2-carboxylic acid;

7-amino-5-{[( E ) -3-(( 6R, 7R ) -2-carboxy-7-{[2-({4-[( E ) -2-carboxytenyl]]-2,5 -Dichlorophenyl} sulfanyl) acetyl] amino} -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-en-3-yl) -2-propenyl] sulfanyl} -1H- [1,2,4] triazolo [1,5-C] pyrimidine-4-isot; And

( 6R, 7R ) -7-{[2-({4-[( E ) -2-carboxycytyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -3-(( E )- 3-{[2- (ethylsulfanyl) -6-methyl-4-pyrimidinyl] sulfanyl} -1-propenyl} -8-oxo-5-thia-1-azabicyclo [4.2.0] oct -2-ene-2-carboxylic acid.

On the other hand, the compound of formula 1 according to the present invention, pharmaceutically acceptable non-toxic salts, physiologically hydrolysable esters, hydrates, solvates or isomers of the compounds of formula 6 in the presence of alkali metal iodide in the solvent Alternatively, the compound may be reacted with a compound represented by the following Chemical Formula 7 in the absence, and, if necessary, may be prepared by removing an acid protecting group or reducing S → oxide before or after the reaction. Therefore, it is another object of the present invention to provide such a manufacturing method.

H-Ar

In the above formula,

R ¹ , R ² , R ³ , R ^{3 '} , Q, n and Ar are as defined above,

L represents halogen, preferably chlorine, fluorine or iodine,

p is 0 or 1,

The dashed line means that the compound is a 2-cefem or 3-cefem compound or a mixture thereof, hereinafter used in the same sense.

In the reaction, any solvent that does not adversely affect the reaction may be used, but preferably, lower organic halogenated alkanes such as lower alkylnitrile such as acetonitrile and propionitrile, chloromethane, dichloromethane and chloroform as organic solvents, Tetrahydrofuran, dioxane, ethers such as diethyl ether, amides such as dimethylformamide, esters such as ethyl acetate, ketones such as acetone, hydrocarbons such as benzene, alcohols such as methanol and ethanol, dimethyl sulfoxide Sulphoxides such as sides or mixtures thereof are used. The reaction temperature is suitably -10 to 80 ° C, preferably 20 to 40 ° C, and 0.5 to 2 equivalents, preferably 1.0 to 1.1 equivalents, of the compound of formula 7 is used with respect to the compound of formula 6.

The conversion of the halogen element represented by L to another halogen element in the compound of formula (6), which is a starting material, can be accomplished by commercial methods. For example, the compound of formula (5) in which L is iodine can be obtained by reacting the compound of formula (5) in which L is chlorine with alkali metal iodide.

In addition, the starting compound may be prepared by activating a compound of formula 8 or a salt thereof with an acylating agent and then reacting with a compound of formula 9 or a salt thereof. However, compounds of formula 6 can also be prepared by reacting the free acid of formula 8 directly with a compound of formula 9 in the presence of a condensing agent such as dicyclohexyl carbodiimide, carbonyl diimidazole.

In the above formula,

R ¹ , R ² , R ³ , R ^{3 '} , Q, n, p and L are as defined above.

Acylated derivatives that are active forms of the compound of formula 8 include acid chlorides, acid anhydrides, mixed acid anhydrides (preferably acid anhydrides formed with methylchloroformate, mesitylenesulfonyl chloride, p-toluenesulfonyl chloride or chlorophosphate) or Activated esters (preferably esters formed in the reaction with N-hydroxy benzotriazole in the presence of a condensing agent such as dicyclohexyl carbodiimide).

The reaction of preparing the compound of formula 6 by coupling the compound of formula 8 with the compound of formula 9 is usually an organic base such as tertiary amine, preferably triethylamine, dimethylaniline, pyridine or inorganic such as sodium bicarbonate or sodium carbonate. Organic solvents that proceed well in the presence of a base and may be used include halogenated hydrocarbons such as methylene chloride, chloroform, or tetrahydrofuran, acetonitrile, dimethylformamide, dimethylacetamide or a mixed solvent of these solvents. The reaction can also proceed in aqueous solution. The reaction temperature is about -50 to 50 DEG C, preferably about -30 to 20 DEG C, and the compound of Formula 8 is used in an equivalent amount or a slight excess, that is, 1.05 to 1.2 equivalent times, relative to the compound of Formula 9.

