NL8601026A - Piperazine-carboxamido-acetamido-cephalosporin derivs. - broad spectrum antibacterials stable towards beta-lactamase - Google Patents

Abstract

Cephalosporins of formula (I) and their salts are new. In (I) R1 is H or protecting gp. R2 is organic gp. bonded via an O or S atom. esp. acetoxy or a substd. 5-(1,2,3,4-tetrazolyl)thio or 2-(1,2,3,4(sic)-thiadiazolyl)thio. R3 is lower alkyl. n is 0-2.A is H or opt. substd. alkyl. B is alkyl, cycloalkyl, cycloalkenyl, cycloalkadienyl or heterocyclyl, all opt. substd. They are prepd. e.g. by reacting a 7-(alpha-aminoacetylamino)cephem with a reactive deriv. of 4-A-2,3-dioxo-piperazine-1-carboxylic acid. (I) are esp. effective against Pseudomonas aeruginosa, Klebsiella pneumoniae and Proteus. They are useful in human or veterinary medicine. A typical cpd. 7 beta- D(-)-alpha-(4-ethyl-2,3-dioxo-1,piperazinecarboxamido)-alph- a-(2-thienyl)-acetamido -7 alpha-methoxy-3 5-(1-methyl,1,2,3,4-tetrazoyl)thiomethyl -DELTA3-- cephem-4-carboxylic acid (Ia) has intravenous LD50 in mice =5g/kg and has ID50 against beta-lactamase from E. coli GN-5482 of 0.82 mu g/ml.

Description

t s'

Novel 7x-methoxycephalosporins and method for their preparation.

The invention relates to new 7a-methoxy-cephalosporins and to processes for the preparation of these compounds.

The compounds of the invention are characterized by a broad antibacterial spectrum against gram-positive and negative bacteria and by their resistance to bacterial O-lactamase. They are therefore very useful for the treatment of various infectious diseases.

Dutch patent application 75.05375 describes cephalosporins with a substituted methyl group in the 3-position and a (4-alkyl-2,3-dioxopiperazin-1-ylcarbonylamino) group in the 70-position. French Pat. No. 2,338,944 describes 70- / 2- (4-alkyl-2,3-dioxo-15-piperazin-1-ylcarbonylamino) -2-phenyl-acetylamino / -7-methoxy-3- (substituted methyl) -3 -cephem-4-carboxylic acid.

J. Antibiotics 29, 554-558 and 973-975 (1976) describe the introduction of a 7a-methoxy group in certain cephalosporins.

It has now been found that new compounds of the general formula I, further described below, in which the cephem ring bears a substituted methyl group in the 3-position and a methoxy group in the 7a-position and in which the amino group in the 70-position is bound to the group of the formula II, wherein A and B have the meanings indicated below, and the salts of these compounds show a surprisingly good antibacterial activity.

An object of this invention is to provide novel 7a-methoxycephalosporins having in their molecule a 2,3-dioxo-1-piperazine carbonylamido group and a.

* * 7 - 2 - group B.

Another object of this invention is to provide novel 7α-methoxycealosporins with a broad antibacterial spectrum, with a high resistance to bacterial β-lactamase and with effective antibacterial activity against clinical isolates of bacteria.

A further object of the invention is to provide a process for the preparation of the new 7 a-10 methoxycephalosporins.

Yet a further object of the invention is to provide a pharmaceutical composition containing the new 7a-methoxycephalosporin or a pharmaceutically acceptable salt thereof as the active ingredient.

Other objects and advantages of this invention will become apparent from the following description.

The compound of the present invention comprises a 7a-methoxycephalosporin of the general Formula 1, wherein represents a hydrogen atom or a carboxyl protecting group, represents an acetoxy group or an optionally substituted 1,2,3,4-tetrazol-5-yl- thio or 1,3,4-thiadiazol-2-ylthio group, A represents an alkyl group having 1 to 5 carbon atoms and B represents a thienyl, furyl, aminothiazolyl or cyclohexadienyl group, or a pharmaceutically acceptable salt thereof.

The above groups R 1 may be substituted by a halogen atom or lower alkyl, phenyl, C 1 alkenyl, hydroxyl, lower alkoxy, lower alkyl-30 thio, nitro, cyano, lower alkylamino, di-lower alkylamino, acylamino, acyl, acyloxy , acyl-lower alkyl, carboxyl, carbamoyl, amino-lower alkyl, N-lower-alkylamino-lower-alkyl, NN-di-lower-alkylamino-lower-alkyl, hydroxy-lower-alkyl, hydroxyimino-lower-alkyl, lower-alkoxy- lower alkyl, carboxy-lower alkyl, sulfo-lower alkyl, sulfo, sulfamoyl-lower alkyl, sulfamoyl, carbamoyl-lower alkyl, carbamoyl- * \ -¾ 'ii * Λ - 3-alkenyl, N-hydroxycarbamoyl-lower alkyl or similar.

When a tautomerie is known to occur in one of the groups represented by B, the tautomers are within the scope of this invention. For example, an aminothiazolyl group exists in tautomeric forms that are in equilibrium, as shown by reaction scheme A «

The pharmaceutically acceptable salt of the 7a-methoxycephalosporin of this invention, represented by the general formula 1, includes those formed on the acidic group and those formed on the basic group, those salts well known in the penicillin and cephalosporin chemistry.

All optical isomers and racemic compounds and all crystal forms and hydrates of the 7a-methoxy-cephalosporin represented by the general formula 1 or a pharmaceutically acceptable salt thereof are within the scope of this invention.

The compounds of general formula 1 or salts thereof are prepared by known methods, such as, for example, those described below.

Preparation Method (1): A method, wherein a compound of the general formula 2, wherein R 9 represents a hydrogen atom, an organic silyl group or an organic phosphorus-containing group, R 1 a represents a hydrogen atom or a conventional carboxy protecting group and R And B have the meanings indicated above, are reacted with a reactive derivative in the carboxyl group of a compound of the general formula 3, wherein A has the meanings indicated above.

Preparation Method (2): A method in which a compound of the general formula 4, wherein R, let R2 and Rg have the meanings indicated above, is reacted with a compound of the general formula 5, wherein A and B are the above have indicated meanings, or a reactive derivative in the carboxyl group of *

* I

- 4 - this connection 5.

Preparation Method (3): A method in which a cephalosporin of the general formula 6, wherein R 1, A and B have the meanings indicated above, is reacted in the presence of methanol with an alkali metal methylate of the general formula 7, which represents an alkali metal, and then reacted with a halogenating agent.

Preparation Method (4): A method, wherein 10 is a 7a-methoxycephalosporin of the general formula 8, where in R? represents an easily replaceable nucleophilic reagent and Rj, A and B have the meanings indicated above, are reacted with a compound of the general formula 9 wherein M2 represents a hydrogen-15 atom, an alkali metal or an alkaline earth metal and R ^ has the meanings stated above.

The organic silyl groups and the organic phosphorus-containing groups represented by Rg in the above general formulas include the groups commonly used amino or carboxyl protecting group in penicillin and cephalosporin synthesis, such as ( CH3) 3Si-v (CH3) 2Si <, pP-, ^> P-, (C2H50) 2P- 25 and (C'H_) -P- which are easily removed by a 1 5 2 treatment with, for example, water or an alcohol .

