CA2015283A1 - Cephalosporin compounds and their use - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE
A cephalosporin compound of the formula (I):

Description

:.1 .,., , 2ol5283 :~ SPECIFICATION
~ ' 'rITLE OF THE INVENTION
sCephalosporin Compounds and Their Use BACKGROUND OF THE INVENTION
.. . .
~1. Field of the Invention .~This invention relates to novel cephalosporin ~,compounds possessing antibacterial activity, their composition and a process for preparing the same.

2. Description of the Prior Arts .Cephalosporin antibiotics generally show a broad ljspectrum of antibacterial activities against Gram-positive and Gram-negative bacteria and hence various cephalosporin compounds have been clinically used as ~ I
~,therapeutic agents for a variety of infectious `diseases. However, there are not many therapeutic agents of cephalosporin compounds which show potent ~ .
j~activity against Pseudomonas strains and Proteus ^~'strains. Also, many cephalosporin compounds have the .~drawbacks that they are unstable for ~-lactamase which :is produced by a resistant strain and are low in anti-.~bacterial activity against Pseudomonas strains -which are presently clinical problem ("Cephalosporins and Penicillins, Chemistry and Biology" edited by E. H. ~ :
:
::
~ ':` 1 :
,;

.'5 . .

2~15283 Flynn, Academic Press, New York, N.Y., 1972, Chapter Il) Among various acyl groups at 7th position of cephem compounds, the following has been disclosed in Japanese Unexamined Publication Nos. 62(1987)-270589 and 63(1988)-99077.
OONa = N - O - CH ~ - OH

, .
SUMMARY OF THE INVENTION
The inventors have extensively studied on the relationship between chemical structures of 7th and 3rd substituents in cephem compounds and their activities.
' As the result, we found that new cephalosporin compounds of the following formula (I) show a broad spectrum of antibacterial activity against Gram-positive and Gram-negative strains, especially a potent activity against Gram-positive strains, Pseudomonas strains and B-lactamase producing strains and are well absorbed with low toxicity.
According to this invention, it provides a cephalosporin compound of the formula ~

~;, ~ , ~ ~ ! 2 :;
, .
., .

. . :.

:.';,' - ' ' :

2 ~ 3 :

.

X ol~
~o~
N'O \Y
,LI ( I ) N ~ '`CON~I ~S~

~ COOI~ ~

.1 ~herein X and Y may be the same or different and are a hydrogen or halogen atom; Z is a hydrogen or halogen atom, ~CH=CH2, -CH=CH-CH3, -OR1, -SR2 or -CH2W in which R1 and R2 are a lower alkyl group and W is a hydrogen atom or a nucleophilic residue, or its pharmaceutically acceptable salt.
' Also, it provides an antibacterial composition ~, comprising an effective amount of the compound (I) or `~, its pharmaceutically acceptable salt and a i~ pharmaceutically acceptable carrier or diluent.
.
PREFERRED EN~30DIMENT OF THE INVENTION
Examples of the pharmaceutically acceptable salts of the compounds (I~ include conventional non-toxic .
~ salts, such as alkali metal salts (e.g., sodium or .' .~ ~'',~, 2~ ~2~3 potassium salt), alkali earth metal salts (e.g., calcium or magnesium salts), ammonium salt and salts with organic bases (e.g., triethylamine salt, pyridine salt, ethanolamine salt, triethanolamine salt !
dicyclohexylamine salt, lysine salt or arginine salt).
The halogen atom in the definitions of X and Y of the compounds (I) includes chlorine, bromine and fluo-rine atoms. Preferably, the halogen atom is chlorine or fluorine atom. More preferably, the halogen atom is chlorine atom.
It is preferable that both of X and Y are hydrogen atom, chlorine atom or fluorine atom, or any one of X
and Y is chlorine atom or fluorine atom and the remaining one is hydrogen atom.
The lower alkyl group of the group -OR1 or -SR2 means a straight-chain or branched-chain alkyl group containing 1 - 4 carbon atoms, specifically methyl, ethyl, propyl, isopropyl, butyl and isobutyl.
The nucleophilic residue of W in the group -CH2W
includes a halogen atom, an optionally substituted arom~tic heterocycle-thio group, a lower alkylthio group, a lower alkoxy group, a primary, secondary or tertiary amino group or a heterocyclic group containing at least one nitrogen group to be bonded to the methylene group via the nitrogen atom. The halogen , .

~ ., .~ ! . .- . ' . - ' . ' 2~ 2~

atoms mentioned in X and Y are applicable to the above halogen atom. The aromatic heterocycle in the Gptionally substituted aromatic heterocyclo-thio group includes a five or six membered heterocycle containing at least one hetero atom of nitrogen, sulfur and oxygen atoms, specifically thiazole, isothiazole, thiadiazole, triazole, tetrazole, pyridine, pyrimidine, pyridazine, oxazole, imidazole, pyrazole, oxadiazole, isoxazole and triazine. Examples of the substituents on the aromatic heterocycle are a lower alkyl group such as methyl, ethyl or propyl, a halogen atom such as chlorine, bromine or fluorine, amino group and cyano group.
Examples of the lower alkylthio group are methyl-thio, ethylthio and propylthio. Examples of the lower alkoxyl groups are methoxyl, ethoxyl and propoxyl.
Examples of the primary, secondary or tertiary amino groups are amino groups substituted by one to three of a lower alkyl or benzyl; specifically methylamino, ethylamino, dimethylamino, triethylamino and methylbenzylamino.

The heterocyclic groups containing at least one nitrogen atom to be bonded to the methylene group via the nitrogen atom include such heterocyclic groups containing nitrogen atom(s1 as mentioned in the above aromatic hetero-cycle. The heterocylic groups may be substituted by a lower alkyl such as methyl or ethyl; a . ' ' :, . '' ' ,. ' " : ' .

2~ ~283 halogen such as chlorine, bromine or fluorine, amino or cyano. Preferably, the neucleophilic residue is (1,2,3-thiadiazol-5-yl)thio, (5-methyl-2-tetrazol-2-yl) ~Ithio or ~N-methylpyridin-4-yl)thio.
~ he compound (I) of this invention can be prepared by reacting a compound of the formula (II):

112~
~.,,1 ( 11, ' COOR, wherein Z is the same meaning as defined in the formula ~'(I), R is a hydrogen atom or a protecting group fo~ the j 1 '~~ carboxyl group, or its reactive derivative at the amino group or salt, with a compound of the formula (III):
.

.
. X OR3 " ~OR:
I ~
N~O `Y
Ll, ( 11~ ) , N~ ` COOII
R2NII S' ... ,,.,: ~ . ~ - . - . . . .
`.`,.,:.:- . ~ :
:::~. . ` . . . , `

c;,r~

2~ ~2~
, ~' .
, wherein X and Y have the same meanings as defined in the formula (1), R2 is a hydrogen atom or a protecting group for amino group and R3 is a hydrogen atom or a protecting group for hydroxyl group, or its reaetive derivative at the earboxyl group or salt.
Examples of suitable reactive derivatives at the amino group of the eompounds (II) are an imino derivative of Schiff's base type formed by the reaction ; of the eompound (II) and a carbonyl compound such as an aldehyde or ketone, or its enamine type isomer, a silyl derivative formed from the compound (II) and a silylating agent such as bis(tri-methylsilyl)acetamide, or a derivative formed from the compound (II) and phosphorus triehloride or phosgene.
Suitable salts of the compounds (II) and ~III) are acid addition salts with organie aeids sueh as acetic aeid, maleic aeid, tartarie aeid, benzenesulfonic aeid and toluenesulfonic acid; or aeid addition salts with inorganic acids such as hydrochloric acid, hydrobromie aeid, sulfuric acid and phosphoric acid; metal salts sueh as alkali metal or alkaline earth metal salts (e.g., sodium, potassium, calcium or magnesium salt);
ammonium salt and salts with organic amines such as triethylamine and dieyelohexylamine.