In a compound of Formula 6, which is a starting material, a compound having n = 1 may also be prepared by performing a conventional Wittig reaction from a compound having n = 0 to obtain an illide, and then reacting the halogenated acetaldehyde therewith. .

After the reaction is completed, the product can be separated and purified by conventional post-treatment methods such as silica gel column chromatography, recrystallization, iontophoresis, ion exchange resin chromatography, and the like.

The compound of formula 1 according to the present invention can be usefully used as an antibacterial agent. Accordingly, the present invention contains, as an active ingredient, a compound of Formula 1, a pharmaceutically acceptable non-toxic salt thereof, a physiologically hydrolysable ester thereof, a hydrate, a solvate, or an isomer thereof, together with a pharmaceutically acceptable carrier. It is another object of the present invention to provide an antimicrobial composition.

The total daily dose to be administered to an adult in a single dose or in separate doses when administering a compound of the present invention for clinical purposes is typically in the range of about 500 to 5000 mg, usually about 150 to 3000 mg per day for intramuscular or intravenous administration. A range of will be sufficient, but higher daily dosages may be required for infection with some strains. In addition, the specific dose level for a particular patient may vary depending on the particular compound to be used, weight, age, sex, health condition, diet, time of administration, method of administration, rate of excretion, drug mixing and the severity of the disease.

The compounds of the present invention can be administered in injectable and oral formulations as desired. The compounds of the present invention may be incorporated into unit dose forms or multidose containers, which, when formulated in unit dose form, preferably contain about 50 to 1500 mg of the compound of formula (1).

Injectable preparations, for example sterile injectable aqueous or oily solutions, suspensions or emulsions, can be prepared using suitable dispersing agents, wetting agents, suspending agents or stabilizers according to known techniques. Solvents that can be used include water, Ringer's solution and isotonic NaCl solution, and sterile fixed oils are conventionally employed as a solvent or suspending medium. Any non-irritating fixed oil may be used for this purpose, including mono- and diglycerides, and fatty acids such as oleic acid may be used in the preparation of injectables. In addition, it may be in the form of dry powder for immediate use, which is, for example, sterile and depyrogenated water is dissolved before use.

The compounds of the present invention 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 agents. Solid dosage forms are prepared by mixing the active compound of formula 1 according to the present invention with a carrier selected from one or more inert diluents such as sucrose, lactose, starch and the like, lubricants such as magnesium stearate, disintegrants and binders.

When the compound of the present invention is to be clinically administered to obtain the desired antimicrobial effect, the active compound of Formula 1 may be administered in combination with one or more components selected from known antimicrobial agents. Antibacterial agents that can be administered with the compounds of the present invention in this manner include penicillin or cephalosporin-based compounds.

Hereinafter, the present invention will be described in more detail with reference to the following Examples and Experimental Examples. However, these Examples and Experimental Examples are only for better understanding of the present invention, and the scope of the present invention is not limited by them in any sense.

Preparation Example 1

4-methoxybenzyl ( 6R, 7R ) -7-{[2-({4-[( E ) -3- ( t -Butoxy) -3-oxo-1-propenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -3-[( Z ) -3-chloro-1-propenyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylate

4-methoxybenzyl ( 6R, 7R ) -7-amino-3- [3-chloro-1-propenyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene 0.79 g (1.823 mmol) of 2-carboxylate hydrochloride and 2-({4-[( E ) -3- ( t -butoxy) -3-oxo-1-propenyl] -2,5-dichlorophenyl} Dissolve 0.66 g (1.823 mmol) of sulfanyl) acetic acid in 15 ml dichloromethane. After cooling the reactor to 30 ° C., 0.37 ml (4.5575 mmol) of pyridine and 0.22 ml (2.369 mmol) of phosphoryloxy chloride were slowly added dropwise. The reactor was stirred for 4 hours while gradually raising the temperature to 0 ° C. Diluted with excess ethyl acetate and washed once with saturated ammonium chloride solution, 5% sodium bicarbonate solution and brine. After drying over anhydrous magnesium sulfate, the filtrate was filtered under reduced pressure. The residue was solidified with dichloromethane and diethyl ether and purified to give 0.6 g (45.0%) of the title compound.