The groups represented by R 1, which can be easily replaced by a nucleophilic reagent, include halogen atoms, such as chlorine, bromine and the like, lower alkanoyloxy groups, such as formyloxy, acetoxy, propionyloxy, butyryloxy, pivaloyloxy and the like, aryl carbonyloxy groups, such as benzoyloxy, naphthoyloxy and the like, arylthiocarbonyloxy groups, such as thiobenzoyloxy, thionaphthoyloxy and the like, arylcarbonylthio groups, such as benzoylthio, naphthoylthio and the like, arylthiocarbonylthio groups, such as thiobenzoylthio, thionaphtho. > 3

». «; "I" -J

* * m - 5 - and the like, a carbamoyloxy group, thiocarbamoyloxy group, pyridine-N-oxide-2-yl group and pyridazine-N-oxide-6-yl group. These groups represented by may further contain substituents such as, for example, a halogen atom, nitro group, lower alkyl group, lower alkoxy group, lower alkylthio group, acyl group and the like.

The compound of the general formula 5 is readily obtained by reaction between an alkali metal salt, an alkaline earth metal salt or an organic base salt of a compound of the general formula 10, wherein B

has the meanings indicated above, and a reactive derivative in the carboxyl group of a compound of the general formula 3 in the presence of an acid binding agent and an inert solvent.

The compounds of general formulas 2 and 4 can be synthesized in a manner known per se, for example, as described in Journal of Synthetic Organic Chemistry, Japan, Vol. 35, 568-574 (1977).

The practical embodiments of the preparation methods (1), (2), (3) and (4) are described below.

Methods (1) and (2) can be performed under almost the same conditions. The compound 2 or 4 is dissolved or suspended in an inert solvent, such as, for example, water, acetone, tetrahydrofuran, dioxane, acetonitrile, dimethylformamide, dimethyl acetamide, methanol, ethanol, methoxyethanol, diethyl ether, diisopropyl ether, benzene, toluene, methylene chloride , chloroform, ethyl acetate or methyl isobutyl ketone, each individually or a mixture of two or more thereof. A reactive derivative in the carboxyl group of compound 3 or compound 5 or a reactive derivative in the carboxyl group of compound 5 is added to the resulting solution or suspension.

The mixture is reacted in the presence or absence of a base at -60 ° C to 80 ° C, preferably -40 ° C to 30 ° C. A reaction time of 5 minutes to 5 hours is generally ► 8.

- 6 -Λ common enough.

The bases used in the above reaction include inorganic bases, such as alkali metal hydroxides, alkali metal hydrogen carbonates, alkali metal carbonates, and alkali metal acetates, tertiary amines, such as trimethylamine, triethylamine, tributylamine, pyridine, N-methyl-methyl-n-methyl-n-methyl-n-methyl-n-methyl-methyl , collidine and the like, and secondary amines, such as dicyclohexylamine, diethylamine, and the like.

When the compound 5 or a salt thereof is used as starting material in method (2), the reaction can be carried out in the presence of a dehydration condensation agent, such as, for example, N, N '-dicyclohexylcarbodiimide, N-cyclohexyl-N1-morpholino- Ethylcarbodiimide, N.N'-diethylcarbodiimide, N.N'-carbonyl bis (2-methylimidazole), trialkylphosphites, ethylpolyphosphate, phosphorus oxychloride, phosphorus trichloride, 2-chloro-1,3,2-dioxa-phospholane, oxazolyl chloride, dimethylchlororforminium chloro dimethylethoxyforminium chloride.

The method (3) is carried out in the following manner: A cephalosporin of the formula 6, knownly obtained / Japanese patent application Kokai (published) 70,788 / 76 and 113,890 / 76 7, is dissolved or suspended in an inert solvent, such as for example, tetra-hydrofuran, dioxane, ethylene glycol dimethyl ether, methylene choride, chloroform, dimethylformamide, dimethyl acetamide, acetonitrile, methanol or the like or a mixture of two or more of these solvents. To the resulting solution or suspension, an alkali metal methylate 30 (7) is added together with methanol. The resulting mixture is reacted and the reaction mixture is then reacted with a halogenating agent. In this reaction, methanol is used in excess and the amount of the alkali metal methylate (7) used is preferably 2-6 equivalents per equivalent of the use of cephalosporin (6). The term "in excess" means * * - 7 - an amount of more than one equivalent per equivalent of the cephalosporin (6). All the above reactions are performed at -120 ° C to -10 ° C, preferably -100 ° C to -50 ° C. A reaction time of 5-30 minutes is sufficient and the reaction is terminated by acidification of the reaction system.

The halogenating agent used in this method is generally known to be a source of procuring a positive halogen atom such as Cl, Br 10 or J +. Examples of such halogenating agents include halogens such as chlorine, bromine and the like, N-haloimides such as N-chlorosuccinimide, N-bromosuccinimide and the like, N-haloamides such as N-chloroacetaraide, N-bromoacetamide and the like, N- Halogen sulfonamides, such as N-chlorobenzenesulfonamide, N-chloro-p-toluenesulfonamide and the like, 1-halo-benzotriazoles, 1-halo-triazines, organic hypohalogen, such as tert-butyl hypochlorite, tert-butyl hypiodide and the like, halohydantoin such as NN-di-20 bromohydantoin, and the like. Of these halogens, tert.-butyl hypochlorite is preferred. The halogenating agent is used in an amount sufficient to provide a positive halogen in an amount equivalent to that of the cephalosporin of general formula 6.

Suitable acids for terminating the reaction are those which, when added to a cold reaction mixture, will not cause solidification of the reaction mixture or freezing of the reaction mixture to a heavy viscous mixture. Examples of suitable acids are 98% formic acid, glacial acetic acid, trichloroacetic acid and methanesulfonic acid.

After the completion of the reaction, the excess halogenating agent can be removed by treatment with a reducing agent, such as trialkyl phosphite, sodium thiosulfate or the like.

* • i - 8 -

In the practice of preparation method (4), when using a compound of the formula 8 other than the compound, wherein the group R 1 represents a heterocyclic aromatic amine N-oxide-thio group 5 with a thio group the carbon atom adjacent to the N-oxide group is this compound reacted with a compound of the formula 10 in an inert solvent such as, for example, water, methanol, ethanol, propanol, isopropylol, butanol, acetone, methyl ethyl ketone, methyl Isobutyl ketone, tetrahydrofuran, dioxane, acetonitrile, ethyl acetate, 2-methoxyethanol, dimethoxyethane, dimethylformamide, dimethylsulfoxide, dimethylacetamide, dichloroethane, chloroform, dichloromethane and the like, alone or in a mixture of two or more thereof.

The above reaction is preferably carried out in a highly polar solvent, such as water. It is advantageous to maintain the pH of the reaction solvent at 2-10 and preferably 4-8. The reaction is effected after the addition of a buffer, such as sodium phosphate, to adjust the pH to a desired value. Although the reaction conditions are not critical, the reaction is generally conducted at 0-100 ° C for several hours to several tenths of hours.

