GB1573614A

GB1573614A - Amidinopenicillanoyloxyalkyl cephalosporanates

Info

Publication number: GB1573614A
Application number: GB7472/76A
Authority: GB
Original assignee: Leo Pharmaceutical Products Ltd AS
Current assignee: Leo Pharma AS
Priority date: 1976-02-25
Filing date: 1976-02-25
Publication date: 1980-08-28
Also published as: FR2342294A1; DK77577A; DE2707577A1; CA1118412A; JPS52102295A; FR2342294B1

Description

(54) AMIDINOPENICILLANOYLOXYALKYL CEPHALOSPORANATES (71) We, LEO PHARMACEUTICAL PRODUCTS LTD. A/S (LIVENS KEMISKE FABRIK PRODUKTION SAKTIESELKAB), a Company organized under the Laws of Denmark, of DK-2750 Ballerup, Denmark, do hereby declare the invention, for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement:- The present invention relates to hitherto unknown esters of the formula I below, to salts of the esters of formula I with pharmaceutically acceptable acids, to methods for producing said new compounds, to pharmaceutical compositions containing said new compounds, and to methods of treating non-human patients suffering from infectious diseases using said new compounds.

The compounds of the invention, which are valuable in human and veterinary practice, have the formula I

in which Rl, R2, and R, each represent an unsub- stituted or substituted aliphatic hydrocarbon radical, a mono- or bicyclic aryl radical, an aralkyl radical, a cycloalkyl radical, a cycloalkylalkyl radical, a heterocyclic radical or a heterocyclically substituted alkyl radical; R, further representing hydrogen; or each of the radicals R1, R2, and R3 can together with one or more of the other radicals form a heterocyclic ring system; R represents hydrogen, or any unsubstituted or substituted alkyl, aralkyl or aryl radical; R represents hydrogen, azido, or an unsubstituted or substituted alkoxy, acyloxy or carbamoyloxy radical or a heterocyclic ring; R can also be Sirs, where R8 stands for an unsubstituted or substituted alkyl, aralkyl or aryl radical or a heterocyclic ring; R, represents an unsubstituted or substituted aliphatic hydrocarbon radical, a mono- or bicyclic aryl radical, an aralkyl radical, a cyclolalkyl radical, a cycloalkylalkyl radical, a heterocyclic radical or a heterocyclically substituted alkyl radical; and R7 represents hydrogen, alkoxy, aralkyloxy, aryloxy, alkylthio, aralkylthio or arylthio.

More particularly, Rl, R2, and R3 may each represent an aliphatic hydrocarbon radical in which the carbon chain can be straight or branched, and saturated or unsaturated, e.g.

methyl, ethyl, propyl, isopropyl, butyl, secbutyl, tert-butyl, pentyl, hexyl, dodecyl, allyl, butenyl, pentenyl or propargyl; a mono- or bicyclic aryl radical, e.g. a phenyl radical or a naphthyl radical; an aralkyl radical, such as a mono- or bicyclic aralkyl radical, e.g. benzyl, phenylethyl or 1- or 2-naphthylmethyl; a cycloalkyl or cycloalkyl-alkyl radical, in which the cycloalkyl group can have 3 to 10 ring members and can be saturated or have one or two double bonds, e.g.

cyclopentyl, cyclohexyl, 1-adamantyl, 1bicyclo(2.2.2.)octyl, cydopentenfl and cyclo hexenyl, cyclopentylmethyl, cyclohexylmethyl, cyclopentenylethyl or cyclohexenylmethyl; a heterocyclic radical or a heterocyclically substituted alkyl radical in which the heterocyclic part can have from 5 to 10 atoms in the ring and can contain oxygen, sulphur, and/or nitrogen atoms, in all of which heterocyclic radicals or heterocyclically substituted alkyl radicals the hetero atoms may be placed in any of the available positions, and all of which may be saturated or unsaturated, e.g. pyridyl, pyrazinyl, pyrimidyl, pyrrolidyl, piperidyl, morpholinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, thiazinyl, furyl, thienyl or quinolyl; each of the radicals R1 to Rs can together with one or more of the other radicals represent heterocyclic radicals having from 5 ta 8 ring atoms and optionally containing other hetero atoms in the ring, such as S, 0 and/or N, forming saturated or unsaturated ring systems, e.g. pyrazolinyl, pyrazolidinyl, 1,2,3, 6-tetrahydropyridazinyl, 1,4,5,6-tetrahydropyridazinyl, hexahydropyridazinyl, piperidyl, morpholinyl, hexahydro - 111 - azepin - 1 - yl, or hexahydro - 1(2H) - azocinyl; and Rs furthermore, as stated above, can be hydrogen.

The radicals R1 to R6 may be further substituted, where it is previously indicated that substitution is possible, with halogen atoms, and with alkyl, haloallryl, hydroxy, alkoxy, aryl, aryloxy, alkylthio, arylthio groups and acyl groups, carboxy, N- cyanocarboximido, carbalkoxy, carbamoyl, carbamido, cyano or organic sulfonyl groups, azido, amino or substituted amino groups.

More particularly R6 stands for alkyl having 1 to 6 carbon atoms, unsubstituted and substituted benzyl, phenoxyalkyl of which the alkyl radical has 1 to 6 carbon atoms, or alkyl having 1 to 6 carbon atoms substituted with 5- or 6-membered heterocyclic ring systems.

In particular, Rl and R2 are selected from alkyl, chloroalkyl and hydroxyalkyl having from 1 to 7 carbon atoms, alkoxyalkyl having from 2 to 7 carbon atoms, carbalkoxyalkyl having 3 or 4 carbon atoms, cyanoalkyl having 2 or 3 carbon atoms, and allyl radicals; phenyl-, chlorophenyl-, bromophenyl-, phen Oxy- or carbobenzoxy-substituted methyl and ethyl radicals; cycloalkyl and cycloalkylmethyl radicals having from 5 to 10 carbon atoms; methyl and ethyl radicals substituted with a 5-membered, unsaturated heterocyclic ring having as the hetero atom an oxygen or sulphur atom, or a 6-membered, unsaturated heterocyclic ring having as the hetero atom a nitrogen atom; or Rl and R2 form a straight or branched alkylene chain which together with the adjacent nitrogen atom form a saturated heterocyclic ring having from 4 to 8 carbon atoms; or R1 and R2 together with the adjacent nitrogen atom form morpholinyl-4, thiomorpholinyl-4, 4-methyl-piperazinyl-1, or 1,2,3,4-tetrahydro-isoquinolyl-2.