¹ HNMR (CDCl ₃ 500 MHz) δ 7.84 to 7.81 (1H, d, J = 16.05 Hz), 7.60 (1H, s), 7.28 to 7.33 (3H.m), 6.87 to 6.85 (1H, d, J = 8.70 Hz), 6.34-6.31 (1H, d, J = 16.05 Hz), 6.23-6.21 (1H, d, J = 10.95 Hz), 5.74-5.72 (2H, m), 5.14 (2H, s), 4.98-4.97 (1H, d, J = 5.05 Hz), 3.95-3.89 (1H, m), 3.79-3.73 (4H, m), 3.45-3.41 (1H, m), 3.25-3.23 (1H, m)

Mass (m / e) 738

Example 1

( 6R, 7R ) -7-{[2-({4-[( E ) -Carboxyethenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -8-oxo-3-[( E ) -3- (1 H -Pyrazolo [3,4- d ] -Pyrimidin-4-ylsulfanyl) -1-propenyl] -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

4-methoxybenzyl ( 6R, 7R ) -7-{[2-({4-[( E ) -3- ( t -butoxy) -3-oxo-1-propenyl] -2,5-dichloro Phenyl} sulfanyl) acetyl] amino} -3-[( Z ) -3-chloro-1-propenyl] -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene- 0.3 g (0.4053 mmol) of 2-carboxylate was dissolved in 10 ml acetone and 0.18 g (1.2159 mmol) of sodium iodide was added. After stirring for 1 hour at room temperature, the solvent was removed by distillation under reduced pressure. The remaining residue was dissolved in 5 ml dimethyl formamide, then 0.074 g (0.4864 mmol) of 4-mercapto-1 H -pyrazolo [3.4- d ] -pyrimidine was added and stirred at room temperature for 24 hours. Diluted with excess ethyl acetate and washed three times with brine and water. After drying over anhydrous magnesium sulfate, the filtrate was filtered under reduced pressure. The residue was purified by diethyl ether and dried under nitrogen.

0.30 g of the solid obtained above was deprotected with trifluoroacetic acid, anisole, and triethylsilane and separated and purified by high pressure preparative liquid chromatography to obtain the title compound (0.030 g, two-step yield: 10.9%).

¹ HNMR (D ₂ O, 400 MHz) δ 8.48 (1H, s), 8.06 (1H, s), 7.52 (1H, s), 7.43-7.39 (1H, d, J = 16.0 Hz), 7.22 (1H, s ), 6.82 to 6.68 (1H, d, J = 16.0 Hz), 6.33 to 6.29 (1H, d, J = 16.0 Hz), 5.96 to 5.92 (1H, m), 5.49 to 5.48 (1H, d, J = 4.4 Hz), 4.98-4.97 (1H, d, J = 4.8 Hz), 4.03-4.01 (2H, m), 3.86-3.74 (2H, Abq, J = 15.6 Hz), 3.44-3.29 (2H, m)

Mass (m / e) 678

Example 2

( 6R, 7R ) -7-{[2-({4-[( E ) -2-carboxyethenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -3-{( E ) -3-[(2,6-diamino-4-pyrimidinyl) sulfanyl] -1-propenyl} -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2- N-2-carboxylic acid

In the same manner as in Example 1, the title compound (two-step yield 11.1%) was obtained.

¹ HNMR (D ₂ O, 400 MHz) δ 7.72 (1H, s), 7.55 to 7.51 (1H, d, J = 16.0 Hz), 7.37 (1H, s), 6.67 to 6.63 (1H, d, J = 16.0 Hz ), 6.42-6.38 (1H, d, J = 16.0 Hz), 5.83-5.79 (1H, m), 5.45-5.44 (1H, d, J = 4.4 Hz), 4.96-4.95 (1H, d, J = 4.4 Hz), 3.90-3.76 (2H, Abq, J = 16.0 Hz), 3.68-3.66 (2H, m), 3.41 (2H, m)

Mass (m / e) 668

Example 3

( 6R, 7R ) -7-{[2-({4-[( E ) -Carboxyethenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -8-oxo-3-[( E ) -3- (4,5,6-triamino-1λ ⁵ -Pyrimidin-1-yl) -1-propenyl] -5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid

In the same manner as in Example 1, the title compound (two-step yield 8.5%) was obtained.

