JPS5811956B2 - New antibacterial compound - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a novel cephalosporin derivative represented by the general formula or a salt thereof.

In the above general formula (I), R1 means a carboxyl group or a cyan group, and R2 means a hydroxyl group, an amine group, or a mono- or di-lower alkylamino group.

Here, examples of the mono- or di-lower alkylamino group for R2 include a methylamine group, ethylamino group, dimethylamine group, diethylamino group, and the like.

Compound (I) provided by the present invention is a completely new cephalosporin derivative in chemical structure in that it has a 1,3-dichetane ring in the side chain at position 7 of the cephalosporin core, and has excellent antibacterial properties. Since it shows activity, it is promising as an antibacterial agent.

The antibacterial activity (minimum effective inhibitory concentration) of compound (I) is shown below in comparison with commercially available cefazolin (CEZ).

In particular, it has been shown to have excellent efficacy against several important pathogenic bacteria belonging to Durham-negative bacteria.

Therefore, the compound [I] of the present invention is suitable for pharmaceuticals, especially antibacterial agents,
It is useful as a feed additive and preservative.

The compound of the present invention is available as it is or as a salt thereof.

Preferred salts include salts with pharmaceutically acceptable non-toxic bases, such as alkali metal salts such as sodium salts and potassium salts; ammonium salts; dicyclohexylamine salts, cyclohexylamine salts;
Examples include salts with organic bases such as trimethylamine salt, triethylamine salt, ethanolamine salt, ornithine salt, and lysine salt.

The compound of the present invention or a salt thereof is administered orally or parenterally as an antibacterial agent.

The dosage varies depending on the symptoms, body weight, etc., but for adults it is usually administered in 3 to 4 doses over an area of about 250 to 3000 m2.

Dosage forms suitable for administration include injections, tablets, capsules, syrups, etc., but when preparing these dosage forms, excipients, preservatives, stabilizers, etc. used in pharmaceuticals must be added. , can be carried out by conventional methods.

According to the present invention, the compound (I) can be produced by the following method.

1st method (In the formula, R1 has the above meaning.

) This method is carried out by treating a cephalosporin compound (n) having a (4-monosubstituted-3-hydroxyisothiazol-5-yl)thioacetamide group in the 7-position with a base.

Suitable bases include weakly basic substances such as sodium hydrogen carbonate, potassium hydrogen carbonate, and sodium carbonate.

This reaction is usually carried out in a solvent at room temperature or under cooling.

The solvent is not particularly limited as long as it does not participate in the reaction, but water or water-miscible methanol, acetone, tetrahydrone, dimethylformamide, etc. are preferably used alone or in appropriate mixtures.

Isolation and purification of the product from the reaction solution is carried out according to conventional methods.
Extraction with organic solvents, crystallization, and separation and purification by column chromatography are used.

Second method This method is shown by the following reaction formula.

(In the formula, R3 means a hydrogen atom or a protecting group for a carboxyl group.

Moreover, R7 and R2 have the above meanings.

) The reaction for producing the target compound [I] of the present invention by the second method is a 7-aminocephalosporin derivative [■]
Alternatively, the compound [v] is prepared by reacting 4-(substituted-methylene)-1,3-dichetane-2-carboxylic acid (IV) or a reactive derivative of the carboxylic acid, and R3 of the compound is a carboxyl group. When it is a protecting group for a group,
This is then done by removing it.

In the reaction between the above compound 1) and [IV,), the compound (I
V) can be converted into a reactive derivative of its carboxylic acid and then subjected to the reaction.

Suitable reactive derivatives include acid halides, mixed acid anhydrides, active esters, active amides, acid anhydrides, acid azides, and the like.

When compound (IV) is used in a free state, a condensing agent may be used.

Suitable condensing agents include N,N'-dicyclohexylcarbodiimide and N,N'-diethylcarbodiimide.

In addition, when R1 in compound (IV) is a carboxyl group or R2 is a hydroxyl group, in order to avoid side reactions between these active groups and compound (1), these active groups are converted into tert-butyl groups in advance. It is advisable to protect it with a protective film such as benzhydryl group or benzhydryl group.