25

When a compound of the general formula 8, wherein the group is a heterocyclic aromatic amine N-oxide-thio group with a thio group on the carbon atom adjacent to the N-oxide group, is used, the compound of the general formula 8 and 30 a compound of the general formula 9 reacted together in one or more of the above inert solvents in the presence of a divalent copper compound. This procedure is particularly useful when the compound of the general formula 9 is an alcohol such as methyl alcohol, ethyl alcohol, propyl alcohol, isopropyl alcohol, n-butyl alcohol, benzyl alcohol, ethylene glycol or the like. In this case, the reaction proceeds easily using an excess of the alcohol serving as the solvent. The divalent copper compounds used in this procedure are inorganic or organic divalent copper compounds, such as, for example, cupric chloride, cupric bromide, cupric fluoride, cupric nitrate, cupric sulphate, cupric acid, cupric phosphate, cupricyanide, cupric citrate, cupric citrate, cupric citrate, cupric citrate and cuprisalicylate. The amount of the divalent copper compound used is preferably 0.5 mole or more per mole of the compound of the general formula 8. Although depending on the types of the compound of the general formula 8 used, the divalent copper compound used and The compound 15 of the general formula 9 used is the reaction temperature in general 0-100 ° C and the reaction time from several minutes to several days.

The conversion of a compound of the general formula 1, which is a carboxyl protecting group, into a compound of the general formula 1, wherein R 1 is a hydrogen atom, or a salt of the latter compound, the reaction of a compound with the general formula 1, wherein is a hydrogen atom, in a salt or in a compound of the general formula 1, wherein R 1 is a carboxyl-protecting group, or the conversion of a salt of a compound of the general formula 1 into the free acid thereof can be carried out in a conventional manner.

In the reaction of a compound in which the group is A, B or reactive, the reactive group can be protected with a protecting group, which is commonly used in the protection of a carboxyl, amino or hydroxy 1 group. After the reaction, such a protecting group can be removed in a conventional manner to recover the group A, B or R2.

The conditions for the preparation are not limited to those described above, but 4-10 can be suitably modified depending on the special type of reagent used.

The isolation of a 7a-methoxycephalosporin (1) or a salt thereof from the reaction mixture can be carried out in a conventional manner.

The methods of preparing a 7a-methoxycephalosporin of the general formula 1 and a salt thereof are not limited to the methods described above. These compounds can also be prepared by other methods.

The thus obtained 7a-methoxycephalosporins of the general formula 1 and their salts are very useful for the therapy of human and animal diseases because of their broad antibacterial spectrum against 15 gram positive bacteria and gram negative bacteria, their excellent antibacterial activity against Pseudomonas aeruginosa , Klebsiella pneumoniae and Proteus species, and their resistance to β-lactamase.

The antibacterial activities of the 20 representative compounds 1b of the 7a-methoxycephalosporins of the invention are shown in Tables A, B and C.

The values, indicated in Tables A, B and C, are the minimum inhibitory concentration (MIC in mcg / ml) of each compound determined according to the method described

in Chemotherapy (Society of Chemotherapy, Japan), Vol. 16, 98-99 (1968): A culture obtained by culturing the test bacteria in a heart infusion medium (Oak Kagaku Co.) was inoculated into a heart infusion agar medium (Oak 30 Kagaku Co.) . After an incubation for 20 hours at 37 ° C

bacterial growth was examined to determine the minimum inhibitory concentration (MIC in mcg / ml). The bacterial inoculation amount was 10 cells / plate (10 cells / ml).

35 (1) Minimum inhibitory concentration (MIC in mcg / ml).

; - ^ ... -5 _ - 11 -

Table A

unit: irtcg / ml

X. Connection B

(lb) CS-I170 S

(control)? ~ ï

. \ r2 «sa ^ ir -SAyJT

N. CH-CBLQQQE

Bacteria \ 2 * - ___ \ ^ _ 1__2_ S ·. aureus F-19 (penicillinase- 1/56 3/13 6/25 forming bacteria) E. coli HIHJ 0/39 40.1 40.1 JC. pneumoniae Y-50 0 ^ 39 0.39 6.25

Prot. vulgaris GF-76 3.13 0.2 0.78

Ps. aeruginosa I> 200 ^ 5 25 rp 0-5445 _- [’___

Ser. marcescens ¥ -8 = 2200 25 50

Acinetobacter τη r nc iriüüHê ^ TA-6 50 12'5 25 C. freundii GrïT-346 (Cephalosporinase- ^ 200 25 50 forming bacteria)

Graft, cloacae IID977> 200 25 25

Citrobacter ¥ -7 25 0.78 1.56

Ser. marcescens,, n n n no ÏTO62S-1-3-313 0) 78 °> 78

Klebsiella sot. T-72 12.5 6.25 25

Ser. τη-arceengns ¥ —35 100 6.25 100 '

Ps. aeruginosa S-68 2i20Q 12.5 50 OCH-: 3 q note ·: CS-1170 means KCCHgSCHgCOMn f | «J— ΪΓ CH9S iT3" c-jo.: ·; ,, 0 C00H (k.

*> - 12 -

Table A (continued)

-CP

H —- ίί ΪΓ— ΪΓ ¥ = ÏT

-0C0CH3 -S -sA ^ ir-CH2CH2IT (CH5) 2 CH2C0M2 CH2CH2OI 6.25 3.13 3.13 - 6.25 <0.1 £ 0.1 - £ 0.1% 1.56 0.39 0, 39 0.78 1.56 0.78 0.39 3.13 50. 12.5 12.5 50 100 25 50 50 25. 6.25 6.25 25 100 12.5 25 50. '12.5 25 25 3.13 3.13' 1.56 3.13 • 3.13 0.2 0.2 0.2 6.25 1.56 1.56 50 12.5 12.5 _ 50 12.5 12.5 _

Table A (continued) - 13 -: Ïü ÏÏT — 1Γ, r, τ Έ - Έ Έ - Ή Λ u 11 - yes .I i tl «

t b I I

CE2C32ïï (öE3) 2 GH = CH2 CS2CE2m2 6.25 6.25 5.15 6.25 ^ 0.1 £ 0.1 <0.1 £ 0.1 0.78 0.59 0.59 0.78 1 .56 0.78 0.78 0.78 50 25 25 12.5 50 25 25 50 | 12.5 12.5 12.5 12.5 25 100 12.5 25 25 100 25 50 6.25 12.5 5.15 5.15 0.59 0.78 0.2 0.2 12.5 1 , 56 '6.25 5.15 50 12.5 12.5 6.25 25 25 12.5 12.5

'-j' l · _ 1. V

Table A (continued) - 14 -

Yy [ó ~

-? ””? F — F F — F ~ F — F

“S- ^ rF -sX ^ F -sXrF -sX ^ Jf

CH0C = FOH I | I

CH ^ C12COCI5 CHgCOFHOH CE ^ 3,13 6,25 _ 25 3,13 1θ, 1 <1 0,1 £ 0,1 £ 0,1 0,78 0,59 1,56 0,78 0,78 0, 78 0.78 3.13 25 '25 12.5 12.5 50 50 100 - 100 12..5 25 1.56 6.25. 50 25 2 200 25 ': 25 50 200 25 6.25 6.25 50 6.25 0.2 0.2 0.78 0.78' 6.25 6.25 100 12.5 25 25 200 50 12.5 25 25 25 ^ '* v * λ, Λ

D? Β SJrn .'yY

Table A (continued) - 15 -

-Q

Έ — Ή Έ — E Ή Έ Έ ΪΓ -sX ^ Jr -s-ï ^ ir -sX ^ ir CH ^ CH2C0M2 GÏ2CH20H ch = ch2 6.25 6.25 6.25 '3.13 £ 0.1 <0 , 1 <0.1 <0.1 0.2 0.2 0.2 0.2 0.78 1.56 1.56 3.13 12.5 12.5 12.5 25 100 12.5 25 25 25 25 25 < 0.1 <0.1 <0.1 <0.1 6.25 _

12.5 ____j_L

25 ·· * '- ..j ^

Table A (continued) - 16 - Ή-r ___________ Ή — Ή Ή — Κ F - Ή -S- ^ KJr -S Λ ή, Ή -0000-SA g Λ CH5 ci ^ cH2com2 3.13 3.13 6, 25 6.25 ^ 0.1 10.1 £ 0.1 <0.1 0.2 0.78 '1.56 5.13 3.13 12.5 12.5 25 25.100 25 12.5 25 25 50 25 1.56 10.1 10.1 0.39 0.339 1.56 6.25 25 '- U -J, ο - 17 -

Overview investigated connections.