¹ HNMR (D ₂ O, 500 MHz) δ 7.82 (1H, s), 7.72 (1H.s), 7.63 to 7.59 (1H, d, J = 16.06 Hz), 7.47 (1H, s), 6.49 to 6.63 (2H m), 5.93-5.89 (1H, m), 5.48-55.4 (1H, d, J = 4.55 Hz), 4.97-4.96 (1H, d, J = 4.6 Hz), 3.95-3.80 (2H, m), 3.53 to 3.41 (4H, m)

Mass (m / e) 653

Example 4

7-amino-5-{[( E ) -3-(( 6R, 7R ) -2-carboxy-7-{[2-({4-[( E ) -2-carboxyethenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-3 -Yl) -2-propenyl] sulfanyl} -1 H- [1,2,4] triazolo [1,5-C] pyrimidin-4-isene

In the same manner as in Example 1, the title compound (two-step yield 10.6%) was obtained.

¹ HNMR (D ₂ O, 400 MHz) δ 7.91 (1H, s), 7.33 to 7.31 (2H, s, m), 6.97 (1H, s), 6.91 to 6.77 (1H, d, J = 16.8 Hz), 6.20 -6.14 (1H, d, J = 16.0 Hz), 5.98 (1H, s), 5.88-5.84 (1H, m), 5.46-5.45 (1H, d, J = 4.0 Hz), 5.00-4.99 (1H, d , J = 4.0 Hz), 3.90-3.87 (4H, m), 3.47-3.41 (2H, m)

Mass (m / e) 694

Example 5

( 6R, 7R ) -7-{[2-({4-[( E ) -2-carboxyethenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -3-(( E ) -3-{[2- (ethylsulfanyl) -6-methyl-4-pyrimidinyl] sulfanyl} -1-propenyl} -8-oxo-5-thia-1-azabicyclo [4.2.0 Oct-2-ene-2-carboxylic acid

In the same manner as in Example 1, the title compound (two-step yield 21.8%) was obtained.

¹ HNMR (D ₂ O, 500 MHz) δ 7.46-7.40 (2H, br m), 7.17 (1H, br, s), 6.81 (1H, br, m), 6.60 (1H, br s), 6.32 (1H, br, m), 5.81 (1H, br, m), 5.56 (1H, br, m), 5.00 (1H, br, m), 3.85-3.76 (4H, br, m), 3.40 (2H, br, m) ), 2.94 (2H, br, m), 2.13 (3H, br, s), 1.21 (3H, br, m)

Mass (m / e) 712

Experimental Example 1

Minimum Inhibition Concentration (MIC)

The usefulness of the compound according to the present invention was evaluated by obtaining a minimum inhibitory concentration (Minimum Inhibitory Concentration) for the standard strain using vancomycin (Vancomycin) showing an excellent effect on the Gram-positive bacteria among the known compounds. The minimum inhibitory concentration was obtained by diluting the test substance by a 2-fold dilution method, dispersing it in Mueller-Hinton agar medium, and inoculating 2 μl of a test strain having 10 ⁷ cfu (colony forming unit) per ml. It was obtained by incubating for 20 hours at ℃ and the results are shown in Table 1 below. As a result of the minimum inhibitory concentration test for the strains, the compound according to the present invention was shown to have excellent inhibitory activity against the main infectious bacteria including the MRSA strain.

Susceptibility test results for the standard strain (㎍ / ㎖) compound S. aureus giorgio S. aureus 77 S. aureus K311 S. epidermidis R005 E faecalis EFS004 Example 1 0.031 0.5 0.5 One One Example 2 0.031 0.5 0.5 One 4 Example 3 0.063 2 One 2 4 Example 4 0.031 0.5 One One 2 Example 5 0.031 One One 2 One Vancomycin One One 2 One One

The compounds of formula 1 according to the present invention, pharmaceutically acceptable non-toxic salts thereof, physiologically hydrolysable esters, hydrates, solvates, and isomers thereof are potent antimicrobial activities against a wide range of pathogenic bacteria, including various Gram-positive bacteria. And a broad antimicrobial spectrum, which is very useful for the prevention and treatment of diseases caused by bacterial infection in animals including humans.