The reaction between compound [■] and compound [■] or its reactive derivative is carried out under heating or cooling, but in order to avoid the 7-position methoxy group undergoing ebimerization during the reaction, low temperature, It is particularly preferable to carry out the reaction at a temperature of -20°C or lower.

At the 7-position thus generated, 4-(substituted-methylene)-1,
The cephalosporin compound [v] having a 3-dichetane-2-carboxamide group can then be led to the target compound CI) by removing the protecting group.

Third Method The object compound CI of the present invention can also be produced by the reaction of the following formula.

(In the formula, R4 represents an acetyl group or a carbamoyl group.

Moreover, R1 and R2 have the above meanings. ]] This method involves the addition of 7β-(4-(substituted-methylene)-51,3-dicetan-2-yl)carboxamide cephalosporin derivatives [■] or its salts to 1-methyl-]]]H-
This is carried out by reacting tetrazole-5-thi or its mercapto group with an alkali metal substituted for hydrogen.

The reaction is carried out at room temperature to elevated temperature, usually in a solvent.

Solvents that do not participate in this reaction include, for example, acetone, dimethylformamide, methanol, ethanol, water, and phosphate buffer, but these may be used in combination if necessary.

The reaction is preferably carried out near neutrality.

As starting material, 1-methyl-1H-tetrazole-5
- When thiol is used in its free state, it is preferably carried out in the presence of a base such as an alkali metal hydroxide, an alkali metal carbonate, an alkali metal hydrogen carbonate, a trialkylamine, pyridine, or dimethylaniline.

In order to isolate the product after the reaction is completed, a reaction solution may be made acidic and a precipitated product may be collected, or a solvent extraction method may be used.

As another production method, other desired products can be obtained by converting the desired product in which R2 of -C0-R2- is an amino group to -CN using a dehydrating agent such as phosphorus pentachloride.

Next, the present invention will be explained in more detail with reference to Examples.

Example 1 7β-(4-carboxy-3-hydroxyisothiazol-5-yl)thioacetamide-7-α-methoxy-
Add 6.0 g of 3-(1-methyltetrazol-5-yl)thiomethyl-△3-cephemu-4-carboxylic acid to 40 ml of methanol and 300 ml of 15% aqueous sodium bicarbonate solution.
1 and stir at room temperature for 5 hours.

This solution was washed with 300 ml of ethyl acetate, then acidified with dilute hydrochloric acid, and 2 equal volumes of 100 ml of each 200 ml of n-butanol-ethyl acetate (volume ratio 1:1) were added.
Extract once.

The organic layers are combined, washed twice with 50 ml each of saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and then the solvent is distilled off under reduced pressure.

Add 50 ml of ether to the residue, collect the resulting precipitate,
After washing with ether and drying, a crude product is obtained.

The crude product was purified by silica gel column chromatography using a mixture of chloroform-methanol-formic acid (volume ratio 100; 20:1.5) as the eluent.

Fractions containing the target product were collected and the solvent was distilled off under reduced pressure to obtain 7β-[4-(carbamoylcarboxymethylene)-1,3-dicetan-2-yl] as a white to pale yellowish white powder.
3.5 g of carboxamide 7α-methoxy-3-(1-methyltetrazol-5-yl)thiomethyl-Δ3-cephemu-4-carboxylic acid are obtained.

Cough magnetic resonance spectrum (DMSOd6) δ (ppm): 3.40 (3H) C7 CH30-3,61 (2
H) -CH2- infrared absorption spectrum of C2 (K
Br) Example 2゜a) 7β-(4-(carbamoyl carboxymethylene)-1,3-dicetan-2-yl]carboxamide 7α-methoxy-3-(1-methyltetrazole-5
-yl)thiomethyl-△3-cephemu-4-carboxylic acid (1.0 g) was suspended in a mixture of 5 Qm of chloroform and 10 ml of acetone, and about 700 ml of diphenyldiazomethane was added. A solution prepared by dissolving this in 5 ml of chloroform is added dropwise.

After stirring at room temperature for 30 minutes, the solvent was distilled off.

