JP3012986B2 - Cephem compound and method for producing the same - Google Patents

Description

The present invention relates to a novel compound having excellent antibacterial activity and a method for producing the same.

[Conventional technology]

formula 7β-[(Z) -2- (5-amino-1,2,
4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3- (1-imidazo [1,2-b] -pyridazinium) methyl-3-cephem-4-carboxylate [hereinafter simply cephem compound (I) It may be called. Are compounds having excellent antibacterial activity described in JP-A-62-149682.

[Problems to be solved by the invention]

The cephem compound (I) or an acid addition salt thereof exhibits excellent antibacterial activity, and exhibits strong antibacterial activity against a wide range of pathogenic bacteria including Gram-positive and Gram-negative bacteria. Therefore, it is useful as a therapeutic drug for infectious diseases. However, crystals of the compound of formula (I) (ie the free form) do not exhibit sufficient solubility for use as injections, and the amorphous solid of the compound of formula (I) has the disadvantage of poor stability. . An object of the present invention is to overcome the above drawbacks and to provide a stable and excellent water-soluble form of the cephem compound (I) or a derivative thereof.

[Means for solving the problem]

The present inventors have conducted intensive studies on the reaction under various conditions using a compound of the formula (I) or a salt containing an acid addition salt thereof in order to achieve the above object. As a result, the compound of the formula (I) or a salt thereof And carbonate in the presence of water, the formula A salt consisting of an anion and a cation represented by the formula [hereinafter sometimes referred to as a cephem compound (II)] or a hydrate thereof is formed, and this compound has a remarkably improved solubility in water as compared with the cephem compound (I). And excellent antimicrobial activity were found, and the present inventors completed further studies and completed the present invention.

That is, the present invention relates to the cephem compound (II), a method for producing the same, a therapeutic agent for infectious diseases and an antibacterial agent containing the cephem compound (II).

The cephem compound (II) may be a hydrate, and such a hydrate is also included in the present invention (hereinafter, a hydrate may be referred to as a cephem compound (II)).

Examples of the cation include alkali metal ions, alkaline earth metal ions, and ammonium ions.
Particularly, an alkali metal ion is preferable. When the cation is an alkali metal ion, the cephem compound (II) can be represented by the following formula.

(Wherein, M represents an alkali metal) As such an alkali metal, sodium and potassium are preferable, and sodium is particularly preferable.

Cephem compound (II) is produced by reacting cephem compound (I) or a salt thereof with carbon dioxide, carbonic acid or carbonate in the presence of water in a basic manner.

Examples of the salt of the cephem compound (I) include a salt with an alkali metal or an alkaline earth metal, and an acid addition salt.

Suitable acid addition salts of cephem compound (I) are non-toxic pharmaceutically acceptable hemi- and mono-acid addition salts purified by cephem compound (I) and a monobasic or polybasic acid, such as hydrochloride, Examples thereof include inorganic acid addition salts such as sulfates and organic acid addition salts such as acetic acid. Among them,
Particularly, the hydrochloride is preferred. Cephem compound (I) or a salt thereof may be subjected to the reaction in the form of a solvate including a hydrate thereof.

Incidentally, the cephem compound (I) is disclosed in the above-mentioned JP-A-62-149682.
Can be produced.

The acid addition salt can be produced according to a conventional method. The acid addition salt is preferably obtained as crystals from the viewpoints of stability, purity and the like. For example, crystals of the hydrochloride of the cephem compound (I) can be obtained, for example, by the method described in Japanese Patent Application No. 63-301923. it can.

The cephem compound (I) or an acid addition salt thereof may be subjected to a reaction with a carbonate as it is in the form of a solvate of the solvent used. However, in the case of a solvate of an organic solvent, the solvent is generally removed by a conventional method. Is preferred.

As the carbonate, any pharmaceutically acceptable salt can be used. Such suitable pharmaceutically acceptable carbonates include, for example, alkali metal bicarbonates such as sodium bicarbonate and potassium bicarbonate, and alkali metal carbonates such as sodium carbonate and potassium carbonate. Particularly preferred carbonates include sodium hydrogen carbonate and sodium carbonate.