(1) Compounds according to Dutch patent application 75.05375 T-I55I (Gefoperazone) o o y_ ^ * (d) __ C2RyN n — CONH-CH CONH —η — γ8> Η Η ‘

cooh ch3 OH

T-1554 0 0 W * (D) - CZHSH N-CONH-CHCQNH —τ- ^^ X— CH ^ OCO CH3

[O) o 'T

V1 CO OH

T-I555

Oo ^ # (0) _ '

m ° COOH

(2) Compounds of the invention: s * r * - 18 - fi.

: R 't r T — 1414 * id)

CjHyN N-CONH-CHCONH -! - / SN ff w l ^ V-c ', sJir: \ = j CO OH CH3 1-1461 * \) -. % (D)? CHi C ^ HSN N— CONH— CH CONH — I yy J = j qJ CHgOCOCHj || ^ = 1 COOH i; i T-1688! | 1 Ο ΛΛϋ i1 ^ * (o) ° cCHa _! QH5-N N-CONH-CH CONH-Jty-Pf

<Y = j q ^ ~~~ N ^ "CHaS I

COOH CH? -C = NOH

TM689. £ Hs I

^ β. y (d) 'OCH 3 C 2 H 5 F 1 N -CONH -CH CONH-j- ^ Sn ^ ^ ^ = V 0 ^ ~ - \ = J COOH CH * COCH 3 Τ-Ί70Ι ° H ° * (D)? CH *'.

C, HrN N-CONH-CHCONH — iπ W.

w h I

^ COOH CHaCH ^ NH ^ T-1658 Λ.

° H * (D) Γ% n-n

GHcN N-CONH-CHCONH —f— [1 | y N — f} = r \ Y —- Nv ^ L-CHjS “* sn ^ N

- J ^ \ j I t) S \ = J COOH CH2C0NH0H

; ; - 19 - - T-1659 o O,. OCH *

M ', (D1 I S N-W

CiM N— CONH — CHCONH —f-S S [| | 1 N 'Y = y / -Nv ^ ~ CH? S ^ n <n

CO = OH CHROME

T-166I

° M ° * (D) ° ch 3

CjHrflN — CONH — CHCONH —ί ^ n, —N

^ CO OH C ^ CHgNfCHa ^ T-I487 V <° * 1 «? CH> N_m C ^ Hdi N-CONH —CHCONH 7 -f ^ N Ij ||

- / ^ = y QJ-Nv ^ Y — CHjS — Vjj | ^ N

w CO OH CH = CHe T-1488 ° j /? * (0)? CHi. _

CsHs-NN-CONH- CHCONH -f - ^ 'S 0 - | ί W f]

\ J COOH CH * CH * OH

T-1482 ° M ° * (p) ° CH>

C 8 H 3 -N N-CONH-CHCONH-f-r 3 S

^, sH <y-V "C * S-VN

C00H CHiCONHj T-1462 ^ β # (θ)? CH3

CzH-jNN-CONH-CHCONH-f-1 f1

O;; j v = J u COOH

-20 - T — 1711 ο Ο ', OCHj -Κ (Ο) = _ C ^ Hj-N Ν - CONH— CHCONH —τ-r'®' ') »ff ^ Λ“ JV-cb'sJ! · » KéJ cooh · ^ Τ-1719 ° ^> _ β «(Dt) ° chi

C HN N-CONH-CHCONH-P-f'5 ''. j f |

^ L JY-A ^ cV-yN

° \ J COOH CHj T-ï 753 $ (DL)? CHl C2HrN n— CONH — CH CONH u f} 1

W / J * H ^ -N

s Q. T I

COOH CH3 nh2 - 21 -:

rt \ vO vO vO 00 O CO O * vO

£ - £ in tn r- o p- λ tn * n vO -> · <- ·> - - ~ "" _Γ

5 O ~ - o O o - <N

H Vil co VO VD vO 03 O 03 03 'fj ίη - u ~> m m p * o p' ^

^ V * «-Γ O *" o ‘O - M

I ”E-> V» _ co and oo co ui O - 04 ”- cm O - r- - o cT o" <no "« no * * o «H Vlf r * o \ er \ vo co co * 0 ^ CO CO CO IA CM O - ^ ^ ^ vO Λ Λ * * ~. * - o OO o COO ^ Ö E- * 'ill co 03 00 PO _ Μ <2 Jf) 1 *

CO - CO Γ "* - CM O - p ~ P '·" V

S · *** «« * «o *“ * _ o OO CO O COO ^ OP ^ H Vil „cooveocnO-OoC'O ^ o _“ co - - o * n r- uo cm <f Λ Λ Λ * · Λ * "* * * - Ο Ο Ο οο d - Ο -

^ VII

3 'I 5-2 S £' ÏÏ S'cm £ -ο

Η 5 ο <ο "ο - ο - Ο - CJ

Vrf irt 00 Ο 03 Ο “0 Ο £ - Ο CM Ο Γ» Ο - “3 CM“ Ο m *> ui «uo« cm ~ «- o o o ^ .co l · * Vl!

5 s s “s onion o S- s I'S

* 7 O ~ O * OM CM -

Η - / V

- a \ CO O MD UI O O

UI — .UÓ CO «Λ Γ-- O UO CM O UO

I / O λ CM * «*> CM« - '- O O CM O - H Vfl ~. 01 onion

CC I

o <U <U <11 ω i-jcoca rj o ü οι oi o -r · I -H - f— / 03 01 -M 01 -fM 2 _

ij kV Ö C <U CJ f “I” r-1 i — l O 'O

5 f-ί O o CJ O O ** M c O I

g g s-ι cm ^ * yes - ta: c o * -j <• rl · Η 3 0 ~ Ά ca Ό «3 * <0 - MM3« J C.

, -j t — i ojuo ai <r g C ε CO Ju - <- <r ·. cd— · o cn O O Cl Cl * M * | --- 11 3 2: CM I -3 o cj o. O.> - J- / M U3 = E H> o «ca -na

«... ai ai · i — i ο-H

cuwiil MüocnfriCi-Mï'Mi ^, - -4 - J '* ^ - 22 - m σν o oo co and m u-ι - - cs. and - O r-- r- ~ - cs fS Μ Λ Λ Λ Λ Λ Λ Λ - ο Ο Ο Ο Ο Ο Ο and

ε! η Mil Vil VII

σν οο ο S3 οο οι. ~ CS | - * - Ο ιΠ Ρ- - ιπ ^ Μ Μ η Α Α Λ Λ Λ Λ - οο.ο ο ο - ο co cs Η vil VU Vil '”_ comco oincntnin - - cs ρ» cs p- ο cs - cs cs fS AAA Λ '«^ Λ Λ Α Λ„ OO'OvOO S3 and S3 S3 cL, ν / ι <u 051 Η * Η w 0 Vi

• Η · Η <U

0 Μ -U

ο cs unovencommcocn 0 ο tg \ ο '- es en - ρ * es cs ρ * - in jjcö

ft ft. ft Λ Λ Λ ft »« Ο JO

- ο 'Λ Ο and O vö oencs 0.

