CA2031008A1 - Cephalosporins - Google Patents

Abstract

Abstract Novel cephalosporins of the formula (I) in which R1 - represents halogen or - represents straight-chain or branched alkyl, alkyl-thio or alkenyl having up to 10 carbon atoms, which is optionally substituted by halogen, straight-chain or branched alkoxy having up to 8 carbon atoms or by a 5-membered unsaturated heterocycle having up to 4 heteroatoms from the group consisting of nitrogen and sulphur, which for its part may be substituted by straight-chain or branched alkyl or alkylaminodi-alkyl having up to 6 carbon atoms, - or which is optionally substituted by a group of the formula -OCOR2, in which R2 - denotes amino or straight-chain or branched alkyl having up to 6 carbon atoms, - represents straight-chain or branched alkoxy having up to 8 carbon atoms, - represents a group of the formula -CH2-S-R3, in which R3 - denotes a 5-membered unsaturated heterocycle having up to 4 heteroatoms from the group consisting of nitrogen and sulphur, which is optionally substituted by straight-chain or branched alkyl or alkylaminodialkyl having up to 6 carbon atoms, and Abstract (continued) X - represents oxygen, sulphur or the radical -NH, and their physiologically acceptable salts.
Novel intermediates are also shown.

Description

2 ~ 0 ~3 The in~ention relates to p-lactam compounds of the general formula (I~

H2N ~ S,~IH-CO-N (I) NH2 ~R 1 COOH
in which R1 - repre~ents halogen or - represents straight-ch~in or branched alXyl, alkyl~
thio or alkenyl ha~ing up ~o 10 carbon atoms, which is optionally substituted by halogen, straight-chain or branched alkoxy having up to 8 carbon atoms or by a 5-membered unsa~ura~ed heterocycle having up to 4 heteroatoms from the series comprising nitrogen or sulphur, which for its part may be substituted by ~-~traight-chain or branched alkyl or alkylaminodi-alkyl having up to 6 carbon atoms, - or which is optionally substituted by a group of the formula -OCOR2, in which R2 _ denotes amino or straight-chain or branched alkyl having up to 6 carbon atoms, ..
- represents straight-chain or branched alkoxy having up to 8 carbon atoms, Le A 27 337 - 1 -. - .. . . .

: ~ : ., .

- .
~ ~3 ;~

- represents a group of the formula -CH2-S-R3, in which R3 - denote~ a 5-membered unsaturated heterocycle having up to 4 heteroatoms from the series comprising nitrogen or sulphur, which is optionally substituted by straight-chaln or branched al~yl or alkylaminodialkyl having up to 6 carbon atoms, X - represents oxygen, sulphur or the radical -NH, and their physiologically ~cceptable salts.
The compounds of the formula 5I) can be present as free acids, esters, as inner salts or as non-toxic pharmaceutically tolerable salts of the acidic carboxyl groups, such as sodium, potassium, magnesium, calcium, aluminUm or a~monium salts, with amines such as di- or tri-lower alkylamines, procaine~ dibenzylamine, N,N~-di-benzylethylenediamine, N-benzyl-~-phenyl-ethylamine, N-methyl- and N-ethylmorpholine, l-ephenamine, dihydro-abietylamine, N,N'-bis-dihydroabietylethylenediamine, N lower alkylpiperidine and other amines which can be used for the formation of salts of penicillins and cephalosporins.
Non-toxic, pharmaceutically tolerable salts of the basic amino groups with inorganic or organic acid radicals which may preferably be mentioned are, for example, chloride, bromide, iodide, sulphate, hydrogen sulphate, phosphate, carbonate, hydrogen carbonate, or sulphonates such as methylsulphonate, ethanesulphonate, toluenesulphonate, benzenesulphonate, naphthalenedi-sulphonate, or carboxylates such as acetate, formate, ',', . ~ ' ~e A 27 337 - 2 -:: : . .
.- ..
-- , .. . .. . .
- : ~ - . -:: - . . - , . , :. , '', i - ~: , : .
. :. . .

0~10~3'~3 oxalate, tartrate, citrate, maleate, fumarate, benzoate, succinate or lactate.
On account of the presence of the as~nmetric carbon atom designated by *, the new ~-lactam antibiotics S o the formula (I) include the D-, L- and D,L-form. The D forms of the compounds of the general formula (I~
according to the invention are preferred.
Both ths diastereomer mixtures and the D-form and L-form of the compounds according to the invention can be employed for the treatment of bactexial infectious diseases.
Preferred compounds of the general formula (I) are those in which R1 - represents fluorine, chlorine or bromine, or - represents straight-chain or branched alkyl, alkyl-thio or alkenyl having up to 8 carbon atoms, which is optionally substituted by fluorine, chlorine, bromine or by straight-chain or branched alkoxy having up to 6 carbon atoms or by tetrazolyl, thiazolyl, triazolyl or thiadiazolyl, which for their part may be substituted by straight-chain or branched alkyl or alkylaminodialkyl having up to 4 carbon atoms, h - or which is optionally substituted by a group of the formula -OCOR2, in which R2 _ denotes amino or straight-chain or branched alkyl having up to 4 carbon atoms, or - represents straight-chain or branched alkoxy having ::
up to 6 carbon atoms, Le_A 27 337 - 3 -- :.
- , ` ~ . - .-. : :..... : ... . -: : :, . - :- .. , : -:
,,: : . : . :
- - - , ,.
, ~ : . : --, ~ - , : : : -:~ : :: ;

~03~

X - represents oxygen or sulphur, and their physiologically acceptable salts.
Particularly preferred compounds of the general formula (I) are those in which R1 - represents chlorine or straight-chain or branched alkyl or alkenyl having ~p to 6 carbon atoms, which is optionally substituted by fluorine, chlorine or straight-chain or branched alkoxy having up to 4 carbon atoms, or - represents straiyht~chain or branched alkoxy having up to 4 carbon atom~, X - represents sulphur, and their physiologically acceptable salts.
In addikion, a process has been found for the preparation of the compounds of the general formula (I) X
H2N~S~H-CO-Nl~f ~ ( I ) ; .
NH2 ( R~
COOH
: .
in which R1 and X havs the abovementioned meanings, ; -20 characterized in that either ~:~
[A] in a 4-component reaction, compounds of the general formulae ~II), (III) and (IV), :~ ~ :
.