In particular, R3 is selected from hydrogen, and an alkyl or alkenyl radical having from 1 to 7 carbon atoms, a phenyl or benzyl radical, a cycloalkyl or a cycloalkylalkyl radical, the cycloalkyl group of which has 5 or 6 carbon atoms and the alkyl group 1 or 2 carbon atoms, a heterocyclic radical consisting of a 5- or 6membered ring containing an oxygen, sulphur or nitrogen atom. Further, R, and Rl when taken together with the N- and C-atoms between them may represent a heterocyclic fiveor six-membered ring system.

In particular, R4 may represent hydrogen, alkyl having 1 to 6 carbon atoms, e.g. methyl, ethyl, propyl, and isopropyl, trichloromethyl, benzyl, phenethyl or phenyL In particular, R, may represent hydrogen, acetoxy, propionyloxy, benzoyloxy, pyridyl, carbamoyloxy, azido, methoxy or SRa in which Rs stands for 1 - methyl - 1,2,3,4 - tetrazol5 - yl, 1,2,3 - (1H) - triazol - 5 - yl, 1,3,4thiadiazol - 2 - yl or 5 - methyl - 1,3,4thiadiazol - 2 - yl.

In particular, R6 may represent methyl, benzyl or substituted benzyl, such as ahydroxy-, a-formyloxy, a-azido, a-amino, acarboxy, a-sulfo, a-sulfoamino-benzyl radicals, a - amino - p - hydroxy-benzyl, and a -aminom - chloro - p - hydroxybenzyl; phenoxymethyl, cyanomethyl, a - amino - 1,4 - cyclohexadienylmethyl, trifiuoromethylthiomethyl, cyanomethylthiomethyl, heterocyclically substituted methyl, such as 2- and 3-thienylmethyl, a - - amino - 2 - he?iiethf, - a amino - 3 thienylmethyl, 1 - (1H) - tetrazo lyl - - methyl, a - methoximino - 2 - furylmethyl, (1,2,3 - oxadiazol - 5 - on - 3 - yl)- methyl or 4-pyridylthiomethyl.

In particular, R, may represent hydrogen, methoxy, and methylthio.

The invention comprises all possible isomeric forms of the compounds of formula I, depending on the different substituents, whereas the 6-aminopenicillanic or 7-aminocephalosporanic acid moieties have the configurations indicated in formula I.

As stated above, the invention also relates to salts of the esters of formula I with pharmaceutically acceptable acids, e.g. hydrochloric and hydrobromic acid, phosphoric acid, sulphuric acid, nitric acid, p-toluenesulphonic acid, methanesulphonic acid, formic add, acetic acid, propionic acid, citric acid, tartaric acid and maleic acid. However, salts with other, in themselves therapeutically active acids, such as penicillins, cephalosporins, and fusidic acid, are also within the scope of the invention.

The compounds of the invention possess valuable in vivo antibacterial activity and their toxicity is low, even when administered parenterally.

It has now been shown that during or after the absorption, the new esters of the invention are hydrolyzed by enzymes present in blood and many tissues with formation of the corresponding free cephalosporins and amidino-penicillanic acids whereby the simultaneous presence of these two types of antibiotics in the organism is obtained. This is particularly advantageous since it is known that cephalosporins and certain 6-amidinopenicillanic acids show a synergistic effect both in vitro and in vivo (E. Grunberg et al.; H. C. Neu; s'). This is probably due to the fact that the two types of antibiotics have a different mode of action. Both interfere with the biosynethesis of the bacterial cell wall, but the target is different (J. T. Park and L.

Burman, Biochem. and Biophys. Research Comm. 51, 863 (1973).

The invention also comprises a method for the preparation of the above described compounds. In one embodiment, an ester of the formula II

in which formula R1, R2, R, and R4 are as defined above, and X is a leaving group, e.g.

a chlorine, bromine or iodine atom or ptoluenesulphonyloxy radical, is reacted with a salt of the formula III

in which R,, R6, and R7 have the above meanings and M+ stands for a cation, e.g. Na+, or or (C2H, )NH+, to form the desired com pound of formula I.

The reaction is performed in a suitable solvent, e.g. N,N-dimethylformamide, acetone, or hexamethylphosphoric acid triamide, usually at a temperature from about 0 C to about 60"C.

In the above embodiment, it is preferred to use a reactant of formula II with X repre senting iodine because this results in a product mainly consisting of a compound in which the double bond of the cephalosporin moiety is in the 3-position. This is due to the speed of the reaction which does not permit the undesired migration of the double bond to the 2-position which would result in compounds containing an inactive cephalosporin moiety.

In a second embodiment of the method of the invention, an ester of the formula II is Presented at the Fifteenth Interscience Conference on Antimicrobial Agents and Chemotherapy. in Washington, D.C., September 24-26th, 1975.

reacted with a compound of formula III in which the S atom is oxidized to the > S30 grouping, to form a compound of formula IV

in which R1 to R7 have the above defined meanings, said compound being subjected to a reduction process to form the desired compound of formula I.

The reaction is performed in a suitable solvent, e.g. methylene chloride or N,Ndimethylformamide, usually at a temperature of from about 0 C to about 600C.

The reduction is performed by methods wellknown from penicillin and cephalosporin chemistry; confer, e.g., J. Org. Chem. 35, (1970), p. 2433.