.~ ~

' ' :-. ,.

2~2~3 Rxamples of suitable reactive derivatives at the carboxyl group of the compounds (III) include the acid halides, acid azide, mixed anhydrides, active amides and active esters. Specifically, the acid halide may be the acid chloride and acid bromide; the mixed acid anhydride may be those formed with an acid such as a substituted phosphoric acid (e.g., a dialkyl phos-phate, dibenzyl phosphate or a halogenated phosphoric acid), dialkyl phosphite, sulfurous acid, thiosulfuric acid, sulfuric acid, an alkyl carbonate (e.g., methyl carbonate or ethyl carbonate), an aliphtic carboxylic acid (e.g., pivalic acid, valeric acid, isovaleric acid, 2-ethylacetic acid or trichloroacetic acid) or an aromatic carboxylic acid (e.g., benzoic acid); the active amide may be the active amide formed with i imidazole, dimethylpyrazole, triazole or tetrazole; the active ester may be the cyanomethyl ester, methoxy-methyl ester, dimethyliminomethyl[(CH3)2N=CH-) ester, vinyl ester, propargyl ester, p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenyl ester, penta-chlorophenyl ester, mesylphenyl ester, phenylazophenyl ester, phenylthio ester, p-nitrophenylthio ester, p-cresylthio ester, carboxymethylthio ester, pyranyl ester, pyridyl ester, piperidyl ester, 8-quinolylthio ester or an ester formed with a N-hydroxy compound : .. _ ... .

2~2~

(e.g., N,N-dimethylhydroxyamine, l-hydroxy-2-(lH)-pyridone, N-hydroxysuccinimide, N-hydroxyphthalimide or l-hydroxy-6-chloro-lH-benzotriazole). The reactive derivative is suitably selected and used, depending upon the kind of the compound (III) as the reactant.

The reaction of the compound (II) and the compound ( III) is usually conducted in a conventional solvent such as water, acetone, dioxane, acetonitrile, chloroform, methylene chloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide or pyridine, or other organic solvent which does not impede the reaction.
The organic solvents as mentioned above may be used as a mixture with water.
When the compound (III) is used in the free acid form or salt form, the reaction is preferably conducted in the presence of a condensing agent, such as N,N'-dicyclohexylcarbodiimide, N-cyclohexyl-N`-morpholino-ethylcarbodiimide, N-cyclohexyl-N'-(4-diethylamino-cyclohexyl)carbodiimide, N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide, N-ethyl-N'-(3-dimethyl-aminopropyl)carbodiimide, N,N'-carbonylbis~2-methyl-imidazole), pentamethyleneketene-N-cyclohexylimine, diphenylketene, N-cyclohexylimine, ethoxyacetylene, 1-alkoxy-l-chloroethylene, trialkyl phosphite, ethyl polyphosphate, isopropyl polyphosphate, phosphorus oxy-.

~. 9 .'` .

-.i:

2~283 chloride, phosphorus trichloride, thionyl chloride,oxal~l chloride, triphenylphosphinej 2-ethyl-7-hydroxybenzisoxazolium salt, 2-ethyl-5-(m-sulfophenyl)-isoxazolium hydroxide inner salt, l-(p-chlorobenzene-sulfonyloxy)-6-chloro-lH-benzotriazole, or Vilsmeyer's reagent which is produced by the reaction of dimethyl-formamide with thionyl chloride, phosgene or phosphorus oxychloride.
The above reaction can be also conducted in the presence of an inorganic or organic base, especially such as an alkali metal hydrogen carbonate (e.g., sodium hydrogen carbonate or potassium hydrogen carbonate), an alkaline earth metal carbonate (e.g., potassium carbonate), a tri-lower alkylamine (e.g., triethylamine or trimethylamine), pyridine, a N-lower alkylmorpholine or a N,N-dilower alkylbenzylamine.
The reaction is usually conducted under cooling or warming, without limitation thereto.
Alternatively, the compound (I) can be prepared by reacting a compound of the iormula (1~):

N/
R2 NH ~5 F~ -.. `. . ~

2~1~2~

wherein A is aeetoxyl group or a halogen atom, and X, ~, Rl, R2 and R3 have the same meanings as defined above, with a nueleophilic compound.
The nueleophilie eompounds to be used inelude e.g., an optionally substi~uted heteroeyelie thiol containing at least one of sulfur, nitrogen and oxygen atoms, a lower alkylthiol, a lower alkanol, a primary, seeondary or tertiary amine or a heteroeyclic compound eontaining at least one nitrogen atom.
When the above reaetion gives a proteeted derivative at the amino, carboxyl and/or phenolic hydroxyl groups of the eompound (1), such protected derivative if necessary is subjected to a removal for protecting group in accordance with a eonventional method. Appropriate method for the removal reaetion is seleeted and used depending upon the kinds of the proteeting groups. For example, the removal for amino-proteeting group is usually eondueted by hydrolysis or reduetion but aeyl group as the amino-protecting group ean be removed e.g., by the treatment with an imino-. .
- halogenating agent and then with an imino-etherizating `~ agent, followed by hydrolysis. A hydrolysis using an aeid is conventional for amino-proteeting group and is ` applicable to remove alkoxyearbonyl, formyl, trityl or :~ .
',, 11 . ~

.

201 ~2,~3 .
like group. Examples of the acids are formic acid, trifluoroacetic acid, hydrochloric acid or like acid, ~3 which is suitably selected upon the kind of amino-protecting group. The removal for amino-protecting `:~
' group can be generally conducted without solvent or in the presence of water, a hydrophilic organic solvent or a mixture thereof. When trifluoroacetic acid is used, . the removal may be conducted in the presence of anisole. Also, the removal for carboxyl-protecting group is usually conducted by a conventional method such as hydrolysis or reduction. A hydrolysis using an acid is conventional and applicable to silyl group, p-methoxybenzyl group, diphenylmethyl group or like group. The removal for phenolic hydroxy-protecting group is also conducted by any conventional method such as hydrolysis or reduction. For the hydrolysis, an acid or base is used. For example, p-methoxybenzyl or diphenylmethyl group can be removed by a hydrolysis using an acid, and acetyl, benzoyl or like acyl group be removed by a hydrolysis using a base.
When th~ compound (I) is obtained in the free form at 3rd carboxyl group, it can be converted into the ;~ corresponding salt in accordance with the conventional - method. On the other hand, a salt of the compound (I) :......................................................................... ' :, :.