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1 4 ΐ S

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Vil Ö oi c 00 - (

0 C U

• H o

f— ^ r — 1 CU

co and σΊ ca o οο σι vo »n £ R w oo - - and p. es m p. en m es ^ r o ^ η n A a a Λ A A A M M Ü ^ - oen o o o -o o - sa 0'H «ι, ι-t C os eL \ II. i — 10 0

^ 0 & 4 O

O

M ··

vO

f ^, CO ’OSCO LACHCOCO O ri‘ 5 co «- -> and - 04 cN - r · * m - £

MM «% #» «l MMMM

- o and O and O and o and es ^ -vin - co and and. o and o vo - - r »- and in - m un m vo * Α μ a μ. ft ft ft es - σ o o and o en - es' H νή Mn ”en en cc ι S! . 0 0 0 0 0 * n en 'ed ei oow eno 53 ι * h * it en oi R oi R w M3eJC C 0 0 RR η 0Ό 2 hoo ooa r> s> e «01 £ E u es υ X yes - 0 * a --1-05 • H -H 3 0 3X 0 vQ 0 X 0 -. 00 for 0 0 r r 0m 0sr £ o E co υ - r p. 0 · - uoi O O Cl c’l · R · I R I 32 RI * 3 o υ o.> H &.> I hr us θ Η ρ ο · ca -hu ·. . . 0 0. R 0 * 1-1

fJ Cdfcrf Pi ca ca cu P * <<R

.Λ \ ..5 ij -, j>, <v '-j - 23 -

Table C

Comparative study between, on the one hand, the compounds according to Dutch patent application 75,05375 and French patent 2,338,944 and the compounds according to the invention.

_T-1551 T-1559 T-1711 T-1719 T-1753 E.coli TK-3 * 25 1.56 0.2 5 0.1 50.1 K. pneumoniae Y-50 0.39 0.78 0, 78 0.2 .0.2 K. pneumoniae Y-4 * 12.5 3.13 6.25 0.78 0.39

Ser.marcescens IID620 0.78 0.78 0.78 5 0.1 <0.1

Al.faecalis I B-l 100 3.13 6.25 3.13 3.13 g 10 cells / ml unit: jxg / mL *: Penicillinase-forming bacteria

Notes to table:

Representative connection of the Dutch patent application 75.05375 T'1551 0_0 * (0)

Ww (j-C0NH-CHC0MHn-N - N

W lAi Y-JL / N

O CT I

T COOH L

OH CHJ

Λ ^ Λ / ’>;> - 24 -

Representative Compound of French Patent 2,338,944 T-1559 0 0. OCHi W * (D) _

N-CONH-CHCONH-f-SNN

W (h 0 ^ No \ CH, S ^ N ^

^ COOH I

CHj

Compounds according to the invention Τ-Ί711 η n OCHi ° y_ ^ ° * (d) / _ C2HrN N-CONH-CHCONH -f — π, 1 w ó COCH iHi T-1719 ° yj. ° * (0L) PCH5

CjHfW N-CONH-CHCONH - ^ - r ^ S N π

^ C00H I

CH3 ^ Λ Λ) __ΐ

* · Ϊ · ✓ i \ J

«V - 25 - T-1753 0 0 * (DL) Ρ *

r ~ i 1 / N-N

C, HN N-CONH-CHCONH-f — S> || - j = y / “N C0C1H CH, MHi *>% \ - 26 - (2) Resistance to β-lac-tamase.

Resistance to β-lactamase was tested by iodometry at 30 ° C using the method of Perret / "cJ Ferret," lodometric assay of penicillinase ", Nature 174, 1012-1013 (1954) except that a 0, 1 M. phosphate buffer solution (pH = 7.0) was used in place of the 0.2 M. phosphate buffer solution (pH = 6.5). The resistance (stability) of each 10 compound is shown in Table D in terms of the relative degree of hydrolysis, whereby the stability of Gephaloridin (CER) to cephalosporinase is assumed to be 100 and the stability of penicillin G (PC-G) to penicillinase is 100.

15 - 27 - Η <μ ο cn o- Η Ο Ο Ο Ο —J> · A · Χ Λ ^ Ι Ο ο ο ο ο

& V ν V

--.- 1 --- * =% ^ S— Ο 03 Ο 'Φ Η ^ <Μ, -, __ / Η η σ> w ° Η 02

CM

------ W

Ο

fc {Ο Ο ΙΑ Η CO I S

§ ° Η Η * „Hf \ ------- η γ *

ε I

«Ώ κ S ρ cm CM Ο ο ο Ο \ 0 Μ- Ρη Η Η Η § ^ 2 £ S-Π +» to ι \ / ° 13 ι 03 ΤίΞ. § 02 ω "ω ® <Μ 3: HO Ο

If g S ο Φ d 2 '

* 3 5 ΙΟ S3 0Λ d CQ

C Η I Φ I Ή Ο]

0) Ρ S ο W S3 Ö YY

Μ Ή Ο! Π · Β Ο 2 μ Ο S μ £ ^ ® 3 3 £ η I 2 - g ° ^ 0 Ο d Ο Ö Φ · Η ffi Λ Ο εΐ Ο- Pi Cd Η

You, · · · · «Ν W

Ο pqj cal PQ1 WI Λ (d ο τΗ Μ 1. ® β

0 - Ο (U

01 «5" r m

Sum S $ • H 2 d Φ Λ

U o S3 -P

o § -η ω 2 2 ft λ ’% 5. ^ 5 * ill ,, P S Ν ή h g. g rj u è *

S I Η H

(¾ SJ d · η

O g Λ O

o> ft. -H

ω S3, o CD a II p * si

* o · - A o V

"". - · J * - · .. "- ν Λ -> * * y - 28 - (3) Remdose relative to β-lactamase.

The inhibit dose (ID <.q) of T-1414 relative to β-lactamase is that indicated in Table E, as-5 as determined by Perret's iodometric method using Cefazolin as substrate.