The reaction is usually performed in a commonly used solvent such as water, acetone, dioxane, acetonitrile, chloroform, dichloromethane, tetrahydrofuran, ethyl acetate, N, N-dimethylformamide, dimethylsulfoxide, and these solvents may be used alone or in combination with water. It may be used as a mixture.

Although the reaction temperature is not particularly limited, it is usually carried out under ice cooling or heating.

The amount of carbonate to be used is usually the amount of the cephem compound (I)
0.5 to 20 equivalents are used per 1 mol, and in the case of alkali hydrogen carbonate, 1 to 10 equivalents are preferable. In the case of an alkali carbonate, 1 to 5 equivalents are preferable. In one embodiment of the present invention, the cephem compound (II) can be produced by reacting the cephem compound (I) with carbon dioxide in the presence of a base. In this case, examples of the base include the above-mentioned carbonates and alkali hydroxides (eg, sodium hydroxide, potassium hydroxide). The reaction can be usually carried out by blowing gaseous carbon dioxide into the cephem compound (II) or by using Dries. Carbon dioxide is conveniently used in excess. The base is used usually in an equivalent amount or more.

The reaction is carried out in an open system or a closed system, preferably in a closed system.

The thus obtained cephem compound (II) can be isolated and purified by known processing means such as column chromatography, precipitation, and recrystallization.

The cephem compound-containing compound (II) provided by the present invention is a novel compound, has a strong antibacterial activity, exhibits a remarkably high solubility in water as compared with the cephem compound (I), and is therefore useful as an antibacterial agent. . The cephem compound (II) of the present invention can be administered parenterally or orally as injections, capsules, tablets, granules and the like, similarly to known penicillins and cephalosporins.

Carriers used as injections include, for example, distilled water, physiological saline and the like. Capsules,
When used as powders, granules, or tablets, they may be used as they are or as known pharmacologically acceptable excipients (eg, starch, lactose, sucrose, calcium carbonate, calcium phosphate, etc.), binders (eg, starch, gum arabic, It is used as a mixture with carboxymethylcellulose, hydroxypropylcellulose, crystalline cellulose, etc.), a lubricant (eg, magnesium stearate, talc, etc.), and a breaking agent (eg, carboxymethylcalcium, talc, etc.).

When the cephem compound (II) of the present invention is used for the treatment of infectious diseases, for example, as an injectable preparation, the cephem compound (II) may be used by dissolving it in distilled water for injection or physiological saline or the like by an ordinary method as appropriate. . Further, as described in the text, a basic mixture of cephem compound (I) or a salt thereof and carbonic acid or carbonate may be prepared by adding distilled water for injection or physiological saline. In this case, distilled water for injection or the like is used in accordance with the usual method of the usual preparation for injection of cephem antibiotics.
You may add and dissolve in several or more times.

The dosage is 0.5 to 80 mg / kg / day for humans and animals infected with pathogenic bacteria including gram-positive and negative bacteria.
Preferably, 1 to 20 mg / day is divided and administered twice to four times a day.

The cephem compound (II) of the present invention has much higher solubility in water than the cephem compound (I).

Distilled water was added to the cephem compound (II) of the present invention at room temperature, and the solubility was measured. As a result, a solubility of 20% w / v or more was shown.

In order to further demonstrate the usefulness of the cephem compound (II),
The in vitro antibacterial activity was examined. The results are shown in Table I.
The in vitro antimicrobial activity was represented by the minimum inhibitory concentration (MIC) in μg / ml according to the following method.

Measurement method: The MIC value of the test compound was determined by agar dilution
ethod). That is, 1.0 ml of a sequentially diluted aqueous solution of the test compound is poured into a petri dish (petri dish),
Next is Trypticase Soi Agar
pour 9.0 ml of soy agar) and mix. The suspension of the test bacteria (about 10 ⁸ CFU / ml) is spread on the mixed agar plate.

