Title of Invention

CEPHALOSPORIN DERIVATIVES

Abstract

wherein Rl denotes hydrogen, or a cation in its free form (Cl-6)alkyl, benzyl, indanyl, phtalidyl, methoxymethyl, (Cl-6)alkanoyloxy(Cl-6)alkyl, (Cl-6) alkoxycarbonyloxy(Cl-6)alkyl, glycyloxymethyl, phenylglycyloxymethyl or (5 - methyl-2-oxo- 1,3 -dioxolen-4-yl)methyl; R2 denotes (C 1-6)alkyl, ar(C 1-6)alkyl, (C2- 6)alkenyl, and R3 denotes hydrogen, (Cl-6)alkyl, ar(Cl-6)alkyl or (C2-6)alkenyl and its pharmaceutically acceptable salts and isomers.

Full Text

The present invention relates to antimicrobial cephalosporins. In one aspect the present invention provides a compound of formula wherein Ri denotes hydrogen or an ester-moiety; e.g. hydrogen; and R2 and R3 independently of each other denote alkyl, e.g. Chalky!, such as Ci.4alkyl; cycloalkyl, aralkyl, e.g. ar(Ci.6)alkyl, such as ar(Ci_4 )alkyl; aryl; alkenyl, e.g. (C2-6)alkenyl, such as (C2-4) alkenyl; or alkinyl; and R3 additionally denotes hydrogen; e.g. R2 denotes alkyl, aikenyl or aralkyl; e.g. R3 denotes hydrogen or alkyl; e.g.in free form and in the form of a salt and/or in the form of a solvate. Ri denotes hydrogen or an ester moiety. An ester-moiety includes alkyl, preferably C1.6alkyl, e.g. Ci.4alkyl; aralkyl, for example benzyl, alkoxybenzyi, such as 4-methoxybenzyl; indanyl, phthalidyl, alkoxymethyl, e.g. methoxymethyl; (Ci.6)alkanoyloxy(Ci.6)alkyl, (C16)alkoxy-carbonyl-oxy(Ci-6)alkyl, glycyloxymethyl, phenylglycyloxymethyl, (5-methyl-2-oxo-l,3-dioxolen-4-yl)methy|; an ester moiety also includes ester moieties which form with the COO-group a physiologically hydrolysable and acceptable ester, e.g. such known to be hydrolysable ester groups in the field of cephalosporins. A compound of formula I may thus be in the form of 1 an physiologically-hydrolysable and -acceptable ester. By physiologically-hydrolysable and -acceptable esters as used herein is meant an ester in which the COO- group is esterified and which is hydrolysable under physiological conditions to yield an acid which is itself physiologically tolerable at dosages to be administered. The term is thus to be understood as defining regular pro-drug forms. An ester moiety may be preferably a group which is easily hydrolysable under physiological conditions. Such esters may be administered preferably orally. Parenteral administration may be indicated if the ester per se is an active compound or, if hydrolysis occurs in the blood. Ri denotes preferably hydrogen. R2 denotes preferably alkyl; such as lower alkyl, e.g. methyl, ethyl, e.g. methyl; aralkyl, e.g. benzyl; cycloalkyl; or alkenyl, e.g. allyl; such as alkyl, aralkyl or alkenyl, e.g. alkyl, such as i methyl. R3 denotes preferably hydrogen or alkyl, e.g. lower alkyl, such as methyl, ethyl. Alkyl in the meaning of R2 and R3 preferably includes unsubstituted alkyl and alkyl substituted e.g. by substituents which are conventional in P-lactam chemistry; such as halogen, hydroxy; and is preferably unsubstituted. Aralkyl preferably includes aralkyl wherein the phenyl group is unsubstituted or substituted, e.g. by substituents which are conventional in p-lactam chemistry; e.g. by one or more, e.g. 1 to 3, substituents; e.g. hydroxy, alkoxy, e.g. lower alkoxy, e.g. methoxy groups. If not otherwise defined herein alkyl includes e.g. (Ci-i2)alkyl, such as (Ci.e)alkyl, e.g. (Ci.4)alkyl. Lower alkyl includes (Ci.4)alkyl. Cycloalkyl includes e.g. (C3.g)cycloalkyl, such as (C4.7)cycloalkyl, e.g. (C5-6)cycloalkyl. Alkenyl includes e.g. (C2.i2)alkenyl, such as (C2-6)alkenyl, e.g. (C2.4)alkenyl. Alkinyl includes e.g. (C2-i2)alkinyI, such as (C2-6)alkinyl, for example (C2-4)alkinyl. Aryl includes phenyl or naphthyl, e.g. phenyl. Aralkyl includes ar(Ci.6)alkyl, such as ar(Ci.4)alkyl, e.g. benzyl. A silyl group includes a silyl protecting group, e.g. a conventional silyl protecting group, such as a trialkylsilyl group, for example a trimethylsilyl group. Any group defined above may be e.g. unsubstituted or e.g. substituted, e.g. by groups which are conventional in P-lactam chemistry. Preferred compounds include e.g. 7-(((5-Amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluormethoxy-imino)acetyl)amino)-3-((imino-l-piperazinylmethyI)methylhydrazono)methyl-3-cephem-4-carboxylic acid; 7-(((5-Amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluormethoxyimino)acetyl)- amino)-3-((imino-l-piperazinylmethyl)ethyIhydrazono)methyl-3-cephem-4-carboxylic acid; 