Le A 27 337 - 4 -: : ~:: . . . ...
. . . -, ~ , ~, . . .
.... . . .

-: .

: .

2~31~.3~3~

X CN~T----_~S~ and R4R5N l N,~ S~--~CHo , ~--N ~ Rl R7 ~II) (III) ~IV) in which R1 and X have the aboveme~tioned meaning~, R4 and R5 are identical or different and repre~nt an amino protecting ~roup, for example tert.-butoxy-carbonyl (Boc), R6 represents a carboxyl pro~ecting group, and 0 R7 - represents an easily removable benzylically or glycosidically bonded radical, such as, for example, 4,4'-dimethoxybenzhydryl or 2,4-dimethoxybenzyl or tetra-O-pivaloylgalactosyl, or - represents a benzylic chiral radical, such as, for example, ~-ferrocenyl-(C2-C6)-alkylamino, are reacted in the presence of a carboxylic acid of the formula (V) RB-COOH (V~

in which 0 RB _ denotes hydrogen, straight-chain or branched al~yl having up to 6 carbon atoms or a radical of the Le A_27 337 - 5 -. . : -. . ~:

2 ~ l3 g formula C6H5` S 1I H-CH2- ,, in inert solvents, if appropriate in the presence of a ca~alyst, to give compounds of the general formula (Ia) R~R5N ~ 5 ~ ~-Co-N ~ ~ ~Ia) N-COR8 C,~I~Rl ~ - :

in which Rl, R4, R5, R6, R7, R8 and X have the abovementioned mean~
ings, then the protecting groups R4, R5, R5 and the radicals R7 and -COR~ are removed with acids, ~ :
or in that [B~ carboxylic acids of the general formula (VI) x ~. :
R4 R5 NJ~s~cH - c ooH ( VI ) NHR

. , , ~ :
,~ , e A 27 337 - 6 -.

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

, 2~3~1,3~

in which R3, R4 and X have the abovementioned meaning and R7 - has the abovementioned meaning of R7 and i8 identi-cal to or different from this, or - represents an amino protecting group, are re~cted, after activation of the carboxyl group by a customary method, with a p-lactam compound of the general formula (VII) Hz ~ ~ -~ Rl (~II) .

in which .
Rl and R6 have the abovementioned meanings/
in iner~ solvents and in the last step the protecting .
groups R4, R5, R6 and R7 are optionally removed. :
The process variants according to the invention can be illustrated by the following equation-~

tA]
soc~ I ~ C ~ ~ IH3 ~:
N ~ s ~ CHo ~ ~ ;
sOc~ C02-CH2 ~ CH3 NH
I z ~ ~ ~ HCOOH
H3C ~ CH3 Le A 2? 337 - 7 -. - . . :.. . . . . .

-' ~: , .
- : ~ , . ;

~3~ 3~

N ~ ~ CONH~ ~ CH3 NCHO
~ ~ C02-CHz~OCH3 ¦ CF3COOH / C6H5ocH3 H2N ~ S ~ CONH ~ S CH3 HNCHO o ~ ~
C02H : . :

¦ HCl, H20, CH30H

H2N ~ CONH ~ ~ CH3 ~ :
NH2 ~ ~J -' "
c02H ~ :

HzN ~ HBoc* ~ CH3 ~.
C02CH2 ~ 0CH3 1.) Activation 2.) Coupling ~ -: ~
, ' -,.
Le A 2? 337 - 8 - ~

::
- :
, . . . .
. - , - :
' ~31~

S~CON S CH 1 NH ~J
Boc* CO2-cH~ocH3 ¦ CF3COOH

H 2N 1N~S~C ON ~ C H 3 *Boc=(CH3)3-C-~-CO-Amino protecting gxoup in the context of the above-mentioned definition in general represents a protecting group custom~ry in ~-lactam chemistry from the ~eries comprisin~: 4-methyoxyphenyl, 4-methoxymethyloxyphenyl, 4-[(2-methoxyethoxy)-methyloxyJphenyl, 3,4-dLmethoxy~
phenyl, benzyl, 2-nitrobenzyl, 4-nitrobenzyl, 4-methoxy~
benzyl, 2,4~dLmethoxyben~yl, 3,4-dimethoxybenzyl, 2,4,6-trLmethoxybenzyl, vinyl, allyl, tert-butoxy-carbonyl, benzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, formyl, acetyl, chloroacetyl, trichloroacetyl, trifluoroacetyl, benzoyl, methoxy-carbonyl, allyloxycarbonyl, 2,4-dLmethoxybenzyloxy-carbonyl, 2,2-diethoxyethyl, methoxycarbonylmethyl, tert-butoxycarbonylmethyl, allyloxymethyl, benzoylmethyl, Le A 27 337 - 9 -. : . ~ .. .. .