Table E

Remdose compared to β-lactamase 10 Cephalosporinase- ID50 ^ Wi / ih1) bacteria-forming E.coli GN-5482 0.82 S. marcescens W-8 0.54 C. freundii GN-346 1.7 15 (4) Anti-infective activity. Groups of four-week-old male mice of the ICR strain, each group consisting of five mice, were injected intraperitoneally with a predetermined amount of a pathogenic bacterium suspended in 5% Vs gum solution. After 1 hour from the injection, the mice were administered subcutaneously the preparation to be tested to determine the anti-infective activity. The results obtained are shown in Table F. The values in Table F indicate the protective

activity in terms of the ED

- 29 -

O

a r — i *> 1 Ώ n O GO H u S Λ O - Ö S9 ---- ® +1 m S3. w OSH -tf * o | oi 5 ω <'^ Βί .. H o "s« * Λ Ο

. - '' -. ~~ ’" 1.1 1 rH

φ Η Ο Λ Φ C— (3 λ Η Ο Ο -Ο <0 + »Η Ο Ο - (- 1 ^ • Η I <Μ Η, η ® εα λ -η £ ο" Λ>. H Ό Χ> S ^ Ρ-} Φ Φ ο \ Η in cvi -η φ ίΰ Μ ^ * »^ ö · η

5-1 i 2 ^ ΰ -S

Ή J5, Λ Φ «Η ö - ^« α φ

ο __ φ tS

go 01 Φ * · ® S Ό - 0J ° * OW β S ö a * 3 S ϋ '\ ο - Ά α

2 φ 8 & Ö5 ωο ο § I

• Η> Φ + J · ~ ί Η Η TS Φ θ 'Ο Φ ® ^ Ο Ο G Φ Ο $ g φ .η-μ% $ 5 "51 ·« «ο · η« a? · Ϊ́Λ. ο ΙΗ Η1 3 Η '33

& V * W Q

• Φ> ^ Ο Β Φ Φ

+ J ι-I

• Η Η OJ

D Φ ö

^ I

--—- · - Ο m ö 0) ο

CQ

ω i ο - * 1 § 0 I J3 S m

01 V

! col sï - 30 -

The 7a-methoxycephalosporins according to the invention have a low toxicity. For example, T-1414 exhibited an LD 1 of 5 g / kg or higher (mice of the ICR strain; intravenous).

The 7a-methoxycephalosporins of the invention of general formula 1 and the pharmaceutically acceptable salts thereof can be administered to humans and animals in the form of the free acid or in the form of a pharmaceutically acceptable salt or a pharmaceutically acceptable ester. The compounds can be formulated into the various dosage forms common to penicillin or cephalosporin preparations, for example, capsules, syrups and injection fluids, and can be administered orally or parenterally.

The invention is further illustrated but not limited by the following examples, wherein the percentages are weight percentages unless otherwise indicated.

15

Example I

(1) 0.80 g of diphenylmethyl-76- / D (-) - a- (4-ethyl-2,3-dioxo-1-piperazinecarboxamide) -a- in a mixture of 20 ml of anhydrous chloroform and 5 ml of anhydrous tetrahydrofuran were dissolved (2-thienyl) -20 acetamidoyl V 5- (1-raethyl-1,2,3,4-tetrazolyl) thiomethyl / & 3-cephem-4-carboxylate. The solution was cooled to -70 ° C and, while maintaining the temperature at -70 ° C, 3.5 ml of a methanol solution of lithium methoxide (1.425 mmol / ml) was added to the solution. After stirring for 3 min, 0.18 ml of tert-butyl hypochlorite was added to the resulting mixture and stirred at the same temperature for 15 min, after which 0.29 ml of acetic acid and 0.1 ml of triethyl phosphite were added in this order were added. The temperature of the reaction mixture was then raised to room temperature. The reaction mixture was placed in 25 ml of a citrate buffer solution (pH 7.0). The organic layer was separated, washed with water, dried over magnesium sulfate and freed from solvent by distillation under reduced pressure. The residue was purified by column chromatography (stationary phase: silica gel; eluent: a 1: 1 mixture of benzene and ethyl acetate) to obtain 0.50 g of diphenylmethyl-76- / D (-) - a- (4-ethyl -2,3-dioxo-1-piperazinecarboxamido) -a- (2-thienyl) -35 acetamido _ / - 7a-methoxy-3 - /> 5- (1-methyl-1,2,3,4-tetrazolyl) thiomethyl 1 __ / -, &3-cephem-4-carboxylate; yield 60.2%.

Λ _ Λ ('? V O Vl · i - 31 - IR (KBr) cm Sv C = O 1775, 1705, 1675.

NMR (CDCl 3) δ values: 1.09 (3H, t, -CH), 3.20-3.65 (611, m,> CH 2 x 3), 3.46 (3H, s, -CH,) , 3.65-4.65 <2H, m,> CH), 3.75 (3H, s, -CH), 4.31 {2H, Abq> CH), 5 4.97 (1H, s, ^ CH), 5.88 (III, d, - ^ CH), 6.80-7.47 (14H, -CgHg x 2, sj, ^ CH), 8.12 C1H, s, 9.72 (1H, d,> NH).

(2) Dissolved 0.3 g of 10 diphenyl-75 - / ^ D (-) -a- (4-ethyl-2,3-dioxo-1-piperazinecarboxainido) -a- (2-thienyl) in 2.0 ral anisole acetaraido [7a] -methoxy-3- [5- (1-methyl-1,2,3,4-tecrazolyl) thio-methyl- [alpha] -cephem-4-carboxylate. 2.0 ml of trifluoroacetic acid were added to the solution under ice-cooling and the mixture was stirred at the same temperature for 30 min. After the completion of the reaction, the solvent was removed by distillation under reduced pressure.

To the residue were added 5 ral ethyl acetate and 5 ml water and the pH of the solution was adjusted to 7.0 with an aqueous sodium hydrogen carbonate solution. The aqueous layer was separated and mixed with 10 ml of fresh raethyl acetate and adjusted to pH 2.0 with 20 2N hydrochloric acid. The organic layer was separated, washed with water, dried over magnesium sulfate and freed from solvent by distillation under reduced pressure. The residue was washed with diethyl ether to give 0.20 g of white 7B - (_ D (-) - α- (4-ethyl-2,3-dioxo-1-piperazine carboxamido) -o (2-thienyl) -acetamido / -7o-raethoxy-3-25 5- (1-methyl-1,2,3,4-tetrazolyl) thiomethyl-A3-cephem-4-carboxylic acid, yield 83.3%.

IR (KBr) cm -1: v C * O 1770, 1705, 1695, 1675.

NMR (DMSO-d.) 6 values: 6 1.12 (3H, t, -CH 3), 3.20-3.70 (6H, ia,> CH 2 x 3), 3.46 (3H, 30 s, -CH3), 3.70-4.35 (4H, m,> CH2 x 2), 3.95 (3H, s, -CH3), 503 (1H, s, ^ CH), 5.80 (1H, d, ^ -CH), 6.95-7.30 (2H, m, ν, Η

SI}, 7.40-7.55 'II

(1H, m, SJ), 9.70-9.90 (2H, m) NH x 2).

35 h; % *** '-5 * - ** - 32 -

The following compounds were obtained in an analogous manner.

70-LD (") - a- (4-ethyl-2,3-dioxo-1-piperazinecarbox-amido) -a- (2-thienyl) -acetamido _ / - 7a-methoxy-3- / 5- {1-carboxymethyl- 1.2.3.4-Tetrazolyl) thiomethyl-A3-cephem-4-carboxylic acid; m.p. = 60 ° C

(dec.).

IR (KBr) cm -1: C> O 1770, 1705, 1670 NMR (DMSO - de): δ values;

O

1.09 (3H, t, -CH3), 3.11-3.72 (6H, m, x 3), 3.41 (3H, s, -CH3), 3.76-4.60 (411 , m,> -Cl2 x 2), 5.04 (1H, s, ^ CH), 5.26 (2H, s, >> CH2), 5.90 (1H, d, ^ CH), 6.92 -7.26 (2H, m,

Η H

7.39-7.50 (1H, m, II), 9.68-9.79 (2H, ra,> NH x 2).

15 70 -, / D (-) - a- (4-ethyl-2,3-dioxo-1-piperazine-carboxamido) -a- (2-thienyl) acetamido _ / - 7a-methoxy-3-acetoxymethyl-A - cefem- 4-carboxylic acid; m.p. 150 ° - 165 ° C (dec.).