7-(((5-Amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluormethoxy-imino)acetyl)amino)-3-((imino-l-piperazinyImethyl)-allylhydrazono)methyl-3-cephem-4-carboxylic acid; 7-(((5-Amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluormethoxyimino)acetyl)amino)-3-((imino-l-piperazinylmethyl)(4-methoxyphenyl)methyl-hydrazono)methyl-3-cephem-4-carboxy!ic acid; 7-(((5-Amino-l,2,4-thiadiazoI-3-yl)-(Z)-(fluormethoxyimino)acetyl)amino)-3-((imino-l-piperazinylmethyl)(3,4,5-methoxyphenyl)-methylhydrazono)methyI-3-cephem-4-carboxyIic acid; 7-(((5-Amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluormethoxy-imino)acetyl)amino)-3-(((N-methyl-imino-l-piperazinylmethyl)methylhydrazono)-methyl-3-cephem-4-carboxylic acid; and 7-(((5-Amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluormethoxy-imino)acetyl)amino)-3-((N-ethyl-imino-l-piperazinylmethyl)methylhydrazono)methyl-3-cephem-4-carboxylic acid. In another aspect the present invention provides the compound 7-(((5-Amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluormethoxy-imino)acetyl)amino)-3-((imino-l-piperazinylmethyI)methyIhydrazono)-methyI-3-cephem-4-carboxyIic acid, e.g. in the form of a salt, such as in the form of a hydrochloride; e.g.and/or in the form of an ester, e.g. a compound wherein Ri in formula I denotes an ester moiety. i t A compound of formula I includes a compound of formula I in any configuration, e.g. in any possible steroisomeric fbrm. Mixtures of stereoisomeric forms may be separated, e.g. as conventional, e.g. by chromatography, fractioned crystallisation. E.g. the configuration of -OCH2F in group -C=N-OCH2F may be syn [(Z)] and anti [(E)] and is preferably, e.g. predominantly, syn [(Z)]; e.g. containing the [(E)] form in an amount of 0 to 5%, e.g. 0 to 2%. A compound of formula I may be in the form of a mixture of the 3(E)-form and 3-(Z)-form, or may be, e.g. predominantly, in the 3(Z)-form, e.g. according to formula Z) or may be, e.g. predominantly, in the 3(E)-form, e.g. according to formula i wherein Ri and R2 are as defined above, and wherein the configuration of the group attached to the nitrogen of the -C=N group in position 3 of the ring system is, e.g. 3(E) and/or 3(Z). A compound of formula I may be, e.g. predominantly, in the 3(E)-form, e.g. containing the 3(Z)-form in an amount of 0 to 5%, e.g. 0 to 2%. In another aspect the present invention provides a compound of formula I predominantly in the 3-(E) form, e.g. 7-(((5-Amino-l,2,4-thiadiazol-3»yl)-(Z)-(fluormethoxy-imino)acetyl)amino)-3(E)-((imino-l-piperazinyImethyl)methylhydrazono)-methyl-3-cephem-4-carboxyIic acid, e.g. in the form of a salt, such as in the form of a hydrochloride and/or in the form of a solvate. ■ i ! ! ' A salt includes any possible salt, e.g. an acid addition salt; such as a hydrochloride, internal salt, metal salt, quaternary salt and an amine salt of a compound of formula I. Metal salts include for example sodium, potassiiim, calcium, barium, zinc, aluminum salts* preferably sodium or potassium salts. Amine salts include for example trialkylamine, procaine, dibenzylamine and benzylamine salts. A salt may preferably.be a pharmaceutical^ acceptable salt of a compdund of formula I. A solvate includes a solvate with an organic solvent and a solvate with water, such as a hydrate. A free form of a compound of formula I may be converted into a salt form and vice versa. A solvate form of a confound of formula I, e.g. in free form or in the form of a salt, may be converted in a non-sofyate form and vice versa. In another aspect the present invention provides a process for the production of a compound of formula I as defined in claim 1, comprising reacting a compound of formula i wherein either a) Rb denotes hydroxy and Rc and R^ together form a bond, or (3) Rd denotes hydrogen, an ester-moiety or a silyl group, and Rb and Rc together denote an oxo group, e.g. in free form or in the form of a salt and/or a solvate; e.g. and/or in a form where reactive groups are protected, e.g. by appropriate groups, such as conventional protecting groups; with a compound of formula wherein R2 and Rj are as defined above, e.g. in free form or in the form of a salt; and/or a i solvate; e.g. and/or in a form wherein reactive groups are protected, e.g. by appropriate groups, such as conventional protecting groups; splitting off a silyl group from the carboxylic acid group in position 4 of the ring system, if present; e.g. splitting off other protective groups if present and if desired; and isolating a compound of formula I; e.g. in free form or in the form of a salt and/or a