In a third embodiment, a compound of formula V

in which R4 is as defined above, and R, is a protective group, e.g. benzyloxycarbonyl, triphenylmethyl, or ss"B"B-trichloroethoxycar- bonyl, is converted into the corresponding iodoalkyl ester by treatment with sodium iodide in acetone, said iodoalkyl ester being reacted with a compound of formula III to yield a compound of formula VI

in which R4 to R7 and R9 have the above defined meaning.By removal of the protective group R9, a compound of formula VII is obtained:

in which R4 to R, are as defined above, and said compound is thereafter reacted with a reactive derivative of an amide or thioamide of the formula VIII

in which Rl, R2 and R3 have the above meanings, and Z stands for oxygen or sulphur, to yield the desired compound of formula I.

As examples of reactive derivatives of a compound of formula VIII, the following nonlimiting types may be given:

Ci-rmlnum chlorides

BFt tmmum ethers

thioamide acetals in which Rl, R2, and R, have the above 'meanings.

The reactions with the said reactive derivatives are well-known to the man skilled in the art for preparing amidinopenicillanic acid derivatives; confer, e.g., British Patent Specifi cation No. 1,293,590 which also describes in detail the meaning of "a refractive derivative of a compound of formula VIII".

The starting materials of formulae II, III and VIII are known or may be prepared by methods analogous to those used for the preparation of known compounds.

The starting material of formula V can, e.g., be prepared as follows: a salt of a Nprotected 6-amino-penicillanic acid is reacted with a compound of formula IX:

where R4 is as previously defined and Y is a leaving group, e.g. a bromine or iodine atom or a p-toluenesulphonyloxy group. The reaction is performed in a suitable solvent, e.g.

dimethylformamide or acetone, and usually at a temperature between 0 C and 60"C.

The reaction products of formula I can be purified and isolated in the usual manner and may be obtained either in the free state or in the form of a salt.

It is a further object of the present invention to provide pharmaceutical compositions which are useful in the treatment of infectious diseases in human and veterinary practice.

With this object in view, the compositions of the invention contain as an active component at least one member selected from compounds of the formula I and salts thereof as defined above, together with solid or liquid pharmaceutical carriers and/or diluents.

In the said compositions, the proportion of therapeutically active material to carrier substance can vary between 1% and 95 /O by weight. The compositions can be worked up to various pharmaceutical forms of presentation, such as tablets, pills, dragees, suppositories, capsules, sustained-release tablets, suspensions, ointments and preparations for injection containing the compounds of formula I or their atoxic salts, mixed with carriers and/or diluents.

Pharmaceutical organic or inorganic, solid or liquid carriers and/or diluents suitable for enteral, parenteral, or topical administration can be used to make up compositions containing the present compounds. Gelatine, lactose, starch, magnesium stearate, talc, vegetable and animal fats and oils, gum, polyalkylene glycol, buffers or other known carriers and/or diluents for medicaments are all suitable.

Furthermore, the compositions may contain other pharmaceutically active components which can appropriately be administered together with the present compounds in the treatment of infectious diseases, such as other suitable antibiotics.

As indicated above, the present compounds may be worked up to pharmaceutical forms of presentation including suspensions and nonaqueous ointments and creams. A pharmaceutical preparation may be in the form of a solution or suspension of at least one of the present compounds as active ingredient, the active ingredient being dissolved or suspended in a non-toxic pharmaceutically acceptable liquid carrier. The preparation may contain from 10 to 100 mg per ml of a non-aqueous vehicle. A further preparation for oral treatment may be in the form of a suspension of at least one compound of formula I as such or of a sparingly soluble salt, said preparation containing from 25 to 100 mg per ml of non-aqueous vehicle.A pharmaceutical preparation for topical treatment may be in the form of a powder or an ointment, preferably non-aqueous ointment, or cream containing at least one compound of formula I or a salt thereof as defined above in an amount of from 1/4 to 10 g per 100 g of preparation.

Another object of the invention resides in the selection of a dose of the compounds of the invention, which dose can be administered so that the desired activity is achieved without simultaneous secondary effects. In human therapy, the present compounds are conveniently administered (to adults) in dosage units containing not less than 25 mg and up to 1000 mg, preferably from 100 to 500 mg, calculated as the compound of formula I.

By the term "dosage unit" is meant a unitary, i.e. a single, dose which is capable of being administered to a patient, and which may be readily handled and packed, remaining as a physically stable unit dose comprising either the active material as such or a mixture of it with solid or liquid pharmaceutical diluents or carriers.

Accordingly, another aspect of the present invention provides a pharmaceutical preparation in dosage unit form for the enteral or parenteral treatment of patients suffering from infectious diseases, which comprises as an active ingredient at least one compound of formula I, or a pharmaceutically acceptable, non-toxic salt thereof, and an atoxic pharmaceutically acceptable carrier, the quantity of the said active compound being between 0.025 g and 1 g.

In the form of a dosage unit, the compound may be administered one or more times a day at appropriate intervals, always depending, however, on the condition of the patient, and in accordance with the prescription made by the medical practitioner.

Thus a daily dose will preferably be an amount of from 1 to 3 g of a compound of formula I or an equivalent amount of a salt thereof as defined before.

For oral treatment, the pharmaceutical preparation may be in the form of a sustainedrelease preparation in dosage unit form of at least one compound of formula I or a pharmaceutically acceptable, non-toxic salt thereof, and an atoxic pharmaceutically acceptable carrier in which the dose of the active compound is from 0.025 g to 1 g.

If the composition is to be injected, a sealed ampoule, a vial or a similar container may be provided containing a parenterally acceptable aqueous or oily injectable solution or dispersion of the active material as the dosage unit.

The dosage unit may be in the form of a parenteral pharmaceutical preparation containing from 0.025 g to 1 g of at least one compound of formula I, or a salt thereof as defined above as dry matter, in an ampoule, vial or other suitable receptacle, for reconstitution.

The parenteral preparations are in particular useful in the treatment of conditions in which a quick response to the treatment is desirable. In the continuous therapy of patients suffering from infectious diseases, tablets or capsules may be the appropriate form of pharmaceutical preparation.