IR (KBr) cm -1: v C = O 1775, 1720, 1705, 1675.

NMR (DMSO- d.): Δ values: 6, 1.07 (3H, t, -CH3), 200 (3H, s, -CH3), 3.19-3.72 (6H, m, '> CH2 x 3), 3.42 (311, s, -CI ^), 3.74-4.12 (2H, m,> CII2), 475 (2H, ABq,> CII2), 5.10 (1H, s . ^ CH), 5.89 (1H, d, ^ CII), 6.89-7.22 (2H, m,

Η H

), 7.40-7.51 (1H, m, JP'1 ^ H), 9.65 (1H, d,> NH), 9.81 (1H, s,

25 S S

> NH).

Example II

1 g of the 70- / D (-) - a (4-ethyl-2,3-dioxo-1-piperazine carboxamido) -a (2-thienyl) -30 acetamido / -7a- were suspended in 15 ml of water. methoxy-3-acetoxymethyl-A-cephem-4-carboxylic acid, obtained in Example 1, and 0.34 g of 5-mercapto-1-carbamoylmethyl-1,2,3,4-tetrazole. 0.28 g of sodium hydrogen carbonate were added to the suspension with stirring. The resulting solution was allowed to react at 55-60 ° C for 20 hours while adjusting the pH of the reaction mixture to 6.0-6.3. After the completion of the reaction, 20 ml of methyl acetate were added to (y · '' i ··· »-λ .λ '·

üov: viU

- 33 - The reaction mixture was added and the pH of the solution was adjusted to 2 by the dropwise addition of 2N hydrochloric acid. The methyl acetate layer was separated and the aqueous layer extracted three times with 15 ml portions of methyl acetate. The organic layers were combined, dried over anhydrous magnesium sulfate and evaporated to dryness. The residue was dissolved in 20 ml of acetone. Diphenyldiazomethane was added dropwise with stirring to the resulting solution until the color remained. The solution was freed from solvent by distillation under reduced pressure and the residue was purified by column chromatography (stationary phase silica gel; eluent: 1: 3 mixture of benzene and ethyl acetate) using diphenylmethyl-7B- / D (-) - a- (4-ethyl-2,3-dioxo-1-piperazinecarboxamido) -a- (2-thienyl) -acetamido _ / - 7a-methoxy-3 - /> 5- (1-carbamoylmethyl-1,2,3,4-tetrazolyl) thiomethyl_ / -A 3 -cephem-4-carboxylate obtained in the form of a slightly yellow powder.

To this compound were added 3 ml of anisole and 3 ml of trifluoroacetic acid under ice-cooling. The mixture was allowed to react with ice for 60 minutes under cooling. After the completion of the reaction, the reaction mixture was freed from solvent by distillation under reduced pressure. To the residue were added 10 ml of ethyl acetate and the precipitated crystals were collected by filtration to obtain 0.32 g of 78- / D (-) - α- (4-ethyl-2'-3-dioxo-1-). piperazinecarbanido) -a- (2-thienyl) -acetamido-7a-methoxy-3- / 5- (1-carbamoylmethyl-1,2,3,4-tetrazolyl) thio-methyl / -Λ-cephem-4-carboxylic acid; m.p. 145 ° - 150 ° C (dec.); yield 27.6%.

25 IR (KBr) cm 1: v C = O 1775, 1670-1720 NMR (DMSO - dg): 5 values: 1.10 (3H, t -CH 3>, 3.20-4.10 (8H, m , "> CH 2 x 4), 3.44 (3H, s, CH 3 O -), 4.34 (2H, ABq,> CH 2), 5.10 (2H, s,> CH 2), 5.19 (1H, s, ^ CH), 5.95 (1H, d, ^ CH), 6.91-7.88 (5H, m, 30 H N_ hJi 1, -CONH2), 9.70 (1H, d,]> NH), 9.05 (1H, d, >NH),

S

The following compounds were obtained in an analogous manner.

35 - 34 - 7β - / _ D (-) -α- (4-ethyl-2,3-dioxo-1-piperazinecarbox-amido) -a- (2-thienyl) acetamido _ / - 7a-methoxy-3- {5- 1- (2-hydroxyethyl) -1,2,3,4-tetrazolyl / thioraethyl} -1-C-em-4-carboxylic acid; m.p. 125 ° - 130 ° C (dec.).

5 IR (KBr) cia'1: v C = O 1770, 1720-1670 NMR (DMSO - d, _): δ values: 6

1.12 (3H, t, -CH), 3.20-4.50 (1411, m,> CH 2 x 7), 3.40 (3H, s, C 13 '' -), 5.00 (1H , s, - ^ CH), 6.05 (1H, d, - ^ CH), 6.05-7.60 (3H, m, Η H

IDH W), 9.70 (1H, d, y NH), 9.86 (1H, s,> NH).

76 ~ LD (-) -cl- (4-ethyl-2,3-dioxo-1-piperazinocarbox-amido) -a- (2-thienyl) -acetamido _ / - 7a-nethoxy-3 - / _ 3- (6-hydroxypyridazinyl -thiomethyl-α-A 2 -cefera-4-carboxylic acid; m.p. 155 ° - 160 ° C (dec.).

15 IR (KBr) enf1: υ C = O 1765, 1700, 1665 NMR (DMSO - d_): 6 values:

O

1.10 (3H, t -CH), 3.20-4.30 (10H, m,> CH, x 5), 3.41 J <(3H, s, CH 3 O -), 5.02 ( 1H, s, CHCH), 5.05 (1H, d, )CH), 6.70-7.50 (5H, m,

Η Η Ηκ— ύ H

(/ (_ 20 H [[N li NrN), 9.70 (1H, d, J> NH), 9.85 (1H, s, J> NH).

S

7β- / d (-) - a- (4-ethyl-2,3-dioxo-1-piperazinecarbox-amido) -a- (2-thienyl) acetamido __ / - 7a-methoxy-3 - /> 5- (2-methyl 1,2,3,4-tetrazolyl) thiomethyl-A-r-cephem-4-carboxylic acid, m.p. 165 ° - 174 ° C (dec.). IR (KBr) cm -1: v C ~ O 1770, 1710, 1670.

NMR (DMSO - d,): values /:

O

1.07 (3H * t, -CH 3), 3.20-3.76 (6H, m,> CH 2 x 3), 3.41 (3H, s, CH 3 O-), 3.76-4.2O ( 4H, m,> Cl 12 x 2), 4.39 (3H, s,> N-CH 3), 5.04 (1H, s,> CH), 5.90 (1H, d, ^ CH), 6, 89-7.25 (2H, ra,

30 H_ H

JT T), 7.35-7.48 (HI, m X JL H), 9.60-9.77 (2H, m,> NH x 2).

S 5 7β- / D (-) - o- (4-ethyl-2,3-dioxo-1-piperazinecarbox-amido) -n- (2-t, hienyl) acetamido _ / - 7a-methoxy-3 - /> 5- (1.3,4-thiadiazolyl) -33 thiomethyl-A-1-cephem-4-carboxylic acid, m.p. 160 ° - 175 ° C (dec.).

c 'j, -j 2, 5 - 35 -? The IR (KBr) cm "1: υ C = O 1780, 1710, 1675 NMR (DMSO - d_): δ values: Θ 1.09 (3H, t, -CH3), 3.21-3.75 ( 6H, m,> CH2 x 3), 3.43 (3H, s, CH30-}, 3.75-4.10 (211, m,> CH2), 4.40 (2H, ABq,> CH2), 5.14 (1H, s, ^ rCH), 5.94 (1H, d, ^ CH), 6.91-7.39 (211, m,

H_j H _ N N

Jr Jj), 7.40-7.53 (1H, m, JQ- H), 9., 53 (1H, s, jf jP H), 9.65-9.84 (2H, ΐί, / ΝΗχ 2 ).