The residue was subjected to silica gel column chromatography using a mixture of chloroform and ethyl acetate (volume ratio 2:1) as an eluent to isolate and purify the desired product.

The fractions containing the target product were collected and the solvent was distilled off under reduced pressure.
β-(4-(Benzhydryloxycarbonyl carbamoylmethylene)-1,3-dicetan-2-yl]
0.8 g of carboxamide 7α-methoxy-3-(1-methyltetrazol-5-yl)thiomethyl-Δ3-cephemu 4-carboxylic acid benzhydryl ester was obtained.

Nuclear magnetic resonance spectrum (CDC15) δ (ppm): 3.52 (5H) CH30- of C2, H and C7
4,35(2H) Ca CH2-8-5,00,5
,03(IH) H port of C6)7β-(4-(benzhydryloxycarbonyl carbamoylmethylene)-
1,3-dichetan-2-yl]carboxamide 7α
-Metho.

1.0 g of xy-3-(1-methyltetrazol-5-yl)thiomethyl-Δ3-cephemu 4-carboxylic acid benzhydryl ester was dissolved in 10 ml of chloroform,
While stirring while cooling with water, add 0.3 ml of pyridine and 0.45 g of phosphorus pentachloride, stir further at room temperature for 1 hour, cool with water, and add 3 mA of water.

The organic layer is separated into an organic layer and an aqueous layer, and the organic layer is washed with 2 ml of water, dried over anhydrous magnesium sulfate, and the solvent is distilled off under reduced pressure.

The residue was subjected to silica gel column chromatography using a mixture of chloroform and ethyl acetate (volume ratio 4:1) as an eluent to isolate and purify the desired product.

Collect fractions containing the target product, remove the solvent under reduced pressure,
7β-(4-(benzhydryloxycarbonylcyanomethylene)-1,3-dicetan-2-yl]carboxamide 7α-methoxy-3-(]-]]]methyltetrazol-5-ylthiomethy△3-cephemu 4 -0.5 g of carboxylic acid benzhydryl ester is obtained.

Nuclear magnetic resonance spectrum (CDC15) δ (ppm) 3.45 (5H) H of C2 and C of C7) (30-4,32(2H) -CH2-5-Ha of C3) 7β
-(4-(benzhydryloxycarbonyl cyanomethylene) -1,3-dicetan-2-yl]carboxamide 7α-methoxy-3-(]-]]]methyltetrazol-5-ylthiomethy△3-cephemu 4- Dissolve 0.5 g of carboxylic acid benzhydryl ester in 2 parts methylene chloride, add 57 of a mixture of trifluoroacetic acid and anisole (volume ratio 4:1) at -10°C, and further dissolve at the same temperature for 30°C.
Stir for a minute.

The solvent is removed under reduced pressure at low temperature.

Add ether to the residue, collect the resulting precipitate, and dry to obtain powdered 7β-[4-(carboxy cyanomethylene).
-1,3-diethan-2-yl]carboxamide-7
α-Methoxy-3-(]-]]]methyltetrazol-5-ylthiomethy Δ3-cephemu-4-carboxylic acid 0.
Obtain 2g.

Nuclear magnetic resonance spectrum (DMSOdo) δ (ppm)
: 3.44 (3H) CH303,62 (2
H) H of C2 4,32(2H) -CH2-85,16(I
H) H of C6 a) (4-bis(tert-butoxycarbonyl)methylene-1,3-dicetan-2-yl]carboxylic acid 50
Dissolve 0IIlfI in 5 ml of methylene chloride,
Next, 226 m9 of pyridine was added, and 360 m~ of phosphorus was further added for 1 hour while cooling with water, followed by stirring for 30 minutes.

This solution was mixed with 7β-amino-7α-methoxy-3-(
-Methyltetrazol-5-yl)thiomethyl-△3-
Cephemu 4-carboxylic acid benzhydryl ester 50
Dissolve 0~ in methylene chloride]Qml -20~-
Add to the solution cooled to 30°C and stir at that temperature for 1 hour.

The reaction solution was washed successively with 10 ml of water, 5 ml of dilute hydrochloric acid, and 5 ml of water, dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure.