In the treatment of infectious diseases, such tablets may advantageously contain other active components. As mentioned hereinbefore, the other active components may be antibiotics, such as penicillanic acid derivative, a cephalosporanic acid derivative, a fusidic acid derivative or a tetracycline derivative. That is to say an antibiotic derived from penicillanic acid, cephalosporanic acid, fusidic acid or a tetracycline.

The present compounds may be used for treating patients suffering from infectious diseases by administering to adult patients from 0.5 to 5 g per day, preferably from 1 to 3 g per day, of at least one compound of the formula I or an equivalent amount of a salt as defined before of a compound of the formula I. For an average human being weighing 70 kg., these ranges are from 7 to 70, preferably 14 to 43, mg/kg body weight/day. Still another object of the invention is to provide a method of treating non-human patients suffering from infectious diseases, the method comprising administering to the patients from 7 to 70, preferably 14 to 43, mg/kg body weight/day of at least one compound of the formula I or an equivalent amount of a salt thereof as defined above.

Preferably, the compound is given in the form of the dosage units aforesaid.

The invention will be further described in the following Examples which are not to be construed as limiting the invention.

Example 1.

Iodomethyl 6 - [(hexahydro - 1H - azepin 1 - yl) - methylene - amino] - penicil lanate.

To a solution of chloromethyl 6 - [(hexahydro - 1H - azepin - 1 - yl) - methylene amino] - penicillanate (0.5 g) in acetone (4 ml) was added sodium iodide (0.8 g). The mixture was stirred at 40 C for 1 hour, cooled and fitered. The filtrate was diluted with ether and washed repeatedly with water. The organic phase was dried and evaporated to leave the desired compound as an oil, which was used in the next step without purification.

The NMR-spectrum (CDCl8) showed signals at #=1.55 (s), 1.70 (s), 4.37 (s), 1.5-2.0 (m), 3.3-3.6 (m), 5.20 (dd, J=1, J=4), 5.50 (d, J=4), 5.90 (d, J=6), 6.03 (d, J=6) and 7.68 (s) ppm.

TMS was used as internal standard.

Example 2.

6 - [(Hexahydro - 1H - azepin - 1 - yl) methyleneamino] - penicillanoyloxy methyl 7 - (D - α - azidophenylacet amido) - cephalosporanate, hydrochloride.

To an ice-cold solution of iodomethyl 6 [(hexahydro - 1H - azepin - 1 - yl)methyleneamino] - penicillanate (1.19 g) in dimethylformamide (20 ml), potassium 7 - (D a - azidophenylacetamido) - cephalosporanate (1.5 g) was added. After stirring for 10 minutes the mixture was diluted with ethyl acetate and washed repeatedly with water.

The organic phase and water were stirred while hydrochloric acid was added to pH=4.

The aqueous phase containing some impurities was discarded and the organic phase was stirred with water while hydrochloric acid was added to pH=2.5. The aqueous phase was freeze-dried to yield the title compound as a colourless powder.

The IR-spectrum (KBr) showed strong bands at 2110, 1780, 1740 and 1680 cm1 The NMR-spectrum (CD,OD, TMS as intemal reference) showed peaks at 3=1.58 (s), 1.73 (s), 1.4-2.2 (m), 2.05 (s), 3.55 (m), 3.6-4.0 (m), 4.63 (s), 4.82 (d, J= 14), 5.08 (d, J=14), 5.10 (d, J=5), 5.15 (s), 5.4-5.9 (m), 5.99 (s), 7.43 (s) and 8.17 (bs) ppm.

Example 3.

6 - [(Hexahydro - 1H - azepin - 1 yl) methyleneamino] - penicillanoyloxy methyl 7 - (D - α - aminophenylacet amido) - cephalosporanate, dihydro chloride.

To a stirred solution of the compound prepared according to Example 2 (0.5 g) in water (25 ml), 10% Pd-C (0.2 g) was added.

Hydrogen was bubbled through the mixture while a pH-value of 3 was maintained by the addition of hydrochloric acid. When the consumption of acid stopped the catalyst was filtered off and the filtrate was freeze-dried to yield the title compound as a colourless powder.

The IR-spectrum (KBr) showed strong bands at 1775, 1740 and 1685 cm-l.

The NMR-spectrum (CD30D, TMS as internal reference) showed peaks at #=1.57 (s), 1.73 (s), 1.5-2.2 (m), 2.03 (s), 3.53 (m), 3.5-4.0 ,m), 4.62 (s), 5.11 (s), 4.85.3 (m), 5.5-6.0 (m), 5.98 (s), 7.50 (s) and 8.20 (s) ppm.

Examples 4-15.

By following the procedures described in Example 2 and/or 3, but replacing potassium 7 - (D - α - azidophenylacetamido)cephalosporanate with another compound of formula III, the compounds of formula I in Table I are prepared in the form of their hydrochlorides.

TABLE I

Ex. No. R1 + R2 R3 R4 R5 R6 R7 4 1,6-hexamethylene H H (5-methyl-1,3,4-thiadiazol- 1-(1H)-tetrazolyl-methyl H 2-yl)-thio 5 do do do (1-methyl-1,2,3,4-tetrazol- α-hydroxybenzyl H 5-yl)-thio 6 do do do H α-amino-p-hydroxybenzyl H 7 do do do acetoxy 2-thienylmethyl H 8 do do do carbamoyloxy 2-thienylmethyl methoxy 9 do do do acetoxy cyanomethyl H 10 do do do acetoxy 4-pyridylthiomethyl H 11 do do do H α;-aminobenzyl H 12 do do do H α-amino-1,4-cyclohexadienyl- H methyl 13 do do do (1-methyl-1,2,3,4-tetrazol- trifluoromethylthio- H 5-yl)-thio methyl 14 do do do carbamoyloxy α-methoximino-2-furyl- H methyl 15 do do do (1,2,3-(1H)-triazol-5-yl)-thio α-amino-p-hydroxybenzyl H Example 16.