`: :~

.i :

20~2~3 when produced can be also converted into the correspollding free form.
TlhP compounds (Il) as produced can be purlfiPd and isolated by the conventional methods, e.g., purifica-tion using ad,orptive resins ~e.g., Amberlite~ XAD-2 (Rohn, & Haas), Diaion~ HP-20 or Sepabeads~ SP207 (Mitsubishi Chem., Japan)], precipitation, crystalli-zation and a combination thereof.
The compounds (I) or their salts of the invention are valuable antibiotics showing potent antibacterial activities against broad pathogenic bacteria including Gram-positive and Gram-negative strains and can be safely used as an antibacterial agent for treating or preventing infections caused by the above bacteria in man or domestic animals.
The compounds (I) or their salts may be used in various preparations such as parenteral preparations (e.g., intravenous or intramascular injections), oral preparations (e.g., capsules, tablets or powders) or rectal preparations (e.g., oily or water-soluble suppositories). Such various preparations can be prepared by the conventional methods, using diluent, excipients, binders, wetting agents, disintegrating agents, surfactants, lubricants, dispersing agents, ' ~
]3 ~

~ `:

2~283 bufering agents, preservatives, solubilizers, anti-septics, correctives and/or soothing agents.
Daily dose of the compound (I) or its salt may depend upon the conditions and weights of the patients, administration routes and the like. For parenteral administration, it is suitably about 250 - 3000 mg for adult in one to four divided doses per day.
The invention will be further illustrated by Reference Examples and Examples.
In these Examples, unless specific attention is given, NMR was measured by use of 90 MHz, ~ value in D2O bases on 4.82 as ~ value for water peak and ~ value in other deutrium solvents bases on TMS as the internal standard. In the formula, PMB means p-methoxybenzyl and Ph3C is trityl.

,. , Reference Example 1 i) 2,5-Dichloro-3,4-dihydroxvbenzoic acid In 200ml of acetic acid were added 50g of 3,4-dihydroxybenzoic acid (protocatechuic acid) and 105.lg of sulfuryl chloride. The mixture was stirred for 10 ,! hours at 50C, and then was cooled to crystallize the product. The crude crystals were recrystallized from ` ethyl acetate and n-hexane to obtain 35g of the title ~-~
compound (mp. 230C). ~ ~
.' :

. ' - .

201~283 ii) P-Methoxybenzyl 2,5-dichloro-3,4-di-P-methoxy-benzyloxybenzoate To a solution of lOg of 2,5-dichloro-3,4-dihydro-xybenzoic acid in 50ml of dimethylformamide (DMF) were added 28g of p-methoxybenzyl chloride and 33g of potassium carbonate. The mixture was stirred for 10 hours at 70 - 80~C. After the completion of the reaction, 200ml of ethyl acetate were added to the reaction mixture and insoluble material was removed by filtration. The filtrate was distilled under reduced pressure to remove the solvent and DMF. To the residue were added ethyl acetate and saline. The organic phase as collected was washed with water, dried over anhydrous MgSO4 and distilled under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography (Wako-gel~ C-200 : 700g, developing solvent : benzene and ethyl acetate (20:1)], to obtain 18g of the title compound.

' ` iii) 2,5-Dichloro-3,4-di-p-methoxybenzyloxybenzyl-alcohol LiAlH4 (2.7g) was suspended in lOOml of anhydrous tetrahydrofuran and ice-cooled. A solution of lOg of p-methoxybenzyl 2,5-dichloro-3,4-di-p-methoxybenzyloxy-benzoate in lOml of anhydrous tetrahydrofuran was ]5 ~. j 2~15~83 s.

dl-opwise added to the above cooled suspension and stirred for 6 hours under ice cooling. After the comp1etion of the reaction, the reaction mixture to which lOml of ethyl acetate were added was stirred for 30 minutes and adjusted to p~ 2.0 by addition of 20%
citric acid under ice-cooling. The mixture was filtered to remove insoluble material, and the filtrate 7 was poured into 300ml of ethyl acetate. The mixture was washed with water, dried over anhydrous MgSO4 and distilled under reduced pressure to remove the solvent.
The residue was purified by silica gel column chromato-, graphy [Wa~o-gel~ C-200: lOOg, developing solvent benzene and ethyl acetate (20:1)] to obtain 7g of the title compound.
~ NMR(CDCl3, ~, ppm) `j 3.85(6H,s), 4.55(2H,bs), 4.70(2H,bs), ' 5.00(2~,s), 6.85-6.90(4H,d), 7.20-7.35(5H,m) iv) Allyl 2-(2-tritYlaminothiazol-4-yl)-2(Z)-[2,5-; dichloro-(3,4-di-p-methoxYbenzyloxybenzyloxy)imino) acetate 2,5-Dichloro-(3,4-di-p-methoxybenzyloxy)bezyl-alcohol (2g) was dissolved in 20ml of methylene ` chloride and cooled to -10C. To the solution was added 573mg of 2,6-lutidine and then dropwise added a . .
, .

, ~.'.. ~ : , ' ' : ' .'?' -~ , . ~ , , `.~'. ~:.......... , ""' : -2o~283 :~ ~
solutiol~ of 632mg of thionyl chloride in 5ml of methylene chloride. The mixture was stirred for an hour at the above temperature, washed with water, dried over anhydrous MgSO4 and distilled under reduced pres-SUI e to remove the solvent.
The resulting residue and lg of K2CO3 were added to a solution of 2g of allyl 2-~2-tritylaminothiazol-4-yl)-2(Z)-hydroxyiminoacetate in lOml of DMF, followed by stirring for 20 hours at room temperature. The ... .
reaction mixture was poured into lOOml of ethyl acetate, and the mixture ~as washed with water, dried , .
over anhydrous MgSO4 and distilled to remove th~
solvent. The residue was purified by silica gel column chromatography ~Wako-gel~ C-200: 30g, developing `, solvent: benzene and ethyl acetate (20:1)] to afford 3.2g of the title compound.
IR ~nujol) 3300 - 3200, 1720cm 1 NMR(CDCl3, ~ value ppm) ~-~
3.8(3H,s), 3.81(3H,s), 5.0(2H,s), 5.05(2H,s), t'.'~.
~ 5.1(2H,s), 5.8(2H,d), 6~1(1H,m), 6.7-6.9 ''7 ;~` (7H,m), 7.3(19H,m) v) 2-(2-Tritylaminothiazol-4-yl)-2(Z)-(2,5-dichloro-3,4-di-p-methoxybenzyloxybenzyloxyimino)acetic acid . ,-.

,~ .
.: ~

Y. .

~ 2~1 5283 To an ice-cooled solution of 900mg of allyl 2-(2-tritylaminothiazol-4-yl)-2(Z)-(2,5-dichloro-3,4-di-p-methoxybenzyloxybenzyloxyimino)acetate in 20ml of methylene chloride were added 183mg of sodium 2-ethyl-hexanoate and 50mg of tetrakistriphenylphosphine-palla-I dium complex, followed by stirring for an hour under ice-cooling. The reaction mixture was adjusted to pH
2.0, washed with water, dried over anhydrous MgS04 and distilled under reduced pressure to obtain 620mg of the title compound.
NMR(CDC13, ~ value ppm) 3.75(3H,s), 3.80(3H,s), 5.0(2H,s), 5.05(2H,s), 5.06, 5.10(2H,ABq,J=12Hz), 6~75-6.90(7H,m), 7.25-7.35(19H,m) ,, .' Reference Example 2 i) Allyl 2-(2-tritylaminothiazol-4-yl)-2(Z)-(3,4-di-p-methoxYbenzyloxybenzyloxyimino)acetate To an ice-cooled solution of 470mg of allyl 2-(2-tritylaminothiazol-4-yl)-2(Z)-hydroxyiminoacetate in 5ml of DMF were added 207mg of calcium carbonate and 500mg of 3,4-di-p-methoxybenzyloxybenzyl chloride. The mixt~re was stirred for 20 hours at room temperature, and then poured into 60ml oE ethyl acetate. The mixture was washed with water, dried over anhydrous MgS04 and distilled under reduced pressure to remove the solvent. The residue was purified by silica gel .,s.:::
., ~
, .. , . -20~ ~2~3 column chromatography IWako-gel~ C-200: 209, developing solvent : benzene and ethyl acetate (20:1)] to obtain 450mg of the title compound.
IR (nujol) 1720cm ~
$ NMR(CDCl3, ~ value, ppm) 3.75(3H,s), 3.80(3H,s), 4.95~2H,s), i 5.00(2H,s), 5.05(2H,bs), 5.80(2H,b,d), 6.00(1~,m), 6.70-6.95(9H,m). 7.30-7.50(9H,m) ' ii) 2-(2-Tritylaminothiazol-4-yl)-2(Z)-(3,4-di-p-methoxybenzyloxybenzyloxyimino)acetic acid To an ice-cooled solution of 780mg of allyl 2-~2- ~;
~, tritylaminothizol-4-yl)-2(Z)-~3,4-di-p-methoxybenzyl-`~ oxybenzyloxyimino)acetate in 20ml of methylene chloride .J were added l90mg of sodium 2-ethylhexanoate and 50mg of ~ tetrakistriphenylphosphine-palladium complex. The .
~ mixture was stirred for an hour under ice-cooling, :
poured into water and adjusted to pH 2Ø The organic phase as collected was washed with water, dried over anhydrous MgS04 and distilled under reduced pressure to remo~e the solvent. The residue was washed with isopropylether and dried to obtain 610mg of the title compound.
IR (nujol) 1600cm 1 NMR(CDC13, ~ value, ppm) '. :
19 ~ ~:

. ~

2~ ~2~,~

3.75(9H,s), 3.80(3H,s), 4.95(2H,s), -.00(2H,s), 5.06, 5.10(2H,ABg, J=2Hz), `; 6.75-6 90(9H ,In) ~ 7.25-7.35(9H,m) .. . .
~ .
' Refelence Example 3 --~
3 i) Methyl 5-chloro-3,4-dihydroxybenzoate A solution of 5g of 5-chloro-3,4-dihydroxybenzoic acid in 40ml of anhydrous methanol was cooled to -20C
to -lO~C, saturted with dry hydrogen chloride gas, and -then allowed to stand for 20 hours at room temperature.
After adding a little amount of benzene, the reaction mixture was concentrated in about 1/3 volume and ice-cooled. The precipitated crystals were washed with a mixture of n-hexane and methanol to obtain the title ~ , ? compound. The mother liquor was also treated to obtain -3 second crystals. Yield 4.9g (92%). mp 202 - 203C
"~s ~ .
(methanol). ~
, . .
ii) Methyl 5-chloro-3,4-di-P-methoxybenzyloxybenzoate To a solution of 4.46g of methyl 5-chloro-3,4-dihydroxybenzoate in 16ml of DMF were added 7g of potassium carbonate and 9.5g of p-methoxybenzyl chloride at room temperature. The mixture was stirred I~ for 20 hours. After the completion of the reaction, `~ ~Oml of ethyl acetate were added to the reaction ; 20 .. ,~ .
;
',, ~3 ~

2ol~52~3 mixture and insoluble material was removed by filtration. To the filtrate were added 30ml of ethyl acetate and 20ml of water, and the mixture was adjusted to pH 7.3 by addition of 7% aqueous sodium hydrogen carbonate under ice-cooling. The organic layer was collected, while the aqueous layer was washed several times with ethyl acetate. The combined orgnaic layers were washed with water, dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was crystallized with ethyl acetate-n-hexane to obtain 8.95g (91%) of the title compound as colorless crystals. mp 122 - 123C (methanol).

!i iii) S-Chloro-(3,4-di-p-methoxybenzyloxy)benzylalcohol Methyl 5-chloro-3,4-di-p-methoxybenzyloxybenzoate (6.42g) as obtained by the above ii) was treated by the same method of Reference Example 1, iii) to obtain -, 5.58g of the title compound (94%). mp 107C

iv) Allyl 2-(2-tritylaminothiazol-4-yl)-2(Z)-(5-chloro-3,4-di-p-methoxybenzyloxybenzyl)oxyiminoacetate A solution of 0.65g of trichloromethyl chloro-; formate in 1 ml of methylene chloride was dropwise added to a mixture of 0.55g of DMF and 40ml of methylene chloride cooled to -15C. The mixture was ,, i 21 .`~' ~, :

: ~ :?~
" ;M'` ' : : ~
; ''~';'. ~` . ':

20~5283 .:

gradually warmed to 20 to 30C, stirred for 30 minutes at the same temperature and again cooled to -20C. To the solution were added 2.6Sg of 5-chloro-3,4-di-p-methoxybenzyloxybenzylalcohol, followed by stirring for 40 minutes at -20 to -10C. The reaction mixture was poured into 20ml of ice-water and stirred. The aqueous layer was extracted with methylene chloride several times. The combined orgnaic layers were washed with water, dried over anhydrous MgSO4 and concentrated under reduced pressure.
The residue was dissolved in a mixture of 4ml of DMF and 6ml of DMSO, to which 3.3g of allyl 2-(2-tritylaminothiazol-4- yl)-2(Z)-hydroxyiminoacetate (mp 187 - 189C) and 1.4g of potassium carbonate were added and stirred for 20 hours at room temperature.
After the completion of the reaction, 30ml of ethyl acetate were added to the reaction mixture. Insoluble material was filtered off, and 30ml of ethyl acetate and 20ml of water were added to the filtrate. The mixture was adjusted to pH 7.3 by adding 7% aqueous sodium hydrogen crabonate under ice-cooling and stirring. The organic layer was collected while the aqueous layer was extracted with ethyl acetate several times. The combined organic layers were washed with water, dried over anhydrous MgSO4 and concentrated under reduced pressure. The ~ ?~

2~ ~52~

~esidue WdS pllriEied by silica gel column chromato-graphy (eluting with ethyl acetate and benezene) to obtain 4.16g (74%) of the title compound as powders.

v) (Z)-2-(2-tritvlaminothiazol-4-Yl)-2(Z)-(5-chloro-3,4-di-p-methoxYbenzyloxybenzyl)oxyiminoacetic acid To a solution of 3g of the allyl ester as obtained in the above iv) in 20ml of methylene chloride was added a solution of 1.5g of potassium 2-ethylhexanoate in 8ml of ethyl acetate under nitrogen gas atmosphere, and then added 90mg of triphenylphsphine and 50mg of tetrakistriphenylphoshine-palladium complex, followed by stirring for 3 hours at room temperature. Then, 50ml of isopropyl ether were added to the mixture, stirred for an hour and ice-cooled. The precipitated crystals were washed with a mixture of ethyl acetate and isopropyl ether. Thus, potassium salt of the title compound was obtained. The potassium salt was mixed with 30ml of methylene chloride and lOml of water and the mixture was adjusted to pH 2.5 by adding citric acid under ice-cooling and stirring. The organic layer was sepearted while the aqueous layer was extracted with ethyl acetate several times. The combined organic layers were washed with water, dried over anhydrous MgSO4 and concentrated under reduced pressure. The ,.

:' !
'S ~ '" " "

2 ~

residue was recrystallized with n-hexane to obtain 2.2g (77%) of the title compound as colorless crystals (mp 122 - 126C).