The residue was subjected to silica gel column chromatography using a mixture of chloroform and ethyl acetate (volume ratio 6:1) as an eluent to obtain 7β-(4-[bis(tert-butoxycarbonyl)methylene, l-1,3- Jichetan-2
-yl)carboxamide-7α-methoxy-3-(1-
Methyltetrazol-5-yl)thiomethyl-△3-cephemu-4-carboxylic acid benzhydryl ester 300
get ~.

Nuclear magnetic resonance spectrum (CDC1a) δ (ppm): 1.50 (18H) t-C4H.

3.60 (5H) CH30- of C7 and - of C2
CH2- 4,40(2H) -CH2-8-5,10(
IH) Co8 0) 7β-(4-[bis(tert-butoxycarbonyl)methylene)-1,3-dicetan-2-yl)carboxamide-7α-methoxy-3-(]-]]methyltetrazole- 5-ylthiomethy△3-cephemu4-
Dissolve carboxylic acid benzhydryl ester 2001"? in a mixture of 4 ml of trifluoroacetic acid and 0.5 ml of anisole, and stir for 1 hour under water cooling.

The solvent was distilled off under reduced pressure, 5 ml of ether was added to the residue, and the resulting precipitate was collected.

It was then washed with ether to give 7β-[4-(dicarboxymethylene)-1,3-dicetan-2-yl]carboxamide 7α-methoxy-3-(]-]]]methyltetrazol-5-ylthiomethyl 3-cephem. 100 m2 of -4-carboxylic acid are obtained.

Nuclear magnetic resonance spectrum (DMSOda) δ (ppm)
: 3°44 (3H) CH30-3,64 of C7 (
2H) C2 -CH2- 4,30 (2H) C3 -CI (2-8 One Example
4゜(a)4 (tert-butoxycarbonyl N, N
-dimethylcarbamoylmethylene) -1,3-dichetane-2-carboxylic acid (500 ~) is dissolved in methylene chloride (19 m/!) and pyridine O (19 m/!). Next, add 163" phosphorus pentachloride on ice and stir for about 5 minutes.

This solution was mixed with 7β-amino-7α-methoxy-3-(1-
Methyltetrazol-5-ylthiomethyl)-△3-cephemu-4-carboxylic acid (500~) in methylene chloride (15m)
1, cooled to -30°C, added to a solution containing 0.3 ml of pyridine, and stirred at that temperature for about 30 minutes.

Approximately 6 r) rnl of chloroform was added to the reaction solution, and 30
Wash with approximately 30 ml of 1-2% hydrochloric acid, then wash three times with 30 ml of fishing rod, and dry with anhydrous magnesium sulfate.

The residue obtained by evaporating the solvent under reduced pressure was subjected to silica gel column chromatography using a mixture of benzene and ethyl acetate (volume ratio 3:2) as an eluent to obtain 7β-((4-te
rt-butoxycarbonyl N,N-dimethylcarbamoylmethylene) -1,3-dicetan-2-ylcarboxamide]-7α-methoxy-3-(]-]]]methyltetrazol-5-ylthiomethyl dfem-4-carboxylic acid 200ml of benzhydryl ester is obtained.

Nuclear magnetic resonance spectrum (CDC12) δ (ppm): 1,48 (9H) tert-C4Hg-2,9
6 (6H) (C! (3) 2NOC-Qin 3.60
(5H) CH30- and C2 -CH2-5,06(
IH) H of C6 6,90 (IH) φ2CH- (O) Dissolve this in a mixture of 10rnl of trifluoroacetic acid and 2ml of anisole under water cooling, and stir at 15°C for about 30 minutes.

The solvent was distilled off under reduced pressure, ether 307 was added to the residue, and the resulting precipitate was collected.

Then, it was washed with ether to obtain 7β-[(4-carboxy
N,N-dimethylcarbamoylmethylene) -1,3-
dichetan-2-ylcarboxamide]-7α-methoxy-3-(J-methyltetrazol-5-ylthiomethyl)-Δ-cephemu-4-carboxylic acid 100.