solvate. An ester moiety in the meaning of Rd may be as defined above in the meaning of Ry. A process according to the present invention may be carried out as follows: A compound of formula II, e.g. in free form or in the form of a salt and/or a solvate and/or e.g. in a form wherein reactive groups, e.g. the amine group attached to the thiadiazolyl ring is protected, e.g. by appropriate groups, such as conventional protecting groups; may be reacted in a solvent which is inert under the reaction conditions, including e.g. water, water mixed with a lower alcohol or dioxane, or a dipolar aprotic solvent, for example dimethylformamide or dimethylsulphoxide, e.g. mixed with an alcohol or water; e.g. at temperatures of -20 to 50°C, with a compound of formula EI, e.g. in free form or in the form of a salt and/or a solvate and/or e.g. in a form wherein reactive groups, e.g. the amine group attached to the -N-R2 group and/or a piperazinyl nitrogen group, are protected, e.g. by appropriate groups, such as conventional protecting groups. Silyl protecting group technology in the presence of a solvent which may be inert towards silylation agents, e.g. a chlorinated hydrocarbon, such as dichloromethane; a nitrile such as acetonitrile, an ether such as tetrahydrofuran, a dipolar aprotic solvent, e.g. N,N-dimethylformamide; or a solvent system, e.g. mixtures of individual solvents, e.g. as described above; may be appropriate for the protection of reactive groups by silyl groups. The pH of the reaction mixture may be adjusted to an pH optimum by addition of a pH-influencing agent, e.g. kn organic or inorganic acid or base. A compound of formula I, e.g. in the form of a salt and/or a Solvate, e.g. predominantly in the 3(E)-form, may be obtained and may be isolated from the reaction mixture, eg. as conventional, e.g by precipitation, e.g. in the presence of an anti-solvent, extraction, chromatography, solvent evaporation. Protecting groups present in a compound of formula II or a compound of formula III may be split off, e.g. during the reaction between a compound of formula II and III under appropriate reaction conditions, e.g. as conventional; or, e.g. protecting groups present may be split off from a compound of formula I obtained from the reaction between a compound of formula II and IQ, e.g. as conventional. A compound of formula I obtained, wherein R1 is hydrogen, e.g. in the form of a salt, may be converted into a compound of formula I, wherein R1 is an ester moiety, and vice versa, e.g. as conventional. A compound of formula I may be isolated from the reaction mixture, e.g. as conventional, e.g. by filtration, extraction, centrifugation, solvent evaporation. A compound of formula I in free form may be converted into a compound of formula I in salt form, and/or solvate form; or vice versa, e.g. as conventional. Compounds of formula II and HI are known or may be produced according to known, e.g. analogous methods, or e.g. according to the present examples. The compounds of formula I, hereinafter designated as "active compound(s) of the invention" exhibits pharmacological activity and surprising low toxicity and are therefore useful as pharmaceuticals. In particular, the active compounds of the invention show antimicrobial, e.g. antibacterial, activity against aerobic and anaerobic growing bacteria, e.g. gram negative and gram positive bacteria such as Enterobacter, e.g. Enterobacter cloacae; Enterococcus, e.g. Enterococcus faecalis, Enterococcus faecium; Moraxella, e.g. Moraxella catarrhalis; Haemophilus, e.g. Haemophilus influenza; Klebsiella, e.g. Klebsiella edwardsii, Klebsiella pneumoniae; Streptococcus, e.g. Streptococcus pyogenes; Staphylococcus, e.g. Staphylococcus aureus MSSA (methicillin sensitive strains); Staphylococcus aureus MRSA (methicillin resistant strains); Escherichia, e.g. Escherichia coli; Proteus, e.g. Proteus mirabilis, Salmonella, e.g. Salmonella typhimurium, Serratia, e.g. Serratia marcescens, Pseudomonas, e.g. Pseudomonas aeruginosa; Pneumococci, e.g. Pneumococcus pneumoniae (penicillin sensitive and mult-drug resistant strains); in vitro in the Agar DilutionTest for bacteria according to National Commitee for Clinical Laboratory Standards (NCCLS) 1993, - Document-M7-A3Vol.l3, No. 25: "Methods for dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically - Third Edition, Approved Standard"; and - Document Ml 1-A3 for anaerobic bacteria in a concentration from about 0.001 to ca. 50 ^ig/ml (MIC), e.g. using strains including Staphylococcus