:. ~ : . . , ~ ~ 3 ~

bis-(4-methoxyphenyl)methyl, methox~nethyl, methyl-thiomethyl, methoxyethox~methyl, 2-(methylthiomethoxy)-ethoxycarbonyl, 2-hydroxy-2-phenylmethyl, methoxy-(4-methoxyphenyl)methyl, trimethyl-, triethyl, triphenyl-silyl, tert-butyl-dLmethylsilyl, tert-butyl-diphenyl-silyl, [2-(trLmethylsilyl)ethoxy]methyl, 1-methyl-2-benzoyl-vinyl, l-methyl-2-ethoxy- or 2-methoxyvinyl, mesyl- and ethylsulphonyl.
Carboxyl protecting group in the context of the abovementioned definition repra~ents the carboxyl pro-tecting group customary in ~-lactam chemistry. ~asily removable groups may preferably be mentioned, ~uch as, for example: tert.butyl, 2,2,2-trichloroethyl, diphanyl-methyl, triphenylmethyl, acetoxymethyl, allyl, benzyl, 4-methoxyphenyl,4-nitrobenzyl, 2-nitrobenzyl, 4-methoxy-benzyl, 2,4-dLmethoxybenzyl, trLmethylsilylethyl, tri-methylsilyl, tert.butyl-dimethylsilyl, acetonyl, l-phenoxyethyl or 2-methyl 2-propenyl. ~ -Suitable solvents for process variant lA~ ;
(4-component reaction) are the customary organic solvents which do not change under the reaction conditions. These preferably include ethers such as diethyl ether, butylmethyl ether, dioxane of tetrahydrofuran, or hydro- -~
carbons such as benzene, toluene, xylene or cyclohexane, or amides such as dimethylformamide or hexamethyl-phosphoramide, or alcohols such as methanol, ethanol, propanol or isopropanol, or chlorohydrocarbons such as methylene chloride, chloroform or carbon tetrachloride, or acetone, acetonitrile or ethyl acetate. It is also possible to use mixtures of the solvents mentioned.

Le A 27 337 - 10 -- ~ ... '. ,.................. : : .................. .

, ~ ~ 3 ~ f) Methanol is preferred.
The reaction can be carried out in a temperature range from OJC to +50~C, preferably at room temperature.
In general, the reaction is carried out at normal pressure. However, it is also possible to work at reduced pressure or at elevated pressure (for example from O.5 to 5 bar).
The carboxylic acids of the general formula (V) are known. Preferably, formic acid or N-dithiocarbo-benzyloxyglycine are employed.
In the case of the 4-component reaction, the carboxylic acid is employed in an amount of 1 to 20 moleq, preferably 10 moles, relative in each case to 1 mole of the compound~ of the general formulae (II), (III) and (IV).
In the case in which R7 in the formula (IV) represents a glycosidically bonded radical, the use of catalysts is necessary in the case of the 4-component reaction [A]. Suitable catalysts are zinc ~alts and titanium complexes, such as, for example, zinc chloride and Ti(4) isopropoxide. The catalyst is employed in an amount of 0.5 to 5 moles, preferably 1 mole, relative ~o 1 mole of the compounds of the formula (IV).
The removal of the amine and carboxyl protecting groups for the processes [A3 and [B] is preferably carried out with trifluoroacetic acid. However, it is also possible to use other methods known from the liter-ature.
The compound~ of the general formula (Ia) are al~o new.
..

~ .

Le A 27 337 : ,' '-: ' '' ' ' , ~ ' ' , ;: : , . . .

~t~ g The subsequent removal of the formyl or acyl group ~rom the c~mpo~nds of the general formula (Ia) is also carried out with trifluoroacetic acid, if R8 doefi not denote hydrogen. In the case in which R8 represents hydrogen, protonic acids, preferably aqueous hydrochloric acid, are employed in a methanol/water mixture.
The removal of the radical R7 (compounds of the formula Ia) i8 facilitated by adding auxiliaries, such as~ for example, anisole, phenol, thiophenol or phenyl thioethers, preferably anisole.
The removals o$ the protecting groups and the acyl/formyl group are in general carried out in a temper-ature range from 0C to +80C, preferably at room temper-ature.
lS The removals can be carried out both at normal pressure and at elevated or reduced pressure (for example 0.5 to 5 bar), preferably at normal pressure.
Suitable solvents for process variant [B] are all solvents which do not change under the reaction condi-tions. These preferably include ethers such as, for example, diethyl ether, dioxane or tetrahydrofuran, or chlorohydrocarbons such as methylene chloride, chloroform or tetrachloromethane, or amides such as dimethyl-formamide or hexamethylphosphoramide, or acetonitrile or acetone. It is also possible to employ mixtures of the solvents mentioned. ~ethylene chloride is preferred.
The condensation is in general carried out in a temperature range from 0C to +60C, preferably at room temperature and at normal prassure.
The removal of the protecting groups is carried ~a A 27 337 - 12 -. , .

, .
:
, , ~, ~ 3 ~ 3 out by the method described under [A].
The activation of the carboxyl group in the compounds of the general formula (VI) is in general carried out by converting into a mixed anhydride with 5 chloroformic acid esters or sulphonic acid derivatives, such as, for example, ethyl chloroformate, isobutyl chloroformate or methanesulphonyl chloride, by converting into the corresponding acid halide, or by con~erting into an activated ester, for example with N-hydroxybenzotri-azole and dicyclohexylcarbodiimide. Reaction with ethyl chloroformate and methanesulphonyl chloride is preferred.
Suitable ~olvent3 for the introduction of the amino protecting groups in process variants lA] and [B]
are all solvents which do not change under the reaction lS conditions. These preferably include chlorohydrocarbons such as methylene chloride, chloroform or carbon tetra-chloride, or amides such as dimethylformamide or hexa-methylphosphoramide, or acetonitrile, formic acid, acetone, pyridine or dimethyl sulphoxide. Pyridine, dimethyl sulphoxide and dLmethylformamide are preferred.
The compounds of the general formula (II) are new and can be prepared by reactin~
compounds of the general formula (VIII) ~
~ ', N - C = N~S~CH O ( VI I I ) -~

in which R9 and R10 are identical ~r different and ~ ;

Le A 27 337 - 13 -.: . ~ ` . :

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

.:

~ 1~ 3 1 ~ 3 ~J

represent straight-chain or branched alkyl having up to 6 carbon atoms or phenyl, reacting by a customary method, for example by thio-cyanation, nitration and then reduction or formylation with s~bsequent oxidation in inert solvents to give compounds of the general formula (IX) R11 ~ (IX) N-c-N ~ 5 H0 in which Rll, for example, represents ~he group -NH2, 0-C-H or -SCN
and R9 and Rl have the abovementioned meanin~s, and then cyclizing in weakly acidic aqueous medium, such -as, for example, wi th Br-CN or in acetic acid/water, to give compounds of the general formula (IIa) X~ (IIa) H2N~S~CHO , in which X has the abovementioned meaning, and in a last step blocking the amine function by intro- :
ducing the protecting groups (R4/~5). ~

~ .