Example 1 A: p-Methoxybenzyl 7-l2-(2-tritylaminothiazol-4-yl)- -2(Z)-t2 5-dichloro-3 4-di-p-methoxybenzyloxybenzyl-oxylmino)acetamidol-3-(1 2 3-thiadiazol-5-yllthio-methyl-3-cephem-4-carboxylate ~C- CONH ~ N~ :

\oCI C02P~lB
L~ OP IIZB
C I

(PMB is p-methoxybenzyl group and Ph3C is trityl group.) 2-(2-Tritylaminothiazol-4-yl)-2(Z)-(2 5-dichloro-` 3,4-d.i-p-methoxybenzyloxybenzyloxyimino)acetic acid ;` (860mg) and p-methoxybenzyl 7-amino-3-(1~2 3-thiadiazol -5-yl)thiomethyl-3-cephem-4-carboxylate (450mg) were dissolved in 20ml of methylene chloride and cooled to -20C. To the solution were added 320ml of pyridine , ~' ~ 1 --`~ 2~2~3 and then dropwise added a solution of 220mg of phos-phorus oxychloride in 5ml of methylene chloride, taking about 5 - 10 minutes. The mixture was stirred for an hour at 0 to 5C and poured into ice-water. Then, the mixture was adjusted to pH 7.0 by addition of 7%
aqueous NaHCO3 solution and stirred for 30 minutes at 5C. The organic phase was washed with water, dried over anhydrous MgSO4 and distilled under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography [Wako-ge C-200: 30g; developing solvent : benzene and ethyl acetate (10:1)] to obtain 750mg of the title compound.
` IR (nujol) 3200, 1780, 1720, 1680, 1620cm NMR(CDC13, o value, ppm) 3.30,3.51(2H,ABq,J=18Hz), 3.80(3H,s,OCH3), 3.82(6H,s), 3.93,4.13(2H,ABq,J=13Hz), 4.95-4.98(6H,m), 5.18(1H,d,J=4.8Hz), 5.36(2H,bI), 5.86(1H,d,d, J=5Hz and lOHz), 6.75-7.10(9H,m), 7.25-7.40(21H,m), 8.40(lH,s) B: Sodium 7-[2-(2-aminothiazol-4-yl)-2(Z)-(2,5-di-chloro-3,4-dihydroxybenzyloxyimino)acetamido]-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylate . .

; A.

. 25 ` !;
;

.r~

,~

20 ~ ,~2~3 N~ 11 CON F~S~S'N

\o Cl CO2Na OH
C I

, To an ice-cooled solution of 200mg of p-methoxy-benzyl 7-12-(2-tritYlaminothiazol-4-Yl)-2(Z)-(2,5-dichloro-3~4-di-p-methoxybenzyloxyimino)acetamidol-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxy-late in 0.5ml of anisole was added 2ml of trifluoro-~ acetic acid. The mixture was stirred for an hour under i ice-cooling. After the completion of the reaction, ~`t~ 50ml of isopropyl ether were added to the reaction l mixture, and the precipitate as trifluoroacetate of the i title compound was collected by filtration. After drying, the trifluoroacetate was suspended in lml of ` water, to which 7% NaHC03 was added to adjust to pH
` 7.2. The resulting solution was subjected to a column~
(Diaio ~ HP-20, 20mi), eluting with 5% aqueous acetone.
~, The eluate was distilled to remove the solvent. The ..
.,.

2~ ~2~3 ., . residue was freeze-dried to obtain 40mg of the title ompound.
IR(nujol) 3400-3200, 1760, 1650, 1600cm ~~
- NMR(DMSO-d6, ~ value, ppm) 3.60(2H,s), 4.40(2H,s), 4.98(1H,d,J=5Hz), 5.05(2H,b,s), 5.56(1H,d,d,J=5Hz and lOHz), 6.74(1H,s), 6.88(1H,s), 7.20(2H,b.s), - 9.04(1H,s), 9.60(1H,d,J=lOHz) .. . .
., ~
Example 2 ~: p-Methoxybenzyl 7-[2-(3-tritylaminothiazo]-4-yl)-2(Z)-(2-chloro-3,4-di-p-methoxybenzyloxybenzyloxy-imino)acetamido]-3-(1,2,3-thiadiazol-5-yl)thio-methyl-3-cephem-4-carboxylate .1 .
`~ C-CONH ~ S~ ~ 'Y\

Ph3CNh ~ ~ N ~ N ~ S-~S,N
~! \ Cl C02PMB
; O ~ PMB

~; , ~, .

~` :
~ .

'`~,~ .
!

. 27 ~ -~
, .
;

.~

2~ ~283 By the same method as in Example 1, the title :~ compound (680mg) was obtained from 825mg of 2-(2-tri-tylaminothiazol-4-yl)-2(Z)-(2-chloro-3,4-di-p-methoxy-benzyloxybenzyloxyimino)acetic acid and 450mg of 7-amino-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid.
IR(nujol) 3200, 1780, 1710, 1680, 1610cm 1 i NMR(CDCl3, ~ value, ppm) 3.23,3.46~2H,ABq,J=18Hz), 3.79(3H,s), ' 3.80(6H,s), 4.06,4.12(2H,ABq,J=14Hz), 4.91(1H,d,J=5Hz), 5.12~4H,m), 5.25~2H,bs), 5.81(lH,dd,J=5Hz and 9Hz), 6.74-6.98(9H,m), 7.25-7.35(22H,m), 8.44~1H,s) ., ~'~ B. Sodium 7-~2-(2-aminothiazol-4-yl)-2~Z)-~2-chloro-:, .
3,4-dihydroxybenzyloxyimino)acetamido]-3-~1,2,3-~, thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylate . . ~
N~ ICI CONH F~ S ¢5~ N

~:, \ Cl CO2Na N ~OH
L<~OH ~ ;~

.`, .
~ :' ,.:

~'.,,~' ~' ' ' ,, ~. '....... . , . ' : ' : .

201~283 By the same method as in Example 1, the title compound (32mg) was obtained from 150mg of p-methoxy-benzyl 7-l2-(2-tritylaminothiazol-4-yl)-2(Z)-(2-chloro-3,4-di-p-methoxybenzyloxybenzyloxyimino)acetamideo]-3~
(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxy-late.
IR (nujol) 3400-3200, 1780, 1680, 1610cm NMR(DMSO-d6, ~ value, ppm) 3.73(2H,s), 4.41(2H,s), 4.98(1H,d,J=SHz), 5.03(2H,s), 5.54(lH,dd,J=4.8Hz and 8.2Hz), 6.70(1H,s), 6.87(1H,d,J=8.6Hz), 7.17(2H,bs), 7.30(1H,d,J=8.6Hz), 9.06(1H,s), 9.55(1H,d,J=8.6Hz) Example 3 A. p-Methoxybenzyl 7-[2-~2-tritylaminothiazol-4-yl)-2(Z)-(2,5-dichloro-3,4-di-p-methoxybenzyloxybenzyl-oxyimino)acetamido]-3-chloromethyl-3-cephem-4-carboxylate N~C--CONH--Ph a CN H 1~ S ~ N ,~ N ~ C I
Cl C02P~lB
O ~, oPllB
~;OP~lB