aureus (ATCC 29213 and ATCC 9144); Enterococcus faecalis (ATCC 29212); Haemophilus influenza (NTCC 49247 and NCTC 11931); Escherichia coli (ATCC 25922 and ATCC 35218); Klebsiella pneumoniae (NCTC 11228); Klebsiella edwardsii (NCTC 10896); Pseudomonas aeruginosa (ATCC 27853 and ATCC 25668); and in vivo in the septicaemia mouse model, in accordance to the method description Nr. 159 A-5, approved by Austrian Health Authorities (MA 58, no. 2968/95 of 12-Oct-1995), e.g. when administerd at dosages from about 0.05 to 50 mg/kg body weight, such as 0.1 to 50 mg/kg body weight (ED50 values). E.g., mice are infected with an ED 95% of Staphylococcus aureus (ATCC 4995), Streprococcus pyogenes (ATCC 29218), Escherichia coli (A 12 NFI culture collection) and are treated 1, 5 and 24 hours after infection. The ED 50% values ranging from ca. 0.2 to 50 mg/kg body weight are calculated by Probit analysis of the administered dosages of compounds. Activity is determined by numbers of surviving animals per group of 8 mice per dosage until day 5 after infection. The active compounds of the invention show an surprising overall activity spectrum. It has, for example, been determined that the MHK (pg/ml) of the compound of Example 1 against, for example Enterococcus faecalis is of ca. 0.1 to 0.4; against Staphylococcus aureus (MSSA) is of ca. 6.4; against multi-drug resistant Pneumococcus is of 0.4. The active compounds of the invention are therefore useful for the treatment of microbial, e.g. bacterial diseases. For this indication, the appropriate dosage will, of course, vary depending upon, for example, the compound of formula I employed, the host, the mode of administration and the nature and severity of the conditions being treated. However, in general, for satisfactory results in larger mammals, for example humans, an indicated daily dosage is in the range from about 0.05 to 5 g, for example 0.1 to about 2.5 g, of an active compound of the invention conveniently administered, for example, in divided doses up to four times a day. ! i I ! An active compound of the invention may be administered by any conventional route, for example orally, e.g. in the form of tablets or capsules, or parenterally in the form of injectable solutions or suspensions, e.g. in analogous manner to cefotaxime. The compound 7-(((5-amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluormethoxyimino)acetyl)amino)-3-((imino-l-piperazinylmethyl)methylhydrazono)methyl-3-cephem-4-carboxylic acid (compound of Example 1) is the preferred compound of the invention for use as an antimicrobial agent. It has, for example been determined that the MHK (pg/ml) of the compound of Example 1 (tested in the form of the hydrochloride) against, for example Haemophilus influenza is ca. A compound of formula I may be administered in the form of a pharmaceutical^ acceptable salt, e.g. an acid addition salt or a base addition salt or in the corresponding free form, if desired in the form of a solvate. Such a salt/solvate may exhibit the same order of activity as the free form. The present invention also provides a pharmaceutical composition comprising a compound of formula I according to claim 1 in the form of a pharmaceutical^ acceptable salt or in free form in association with at least one pharmaceutical carrier or diluent. Such compositions may be manufactured in conventional manner. Unit dosage form may contain, for example 10 mg to about 1 g, for example 10 mg to about 700 mg, such as to about 500 mg. As medicaments, the active ingredients according to the invention may be administered alone or in suitable medicinal forms together with inorganic or organic, pharmacologically inert excipients. For example, they are used as a constituent of capsules, or injection or instillation preparations, which contain a quantity of active compounds that is sufficient to attain an optimum blood level, that is, ca. 10 to 500 mg per capsule. For this application, the dosage to be administered depends on the compound used and the type of administration, as well as the type of treatment. With larger mammals, satisfactory results may be obtained when administering a daily dose of ca. 0.5 to 6 g. If required, this amount may be givdh in correspondingly smaller doses two to four times daily, or in sustained release form. In another aspect the present invention provides a compound of formula I or a composition comprising a compound of formula I in the form of a pharmaceutical^ acceptable salt or in free form in association