Claims (4)

The cephalosporin compound of formula 1, a pharmaceutically acceptable non-toxic salt thereof, physiologically hydrolysable esters, hydrates, solvates, and isomers thereof: [Formula 1] In the above formula R ¹ and R ² each independently represent hydrogen, halogen, C ₁ -C ₆ -alkyl, C ₁ -C ₆ -alkylthio, aryl or arylthio, or one or two heteros selected from nitrogen and oxygen atoms C ₅ -C ₆ -heteroaryl containing atoms, R ³ and R ^{3 ′} each independently represent hydrogen or a carboxy protecting group, Q represents O, S, NH, NCH ₃ or CH ₂ , n represents an integer of 0 or 1, Ar represents heteroaryl of the following structural formula, From here X, Y, Z, A, B, D, E, G and I each independently represent N or C (or CH), except that the 6-membered ring forms a pyrimidine structure, R ⁴ represents hydrogen or C ₁ -C ₄ -alkyl or is unsubstituted or substituted by a substituent selected from hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkyl, aryl and pyridyl Indicates, R ⁵ and R ⁶ each independently represent hydrogen, hydroxy, C ₁ -C ₄ -alkyl or C ₁ -C ₆ -alkylthio, or hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -Amino substituted or unsubstituted by a substituent selected from alkyl, aryl and pyridyl, R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ each independently represent hydrogen or C ₁ -C ₆ -alkyl, or hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkyl, Amino substituted or unsubstituted by a substituent selected from aryl and pyridyl, R ¹² , R ¹³ , R ¹⁴ and R ¹⁵ each independently represent hydrogen, hydroxy or C ₁ -C ₄ -alkyl, or hydroxy-C ₁ -C ₄ -alkyl, C ₁ -C ₄ -alkyl, Amino substituted or unsubstituted by a substituent selected from aryl and pyridyl, Olefins in the C-3 position may exist in cis or trans form Represents a double bond or a single bond. The method of claim 1, ( 6R, 7R ) -7-{[2-({4-[( E ) -carboxyethenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -8-oxo-3-[( E ) -3- ( 1H -pyrazolo [3,4- d ] -pyrimidin-4-ylsulfanyl) -1-propenyl] -5-thia-1-azabicyclo [4.2.0] oct-2- En-2-carboxylic acid; ( 6R, 7R ) -7-{[2-({4-[( E ) -2-carboxycytyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -3-{( E )- 3-[(2,6-diamino-4-pyrimidinyl) sulfanyl] -1-propenyl} -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene- 2-carboxylic acid; ( 6R, 7R ) -7-{[2-({4-[( E ) -carboxyethenyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -8-oxo-3-[( E ) -3- (4,5,6-triamino-1λ ⁵ -pyrimidin-1-yl) -1-propenyl] -5-thia-1-azabicyclo [4.2.0] oct-2-ene- 2-carboxylic acid; 7-amino-5-{[( E ) -3-(( 6R, 7R ) -2-carboxy-7-{[2-({4-[( E ) -2-carboxytenyl]]-2,5 -Dichlorophenyl} sulfanyl) acetyl] amino} -8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-en-3-yl) -2-propenyl] sulfanyl} -1H- [1,2,4] triazolo [1,5-C] pyrimidine-4-isot; And ( 6R, 7R ) -7-{[2-({4-[( E ) -2-carboxycytyl] -2,5-dichlorophenyl} sulfanyl) acetyl] amino} -3-(( E )- 3-{[2- (ethylsulfanyl) -6-methyl-4-pyrimidinyl] sulfanyl} -1-propenyl} -8-oxo-5-thia-1-azabicyclo [4.2.0] oct A compound selected from the group consisting of -2-ene-2-carboxylic acid. The compound of formula 6 is reacted with a compound of formula 7 in the presence or absence of an alkali metal iodide in a solvent, and if necessary, the acid protecting group is removed before or after the reaction or the S → oxide is reduced. To prepare a compound of Formula 1: [Formula 6] [Formula 7] H-Ar In the above formula, R ¹ , R ² , R ³ , R ^{3 ′} , Q, n and Ar are as defined in claim 1, L represents halogen, p is 0 or 1, The dashed line means that the compound is a 2-cefem or 3-cefem compound or a mixture thereof. An antimicrobial composition comprising a compound of formula 1 as defined in claim 1 together with a pharmaceutically acceptable carrier.