Chloromethyl 6-(N-triphenylmethylamino ) penicillanate.

To a solution of 6 - (N - triphenylmethylamino) - penicallanic acid (5 g) in dimethylformamide (50 ml) was added triethylamine (1.53 ml) and chloroiodo-methane (5 ml).

After stirring at room temperature for 3 hours, the mixture was diluted with ethyl acetate (200 ml) and washed with water (4 X 50 ml). The organic phase was dried and evaporated to leave chloromethyl 6 - (N - triphenylmethylamino) - penicillanate as a yellow oil.

The NMR-spectrum (CDCls) showed signals at 6=1.38 (s), 1.58 (s), 3.18 (d, J=11), 4.38 (s), 4.55 (m), 5.60 (d, J=6, 5.75 (d, J=6) and 7.1-7.7 (m) ppm. TMS was used as internal standard.

Example 17.

Iodomethyl 6- (N-triphenylmethylamino ) penicillanate.

To a solution of chloromethyl - 6 - (Ntriphenylmethylamino) - penicillanate (0.65 g) in acetone (4 ml) was added sodium iodide (0.8 g). The mixture was stirred at 400C for 3 hours, cooled and the precipitated sodium chloride filtered off. The filtrate was diluted with ether and washed repeatedly with water. The organic phase was dried and evaporated to leave a yellow oil, which was used in the next step without purification. The NMR-spectrum (CDCl,) showed signals at =1.38 (s), 1.57 (s), 3.15 (m), 4.33 (s), 4.45 (m), 5.77 (d, J=6), 5.92 (d, J=6) and 7.1-7.7 (m) ppm. TMS was used as internal standard.

Example 18.

6 - (N - triphenylmethylamino) - penicillan oyloxymethyl 7 - [1 - (1H) - tetrazoyl acetamido] - 3 - [(5 - methyl - 1,3,4 thiadiazol - 2 - yl) - thiomethyl] - 3 cephem - 4 - carboxylate.

To an ice-cold solution of iodomethyl 6 (N - triphenylmethylamino) - penicillanate (0.29 g) in dimethylformamide (6 ml), sodium 7 - [1 - (1H) - tetrazolylacetamido] 3 - [(5 - methyl - 1,3,4 - thiadiazol - 2 - yl) thiomethyl] - 3 - cephem - 4 - carboxylate (cefazoline sodium) (0.23 g) was added.

After stirring for 10 minutes the mixture was diluted with ethyl acetate and washed several times with water. The organic phase was dried and evaporated to leave the desired compound as an oil.

Thin-layer chromatography (SiO2-plates, solvent system: ethyl acetate) showed after 2 consecutive runs one spot with a Rf-value of 0.53.

Example 19.

6 - Amino - penicillanoyloxymethyl 7 - (1- (1H) - tetrazolylacetamido] - 3 - [(5 methyl - 1,3,4 - thiadiazol - 2 - yl) thiomethyl] - 3 - cephem - 4 - carb oxylate.

A solution of the compound prepared in Example 18 (0.1 g) in acetone (2 ml) was treated with p-toluenesulphonic acid (21 mg) at room temperature for 15 minutes. Then aqueous sodium bicarbonate and ethyl acetate were added. The organic phase was separated and washed with water, dried and evaporated to yield an oil which was used in the next step without further purification.

Example 20.

6 1 [(Hexahydro - 1H- azepin - 1 - yl) methyleneamino] - penicillanoyloxy methyl 7 [1 - (1H) - tetrazolylacet amido] - 3 - ([(5 - methyl - 1,3,4 thiadiazol - 2 - yl) - thiomethyl] - 3 cephem - 4 - carboxylate, hydrochloride.

A solution of 6 - amino - penicillanoyloxymethyl 7 - [1 - (1H) - tetrazolylacetamido] 3 - [(5 - methyl - 1,3,4 - thiadiazol - 2yl) - thiomethyl] - 3 - cephem - 4 - carboxylate (1.75 g) in alcohol-free chloroform was cooled to -400C and pyridine (0.43 ml) was added. A solution of chlorohexamethyleneformiminium chloride (0.95 g) in alcoholfree chloroform was added dropwise and the mixture was stirred for 0.5 hour at --200C.

After stirring for a further 15 minutes at 0 C, the solvent was evaporated and the residue was treated with ethyl acetate and aqueous sodium bicarbonate. The organic phase was separated and washed with water.

The organic phase was stirred with water while hydrochloric acid was added to pH= 2.5. The aqueous phase was separated and freeze-dried to yield the desired compound which was identical with that prepared in Example 4.

The compound was homogeneous according to thin-layer chromatography and a bioautogram showed only one active compound.

The NMR-spectrum (CD3OD, TMS as internal reference) showed peaks at 6=1.60 (s), 1.73 (s), 1.4-2.1 (m), 2.75 (s), 3.60 (m), 3.6-3.9 (m), 4.65 (s), 4.20 (d, J= 14), 4.65 (d, J=14), 5.16 (d, J=6), 5.46 (m), 5.4-5.9 (m), 6.02 (m), 8.24 (s) and 9.18 (s) ppm.

Examples 21-26.

By following the procedures described in Examples 18-20 but replacing cefazoline sodium with another compound of formula III and chlorohexamethylene-formiminium chloride with another reactive derivative of a compound of formula VIII the compounds of formula I in Table II and Table III are prepared in the form of their hydrochlorides. TABLE II

Ex No. R1 + R2 R3 R4 R5 R6 R7 21 1,6-hexamethylene methyl H acetoxy cyanomethyl H 22 1,5-pentamethylene H H (1-methyl-1,2,3,4-tetrazol- α-hydroxybenzyl H 5-yl)-thio 23 1,7-heptamethylene H H (5-methyl-1,3,4-thiadiazol- 1-(1H)-tetrazolylmethyl H 2-yl)-thio TABLE III

Ex. No. R1 R2 R3 R4 R5 R6 R7 24 methyl methyl H H (5-methyl-1,3,4-thiadiazol- 1-(1H)-tetrazolylmethyl H 2-yl)-thio 25 ethyl ethyl phenoxy- H (1-methyl-1,2,3,4-tetrazol- α-hydroxybenzyl H methyl 5-yl)-thio 26 methyl methyl benzyl H carbamoyloxy 2-thienylmethyl methoxy Example 27.