with at least one pharmaceutical carrier or diluent for use as a pharmaceutical, e.g. as an antibiotic; and The use of a compound of formula I, or use of a composition comprising a compound of formula I in the form of a pharmaceutical^ acceptable salt or in free form in association with at least one pharmaceutical carrier or diluent as a pharmaceutical. In a further aspect the present invention provides a method of treatment of microbial diseases, e.g. caused by bacterias selected from Pseudomonas, Enterobacter, Enterococcus, Moraxella, Haemophilus, Klebsiella, Streptococcus, Staphylococcus, Escherichia, Proteus, Salmonella, Serratia or Pneumococci, which comprises administering to a subject in need of such treatment, an effective amount of a compound of formula I; e.g. in the form of a pharmaceutical composition according to the present invention; and A compound of formula I for use in the preparation of a medicament for the treatment of microbial diseases, for example of diseaeses caused by bacterias selected from Pseudomonas, Enterobacter, Enterococcus, Moraxella, Haemophilus, Klebsiella, Streptococcus, Staphylococcus, Escherichia, Proteus, Salmonella, Serratia or Pneumococci. In another aspect the present invention provides compounds of formula IP wherein Ri denotes hydrogen, an ester moiety or a cation, in free form, or, where such forms exist, in the form of acid addition salts, inner salts, quaternary salts or hydrate. In the following examples; which illustrate the invention more fully but should in no way limit its scope, all temperatures are given in degrees Celsius. 'H-NMR: 200MHz, DMSO-d6. Example 1 7-(((5-anrino-l,2,4-tMadiazol-3-ylM piperazinylmethyl)methylhydrazono)methyl-3hem4-carboxylicacid a) N-(1.4Ja.6-tetrahvdro-3-hvdroxv-1.7-dtoxo-3H.7H-aceto(2.1-b)furo(3.4-d)(1.3)"thiazin- 6-Yl)-2-(5-amino-l .2.4-thiadiazol-3-yn-(Z)-2-(fluoromethoxyimino)acetic acid amide (hydroxylactone of 7-(((5-amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluoromethoxyimino)acetyl) amino)-3-formyI-3-cephem-4 A suspension of 10 g of; 7-amino-3-formyI-3-cephem-4-carboxylic acid in a mixture of 220 ml of 1 methylene chloride and 80 ml of acetonitrile is stirred at 0° with 43 ml of N,Obis(trimethylsiIyl)- 1 1 acetamide. 15.7 g of (5-amino-l,2,4-thiadiazol-3-yl)-(Z)-2-fluoromethoxyimino-acetic acid chloride are added to the clear solution obtained and the reaction mixture is stirred for ca. one hour at ca. 0°. The mixture is diluted with 1250 ml of acetonitrile which contains 70 ml of water. 12% aqueous ammonia is added to the mixture to adjust a pH value of 3.5. The mixture is diluted with 2.5 litres of water and extracted with ethyl acetate. The ethyl acetate phase is dried and concentrated. The concentrate is stirred for one hour at 20° with 100 ml of acetonitrile. N- (l,4,5a,6-tetrahydro-3-hydroxy-lJ-dioxo^ amino-l,2,4-thiadiazol-3-yl)-(Z)-2-(fluoromethoxyimino)acetic acid amide precipitates in crytalline form, is filtrated off and dried. Vl-NMR: 3.65 (m, 2x AB quartet, 2H, SCH2); 5.18 (d, J = 5 Hz, 1H, lMactam-H); 5.83 (d, J = 55 Hz, 2H, CH2F); 6.03 (dd, J = 5 and 8.3 Hz, 1H, 6-lactam-H); 6.24 and 6.30 (s, 1H, O-CH-O); 8.25 (broad singulet, 2H, NH2); 9.89 and 9.87 (d, J = 8,3 Hz, 1H, NH). b) 7-(((5-amino-l,2.4-thiadiazol-3-vl)-(Z)-(fluoromethoxvimino)acetvnamino)-3(E)-(imino-l- piperazinylmethvl)methvlhvdrazono)methyl-3-cephem-4-carboxvlic acid 3.77gofN-(l,4,5a,6-tetrahydro-3-hydroxy-l,7-dioxo-3H,7H-aceto(2,l-b)furo(3,4-d)(l,3)-thiazin-6-yl)-2-(5-amino-l,2,4-thiadiazoI-3-yl)-(Z)-2-fluoromet]hoxyimino)acetic acid amide are suspended in a mixture of 75 ml of acetonitrile and 11 ml of water and treated with a solution of 2 g of l-(l-methylhydrazino)iminomethyl)piperazine in the form of a dihydrochloride in 4.5 ml of 2N HCI. The reaction mixture is stirred for ca. one day at room temperature and poured into 600 ml of acetonitrile under stirring. 