After culturing (incubation) at 37 ° C for 18 hours, the minimum concentration of the test compound that completely inhibits the growth of the test bacterium is defined as the minimum inhibitory concentration (MIC).

The above test results show that the cephem compound (II) of the present invention has excellent antibacterial activity.

〔The invention's effect〕

As described above, the cephem compound (II) or a hydrate thereof of the present invention has high solubility in water and has excellent antibacterial activity.

〔Example〕

Next, the present invention will be described in more detail with reference to Reference Examples and Examples, but the present invention should not be limited to these Examples and Reference Examples.

Elution in column chromatography in Reference Examples and Examples was performed by TLC (Thin Layer Chromatography) and HPLC (High Performance Liquid Chromatography)
Under observation by In TLC observation, Kieselgel 60F ₂₅₄ (70) (manufactured by Merck) was used as a TUC plate.
230230 mesh), a solvent used as an elution solvent in column chromatography as a developing solvent, and a UV detector as a detection method. As the silica gel for the column, Kieselgel 60 (70 to 230 mesh) manufactured by Merck was also used. HPLC used Shimadzu LC-5A type device, Nucleosil _C18 250 × 4mm (particle system 5μm) as column, detection connected with Shimadzu Chromatopack 3R to UV detector,
Performed at 254 nm. The NMR spectrum shows proton NMR, and the CMR spectrum shows C ¹³ NMR. These are VARIAN E using tetramethylsilane as internal or external reference.
Measured with M390 (90MHz type spectrometer) etc.
The values are shown in ppm. In the mixed solvents, the numerical values shown in parentheses are the volume mixing ratios of the respective solvents.

The abbreviations used in the examples have the following meanings.

s: singlet, br: broad (broad), d: doublet, dd: doublet, ABq: AB quarlet, m: multiplet, J: coupling constant, Hz: hertz%: weight% Means 15-25 ° C.

Reference Example 1 7β-[(Z) -2- (5-amino-1,2,4-thiazi
Azol-3-yl) -2-methoxyiminoacetami
Do] -3-[(1-imidazo [1,2-b] pyridaziniu
M) Methyl] -3-cephem-4-carboxylate 7β-[(Z) -2- (5-amino-1,2,4-thiazi
Azol-3-yl) -2-methoxyiminoacetami
Do] -3-[(3-oxobutyryloxymethyl) -3
-Cephem-4-carboxylic acid 1.1 g, imidazo [1,2-b]
Pyridazine 1.0 g, potassium iodide 1.1 g
Dissolve in 30 ml of nitrile and stir at 60-70 ° C for 2 hours. cold
After the reaction, the reaction mixture was subjected to silica gel column chromatography.
And elute sequentially with acetone and water-containing acetone.
The fractions containing are combined and the solvent is concentrated under reduced pressure. MCI to residue
Gel CHP20P Column chromatography with resin (Mitsubishi Kasei)
And eluted with water and then hydrous alcohol
The fractions containing the target compound are combined and concentrated under reduced pressure.
Lyophilization gives the title compound.

Elemental _{analysis: C 19 H 17 N 9 O} 5 S 2 · 3H 2 O Calculated (%): C, 40.07; H, 4.07; N, 22.13 Found (%): C, 39.75; H, 3.51; N, 21.89 IR spectrum ν ▲ ^KBr _max ▼ cm ^-1 : 1765,1660,1610,1520. NMR spectrum (d ₆ -DMSO) δ: 3.03 and 3.44 (2H, ABq, J =
18Hz), 3.86 (3H, s), 4.99 (1H, d, J = 4.5Hz), 5.27 and 5.
51 (2H, ABq, J = 14Hz), 5.63 (1H, dd, J = 4.5Hz and 8Hz),
7.8−8.32 (1H, m), 8.12 (2H, br.s), 8.76 (2H, S), 9.0
4 (1H, d, J = 4Hz), 9.31 (1H, d, J = 9Hz), 9.44 (1H, d, J
Reference Example 2 700 g of the freeze-dried product obtained in Reference Example 1 was dissolved in 400 ml of water, and stirred at room temperature for 1.5 hours to crystallize, yielding 7β-[(Z)
-2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-[(1
-Imida [1,2-b] pyridazinium) methyl] -3-
Cephem-4-carboxylate is obtained as crystals.