Enzymatic hydrolysis of 6 - [(hexahydro 1H - azepin -1 - yl) - methyleneamino] penicillanoyloxymethyl 7 - [1 - (1H)-.

tetrazolylacetamido) - 3 - [(5 - methyl- 1,3,4 thiadiazol - 2 - yl) - thiomethyl] 3 - cephem - 4 - carboxylate, hydro chloride (A).

(A) was added to heparinized human blood at a concentration of 0.5 mg/ml and the mixture was incubated at 370C for 30 mintues.

After ultrafiltration the ultrafiltrate was subjected to paper chromatography together with standard solutions of cefazoline sodium (B), 6 - [hexahydro - 1H - azepin - 1 - yl)- methyleneamino] - penicillanic acid (C) and (A).

Solvent systems: a) butanol: acetic acid: water (4:1:5).

b) ethyl acetate (saturated with acetate buffer, pH=5.6).

The paper chromatograms from a) and b) were placed on agar plates seeded with E.

coli and Staph. aureus respectively. After incubation overnight at 370 C both plates showed no zone of inhibition in the ultrafiltrate corresponding to (A).

The E. coli plate showed a zone of inhibition in the ultrafiltrate with the same Rfvalue as that of (C). The Staph. plate showed a zone of inhibition in the ultrafiltrate with the same Rf-value as that of (B).

These results indicate that the title compound (A) is hydrolyzed into (B) and (C) by enzymes present in human blood.

Example 28.

By using the technique described in Example 27, the compounds described in Examples 215 and 2()-'26 under the influence of enzymes present in human blood are cleaved into the corresponding free cephalosporins and amidinopenicillanic acids.

Example 29.

Ingredients for 1000 tablets: 6 - [(Hexahydro - lH - azepin - 1 - yl)- methyleneamino] - penicillanoyloxymethyl 7 [ 1 - (1H) - tetrazolylacetamido] - 3 - [(5- methyl - 1,3,4 - thiadiazol - 2 - yl) - thio methy] - 3 - cephem - 4 - carboxylate, hydro chloride (A) 250 g Polyvinylpyrrolidone 10 g Isopropanol 10 ml Microcrystalline cellulose 165 g Corn starch 71 g Magnesium stearate 4 g Sieve A through a screen with 1 mm openings and wet the powder with a solution of polyvinylpyrrolidone in isopropanol. Dry the moist mass at 30"C and pass it through a sieve with 0.7 mm openings.

Mix the granules with microcrystalline cellulose, corn starch and magnesium stearate.

Press tablets of 0.500 g weight using 12 mm punches and dies to yield tablets each containing 250 mg of A.

WHAT WE CLAIM IS:- 1. A compound of the formula I

in which R1 K2, and R2 each represent an unsubstituted or substituted aliphatic hydrocarbon radical, a mono- or bicyclic aryl radical, an aralkyl radical, a cycloalkyl radical, a cycloalkylalkyl radical, a heterocyclic radical or a heterocyclically substituted alkyl radical; R2 further representing hydrogen; or each of the radicals Rl, R2, and R3 can together with one or more of the other radicals form a heterocyclic ring system; R4 represents hydrogen, or an unsubstituted or substituted alkyl, aralkyl or aryl radical; R5 represents hydrogen, azido, or an unsubstituted or substituted alkoxy, acyloxy or carbamoyloxy radical, or a heterocyclic ring; R5 can also be Sirs, where R8 stands for an unsubstituted or substituted alkyl, aralkyl or aryl radical or a heterocyclic ring; K6 represents an unsubstituted or substituted aliphatic hydrocarbon radical, a mono- or bicyclic aryl radical, an aralkyl radical, a cycloalkyl radical, a cycloalkylalkyl radical, a heterocyclic radical or a heterocyclically substituted alkyl radical; and R7 represents hydrogen, alkoxy, aralkyloxy, aryloxy, alkylthio, aralkylthio or arylthio; and salts thereof with pharmaceutically acceptable acids.

2. A compound according to claim 1, in which Rl and R2 are selected from alkyl, chloroalkyl and hydroxyalkyl having from 1 to 7 carbon atoms, alkoxyalkyl having from 2 to 7 carbon atoms, carbalkoxyalkyl having 3 or 4 carbon atoms, cyanoalkyl having 2 or 3 carbon atoms, and allyl radicals; phenyl-, chlorophenyl-, bromophenyl-, phenoxy- or

**WARNING** end of DESC field may overlap start of CLMS **.

Claims

**WARNING** start of CLMS field may overlap end of DESC **.

Example 27.

Enzymatic hydrolysis of 6 - [(hexahydro 1H - azepin -1 - yl) - methyleneamino] penicillanoyloxymethyl 7 - [1 - (1H)-.

tetrazolylacetamido) - 3 - [(5 - methyl- 1,3,4 thiadiazol - 2 - yl) - thiomethyl]

3 - cephem - 4 - carboxylate, hydro chloride (A).

(A) was added to heparinized human blood at a concentration of 0.5 mg/ml and the mixture was incubated at 370C for 30 mintues.

After ultrafiltration the ultrafiltrate was subjected to paper chromatography together with standard solutions of cefazoline sodium (B), 6 - [hexahydro - 1H - azepin - 1 - yl)- methyleneamino] - penicillanic acid (C) and (A).

Solvent systems: a) butanol: acetic acid: water (4:1:5).

b) ethyl acetate (saturated with acetate buffer, pH=5.6).