Le A 27 337 - 14 - ~

~ ~ .

,, :

The introduction of the radical Rll is carried out by methods known from the literature.
Suitable solvents for the introduction of the r~dic~l Rll are the solvents mentioned under process [A], 5 Alcohols or chlorohydrocarbons, such as, ~or example, methanol, ethanol, propanol, isopropanol or methylenechloride, are preferre~.
The reaction proceeds in a temperature range from O~C to ~80C, preferably at room temperature and at normal pressure.
The cycli~ation is carried out in a temperature range from 40C to 150C, prefexably at 50C to 80~C and at normal pressure.
The compounds of the general formula tVI) are new and can be prepared by reacting compounds of the general formula (II) x il ll 5II) R4R5N, ~5~CHO
:, in ~7hich R4, R5 and X have the abovementioned meanings, with alkali metal cyanides, such as, for example, potas~
sium cyanide, or trimethylsilyl cyanide and amines of the formula (IV) 1~2N ,!
I (IV) Le A 27 ~7 - 15 -,. , ... , ,: : , .

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

in which R7 has the abovementioned meaning, or with ammonium acetate or ammonia to give compounds of the general formula (X) s R R N~--N15 C~NH ( X ) in which R4 and R5 have the abovementioned meanings, and Rl2 - has the abovementioned scope of meaning of R7 and in the case of ammonia also denotes hydrogen, and then removing the pro~ecting groups R4, R5 and R12 (Rl2 $ H) and hydr~lyzing the CN-group with acids, for example :
hydrochloric acid, and, if appropriate, introducing another of the abovementioned amino protecting groups.
15The reaction with the cyanides in general pro-ceeds in a temperature range from O~C ~o +5 0 C, prefer- :
ably at room temperature and normal pressure.
The removal of the amine protecting groups is ~ ~ :
carried out by the abovementioned method in a temperature 20range from +60C to +150C, preferably at +70c to ~90c at normal pressure.
The compounds of the general formulae (VIII) and (I~) are known per se or can be prepared by processes which are known per se. ~

: ~' Le_A ?7 33? _ 16 -: , ~ .. : .

2V~ LO~

The compounds of the general fo~nula (VII) are known per se or can be prepared by a customary method [compare DE-OS (German Offenlegungsschrift) 3,402,642, US Patent 4,639,448].
The compounds of the gener~l formula (III) are kn~wn in ~ome cases or are new and can be prepared in both cases in analogy to methods known from the litera-ture [compare DOS (~erman Offenlegungsschrift) 2,337,107].
The amines of the general formula (IV) are known per se or ~an be prepared by a customary method.
The stereochemically uniform D- or L-form of the compounds of the formula (I) according to the invention is obtained when the diastereomer mixture is isolated by preparative HPLC separa~ion methods.
On the other hand, the pure D- or L-form (prefer-ably the D-form~ i8 obtained when chemical resolution of the racemate, for example with dehydroabietylamine, phenylethylamine or camphorsulphonic acid, or resolution of the racemate, for exa~ple, via N-acetyl-amino acid derivatives, for example with subtilisin, penicillin acylase or porcine renal acylase, is carried out already ~- -at the stage of the protected or unprotected racemic amino acid of the formula (VI) and the stereochemically uniform D- or L-forms of ;~
the compounds of the formula (VI) are then reacted in the manner indicated, or by separating directly into the individual diastereomers a~ the stage of the formula (X) in which R12 represents one of the abovementioned chiral radicals.
The compo~nds of ~he general formula (I) Le A 27 337 - 17 -., , : ', :: ,: ' ' ' :

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

.,~ . . . . .

according to the invention have a wide antibacterial spectrum against gram-positive and gram-negative micro-organisms together with low toxicity. These properties make possible their use as chemotherapeutic active compounds in human and veterinary medicine.
The compounds according to the invention are active against a very wide spectrum of microorgani~ms.
With their aid, gram-negative and gram-positive bacteria and bacteria-like microorganisms can be controllad and the disorders caused by these pa~hogens can be prevented, improved and~or cured.
The compound6 according to the invention are particularly active against bacteria and bacteria-like microorgani~ms. They are therefore particularly highly suitable in human and veterinary medicine for the prophy-laxis and chemotherapy of local and 6y6temic infections which are caused by such pathogens.
The minLmum inhibitory concentrations (~IC) were determined by the serial dilu~ion method on Iso-Sensitest agar (Oxoid). For each test substance, a ~eries of agar plates were prepared which contained decreasing con-centrations of the active compound at twice the dilution in each case. The agar plates were inoculated using a multi-point inoculator (Denley). Overnight cultures of the pathogens which had previously been diluted in such a way that each inoculation point contained about 104 colony-forming particles were used for inoculating. The inoculated agar plates were incubated at 37C, and the growth of microorganism was read off af~er about 20 hours. The MIC value (~g/ml) indicates the lowest Le A 27 337 - 18 -_ ~' - `, ' , ~:

,.. . - , . ~ , :

& ~

active compound concentration at which it was not pos-sible to detect any growth of microorganism using the naked eye.
Use Example MIC values: agar dilution/multi-point Compound of Exam~le 13a:
~ .
Strain MIC (~g/ml) _ lO E coli A 261 4 Klebs. 6097 2 Staph. aur. 25455 ~ 0.25 ` `
Staph. epi. 25185 ~ 0.25 Strept. faec. 27101 16 Enterococc. 27185 The present invention includes pharmaceutical preparations which contain one or more compounds accord-ing to the invention in addition to non-toxic inert pharmaceutically suitable excipients or which are com-posed of one or more active compounds according to the invention and processes for the production of ~hese preparations.
The active compound(s) may optionally also be present in microencapsulated form with one or more o~ the abovementioned excipients.
The therapeutically active compounds should preferably be present in the abovementioned pharma-ceutical preparations in a concentration of about , ~, Le A 27 337 - l9 - ~ ~

.
.. . . .