Reference Example 3 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamide] -3-[(1- Imidazo [1,2
-B] pyridazinium) methyl] -3-cephem-4-
After dissolving 11.3 g of carboxylate in 20 ml of 1N hydrochloric acid, 77 ml of acetone is gradually added with stirring, and the mixture is stirred at room temperature for 7 hours. The crystallized crystals were collected, washed with 20 ml of a mixed solution of acetone: water (6: 1), further washed with 40 ml of acetone, and dried to obtain 7β-[(Z) -2- (5-amino-1,2,2, 4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-[(1-imidazo [1,2-b] pyridazinium) methyl] -3-cephem-4-carboxylate hydrochloride (acetone solvate) ) 7.6 g are obtained.

Reference Example 4 5.0 g of the compound (acetone solvate) obtained in Reference Example 3 was placed on a glass filter, passed through an aqueous layer at 10 ° C., humidified air was sent through the filter (flow rate 1 / min), and the solvent was removed. Dry under reduced pressure and dry 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamide] -3- (1-imidazo [1,2-
b] Pyridazinium) methyl-3-cephem-4-carboxylate.hydrochloride 4.85 g was obtained. The product contained 8.2% water and the residual acetone was less than 0.2% by NMR.

Elemental analysis: C ₁₉ H ₁₈ N ₉ ClO ₅ S ₂・ 2.5H ₂ O Calculated: C, 38.22; H, 3.88; N, 21.11; Cl, 5.94 Found: C, 38.17; H, 3.56; N, 21.02; Cl, 5.96 Example 1 7β-[(Z) -2- (5-carboxylateamino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamide] -3-[(1- Imidazo [1,2-
b] pyridazinium) methyl] -3-cephem-4-carboxylate sodium 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) obtained in Reference Example 4 -2-methoxyiminoacetamide] -3-[(1-imidazo [1,2
-B] pyridazinium) methyl] -3-cephem-4-
0.5g carboxylate / hydrochloride and 150mg sodium carbonate
Then, 1.7 ml of water was added to the mixture to dissolve, the mixture was sealed, and the mixture was left at room temperature for 5 hours. The reaction solution is subjected to column chromatography on Sephadex LH-20 (Pharmacia), and eluted with water. When the fraction containing the target substance is thickened and freeze-dried,
73 mg of the title compound are obtained.

IR spectrum ν ▲ ^KBr _max ▼ cm ^-1 : 1765,1670,1615,1535,1
350,1290. NMR spectrum (D ₂ O) δ: 3.20 and 3.60 (2H, ABq, J = 17.6H
z), 4.07 (3H, s), 5.25 (1H, d, J = 4.6Hz), 5.39 and 5.41
(2H, ABq, J = 15.8Hz), 7.90 (1H, dd, J = 4.6Hz and 9.5H
z), 8.26 (1H, d, J = 2.3Hz), 8.43 (1H, d, J = 2.3Hz), 8,
68 (1H, dd, J = 9.5Hz and 1.4Hz), 8.94 (1H, dd, J = 4.6Hz
1.4Hz) Example 2 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-[(1-imidazo [1,2-b ] Pyridazinium) methyl] -3-cephem-4-carboxylate.
Water in a mixture of 0.4 g of hydrochloride and 250 mg of sodium hydrogen carbonate
Add 1.5 ml to dissolve, stopper tightly and leave at room temperature for 15 hours. The reaction mixture is subjected to Sephdex LH-20 column chromatography, and eluted with water. The fractions containing the target compound were collected and lyophilized to give 340 mg of the same compound as in Example 1.
can get.

The IR, NMR and CMR spectra were all consistent with the compound obtained in Example 1.