2 ~ 3 By the same method of Example 1, the title compound (850mg) was obtained from 860mg of 2-(2-'ri-~ tylaminothiazol-4-yl)-2(Z)-~2,5-dichloro-3,4-di-p-i methoxybenzyloxybenzyloxyimino)acetic acid and 370mg of p-methoxybenzyl 7-amino-3-chloromethyl-3-cephem-4-carboxylate.
IR (nujol) 3300, 1780, 1680, 1610cm 1 NMR~CDC13, ~ value, ppm) 3.36,3.58(2H,ABq,J=18Hz), 3.79(3H,s), 3.82(6H,s), 4.32,4.53(2H,AB~,J=12Hz), 5.92-5.00(6H,m), 5.18(1H,d,J=4.8Hz), , 5.37(2H,bs), 5.91(1H,dd,J=4.8Hz and 9Hz), 6.75-6.90(9H,m), 7.30-7.40(21H,m) ~, , B. p-Methoxybenzyl 7-~2-(2-tritylaminothiazol-4-yl)-2(Z)-(2,5-dichloro-3,4-di-p-methoxybenzyloxybenzyl-oxyimino)acetamido]-3-(N-methylpyridin - 4-yl)thio-methyl-3-cephem-4-carboxylate-iodide N C-CONH ~ S~
~ ~ ~ N ~ S ~ N-CH3 PH3CHII S N O ~ l~
' \ C l CO 2 P~B
~_~OPMB
~ ~ OPMB
`~ Cl :

: , ~
. : :

. ~, ':

', ' ' ' ' ' ~ :` ' ' 2a~283 p-Methoxybenzyl 7-[2-~2-tritylaminothiazol-4-yl)-2(Z)-(2,5-dichloro-3 r 4-di-p-methoxybenzyloxybenzyloxy-imino)acetamidel-3-chloromethyl-3-cephem-4-carboxylate (150mg) as obtained by the above A was dissolved in 5ml of tetrahydrofuran and ice-cooled. To the solution were added 20mg of sodium iodide and 20mg of N-methyl-4-pyridothione, followed by stirring for 4 hours under nitrogen gas atmosphere and under ice-cooling. After the completion of the reaction , the reaction mixture was poured in to 50ml of methylene chloride. The resulting mixture was washed with water, dried over anhydrous MgSO4 and distilled under reduced pressure to remove the solvent. The residue was washed with isopropyl ether and then ethyl acetate to obtain 15Qmg of the title compound.
IR (nujol) 1760cm 1 C. 7-[2-(2-aminothiazol-4-yl)-2~Z)-(2,5-dichloro-3,4-di hydroxybenzyloxyimino)acetamido]-3-(N-methylpyri-din-4-yl)thiomethyl-3-cephem-4-carboxylic acid , C,,, ~

2~ ~2~3 CI~ CONH F''`~' S ~N--CH a OL~Ho~

I ~ suspension of 150mg of p-methoxybenzyl 7-[2-(2- ~-tritylaminothiazol-4-yl)-2(Z)-~2,5-dichloro-3,4-di-p-methoxybenzyloxybenzyloxyimino)acetamido]-3-(N-methyl-pyridin-4-yl)thiomethyl-3-cephem-4-carboxylate-iodide .
~as obtained by the above B ) in 0.2ml of anisole was ice-cooled, to which 2ml of trifluoroacetic acid were added and stirred for an hour at the same temperature.
After the c~mpletion of the reaction, 50ml of isopropyl ether were added to the reaction mixture to afford trifluoroacetate of the title compound as powders. The powders were washed with isopropyl ether, dried and suspended in 5ml of water. The suspension was adjusted to pH 7.2 by addition of 7% NaHCO3. The resulting solution was subjected to a column of DIAIO ~ HP-20 (20ml) eluting with 20% aqueous acetone. The eluate .' ' ' . ~.

~ .
'' i. .

2~ ~2~3 was concentrated under reduced pressure and freeze-dried to obtain 22rng of the title compound.
IR (nujol) 3300-3200, 1760, 1680, 1620cm 1 NMR(DMSO-d6, ~ value, ppm) , 3.7392H,bs), 4.10~3H,s), 4.40,4.56(2H,ABq,J=14Hz), 4.95(1H,bs), 5.03(lH,d,J=4.8Hz), 5.29~lH,dd,J=4.8Hz and 9Hz), 6.63(1H,s), 6.74(1H,s), 8.28,8.58(4H, ~j ABq,J=7Hz), 9.58(1H,d,J=9Hz) Example 4 A. p-Methoxybenzyl 7-~2-(2-tritylaminothiazol-4-yl)-2(Z)-(2-chloro-3,4-di-p-methoxybenzyloxybenzyloxy-imino)acetamido]-3-chloromethyl-3-cephem-4-carboxylate N~~r--C-CONHI i~S~

; Ph3CNH ~ S~ N O ~
\ Cl CO2PMB
~ OPM8 ~; ~ OPMB

:
:' ' :

' 2 ~

By the same method as in Example 1, the title compound (920mg) was obtained from 825mg of 2-(2-tri-tylaminothiazol 4-yl)-2(Z)-(2-chloro-3,4-di-p-methoxy-benzyloxybenzyloxyimino)acetic acid and 37Omg of p-methoxybenzyl 7-amino-3-chloromethyl-3-cephem-4-carboxylate.
IR (nujol) 3300, 1770, 1720, 1680, 1610cm 1 NMR(CDC13, ~ value, ppm) 3.25,3.52(2H,ABq,J=18Hz), 3.80(9H,s), 4.40,4.50(2H,ABq,J=12Hz), 4.93(1H,d,J=5Hz), 5.2-5.25(8H,m), 5.84~1H,dd,J=5Hz and 9Hz), ;
6.70-6.90(9H,m), 7.25-7.35(21H,m) . ~.
!

Example 5 A. p-Methoxybenzyl 7-t2-(2-tritylaminothiazol-4-yl)-2(Z)-(3,4-di-p-methoxybezyloxybenzyloxyimino)-acetamido]-3-(1,2,3-thiadiazol-5-yl)thiomethy-3-cephem-4-carboxylate ~ -C-CONH I ~S~ ~ N~ -k N~s~<s N
Ph~CNH S N O
\ CO2PMB
~_~OP~8 ~ OPMB

' .
' '.'" .

, ~ - , . : . . . ~ ~

2~2~

2-(2-Tritylaminothiazol-4-yl)-2(Z)-(3,4-di-p-methoxybenzyloxybenzyloxyimino)acetic acid (800mg) and p-methoxybenzyl 7-amino-3-(1,2,3-thiadiazol-5-yl)thio-methyl-3-cephem-4-carboxylate (440mg) were dissolved in 20ml of methylene chloride and cooled to -20C. To the cooled solution were added 320mg of pyridine and then drop- wise added a solution of 240mg of phosphorus oxychloride in 5ml of methylene chloride. The mixture was stirred for an hour under ice-cooling and then poured into ice-water. The mixture was adjusted to pH
7.0, washed with water and then dried over anhydrous MgSO4. After removing the solvent under reduced pressure, the residue was purified by silica gel column chromatography [Wako-ge ~ C-200: 30g, developing solvent : benzene and ethyl acetate (20:1)] to obtain 920mg of the title compound.
IR (nujol) 3300-3200, 1760, 1720, 1680, 1610cm 1 NMR(CDCl3, ~ value, ppm) 3.15,3.42(2H,ABq,J=18Hz), 3.82(6H,s), 3.86,4.16(2H,ABq,J=14Hz), 4.90(1H,d,J=5Hz), 5.10(2H,bs), 5.05(2H,bs), 5.25(2H,bs), 5.85(1H,dd,J=5Hz and 9Hz), 6.78-7.05(9H,m), 7.30-7.45(19H,m), 8.40(1H,s) B. Sodium 7-l2-(2-aminothiazol-4-yl)-2(Z)-(3,4-, .

; 35 ~

~,., ~ . .