7-(((5-amino-l,2,4-thiadiazol-3-yl)-(Z)-(fluoromethoxy-imino)acetyl)amino)-3(E)-(imino-l-piperazinylmethyl)methylhydrazono)methyl-3-cephem-4-carboxylic acid in the form of a trihydrochloride precipitates, is filtrated off, washed with acetonitrile and dried. c) 7-(((5-amino-1.2 piperazinylmethvl)methylhvdrazono)methvl-3-cephem-4-carboxvlic acid 0.65 g of crude 7-(((5-amino-l,2,4-thiadiazoI-3-yl)-(Z)-(fluoromethoxy-imino)acetyI)amino)-3(E)-(imino-l-piperazinylmethyl)methylhydrazono)methyl-3-cephem-4-carboxylic acid in the form of a trihydrochloride obtained in step b) are dissolved in 2 ml of water and filled into a column which is filled with 50 g of RP-18R (LiChroprep RP-18R, grain size 40-63 \im9 Merck) and eluated with water (flow rate 20ml/min). Fractions are examined by means of analytical HPLC and the fractions which contain 7-(((5-amino-l,2,4-thiadiazol-3-ylHZ)-(fluoromethoxyimino)acetyl)-amino)-3(E)-(imino-l-piperazinylmethyl)methylhydrazono)methyl-3-cephem-4-carboxylic acid in the form of a monohydrochlorid are determined (HPLC), combined and lyophilised. As described in Example 1 but using a corresponding compound of formula HI wherein R2 and R3 have the meaning given in TABLE 1 below the compounds of formula I, wherein Rt is hydrogen listed in TABLE 1 below are obtained, e.g. in the salt form described: Aminoguanidines used as starting material may be produced as follows: Example A l-(l-Methylhydrazino)iminomethyl)piperazine a) S-Methvl-2-methvl-isothiosemicarbazide A solution of 239.8 g of S-methyl-2-methyIisothio-semicarbazide in the form of a hydriodide in 100 ml of water is placed on a column filled with 1500 ml of a strong basic ion exchanger in chloride form (Amberlite IRA 420R), and eluted with water. The fractions containing S-methyl-2- methylisothio-semi lyophilisate is treated with ether, isolated by filtration and dried. S-methyl-2-methyl-isothiosemicarbazide in the form of a hydrochloride is obtained in the form of a white solid. M.p.: 116° (isopropanol). b) Benzvlidene derivative of 4-formvl-l-((l-methvlhvdrazino)imino-methvl)piperazine A solution of 40.9 g of S-methyI-2-methyl-isothiosemicarbazide in the form of a hydrochloride in 350 ml of ethanol is mixed with 30 g of freshly distilled formylpiperazine and heated under reflux for ca. 39 hours. The reaction mixture is cooled to room temperature, mixed with 26.4 ml of benzaldehyde and stirred for ca. 24 hours. The precipitate obtained is filtrated off, washed with ethanol and dried. The benzylidene derivative of 4-formyl-l-((l-methylhydrazino)imino~ methyl)piperazine in the form of a hydrochloride is obtained. c) l-((l-Methylhvdrazino)iminomethyl)piperazine From 10 g of the benzylidene derivative of 4-formyl-l-((l-methylhydrazino)iminomethyl)-piperazine in the form of a hydrochloride the benaldehyde is split off by steam distillation under addition of 48 ml of 2N HC1. The aqueous slurry obtained is concentrated and an oily residue is obtained which is treated with boiling ethanol. The ethanolic phase is concentrated in vacuum. 1-((l-Methylhydrazino)iminomethyl)piperazine in the form of a dihydrochloride is obtained in the form of a white solid. *H-NMR: 3.16 (m, 7H, N-CH3 and -CH2-N-CH2-); 3.61 (m, 4H, -CH2-N+-CH2-); 6.0 (broad singulet, 3H, N+H3); 8.3 (broad singulet, 1H, NH); 10.0 (broad singulet, 2H, N+H2). Example B l-[(l-Ethylhydrazino)iminomethyl]piperazine a) Benzvlidene derivative of l-(hydrazinoiminomethvl)piperazine The pH of a solution of 10.7 g of the benzylidene derivative of l-(hydrazinoiminomethyl)- piperazine in the form of a dihydrochloride in 100 ml of water is adjusted to 10 by addition of 1 8N NaOH. The mixture obtained is extracted with ethyl acetate. The ethyl acetate phase is dried and the solvent is evaporated off. The benzylidene derivative of 1-(hydrazinoiminomethyl)piperazine is obtained in the form of an amorphous powder. b) Benzvlidene derivative of l-formvl-4-(hydrazinoiminomethyl)piperazine 12.7 ml of acetic acid anhydride are added dropwise to 42 ml of ice-cooled formic acid, the mixture is stirred for ca. 1 hour and 16 g of the benzylidene derivative of l-(hydrazinoimino-methyl)piperazine in 42 ml of formic acid are added dropwise. The mixture is left for ca. 2 hours at 0° and the solvent is evaporated off. The residue is treated with water and the pH of the mixture obtained is adjusted to pH 11 by addition of ION KOH. The mixture is extracted with dichloromethane, the dichloromethane phase is dried and the solvent is evaporated off. The benzylidcnc derivative pf l-formyI-4-(hydrazinoiminomethyi)piperazine is obtained in the form of a white powder. i c) Benzvlidene derivative of l-[(l-ethylhydrazino)iminomethvl1^-formylpiperazine An ice-cooled solution of 2 g of the benzylidene derivative of l-formyl-4-(hydrazino- iminomethyl)piperazine in 40 ml of dry tetrahydrofurane is treated with 9.3 ml of bis-(trimethyl- silyl)-lithiumamid (1M solution in tetrahydrofurane) and stirred for ca. 1 hour at 0°. 2.4 g of ethyl iodide are added to the reaction mixture and the mixture is stirred overnight at room temperature. The solvent is evaporated off and the residue is purified via "Dry-column-flash- chromatography": Eluent: 1. methanol; 2. 