, - ~ .

2n'3~3~

0.1 to 99.5, preferably of about 0.5 to 95~by weight of the total mixture.
The abovementioned pharmaceutical preparations may also contain pharmaceutical active coMpounds in addition to the compounds according to the invention.
In general, i~ has proYed advantageous both in human and in veterinary medicine to administer the active compound(s) according to the invention in total amounts of about 0.5 ~o about 500, preferably 5 to 100 mg/kg of body weigh~ every 24 hours, if appropriate in the form of several individual doses, to achieve the desired results.
An indi~idual dose preferably contains the active com-pound(s) according to the invention in amo~nts of about 1 to about 80, in particular 3 to 30,mg/kg of body weight~
The new compounds can be combined in the cus-tomary concentrations an~ preparations, together with the food or lactamase inhibitors, for example with penicil-lins which are particularly resistant to penicillinase and with clavulanic acid. Such a combination would be, for example, that with oxacillin or dicloxacillin.
The compounds according to ths invention can also be combined with aminoglycoside antibiotics, such as, for example, gentamicin, sisomicin, kanamycin, amikacin or tobramycin for the purpose of extending the spectrum of action and in order to achieve an increase in action.

Le A 27 337 - 20 -, .

: ~ ' ' : ' ' '~ 3 ~

Startinq compounds Example l 5-Dimethylaminomethyleneamino-4-thiocyanato-2-thiophene-carboxaldehyde NCS~
( H3C ) 2N--~S~CHO

10.O ml (194 mmol) of bromine are 810wly added dropwise at room temperature to a solution of 32 . O g (176 mmol) of 5-dimethylaminomethyleneamino-2-thiophene-carboxaldehyde and 40.1 g of ammonium thiocyanate (527 mmol) in 2.2 1 of methanol. The mixture is stirred at room temperature for l h, about 2 l of an ice/water mixture are added, the mixture is subseguently stirred for 45 minutes, and the precipitate formed is filtered off with suction at 5C, washed with water and dried in vacuo.
Yield: 24.4 g (58% of theory) ~-NMR (CD3SOCD3): ~ 3.12 and 3.20 (2s, each 3H, 5'-CH3); 8.00 (s, lH; 3-H); 8.20 (s, lH, N-CH=N); 9.63 (s, lH; :~
CHO).
Example 2 2-Amino-5-thieno[2,3-d]thiazolecarboxaldehyde ~ :

~ " ~, .
H2N 5~A`~HO
,. , Le A 27 337 - 21 -: .: ::: . ,: - ::

-: , ~

., `" ': . ~ :' . . . . . .

24.4 9 (102 mmol) of the compound from Example 1 are suspended in 170 ml of 90~ strength acetic acid and the mixture is heated to 60-65C, whereupon the aldehyde goes into solution. After 24 h at 60-65C, the mixture is allowed to cool to room temperature. After a further 24 h, the black crystals are filtered off with suction, washed with toluene and dried in vacuo.
Yield: 15.4 g ~82% of theo~y~
~H-NMR (CD3SOCD3): ~ = 8.05 (s, lH, 6-H); 8.22 (s, 2H, NH2); 9-74 (~, lH, CHO)-Example 3 2-Ace~amido-5-thieno[2,3-d]thiazole-carboxaldehyde o s Il 1~
H3C-CNH~5~CHO

A solution of 58.6 ml (824 mmol) of acetyl chloride in 660 ml of anhydrous chloroform is added dropwise at 15-20C in the course of 1 h to a solution of 15.4 g (84 mmol) of the compound from Example 2 in 660 ml of anhydrous pyridine. After 30 minutes, the mixture is poured into a mixture of 1 1 of water and 1 1 of chloroform, ~he phases are separated and the organic phase is washed several times with water. The organic ~ -~
phase is filtered through a layer of silica gel a few centimeter9 thick and then concentrated in vacuo. The residue thus obtained is stripped off several times with toluene until it crystallizes. The red-brown crystals are stirred overnight with 450 ml of methanol, filtered off , Le A 27 337 - 22 -. : .. .. .

- :

with sucti.on, washed with methanol and drisd in vacuv.
Yield: 11.2 9 (5~ of theory) H-NMR (CD3SOCD3): ~ 2.23 (s~ 3H, CH3CO); 8.30 ts, lH, 6-H); 9.91 (s, lH, CHO);
12.65 (s, lH, NH).
Example 4 2-Formamido-5-thieno[2/3-d]thiazole-carboxaldehyde HCONH~CHO

20 ml (212 mmol~ of acetic anhydride are added dropwise at 15-20C to 1.84 g (10 mmol) of the compound from Example 2 in 100 ml of formic acid. The mixture i5 -stirred overnight at room temperature, then 500 ml of ice :
water are added and the precipitate which has deposited is filtered off with suction. The crystals are stirred with ethyl acetate, filtered off with suction and dried -~
in vacuo.
Yield: 1.10 g (52% of theory) H-NMR (CD3SOCD3): ~ = 8.33 (s, lH, 6-H); 8.64 (s, lH, : :
N-CHO); 9.92 (s, lH, C-CHO);
12.85 (s, lH, NH).
Example 5 2-(N,N-Di-t-butyloxycarbonyl-amino)-5-thieno[2,3-d]-thiazolecarboxaldehyde s Boc\N ~
Boc~ S~CHO

Le A 27 337 - 23 -.. . . . .

: . : -,: - , , :: :: - :: . . . , ~ :
.
-, :: : . -:, . ,, : ,.
. , ., , .. ~ ' :' . . ~ . - "
: , ' :- : : : .