2~152~3 dihydroxybenzyloxyimino)acetamido]-3 (1,2,3-thia-diazol-5-yl~thiomethyl-3-cephem-4-carboxylate , ' N \\ C-CONH I ~S~ ~ N~ :

`, H2N ~ S~ N ~ N ~ S--~S,N
CO 2 N a ,, . .

. ~ ..
A solution . of 150mg of p-methoxybenzyl 7-[2-(2-tritylaminothiazol-4-yl)-2(Z)-(3,4-di-p-methoxybenzyl-oxybenzyloxyimino)acetamido]-3-(1,2,3-thiadiazol-5-yl) thiomethyl-3-cephem-4-carboxylate (as obtained by the above A) in 2ml of anisole was ice-cooled, to which 2ml of trifluoroacetic acid were added and stirred for an hour at 5~C. To the reaction mixture were added 50ml of isopropyl ether, by which trifluoroacetate of the title compound were precipitated. The precipitate as collected by filtration was suspended in lml of water .
,which was adjusted to pH 7.2 by addition of 7% NaHC03.

'. The resulting solution was subjected to a column of `. DIAIO ~ HP-20 (20ml) eluting with 10% aqueous acetone.
.
ii The eluate was concentrated under reduced pressure to `~'' ' ' r, .' ~:
.,, ~"~:
~ :' . .

20~2~3 remove acetone and then ~reeze-dried lo obtain 42mg of ~he title compound.
IR (nujol) 3300-3200, 1760cm 1 NMR(D20, ~ value, ppm) 3.25,3.65(2H,ABq,J=18Hz), 4.10,4.35~2H,ABq,J=14Hz), 5.0511H,d,J=5Hz), 5.15(2H,s), 5.72(1H,d,J=5Hz), 5.85-7.05(3H,m), 8.70(lH,s) Example 6 Sodium 7-l2-(2-aminothiazol-4-Yl)-2(Z)-(3,4-di-hydroxybenzyloxyimino)acetamido]-3-(5-methyl-2-tetrazol-2-yl)thiomethyl-3-cephem-4-carboxylate A
,i ' O

NH2 ~ ~ ~H ~ ~ t ~ CH
\O /OH COONa CH2 ~ OH

. '. -' .' ``By the same method as in Example 5, the title .compound (18mg) was obtained from 200mg of 2-(2-trityl-., .`` ~.

' .

,, ' ... .

.:,5~, .. :-, - . . . .

201~283 .. .
aminothiazol-4-yl)-2(Z)-(3,4-dimethoxybenzyloxybenzyl-oxyimino)acetic acid and 120mg of diphenylmethyl 7-amino-3-~5-methyl-2H-tetrazol-2-yl)thiomethyl-3-cephem-4-carboxylate.
IR ~nujol) 3300-3200, 1770cm 1 I NMR~D20, o value, ppm) 2.4~3H,s), 3.4~2H,bv.s), 5.0~1H,d,J=6Hz), 5.1(2H,s), 5.6~1H,d,J=5Hz), 5.8(2H,A}3q), 5.9-7.05(3H,m) .. , Example 7 A: p-Methoxybenzyl 7-[2-(2-tritylaminothiazol-4-yl)-i 2(Z)-~5-chloro-3,4-dl-p-methoxybenzyloxybenzyloxy-imino)acetamido]-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylate .,; , . . .

- Ph CNH ~ S ~ ~N
O COOPMB
OPMB
OPMB
:`
, CO

~; ~
~ ~8 ,'' ~:

2 0 ~ 3 ` 2-(2-Tritylamonithiazol-4-yl)-2(Z)-(5-chloro-3,4-di-p-methoxybenzyloxybenzyloxyimino)acetic acid (466mg) and p-methoxybenzyl 7-amino-3-(1,2,3-thiadiazol-5-yl) thiomethyl-3-cephem-4-carboxylate (260mg) were dis-solved in 30ml of methylene chloride and cooled to -20C, to which 180mg of pyridine and a solution of 100mg of phosphorus oxychloride in 2ml of methylene chloride were added. The mixture was stirred for 30 minutes at -20c and for further 30 minutes at -10C, ~' and then poured into 20ml of ice-water. The mixture was adjusted to pH 7.3 by addition of 7% aqueous sodium hydrogen carbonate under stirring. The organic layer was washed with water, dried over anhydrous MgSO4 and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (Wako-ge C-200, 50g; developing solvent: benzene-ethyl acetate) to obtain 631mg (91~) of the title compound.
~ MS m/z: 1258 (M+H) ;l IR (nujol): 3400, 1786, 1719, 1684, 1586cm 1 NMR (CDCl3)~: 3.25, 3.50(2H, ABq, J=18Hz, 2-H), ~;
3.76(6H, bs, OCH3 x 3), 3.95, 4.15 (2H, ABq, J=13Hz, 3'-H~, 4.85(1H, ; .
d, J=5Hz, 6-H), 4.95(2H, S, OCH2), 5.05- 5.20(6H, m, OCH2 x 3), . " .
5.82(1H, dd, J=5Hz, 8Hz, 7-H), -` 6.75-7.40(32H, m, .'i; ~

2~1~52~3 arom, NH x 2, thiazole 5-H), 8.35 (lH, s, thiadiazole 4-H).

B. Sodium 7-l2-(2-aminothiazol-4-Yl)-2(Z)-(5-chloro-3,4-dihydroxybenzyl)oxyimino)acetamido]-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylate Ig CONHF~ S--¢~N

O COONa ~ OH
.^ C~

., .

The ester (610mg) as obtained by the above A was dissolved in 2ml of anisole and ice-cooled. Trifluoro-acetic acid (4ml) was added to the solution and stirred for 2 hours at 0 to 5C. After adding 50ml of iso-propyl ether, the mixture gave a precipitate which was collected by centrifugation and washed with isopropyl ethyl. The product was dissolved in a solution of llOmg of sodium hydrogen carbonate in 5ml of water, and the solution was adjusted to pH 7.5 - 7.8 by addition of O.lN-hydrochloric acid. The mixture was treated .~ , .. .

.,'Y.',., ~ ~. , . : . : . , ~ ': ~ ' ' ': ' ' .
.

2~ ~2~3 with 60ml of Diaio ~ HP-20, which was washed with water , and eluted with 20% aqueous acetone. The ~luate was concentrated under reduced pressure to remove acetone `~ and the residue was freeze-dried to obtain 231mg (71~) o~ the title compound ~sodium salt).
IR (nujol): 3400-3800, 1762, 1718, 1670cm ' NMR(DMSO-d6)~: 3.25, 3.40(2H, ABq, J=18Hz, 2-H), 4.40(2H, bs, 3-H), 4.90(2H, s, ' OCH2), 4.95(1H, d, J=5Hz, 6-H), S.55(1H, dd, J-5Hz, 8Hz, 7-H), 6.65-6.75(2H, m, arom), 6.70(1H, s, thiazol 5-H), 9.05(lH, s, thiadiazole 4-H), 9.50(1H, d, J=8Hz, CONH).

Example 8 .~
Sodium 7-~2-(2-aminothiazol-4-yl)-2(Z)-(2,5-difluoro-3,4-dihydroxybenzyloxyimino)acetamidol-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylate , ',~

: ':

. ~ .:
'~

, ,.s 2~1 ~2~

C - cO~II -F~, \O F COO~la ,'~\~ 0~1 , F

. .