Nuclear magnetic resonance spectrum (DMSOdo) δ (ppm)
: 2.87 (6H) (CH3)2N0C-3,4
3(3H) CH30- (a) 7β-amino-7α-methoxy-3-(]-]
]] Methyltetrazol-5-ylthiomethy △3-cephemu-4-carboxylic acid benzhydryl ester 370~
, dicyclohexyl-rubodiimide] 50 m9 and 4-(carbamoylcyanomethylene)-1,3-dichetane-2=carboxylic acid 150 m (I) is dissolved in 12 ml of tetrahydrofuran and stirred at room temperature for 2 hours.

The formed precipitate was washed away, the solvent was distilled off under reduced pressure, and the resulting residue was subjected to silica gel column chromatography.
Elution with a mixed solvent of chloroform-isopropanol (volume ratio 9:J) yielded 7β-(4-(carbamoyl cyanethylene)-1,3-dicetan-2-ylcarboxamide]-7α-methoxy-3-(''- 190 mg of methyltetrazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid benzhydryl ester are obtained.

Nuclear magnetic resonance spectrum (CDC13) δ (ppm): 3.55 (3H, 2H) -CH2- of CH30 and C2
4,32(2H) Ca -CH2- 6,92(IH) φ2 CH- 7,30(IOH) (C6H,)2-(b) 160 mg was poured into methylene chloride JOmlFC, and anisole 0. Add 5 ml and cool to -20°C.

Add 2.5 ml of trifluoroacetic acid to this at -20° to -1
After dropping at 0°C, stir at -10° to 0°C for 1 hour.

15 ml of ether was added to the residue obtained by distilling off the solvent under reduced pressure.
Add and stir for 20 minutes.

Then, the precipitate obtained by filtration under reduced pressure was thoroughly washed with ether and dried under reduced pressure to obtain 7β-(4-(carbamoyl cyanomethylene)-1,3-dicetan-2-ylcarboxamide]-7α-methoxy-3 80 mg of -(1-methyltetrazol-5-yl)thiomethyl-Δ3-cephemu-4-carboxylic acid are obtained.

Nuclear magnetic resonance spectrum (DMSO-d6) δ (ppm)
: 3.44 (3H) CH30- 3,84(2H) -CH2- of C2 4,32(2H) -CH2- of Ca 5,17(IH) H of C6 Example 6゜(a) 7β-amino -7α-methoxy-3-(]-]
]] Methyltetrazol-5-ylthiomethy △3-cephemu-4-carboxylic acid 500 ~ tetrahydrofuran 3
4-(tert-butoxycarbonylmethylcarbamoylmethylene)-1,3-dicetan-2-carboxylic acid (500~) and dicyclohexylcarbodiimide (DCC) (approximately 400~) were added to the solution dissolved in No. 57, and the mixture was heated at room temperature for 3 hours and 30 minutes. Stir to react.

The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography using chloroform-ethyl acetate (volume ratio 4:1) as an eluent to give 7β-[4-(tert-butoxycarbonyl methylcarbamoylmethylene) -
1,3-dichetan-2-yl]carboxamide 7α
300 mg of -methoxy-3-(1-methyltetrazol-5-yl)thiomethyl-Δ3-cephemu 4-carboxylic acid benzhydryl ester was obtained.

NMR spectrum (CDC1a) δ (ppm): 1.52 (9H, t C4H9) 2.83 (3H, l (3CH3NOC-) 3.60
(5H, CH30- and C2 -CH,,-)5.
08 (IH, H of C6) 6.93 (IH, -CHφ2) (O) Trifluoroacetic acid] □ml and anisole 2r11
7! 7β-(4(tert-butoxycarbonyl methylcarbamoylmethylene)-1,3-dicetan-2-yl)carboxamide 7α-methoxy-3-(J-methyltetrazol-5-yl)thiomethyl- to a mixture of 7α-methoxy-3-(J-methyltetrazol-5-yl)thiomethyl- △3-cephemu 4-carboxylic acid benzhydryl ester 300~ is dissolved and stirred at 15°C for about 1 hour.