90% methanol /10 % acetic acid. Fractions containing the benzylidene derivative of l-[(l-ethylhydrazino)iminomethyl]-4-formylpiperazine (analytical HPLC determination) are combined, the solvent is evaporated off and the benzylidene derivative of l-[(l-ethylhydrazino)iminomethyl]-4-formylpiperazine is obtained in the form of a white powder. d) l-f(l-Ethvlhvdrazino)iminomethvl]piperazine 2.7 g of the benzylidene derivative of l-[(l-ethylhydrazino)iminomethyl]-4-formylpiperazine dissolved in 11.6 ml of 2N HCl are treated by steam distillation. After evaporation of the water from the mixture obtained and drying of the residue l-[(l-ethylhydrazino)iminomethyl]piperazine in the form of a dihydrochloride is obtained in the form of a white solid. * 1H-NMR-spectrum:l.22, tj=5 Hz, 3H, CH3; 3.16, b, 4H, N-CH2; 3.45, q, J=5 Hz, 2H, CH2; 3.65, b, 4H, N-CH2; 10.14, b, 2H, NH In the manner as described in Example B, but using the corresponding reactants the following compounds may be obtained: l-[(l-AHvlhvdrazino)iminomethvl]piperazine (in the form of a dihydrochloride) 'H-NMR-spectrum: 3.14, b, 4H, N-CH2; 3.68, b, 4H, N-CH2; 3.98 - 4.18, m, 2H, CH2-C; 5.16 -5.48, m, 2H, CH2=C; 5.80 - 6.10, m, 1H, CH=C; 10.30, b, 2H, NH l-[[l-(4-Methoxvbenzvl)hvdrazino]iminomethvl1piperazine (in the form of a dihydrochloride) 'H-NMR-spectrum: 3.19, b, 4H, N-CH2; 3.67, b, 4H, N-CH2; 3.77, s, 3H, 0-CH3; 4.59, s, 2H, N-CH2; 6.90 - 7.02 u. 7.25 - 7.38, m, each 2H, CH-arom.; 10.02, b, 2H, NH l-[[l-(3,4.5-Trimethoxvbenzvl)hvdrazino1iminomethyllpiperazine (in the form of a dihydrochloride) 'H-NMR-spectrum: 3.20, b, 4H, N-CH2; 3.67, b, 7H, 4H of N-CH2 and 3H of 0-CH3; 3.81, s, 6H, O-CH3; 4.59, s, 2H, N-CH2; 6.69, s, 2H, CH-arom.; 9.96, b, 2H, NH Example C 1-[(1-Methylhydrazino)(methylinrino)methyl]piperazine a) Benzylidene derivative of l-formvl-4-[hydrazino(methvlimino)mcthyl1pipcrazinc 37 g of l-formyI-4-[hydrazino(methylimino)methyl]piperazine in the form of a hydrochloride dissolved in a mixture of 80 ml of acetonitrile and 185 ml of water are treated with 30 g of benzaldehyde. The mixture is stirred for ca. 3 hours at room temperature and extracted with diethyl ehter. The water of the aqueous phase is evaporated. The residue is treated with water and a pH of 11 of the mixture is adjusted with 2N NaOH. The mixture is extracted with dichloromethane, the organic phase is dried, the solvent evaporated and the residue is dried. The benzylidene derivative of l-formyl-4-[hydrazino(methylimino)methyl]piperazine is obtained in the form of a white powder. b) Benzylidene derivative of l-formyM-[(l"methvlhvdrazino)(methvlimino)methvl]piperazine A solution of 1,62 g of the benzylidene derivative of l-formyl-4-[hydrazino(methylimino)- methyl]piperazine in 3Q ml of acetonitrile is treated with 4,56 g of methyl iodide and the mixture is refluxed overnight. The solvent is evaporated off and the residue is stirred with 20 ml of water and 10 ml Amberlite IJlA-400 (C1)R (ion exchange resin) for ca. 1 hour at room temperature. The mixture is filtrated. The aqueous solution is adjusted to a pH of 11 with 2N NaOH and extracted with dichloromethane. The organic phase is dried and concentrated by solvent evaporation. For purification the concentrate is treated in the manner described in Example B, c). The benzylidene derivative of l-formyl-4-[(l-methylhydrazino)(methylimino)methyl]piperazine is obtained in the form of a white powder. c) l-[(l-Methvlhvdrazino)(methylimino)methvl]piperazine 1.14 g of the benzylidene derivative of lnformyl-4-[(l-methylhydrazino)(methylimino)methyI]-piperazine dissolved in 6 ml of 2N HC1 are treated as described in Example B, d). l-[(l-MethyIhydrazino)(methylimino)methyl]piperazine in the form of a dihydrochloride is obtained in the form of a white solid. 1H-NMR-spectrum: 2.84, s, 3H, CH3; 3.18, b, 7H, 4H of N-CH2 and 3H of CH3; 3.63, b, 4H, N-CH2; 10.13, b, 2H,NH In the manner as described in Example C but using the corresponding reactants l-[(l-methylhydrazino)(ethylimino)methyl]piperazine (in the form of a dihydrochloride) may be obtained. 1H-NMR-spectrum: 1.20, t, J=5 Hz, 3H, CH3; 3.19, b, 9H, 4H of N-CH2 and 3H of CH3 and 2H of CH2; 3.64, b, 4H, N-CH2; 10.12, b, 2H, NH. 'H-NMR-Spectra of the compounds of the Examples Ex. 1 3.25 (broad, 4H, -CH2-N-CH2-); 33 (s, 3H, N-CH3); 3.60 and 4.28 (Ab quartet, j = is m, 2H, SCH2); 3.74 (broad, 4H, -CH2-NH+-CH2-); 5.28 (d, J = 5 Hz, 1H, G-lactam-H); 5.78 (d, J = 55 Hz, 2H, CH2F); 5.91 (dd, J = 5 and 8.3 Hz, 1H, G-lactam-H); 8.1 (s, 1H, CH=N); 9.04 (broad singulet, 1H, NH); 9.35 (broad singulet, 1H, NH); (9.81 (d, J = 8.3 Hz, 1H, NH), 9.9 (broad singulet, 2H, NH2). 