~ ~3 ~ 3 S~

21.8 g (100 mmol) of di-t-butyl pyrocarbonate are added at room temperature to a fiolution of 1,84 g (10 mmol) of the compound from Example 2 and the mixture is then stirred overnight at the same temperature. 3 ml of water are added dropwise, the mixture is concentrated in vacuo and the residue is stripped off several times with toluene. Chromatography on silica gel using toluene/ethyl acetate (97:3) yields 1.30 g (34% of theory) of the desired aldehyde.
lH-NMR (CD3SOCD3): ~ = 1.55 (s, 18H, Boc); 3.35 ts, lH, 6-H); 9.96 (s, lH, CHO).
Example 6 2-Amino-2-(2-amino-5-thieno[2,3-d]thiazolyl)-acetic acid I ~ ~CO2H
H2N~ ~NH2 :. , 360 mg (5.5 mmol3 of potassium cyanide and 1.23 g (16.0 mmol) of ammonium acetate are added to a suspension of 904 mg (4.0 mmol) of the compound from Example 3 in 25 ml of ethanol and the mixture is stirred at room temperature for 72 h. The reaction mixture is concen-trated to dryness, the residue is taken up in 100 ml of 6 N hydrochloric acid and the solution is heated under reflnx for 2 h. The solution is concentrated to dryness in vacuo at a bath temperature of 50C. The residue is dissolved in 200 ml of water and the solution is then adjusted to pH 5 with sodium acetate. The suspension is boiled and filtered hot. The almost colorleSs filtrate is concentrated to dryness and chromatographed on RP-8 Le A 27 337 - 24 -. . , -, .
~ , .. .

- ,, .. - . : - :: :: - --, . , ., : . - , . -: , ~" ~ rS ~ J

using water.
Yield: 29.5 mq (3~ of theory) -NMR (DCOOD): ~ = 5.78 ~6; lH, 2 H); 7.58 (s; lH, 6'-H).
Example 7 p-Methoxybenzyl (6R, 7R)-7-formamido-3-[(Z)-l-propenyl]-3-cephem-4-carboxylate ~ }I ' HCON~ ~ CH3 ~1 , C2 - CH2~3~CH3 A solution of 45 ~ (125 mmol) of p-methoxybenzyl 10~6R, 7R)-7-amino-3-[(Z)-l-propenyl]-3-cephem-4-carboxyl-ate, 9.5 ml (252 ~mol) of formic acid and 53.8 g of dicyclohexylcarbodiimide (261 mmol) in 750 ml of acetonitrile is stirred overnight at room temperature.
'rhe precipitate which has deposited is separa~ed off, the filtrate is concentrated in vacuo and the residue is chromatographed on silica gel using toluene/ethyl acetate (3~
Yield: 38 g (78% of theory) 1H-NMR (CDCl3): ~ = 1.57 (dd, J=6Hz, J = lHz, 3H, CH3-C);
3.36 and 3.50 (2d, J = 19 Hz, each lH, 2-H); 3.82 (s, 3H, CH30); 5.02 (d, J = 6 Hz, lH, 6-H); 5.17 (s, 2H, CH2); 5.68 (dq, J = 11 Hz, J = 5 Hz, lH, 3'-H); 5.88 (dd, J = 9 Hz, L~ A 27 337 - 25 -:: : .

~, . .

3 ~

J = 6 Hz, 1~l, 7-~1); 6.12 (d, J = 11 Hz, 1~1, 3'-H); 6.5B (d, J = 9 Hz, lH, NH); 6.8B and 7.32 ( 2d , J = 8 Hz , each 2H , Ar-H ~; 8 . 2 8 ( s, lH, NCHO ) .
Example 8 p-Methoxybenzyl (6R, 7R)-7-isocyano- 3 -[(Z)-1-propenyl]-3-cephem-4-carboxylate H H
Cl~ CH3 C 2 - CH 2~)C H 3 2.4 ml t20 mmol) of trichloromethyl chloroormate are added dropwise at -78C to a ~olution of 7.8 g (20 mmol) of the compound from Example 7 and 6.9 ml ~85 mmol) of pyridine in 120 ml of anhydrous methylene chloride. After lO minutes, the mixture is allowed ~o warm to 0C, ice water is added, the organic phase is separated off and the aqueous phase is extracted ~wice with methylene chloride. The combined organic pha~es are washed with water, dried over sodium sulphate and con-centrated. Chromatography on silica gel u~ing methylene chloride yields 3.1 g (42% of theory) of the title compound.
H-NMR (CDCl3): ~ = 1.63 (dd, J = 6 Hz, 3 = 1 Hz, 3H, C-CH3); 3.42 and 3.48 (2d, J = 18 Hz, each lH, 2-H); 3.80 (s, 3H, GCH3);

Le A 27 337 - 26 -- : . :. : - , ~: , :. : . :- ,, ;

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

:.. .:; : ::,: . , :: :

,f,'3 ~ J

4.97 (d, J = 6 Hz, lH, 6-H); 5.17 (m, 3H, 7-H and 4'-CH2l; 5.75 (dql J = 11 Hz, J = 6 Hz, lH, 3'-H); 6.26 (d, J = 11 Hz, lH, 3'-H); 6.90 and 7.32 (2d, J = 8 Hz, each 2H, Ar-H).
ExamPle-9 p-Methoxybenzyl 7-{2-[2-(N,N-di-t-butyloxycarbonyl-amino)-5-thienol2,3-d]thia~olylJ-2 [N-(4,4'-dimethoxy-benzhydryl)-N-formamido]-acetamido}-3-[(Z)-1-propenyl]-(6R, 7R)-3-rephem-4-carboxylate s~c~ ~ H H
~oc~ N ~ CO ~ S~ CH3 NCHO
~ co2-CH2 H3~ CH3 ~:

~ solution of 1.30 g (3.5 mmol) of the compound from Example 8, 1.35 g (3.5 mmol) of the compound from Example 5, 0.85 g (3.5 mmol) of 4,4'-dLmethoxybenzhydryl- :
amine and 1.3 ml (35 mmol) of formic acid in 13 ml of methanol and 1.3 ml of dLmethylformamide is stirred at room temperature for 5 days. The solution is ccncen-trated; the residue is chromatographed on silica gel using toluene/ethyl acetate (8:2). :
Yield: 0.25 g (7% of theory) Rf = 0.31 (silica gel 60, F 254, Merck No. 5549;
toluene/ethyl acetate 3:1) Le A 27 337 - 27 -: . ,: , : , .