After distilling off the solvent under reduced pressure, add about 50 ml of ether;
After stirring, remove the resulting precipitate and wash with ether.
7β-(4,-(carboxymethylcarbamoylmethylene)-1,' 3-dicetan-2-yl]carboxamide 7α-methoxy-3-(1-methyltetrazol-5-yl)thiomethyl-Δ3-cephemu4- Carboxylic acid 170~ was obtained.

NMR spectrum (DMSO-d6) δ (ppm): 2.68 (3H,H'3CHNOC-) 3.42
(3H, CH30-) Example 7゜i) 2-Carbamoyl acetic acid tert-butyl ester 1
Dissolve 9 g in 380 mJ of tetrahydrofuran, add 5.7 g of 50% sodium hydride under water cooling, and stir for 5 minutes.
Next, 9.2 g of carbon disulfide was added and the mixture was heated for 10 minutes, then for an additional 5
Add 8.0 g of 0% sodium hydride and stir for 20 minutes.

Next, 15.5 g of dichloroacetic acid was added and stirred for 60 minutes, followed by stirring at room temperature for 90 minutes.

After the reaction, the reaction mixture is dispersed in a mixture of 2 t of ice water and 12 ml of concentrated hydrochloric acid, and extracted with 1 t of ethyl acetate.

The ethyl acetate layer was washed twice with 450% 20% aqueous sodium chloride solution and then concentrated.

90 ml of benzene was added to the resulting residue and stirred for 2 hours, and the precipitated crystals were collected, washed with 90 ml of benzene, and dried under reduced pressure to give 4(tert-butoxycarbonylcarbamoylmethylene)-1,3,-dichetane. 19.9 g of -2-carboxylic acid are obtained.

Nuclear magnetic resonance spectrum (DMSO-da) δ (pp
m): (O)7β-amino-7α-methoxy-3-(]-]]
] Methyltetrazol-5-ylthiomethy △3-cephemu-4-carboxylic acid benzhydryl ester 1.00g
was dissolved in methylene chloride], cooled to -50°C, and 0.62+717 of pyridine was added.
t-butoxycarbonylcarbamoylmethylene)-1,
0.67 g of 3-dichetane-2-carboxylic acid was suspended in 10 ml of methylene chloride, and after cooling to -15°C, pyridine 0.
．． 18 rni and 0.48 g of phosphorus pentachloride were added and stirred at the same temperature for 30 minutes, then under water cooling for 30 minutes, and an acid chloride solution prepared was added dropwise at -50°C to -40°C.

After stirring for 20 minutes at the same temperature, 20ml of methylene chloride
After washing with 40 ml of IN-hydrochloric acid and then 40 ml of water, the organic layer is dried over magnesium sulfate.

After adding 1.24 ml of anisole to the organic layer obtained by filtration, the mixture was cooled on ice, and 0.76 g of aluminum chloride was added thereto, followed by stirring for 3 hours.

After the reaction, collect the precipitated powder and add 30 ml of ethyl acetate,
[Improper Tool] QmJ and IN-Disperse in a mixture of 25 ml of hydrochloric acid, stir at 10°C for 30 minutes, and then extract.

The organic layer is washed with 20 ml of 20% aqueous sodium chloride solution and concentrated.

The obtained residue was subjected to silica gel column chromatography using chloroform-methanol-formic acid (volume ratio 8:2:0.2).
), 7β-[4-(carbamoyl carboxymethylene)-1,3-dicetan-2-yl]carboxamide 7α-methoxy-3-(1-methyltetrazol-5-yl)thiomethyl-Δ3-cephemu 0.77 g of 4-carboxylic acid is obtained.

The nuclear magnetic resonance spectrum was consistent with the target product of Example 1.

Claims (1)

[Claims] Formula 1 (wherein RI represents a carboxyl group or a cyan group, R2
means a hydroxyl group, an amino group or a mono- or di-lower alkylamino group. ) or its salt with a pharmaceutically acceptable non-toxic base. 27β-(4-(carbamoyl carboxymethylene)
1.3-dichetan-2-yl]carboxamide 7α
-methoxy-3-(1-methyltetrazol-5-yl)thiomethyl-Δ3-ceraemu-4-carboxylic acid.