2 1.17, t, J=5 Hz, 3H, CH3; 3.28, b, 4H, N-CH2; 3.60 and 4.21, AB-quartet, J=18 Hz, 2H, S-CH2; 3.67, b, 4H, N-CH2, 3.91, m, 2H, CH2; 5.22, d, J=5 Hz, 1H, G-lactam-H, 5.82, d, J=55 Hz, 2H, CH2F; 5.85, dd, J= 5Hz and 8 Hz, 1H, 6-lactam-H; 8.35, b, 3H, 1H CH=N and 2H, NH; 9.78, d, J=8 Hz, 1H, NH. 3 1.18, t, J=5 Hz, 3H, CH3; 3.30, b, 9H, 4H of NCH2 and 2H of CH2 and 3H of CH3; 3.70, m, 5H, 4H of NCH2 and 1H of S-CH2; 4.10, part of AB-quartet, J=18 Hz, 1H, SCH2; 5.32, d, J=5 Hz, 1H, G-lactam-H; 5.82, d, J=55 Hz, 2H, CH2F; 5.95, dd, J=5 Hz and 8Hz, 1H, 12-lactam-H; 8.08, s, 1H, CH=N; 8.32, b, 1H, NH; 9.82, d, J=8 Hz, 1H, NH. 4 3.30, b, 4H, N-CH2; 3.58 and 4.25, AB-quartet, J=18 Hz, 2H, S-CH2; 3.73, b, 4H, N-CH2; 4.30, m, 2H, N-CH2; 5.26, m, 3H, 1H G-lactam-H and 2H of CH2=C; 5.64, part of dublet, 1H, CH2F; 5.90, m, 4H, 1H of CH2-F and 1H of CH=C and 1H G-lactam-H; 8.11, s, 1H, CH=N; 9.81, d, J=8 Hz, 1H, NH. 5 2.90 and 3.03, 2s (2:1), 3H, N-3-CH,; 3.33, b, 7H, 4H of N-CH2 and N-2-CH3, 3.64, b, 5H, 4H of N-CH2 and 1H of S-CH2; 4.15, part of AB-quartet; J=18 Hz, 1H, S-CH2; 5.21, d, J=5 Hz, 1H, G-lactam-H; 5.81, d, J=55 Hz, 2H, CH2F; 5.83, dd, J=5Hz and J=8 Hz, 1H, G-lactam-H; 8.32, 3H, 1H of CH=N and 2H of NH; 9.79, d, J=8 Hz, 1H, NH. 6 3.31, b, 4H, N-CH2; 3.52 and 4.18, AB-quartet, J=18 Hz, 2H, S-CH2, 3.72, b, 7H, 4H of N-CH2 and 3H of OCH3; 4.95, AB-quartet, J=17 Hz, 2H, CH2; 5.14, d, J=5 Hz, 1H, G-lactam-H; 5.78, d, J=55 Hz, 2H, CH2F; 5.77, dd, J=5 Hz and 8*Hz, 1H, G-lactam-H; 6.86 - 6.91, m, 2H, CH- arom.; 7.15 - 7.19, m, 2H, CH-arom.; 8.26, b, 2H, CH=N and NH; 8.40, b, 1H, NH; 9.74, d, J=8 Hz; 1H, NH. 7 3.34, b, 4H, N-CH2; 3.57 and 4.23, AB-quartet, J=18 Hz, 2H, S-CH2; 3.64, s, 3H, OCH3; 3.79, b, 10H, 4H of N-CH2 and 6H of OCH3; 5.03, AB-quartet, J= 17 Hz, 2H, CH2; 5.27, d, J=5 Hz, 1H, G-lactam-H; 5.81, d, J=55 Hz, 2H, CH2F; 5.92, dd, J=5 Hz and 8 Hz, 1H, G- lactam-H; 6.53, s, 2H, CH-arom.; 8.14, s, 1H, CH=N; 8.30, b, 2H, NH; ^.83, d, J=8 Hz, 1H, NH. 1. A compound of formula I wherein Ri denotes hydrogen or an ester-moiety; and R2 and R3 independently of each other denote alkyl, cycloalkyl, aralkyl, aryl, alkenyl or alkinyl and R3 additionally denotes hydrogen. • 7-(((5-Amino-l,2,4-thiadiazoI-3-yl)-(Z)-(fluormethoxyimino)acetyl)amino)-3-((imino*l-piperazinylmethyl)methylhydrazono)methyl-3-cephem-4-carboxylic acid. !. A compound of any preceding claim in in the form of a salt and/or in the form of a solvate. A compound of any preceding claim which is predominantly the 3(E)-isomer. i. A process for the production of a compound of formula I as defined in claim 1, comprising reacting a compound of formula wherein either a) Rb denotes hydroxy and Rc and Rd together form a bond, or P) Rd denotes hydrogen, an ester-moiety or a silyl group, and Rb and Rc together denote an oxo group, with a compound of formula wherein R2 and R3 are as defined in claim 1, splitting off a silyl group from the carboxylic acid group in position 4 of the ring system, and isolating a compound of formula I. 6. A pharmaceutical composition comprising a compound of formula I according to claim 1 in the form of a pharmaceutical^ acceptable salt or in free form in association with at least one pharmaceutical carrier or diluent. 7. A compound of claim 1 or a composition of claim 7 for use as a pharmaceutical. 8. A method of treatment of microbial diseases which comprises administering to a subject in need of such treatment an effective amount of a compound of formula I. 9. Compounds of formula wherein i 1 RT denotes hydrogen, an ester moiety or a cation, in the free form, or, where such forms exist, in the form of acid addition salts, inner salts, quaternary salts or hydrate. i 1 10. A compound substant ially as herein described and exemplified. 11. A process for the production of a compound substantially as herein described and exemplified.

Documents:

511

abs-511-mas-1998.jpg

mas-511-1998-abstract.pdf

mas-511-1998-assignment.pdf

mas-511-1998-calims duplicate.pdf

mas-511-1998-claims original.pdf

mas-511-1998-correspondance others.pdf

mas-511-1998-correspondance po.pdf

mas-511-1998-description complete duplicate.pdf

mas-511-1998-description complete original.pdf

mas-511-1998-form 1.pdf

mas-511-1998-form 13.pdf

mas-511-1998-form 26.pdf

mas-511-1998-form 3.pdf