: .: .

~ h~ 3 Exam~
Benzhydryl (6R, 7R)-7-formamido-3-[~Z)-l-propenyl]-3-cephem-4-carboxylate HCON~ C H 3 c02CH ( C6H5 ~ Z

In ~nalogy to the procedurs of Example 7, 8.0 g 1g2% of theory) of the title compound are obtained from 8.1 g (20 mmol~ of benzhydryl (6R, 7R)-7-amino-3-[(Z)-l-propenyl~-3-cephem-4-carboxylate, 1.5 ml (40 mmol) of~
formic acid and 4.9 g (23.7 mmol) of dicyclohexyl-carbodiLmide in 70 ml of acetonitrile.
H-NMR (CDCl3/CD3SOCD3): ~ = 1.44 (dd, J = 6 Hz, J = 1 Hz, 3H, CH3); 3.40 and 3.50 (2d, J = 18 Hz, each lH, 2-H); 5.08 (d, J = 6 Hz, lH, 6-H); 5.56 (dq, J = 11 Hz, J = 6 Hz, lH, 3'-H); 5.83 (dd, J = 9 Hz, J = 6 Hz, lH, 7-H); 6.12 (d, J = 11 Hz, lH, 3'-H); 6.92 (s, lH, Ph2CH); 7.25 - 7.43 .;
(mt 10H, Ph-H); 8.24 (s, lH, CHO); 8.87 (d, J = 9 Hz, lH, N-H).

L~ ~ 27 33? - 28 -: , : .. . .

~, , ,~ f'L ~3 ~3 ~

Example 11 senzhydryl (6R,7R)-7 isocyano-3-[(Z)-l-propenyl]-3-cephem-4-carboxylate H H
C~ 3 co2CH(c6Hs) 2 In analo~y to the procedure of Example 8, 2.05 g (49% of theory) of the title compound are obtained from 4.35 g ~10 mmol) of the compound from Example 10, 3.5 ml (43 mmol) of pyridine and 1.2 ml (10 mmol) of trichloro-methyl chloroformate in 60 ml of anhydrous methylene chloride.
H-NMR (CDCl3): ~ = 1.53 (dd, J = 6 Hz, J = 1 Hz, 3H, CH3); 3.43 and 3.46 (2d, J = 18 Hz, each lH, 2-H); 5.01 (d, J = 6 Hz, lH, 6-H); 5.20 (d, J = 6 ~z, lH, 7-H); 5.69 (dq, J = 11 Hz, J = 6 Hz, lH, 3'-H); 6.29 ~d, J = 11 Hz, lH, 3 -~t; ~-95 (s, lH, CHPh2);
7.28 - 7.42 (m, lOH, Ph-H).
Example 12 20 Benzhydryl 7-{2-[2-(N,N-di-t-butyloxycarbonyl amino)-5-thienot2,3-d]thiazolyl]-2-[N-(tetra-O-pivaloyl-l-galactosyl)-N-formamido]-(2S)-acetamido}-3-[(Z)-l-propenyl]-(6R, 7R)-3-cephem-4-carboxylate Le A 27 337 - 29 -,. . : . - - . ; : .: : .: ~ -:' `; ' ~: ' ' ' `' ' ' '' .
.: :, . . .
. . ~ , , . ~ , :, . .

~ ç a~, s 1~¦ H H
Boc-N~N~`5~CO-Nt~S~ 1~13 Boc - 0~

tBU-Co-oll2c - o~N-~::Ho co2C~l(C6H5)2 L Bu - C O - C O - CO - t ~u O-CO- tBu 0.6 ml (2.0 mmol) of titanium(IV) isopropoxide i~
added to a BOlutiOn of 1 . O g ( 2.6 mmol) of the compound from Example 5, 1.29 g (2.5 mmol) of tetra-0-pivaloyl-galactosylamine, 0.92 g ~2.2 mmol) of the compound fromExample 11 and 1.0 ml ~26.5 ~mol) of formic acid in 30 ml of anhydrous tetrahydrofuran and the mixture is stirred at room temperature for 5 days. After stripping off the solvent, the residue is chromatographed on silica gel u~ing toluene~ethyl acetate (3 Yield: 0.31 g ~10~ of theory) Rf = 0.28 (silica gel 60, F 254, Merck No. 5549, toluene/ethyl acetate 3:1) Example 13 7-[2-(2-Amino-5-thienot2,3-d]thiazolyl)-2-formamido-acetamido]-3-[(Z)-l-propenyl]-(6R,7R)-3-cephem-4-carboxylic acid (trifluoroacetate) Le A 27 337 - 30 --~ : . , : .
. :: ., , , ~ - . .

f~

H H
H2N~ 5~CONH~S~ CH3 NHCHO o~N~
C02H x CF ~C02}3 250 mg tO.24 mmol) of the compound from Example 9 are trea~ed at room temperature for 1 h with a mixture of 1 ml of anisole and 10 ml of trifluoroacetic acid. After S concentrating, the product is precipitated with ether, filtered off with suction, washed wi~h ether and dried in vacuo. , :
Yield: 110 mg (76% of theory) Example 13a (general formula (I)) 7-[(2S)-2-(2-amino-5-thieno[2,3-d]thiazolyl)-2-amino-acetamido]-3-[(Z)-l-propenyl]-t6R,7R)-3-cephem-4-carboxylic acid s H2N~I,~CON~ CH3 ;
NH2 ~wJ

: ~ ' 107 mg (0.18 mmol) of the compound from :~
lS Example 13 are dissol~ed in 10 ml of methanol. 1 ml of concentrated hydrochloric acid is added and the mixture is allowed to stand at room temperature for 48 h. It is , Le A 27 33? - 31 -'~;, ~''.