The paper chromatograms from a) and b) were placed on agar plates seeded with E.

coli and Staph. aureus respectively. After incubation overnight at 370 C both plates showed no zone of inhibition in the ultrafiltrate corresponding to (A).

The E. coli plate showed a zone of inhibition in the ultrafiltrate with the same Rfvalue as that of (C). The Staph. plate showed a zone of inhibition in the ultrafiltrate with the same Rf-value as that of (B).

These results indicate that the title compound (A) is hydrolyzed into (B) and (C) by enzymes present in human blood.

Example 28.

By using the technique described in Example 27, the compounds described in Examples 215 and 2()-'26 under the influence of enzymes present in human blood are cleaved into the corresponding free cephalosporins and amidinopenicillanic acids.

Example 29.

Ingredients for 1000 tablets:

6 - [(Hexahydro - lH - azepin - 1 - yl)- methyleneamino] - penicillanoyloxymethyl 7 [ 1 - (1H) - tetrazolylacetamido] - 3 - [(5- methyl - 1,3,4 - thiadiazol - 2 - yl) - thio methy] - 3 - cephem - 4 - carboxylate, hydro chloride (A) 250 g Polyvinylpyrrolidone 10 g Isopropanol 10 ml Microcrystalline cellulose 165 g Corn starch 71 g Magnesium stearate 4 g Sieve A through a screen with 1 mm openings and wet the powder with a solution of polyvinylpyrrolidone in isopropanol. Dry the moist mass at 30"C and pass it through a sieve with 0.7 mm openings.

Mix the granules with microcrystalline cellulose, corn starch and magnesium stearate.

Press tablets of 0.500 g weight using 12 mm punches and dies to yield tablets each containing 250 mg of A.

WHAT WE CLAIM IS:- 1. A compound of the formula I

in which R1 K2, and R2 each represent an unsubstituted or substituted aliphatic hydrocarbon radical, a mono- or bicyclic aryl radical, an aralkyl radical, a cycloalkyl radical, a cycloalkylalkyl radical, a heterocyclic radical or a heterocyclically substituted alkyl radical; R2 further representing hydrogen; or each of the radicals Rl, R2, and R3 can together with one or more of the other radicals form a heterocyclic ring system; R4 represents hydrogen, or an unsubstituted or substituted alkyl, aralkyl or aryl radical; R5 represents hydrogen, azido, or an unsubstituted or substituted alkoxy, acyloxy or carbamoyloxy radical, or a heterocyclic ring; R5 can also be Sirs, where R8 stands for an unsubstituted or substituted alkyl, aralkyl or aryl radical or a heterocyclic ring; K6 represents an unsubstituted or substituted aliphatic hydrocarbon radical, a mono- or bicyclic aryl radical, an aralkyl radical, a cycloalkyl radical, a cycloalkylalkyl radical, a heterocyclic radical or a heterocyclically substituted alkyl radical; and R7 represents hydrogen, alkoxy, aralkyloxy, aryloxy, alkylthio, aralkylthio or arylthio; and salts thereof with pharmaceutically acceptable acids.
2. A compound according to claim 1, in which Rl and R2 are selected from alkyl, chloroalkyl and hydroxyalkyl having from 1 to 7 carbon atoms, alkoxyalkyl having from 2 to 7 carbon atoms, carbalkoxyalkyl having 3 or 4 carbon atoms, cyanoalkyl having 2 or 3 carbon atoms, and allyl radicals; phenyl-, chlorophenyl-, bromophenyl-, phenoxy- or

carbobenzoxy-substituted methyl and ethyl radicals; cycloalkyl and cycloalkylmethyl radicals having from 5 to 10 carbon atoms; methyl and ethyl radicals substituted with a 5-membered, unsaturated heterocyclic ring having as the hetero atom an oxygen or sulphur atom, or a 6-membered, unsaturated heterocyclic ring having as the hetero atom a nitrogen atom; or R1 and R2 form a straight or branched alkylene chain which together with the adjacent nitrogen atom form a saturated heterocyclic ring having from 4 to 8 carbon atoms; or R1 and 2 together with the adjacent nitrogen atom form a morpholinyl-4, thiomorpholinyl-4, 4-methyl-piperazinyl-1, or 1,2,3-4-tetrahydro-isoquinolyl-2 radical; R3 is selected from hydrogen, and an alkyl or alkenyl radical having from 1 to 7 carbon atoms, a phenyl or benzyl radical, a cycloalkyl or a cycloalkylalkyl radical, the cycloalkyl group of which has 5 or 6 carbon atoms and the alkyl group 1 or 2 carbon atoms, a heterocyclic radical consisting of a 5 or 6membered ring containing an oxygen, sulphur or nitrogen atom; or R, and Rl when taken together with the N- and C-atoms between them may represent a heterocyclic five- or sixmembered ring system; R4 may represent hydrogen, alkyl having from 1 to 6 carbon atoms, trichloromethyl, benzyl, phenethyl or phenyl; R, may represent hydrogen, acetoxy, propionyloxy, benzoyloxy, pyridyl, carbamoyloxy, azido, methoxy or SR, in which R, stands for 1 - methyl - 1,2,3,4 - tetrazol - 5 - yl, 1,2,3 - (1H) - triazol - 5 - yl, 1,3,4 - thiadiazol - 2 - yl or 5 - methyl - 1,3,4 - thiadiazol - 2 - yl;; R6 may represent methyl, benzyl or substituted benzyl, phenoxymethyl, cyanomethyl, a - amino - 1,4 - cyclohexadienylmethyl, trifiuoromethylthiomethyl, cyanomethylthiomethyl, 2- and 3-thienylmethyl, a-ammo - 2thienylmethyl, a - amino - 3 - thienylmethyl, 1 - (1H) - tetrazolyl - methyl, a methoximino -2-furylmethyl, (1,2,3 - oxadiazol5 - on - 3 - yl) - methyl or 4 - pyridylthiomethyl and R, may represent hydrogen, methoxy or methylthio; and salts thereof with pharmaceutically acceptable acids.
3. A compound according to claim 2, in which R6 is selected from a-hydroxy-, a-formyloxy-, a-azido, a-amino-, a-carboxy-, a-sulpho- and a-sulphamino-benzyl radicals, a - amino - p - hydroxybenzyl and a - aminom - chloro - p - hydroxybenzyl radicals.
4. -6 - [(Hexahydro - 1H - azepin - 1yl) - methyleneamino] - penicillanoyloxymethyl 7 - [1 - (1H) - tetrazolylacetamide]- 3 - [(5 - methyl - 1,3,4 - thiadiazol - 2 - yl)thiomethyl] - 3 - cephem - 4 - carboxylate; and salts thereof with pharmaceutically acceptable acids.
5. 6 - [(Hexahydro - 1H - azepin - 1yl) - methyleneamino] - penicillanoyloxymethyl 7 - (D - a aminophenylacetamido)cephalosporanate; and salts thereof with pharmaceutically acceptable acids.
6. 6 - [(Hexahydro - 1H - azepin - 1yl) - methyleneamino] - penicillanoyloxymethyl 7 - (D - a -azidophenylacetamido) - cephalosporanate; and salts thereof with pharmaceutically acceptable acids.
7. Method for producing a compound of formula I of claim 1 in which a) an ester of the formula II