By the same method of Example 1-A, p-methoxybenzyl `l 7-[2-(2-tritylaminothiazol-4-yl)-2(Z)-(2,5-difluoro-3,4-di-p-methoxybenzyloxybenzyloxyimino)acetamido]-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxy-late was prepared from p-methoxybenzyl 7-amino-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxy-late and 2-(2-tritylaminothiazol-4-yl)-2~Z)-(2,5-l~ difluoro-3,4-di-p-methoxybenzyloxybenzyloxyimino)acetic b acid. (The latter was prepared by firstly converting 2,5-difluoro-3-methoxy-4-hydroxybenzaldehyde (cf. J.
` Org. Chem. 46(1981), 203) into 2,5-difluoro-3,4-di-p-i methoxybenzyloxybenzyl chloride via four steps and then ;
`~ reacting the benzyl chloride with allyl 2-(2-trityl-~ aminothiazol-4-yl)-2(Z)-hydroxyiminoacetate. -~

.' r~ .
' , ,' ' ~2 ~ 3 The above p-methoxybenzyl ester compound was treated by the same way as in Example l-B to obtain the title compound.
IR(nujol) 3400-3200, 1760, 1650, 1600cm 1 NMR(DMSO-d6, ~ value, ppm) 3.60(2H,s), 4.40(2H,s), 4.98(1H,d,J=5Hz), 5.05(2H,b,s), 5.56(1H,d,d,J=5Hz and lOHz), 6.74(1H,s), 6.88(1H,s), 7.20(2H,b.s), ~ 9.04(1H,s), 9.60(1H,d,J=lOHz) ;~

; Example 9 Sodium 7-l2-(2~aminothiazol-4-Yl)-2(Z)-(5-fluoro-3,4-dihydroxybenzyldxyimino)acetamido]-3-(1,2,3-thiadia- -~
` zol-5-yl)thiomethyl-3-cephem-4-carboxylate i ~

INI F~`N
\O COONa :, 1~ o~

:

.. , ~

:

:

.,.,.-~ :

2~1~283 By the same method of Example 1-A, p-methoxybenzyl 7-[2-(2-tritylaminothiazol-4-yl)-2(Z)-(5-fluoro-3,4-p-methoxybenzyloxybenzyloxyimino)acetamido]-(1,2,3-thia-zol-5-yl)thiomethyl-3-cephem-4-carboxylate was prepared from p-meth~xybenzyl 7-amino-3-(1,2,3-thiadiazol-5-yl) thiomethyl-3-cephem-4-carboxylate and 2-(2-tritylamino-thiazol-4-yl)-2(Z)-(5-fluoro-3,4-di-p-methoxybenzyloxy-benzyloxyimino)acetic acid. IThe latter 5-fluoro compound was prepared from 3-methoxy-4-hydroxy-5-fluorobenzaldehyde [J. Org. Chem. 51 (1986), 4073]}
The above methoxybenzyl ester compound was treated in the same way as in Example 1-B to obtain the title compound.
IR (nujol): 3400-3800, 1762, 1718, 1670cm 1 ;NMR(DMSO-d6)~: 3.25, 3.40(2H, ~3q, J=18Hz, 2-H), 4.40(2H, bs, 3-H), 4.90(2H, s, OCH2), 4.95(1H, d, J=5Hz, 6-H), 5.55(1H, dd, J-5Hz, 8Hz, 7-H), 6.65-6.75(2H, m, arom), 6.70(lH, s, thiazol 5-H), 9.05(1H, s, thiadia-zole 4-H), 9.50(1H, d, J=8Hz, CONH).

`,"`
44 ~

'' ''' 2~ 52~

The following Table 1 show minimum inhibitory concentrations (MIC) against Gram-positive and Gram-negative bacteria of representative compounds of the invention. MIC was measured by agar dilution method.
~ , 2 ~
~ ,n ~
,, o o o o ~ ~ co D ' ~
- O 1` L l O O O O ~`1 ' ~1 o o ~I v v v v ~ .,- .
_ .' In Ll~ In ~ O CO ~D O O 11~
E~ ~ ~ l ~ O O O O
X O O ,/ ~D
1~ V V V V
. . _ - . _ ~ ' ~ ~ ~ n In ,, ~ O O ~D O O O O ~ ~ ' L~t . . ~ :, :
O O O O
_ ~ V V V V ~ . .' ~ In a~ a~ oo ~ ~ ~ ~ o ~0 ~ ~ ~ 1_ o o O ~ ~
., . ~ O O o O O O O O ~::
.. d ~:, Q~ ~ ~
a~ ~ o o o co In O O 00 ~ ~ ~ ~ ~ ~ O ~ ~ ~
a) o ~ o o o o o o o o .q ~ ~
o ~1 o o ~ o o ~ ~ ~ ~_ o o O --~ ~
X O O O O O O O O
,.

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Claims (7)

1. A cephalosporin compound of the formula (I):

(I) wherein X and Y may be the same or different and are a hydrogen or halogen atom; Z is a hydrogen or halogen atom, -CH=CH2, -CH=CH-CH3, -OR1, -SR2 or -CH2W in which R1 and R2 are a lower alkyl group and W is a hydrogen atom or a nucleophilic residue, or its pharmaceutically acceptable salt.

2. A compound of claim 1 in which both of X and Y are chlorine or fluorine atom.

3. A compound of claim 1 in which any one of X and Y
is chlorine or fluorine atom and the remaining one is hydrogen atom.

4. A compound of lciam 1 in which both of X and Y are hydrogen atom.

5. A compound of claim 1 in which Z is (1,2,3-thia-diazol-5-yl)thiomethyl, (5-methyl-2H-tetrazol-2-yl)thio-methyl or (N-methylpyridin-4-yl)thiomethyl.

6. A compound of claim 1 which is 7-[2-(2-aminothia-zol-4-yl)-2(Z)-(2,5-dichloro-3,4-dihydroxybenzyloxy-imino)acetamido]-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid, 7-[2-(2-aminothiazol-4-yl)-2(Z)-(2-chloro-3,4-dihydroxybenzyloxyimino)acetamido]-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxy-lic acid, 7-[2-(2-aminothiazol-4-yl)-2(Z)-(2,5-di-chloro-3,4-dihydroxybenzyloxyimino)acetamido]-3-(N-methylpyridin-4-yl)thiomethyl-3-cephem-4-carboxylic acid, 7-[2-(2-(aminothiazol-4-yl)-2(Z)-(3,4-dihydroxy-benzyloxyimino)acetamido]-3-(1,2,3-thiazol-5-yl)thio-methyl-3-cephem-4-carboxylic acid, 7-[2-(2-amino-thiazol-4-yl)-2(Z)-(3,4-dihydroxybenzyloxyimino)aceta-mido]-3-(5-methyl-2H-tetrazol-2-yl)thiomethyl-3-cephem-4-carboxylic acid, 7-[2-(2-aminothiazol-4-yl)-2(Z)-(5-chloro-3,4-dihydroxyphenzyloxyimino)acetamido]-3-(1,2,3-thiadiazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid, 7-[2-(2-aminothiazol-4-yl)-2(Z)-(2,5-difluoro-3,4-dihydroxybenzyloxyimino)acetamido]-3-(1,2,3-thia-diazol-5-yl)thiomethyl-3-cephem-4-carboxylic acid or

7-[2-(2-aminothiazol-4-yl)-2(Z)-(5-fluoro-3,4-dihydro-xybenzyloxyimino)acetamido[-3-(1,2,3-thidiazol-5-yl) thiomethyl-3-cephem-4-carboxylic acid, or a salt thereof.
7. An antibacterial composition comprising an effective amount of a compound of claim 1 or its salt and a pharmaceutically acceptable carrier or diluent.