~ ~ z~

then diluted with 200 ml of water and ad~usted to pH 5 with sodium acetate. Chromatography on HP-20 u6ing water/acetonitrile yields the pure D-isomer.
Yield: 7 mg (9% of theory) lH-NMR ~DCOOD): ~ = 1.66 (dd, J = 7 Hz, J = 1 Hz, 3H, CH3); 3.44 and 3.54 (2d, J = 18 Hz, each lH, 2-H~; 5.28 (d, J = 6 Hz, lH, 6-H); 5.84 (d, J = 6 Hz, lH, 7-H); 5.80 (dq, J = 11 Hz, J = 7 Hz, lH, ~'-H); 5.93 (8, lH, N-CH-CO);
6.23 (d, J = ll Hz, lH, 3'-H); 7.64 (~, lH, Ar-H).

It is understood that the specification and examples are illustrative but not limitative of the present ihvention and that other embodiments within the spirit and scope of the invention will suggest themselves to those skilled in the art.

Le A 27 337 - 32 -.,~ . , -, . : : ,- : : , ,

Claims (10)

1. A compound of the formula (I) in which R1 - represents halogen or - represents straight-chain or branched alkyl, alkyl-thio or alkenyl having up to 10 carbon atoms, which is optionally substituted by halogen, straight-chain or branched alkoxy having up to 8 carbon atoms or by a 5-membered unsaturated heterocycle having up to 4 heteroatoms from the group consisting of nitrogen and sulphur, which for its part may be substituted by straight-chain or branched alkyl or alkylaminodi-alkyl having up to 6 carbon atoms, - or which is optionally substituted by a group of the formula -CCOR2 in which R2 _ denotes amino or straight-chain or branched alkyl having up to 6 carbon atoms, represents straight-chain or branched alkoxy having up to 8 carbon atoms, - represents a group of the formula -CH2-S-R3, in which R3 - denotes a 5-membered unsaturated heterocycle having up to 4 heteroatoms from the group consisting of nitrogen and sulphur, which is optionally substituted by straight-chain or branched alkyl or alkylaminodialkyl having up to 6 carbon atoms, X - represents oxygen, sulphur or the radical -NH, or a physiologically acceptable salt thereof.

2. A compound or salt thereof according to Claim 1, in which R1 - represents fluorine, chlorine or bromine, or - represents straight-chain or branched alkyl, alkyl-thio or alkenyl having up to 8 carbon atoms, which is optionally substituted by fluorine, chlorine, bromine or by straight-chain or branched alkoxy having up to 6 carbon atoms or by tetrazolyl, thiazolyl, triazolyl or thiadiazolyl each of which is optionally substituted by straight-chain or branched alkyl or alkylaminodialkyl having up to 4 carbon atoms, - or which is optionally substituted by a group of the formula -OCOR2, in which R2 - denotes amino or straight-chain or branched alkyl having up to 4 carbon atoms, - or represents straight-chain or branched alkoxy having up to 6 carbon atoms, and X - represents oxygen or sulphur.

3. A compound or salt thereof according to Claim 1, in which R1 - represents chlorine or straight-chain or branched alkyl or alkenyl having up to 6 carbon atoms, which is optionally substituted by fluorine, chlorine or straight-chain or branched alkoxy having up to 4 carbon atoms, and X - represents sulphur.

4. A compound according to Claim 1, wherein such compound is 7-[(2S)-2-(2-amino-5-thieno[2,3-d]thiazolyl)-2-amino-acetamido]-3-[(Z)-1-propenyl]-(6R,7R)-3-cephem-4-carboxylic acid of the formula or a physiologically acceptable salt thereof.

5. An antibiotically active composition comprising an antibiotically effective amount of a compound or salt according to claim 1, and a physiologically acceptable diluent.

6. A method of combating bacteria in a patient in need thereof which comprises administering to such patient an antibacterially effective amount of a compound or salt according to claim 1.

7. A method of combating bacteria in a patient in need thereof which comprises administering to such patient an antibacterially effective amount of a compound or salt according to claim 4.

8. A compound of the formula (Ia) in which R - represents halogen or - represents straight-chain or branched alkyl, alkyl-thio or alkenyl having up to 10 carbon atoms, which is optionally substituted by halogen, straight-chain or branched alkoxy having up to B carbon atoms or by a 5-membered unsaturated heterocycle having up to 4 heteroatoms from the group consisting of nitrogen and sulphur, which for its part may be substituted by straight-chain or branched alkyl or alkylaminodi-alkyl having up to 6 carbon atoms, - or which is optionally substituted by a group of the formula -OCOR2, in which R2 - denotes amino or straight-chain or branched alkyl having up to 6 carbon atoms, - represents straight-chain or branched alkoxy having up to 8 carbon atoms, - represents a group of the formula -CH2-S-R3, in which R3 - denotes a 5-membered unsaturated heterocycle having up to 4 heteroatoms from the group consisting of nitrogen and sulphur, which is optionally substituted by straight-chain or branched alkyl or alkylaminodialkyl having up to 6 carbon atoms, X - represents oxygen, sulphur or the radical -NH, R4 and R5 are identical or different and represent an amino protecting group, R6 represents a carboxyl protecting group, R7 - represents an easily removable benzylically or glycosidically bonded radical, or - represents a benzylic chiral radical, and R8 _ denotes hydrogen, straight-chain or branched alkyl having up to 6 carbon atoms or a radical of the formula

9. A compound of the formula (II) in which R4 and R5 are identical or different and represent an amino protecting group.

10. A compound of the formula (VI) in which R4 and R5 are identical or different and represent an amino protecting group, and R7 is an easily removable benzylically or glycosidically bound radical, a benzylic chiral radical, or an amino protecting group.