in which formula R,, R2, R, and R4 are as defined in claim 1, and X is a leaving group, is reacted with a salt of the formula III

in which R,, R6, and R7 have the meanings defined in claim 1 and M+ stands for a cation, to form the desired compound of formula I; or b) an ester of formula II above is reacted with a compound of Formula III above in which the
S atom is oxidized to the > SoO grouping to form a compound of formula IV.

in which R1 to R, have the meanings defined in claim 1, and the compound of formula IV is subjected to a reduction process to form the desired compound of formula I; or c) a compound of formula V

in which R, is as defined in claim 1, and R9 is a protective group is converted into ihe corresponding iodoalkyl ester by treatment with sodium iodide in acetone, said iodoalkyl ester being reacted with a compound of formula III to yield a compound of formula VI

in which R to R, have the meanings defined in claim 1 and R9 has the above defined meanings; and by removal of the protective group R9, a compound of formula VII is obtained:

in which K, to K7 are as defined in claim 1, and said compound is thereafter reacted with a reactive derivative of an amide or thioamide of the formula VIII

in which K1, R, and R have the meanings defined in claim 1, and Z stands for oxygen or $t, to yield the desired compound of form - l! 8. A method ab claimed iI1 claim 7, in which X is a chlorine, bromine or iodine atom, or a p-toluenesulphonyloxy radical.
9. A method as claimed in claim 7, in which the cation M stands for Na+, K+ or ( C2Ha ) 3NH+.
10. A method as claimed in claim 7, in which R9 is a benzyloxycarbonyl, triphenylmethyl or .R,P,P - trichloroethoxycarbonyl radical.
11. A pharmaceutical preparation in dosage unit form for the enteral or parenteral treatment of patients suffering from infectious diseases, which comprises as an active ingredient at least one compound of formula I, or a pharmaceutically acceptable, non-toxic salt thereof, and an atoxic pharmaceutically acceptable carrier, the quantity of the said active compound being between 0.025 g and 1 g.
12. A preparation as claimed in claim 11 wherein the dosage unit contains from 0.1 g to 0.5 g of at least one compound of formula I or a salt thereof as defined in claim 1.
13. A preparation as claimed in claim 11 or 12 wherein the dosage unit is in the form of a tablet.
14. A preparation as claimed in claim 11 or 12 wherein the dosage unit is in the form of a capsule.
15. A parenteral pharmaceutical preparation in dosage unit form, containing from 0.025 g to 1 g of at least one compound of formula I of claim 1 or a salt thereof as defined in claim 1 as dry matter, in an amopule, vial or other suitable receptacle, for reconstitution.
16. A preparation according to claim 11, wherein the active ingredient is dissolved or suspended in a non-toxic, pharmaceutically acceptable liquid carrier
17. A pharmaceutical preparation for oral treatment in the form of a sustained-release preparation in dosage unit form of at least one compound of formula I claim 1 or a pharmaceutically acceptable, non-toxic salt thereof and an atoxic pharmaceutically acceptable carrier, in which the dose of the active compound is from 0.025 g to 1 g.
18. A preparation for oral treatment in the form of a suspension of at least one compound of formula I of claim 1 as such or a sparingly soluble salt, said preparation containing from 25 to 100 mg per ml of a non-aqueous vehicle.
19. A preparation for topical treatment in the form of a powder or an ointment or cream containing at least one compound of formula I or a salt as defined in claim 1 in an amount of from 1/4 g to 10 g per 100 g of the preparation.
20. A preparation as claimed in any one of claims 10 to 18 in which the dosage unit additionally contains other antibiotics.
21. A preparation according to claim 20 which in addition to at least one compound of formula I or a salt thereof as defined in claim 1 contains a penicillanic acid derivative.
22. A preparation according to claim 20 which in addition to at least one compound of formula I or a salt thereof as defined in claim 1 contains a cephalosporanic acid derivative.
23. A preparation according to claim 20 which in addition to at least one compound of formula I or a salt thereof as defined in claim 1 contains a fusidic acid derivative.
24. A preparation according to claim 20 which in addition to at least one compound of formula I or a salt thereof as defined in claim 1 contains a tetracycline derivative.
25. A method of treating non-human patients suffering from infectious diseases, comprising administering to patients from 7 to 70 mg/kg body weight/day of at least one compound of formula I of claim 1 or an equivalent amount of a salt thereof as defined in claim 1.
26. A method according to claim 24 in which the dose is from 14 to 43 mg/kg body weight/day.
27. A method according to claim 25 or 26, in which the compound or salt is administered in a preparation according to any one of the claims 11 to 24.
28. A compound of the formula I defined in claim 1 substantially as hereinbefore described in any one of the foregoing Examples.