Example 3 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamide] 3-[(1-imidazo [1,2-b] Pyridazinium) methyl] -3-cephem-4-carboxylate.
Add 5 g to 2 ml of water, dissolve at 60 ° C, and add sodium hydrogen carbonate 2
After adding 100 mg, the mixture is sealed and left at room temperature for 10 hours. The reaction solution was subjected to Sephedex LH-20 column chromatography,
Elute with water. The fractions containing the desired product are collected and lyophilized to give 230 mg of the compound obtained in Example 1.

Example 4 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-[(1-imidazo- [1,2, -B] pyridazinium) methyl] -3-cephem-4-carboxylate hydrochloride (1.09 g) and sodium carbonate (200 mg) in water (10 ml)
And sealed and left at room temperature for 8 hours. The reaction product is subjected to column chromatography on Sephadex LH-20 (Pharmacia), and eluted with water. The fractions containing the target compound were collected and freeze-dried to obtain 68 mg of the target compound described in Example 1.
can get.

Example 5 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoisiminoacetamido] -3-[(1-imidazo- [1,2 -B] pyridazinium) methyl] -3-cephem-4-carboxylate.hydrochloride (1.09 g) and sodium carbonate (200 mg) in water (2 ml)
5 minutes after sealing, 8 ml of water is further added, and the mixture is left at room temperature for 5 hours. The reaction solution was subjected to Sephadex column chromatography, and eluted with water.
0 mg is obtained.

Example 6 7β-[(Z) -2- (5-carboxylateamino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamide] -3-[(1-imidazo- [1, Two
-B] pyridazinium) methyl] -3-cephem-4-
Carboxylate potassium 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-[(1-imidazo- [1,2 -B] pyridazinium) methyl] -3-cephem-4-carboxylate hydrochloride (0.4 g) and potassium hydrogen carbonate (300 mg) in water
Dissolve by adding ml, stopper tightly and leave at room temperature for 15 hours. The reaction solution is subjected to column chromatography on Sephadex LH-20, and eluted with water. The fractions containing the target compound are collected and freeze-dried to obtain 390 mg of the title compound.

Example 7 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) 2-methoxyiminoacetamido] -3-[(1-imidazo [1,2-b] Pyridazinium) methyl] -3-cephem-4-carboxylate.
To a mixture of 0.5 g of the hydrochloride and 120 mg of potassium carbonate was added 2 ml of water to dissolve, sealed, and allowed to stand at room temperature for 8 hours. The reaction is subjected to column chromatography on Sephadex LH-20 and eluted with water. The fractions containing the target compound are collected and freeze-dried to obtain 90 mg of the target compound described in Example 6.

Example 8 7β-[(Z) -2- (5-amino-1,2,4-thiadiazol-3-yl) -2-methoxyiminoacetamido] -3-[(1-imidazo [1,2-b ] Pyridazinium) methyl] -3-cephem-4-carboxylate
Dissolve a mixture of 5 g and 100 mg of sodium carbonate in 5 ml of water,
Add 2.0 g of dry ice and leave in a sealed tube at room temperature for 5 hours. The reaction is subjected to Sephadex LH-20 column chromatography, eluting with water. The fractions containing the target compound were collected and lyophilized to give 180 mg of the target compound described in Example 1.
can get.

Continuation of the front page (56) References JP-A-1-110690 (JP, A) JP-A-62-149682 (JP, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) C07D 501 / 00-501/62 A61K 31/00-31/80 CA (STN) REGISTRY (STN)

Claims (6)

(57) [Claims] (1) Expression A salt comprising an anion and a cation represented by 2. The salt according to claim 1, wherein the cation is an alkali metal ion, an alkaline earth metal ion or an ammonium ion. 3. The salt according to claim 1, wherein the cation is an alkali metal ion. (4) 2. The method for producing a salt or a hydrate thereof according to claim 1, wherein the compound represented by the formula (1) or a salt thereof is reacted with carbon dioxide, carbonic acid or carbonate in the presence of water. (5) A therapeutic agent for an infectious disease comprising the salt according to (1). 6. An antibacterial agent comprising the salt according to claim 1.