10 78- / D (-) - a- (4-ethyl-2,3-dioxo-1-piperazinecarboxamido} -a- (2-thienyl) acetamido _ / - 7a-methoxy-3- / 5- (1-vinyl -1.2.3.4-tetrazolyl) thiomethyl-1-cephem-4-carboxylic acid, mp 150 ° -153 ° C (dec.).

IR (KBr) cm-1: υ CO1770, 1710, 1700, 1665.

NMR (DMSO - d.): Δ values: o 15 1.10 (3H, t, -CH3), 3.20-3.70 (6H, m,> CH2 x 3), 3.43 (3H, s, -CH,), 3.80-4.55 (4H, ra,> CII, x 2), 5.10 (1H, s, CHCH), 5.46 µL.

(1H, d, = CH), 5.03 (1H, d, = CH), 5.94 (11h, d, ^ CH), 6.90-7.52 (4H, m,

II H

j [, = CH-), 9.70 (1H, d,> NII), 9.86 (1H, s, NHNH).

20 ^ 7S - / _ D (-) - a- (4-ethyl-2,3-dioxo-1-piperazinecarbox-amldo) -a- (2-thienyl) acetamido __ / - 7a-methoxy-3 - / _ S - / _ 1- (2-Hydroxyiminopropyl) -1.2.3.4-tetrazolyl / thiaraethyl / -A-1-cfera-4-carboxylic acid, m.p. 165 ° -172 ° C (dec.).

IR (KBr) cm -1: C-O 1775, 1705, 1675.

25 NMR (DMSO - d -): δ values: o 1.09 (3H, t, -CH3), 1.58 (1.7h, s, -CH. ^, 176 (1.3H, s, -CH3) , 3.18-3.69 (6H, m,> CH2 x 3), 3.49 (3H, s, -CH3), 3.77-4.20 (4H, m,> CH2 x 2), 5 .05 (3H, m, ^ CH, .857 (1H, d, - ^ CH), 6.85-7.25

(2H, m, Η H

30 µf), 7.40-7.44 (1H, m, JQL H), 9.60 (1H, d,> NH), 9.65 (1H, s> NH).

78- / 0 (-) - a- (4-ethyl-2,3-dioxo-1-piperazinecarbox-araido) -a- (2-thienyl) acetamido / - 7a-methoxy-3V 5- (1-acetylmethyl-1. 2.3.4-35 tetrazolyl) thiomethyl-A-1-cephem-4-carboxylic acid, m.p. 164 ° - 175 ° C (dec.).

a

IR (KBr) ctn'1: v C = 0 1775, 1720, 1705, 1675.

NMR (DMSO - d,): 6 values: 6 1.10 <3H, t, -CH), 2.27 (3H, s, -CH), 3.10-3.70 (6H, m,> CH2 x 3), 3.43 (3H, s, -CH), 3.75-4.20 (4H, m, ^ CH, x 2), 4.40 5 (2H, a, ^ CH), 5 .00 (1H, s, ^ ai), 5.45 (1H, d, ^ CH), 6.95-7.32 (2H, m,

H_ H

] iJJ), 7.40-7.47 (1H, m, oH), 9.45-9.67 (2H, m, NHx2).

^ 7β - / _ D (-) - a- (4-ethyl-2,3-dioxo-1-piperazinecarbox-araido) -a- (2-thienyl) acetamido _ / - 7a-methoxy-3 - {- 5 -, / 1 - (hydroxyaminocarbo-10nylraethyl) -1.2.3.4-tetrazolyl / thiomethyl} -A'-cephem-4-carboxylic acid; slaps.

160 ° - 170 ° C (dec.).

IR (KBr) cm -1: v C = O 1770, 1710, 1700, 1665.

NMR (DMSO - d.): 6 Values: 6 V.

1.11 (3H, t, -CH), 3.20-3.70 (6H, m,> CII2 x 3), 3.40 15. (3H, s, -CH3), 3.80-4, 65 (411, m,> CH, x 2), 5.07 (III, s, CCII), 5.28 (2H, s, >CH2), 590 (111, d, CICl1), 6, 90-7.50 (3H, m,

Η H

HI), 9.67 (1H, d,> NH), 9.01 (lil, s,> NH).

S

Example III

1 g of TQ - / - D (-) - a- (4-ethyl-2,3-dioxo-1-piperazine carboxamido) -a (2-thienyl) was dissolved in 10 ml of an aqueous solution containing 0.27 g of sodium hydrogen carbonate. acetamido-7a-methoxy-3-acetoxymethyl-A-cephem-4-carboxylic acid and 0.17 g of 5-mercapto-1,2,3,4-25 tetrazole. After the pH of the solution was adjusted to 6.4, the solution was reacted at 60 ° C for 20 hours. 10 ml of raethyl acetate and 5 ml of ethyl acetate were added to the reaction mixture. After its pH was adjusted to 2.2 with 2N hydrochloric acid, the organic layer was washed twice with 10 ml portions of water, dried over magnesium sulfate and freed from solvent by distillation under reduced pressure. The residue was purified by column chromatography (stationary phase: silica gel; eluent: a 1: 2 mixture of benzene and ethyl acetate) to obtain 0.39 g of TQ - / - D (-) - a- (4-ethyl-2,3-dioxo- 1-piperazine-carboxamido) -a- (2-thienyl) acetamido-7a-methoxy-3- / 5- (1,2,3,4-tetrazo-35-lyl) thiomethyl-1-alpha-cephem-4-carboxylic acid; m.p. 178 ° - 183 ° C (dec.); * - ij Z Z, j 1 1 - - - IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR IR V C C C 70 70 70 70 70 70 1770, 1690, 1670. Yield 36.48%.

NMR (DMSO - d-): δ values:

O

1.08 (3H, t, -CH 3), 3.18-3.70 f6H, m,> CH 2 x 3), 3.40 {3H, s, C 11 ^ O-}, 3.70-4, 10 (4K, m,> »CH2 x 2), 5.07 (lil, s, ^ -CH), 5.89 5Γ (1H, d, ^ CII), 5.87-7.20 (2H, m ,

Η H

X? 7.39-7.50 (1H, m, H), 9.65 (1H, d,> NH), 9.81 (1H, s,> NH).

jq The following compound was obtained in an analogous manner.

70- / D (-) - a- (4-ethyl-2,3-dioxo-1-piperazinecar-boxamido) -a- (2-thienyl) acetamido __ / - 7a-methoxy ~ 3 - / _ 5- (1.2.3.4 -triazolyl) -thionethyl-A-1-cephem-4-carboxylic acid, m.p. 178 ° - 190 ° C (dec.).

15; IR (KBr) ca v C = O 1770, 1695, 1680.

NMR (DMSO - dg) 0 values :.

1.10 (3H, t-CH-j), 3.20-3.70 (6H, ra,> CH2 x 3), 3.40 (3H, s, C 11-10), 3.80-4.20 (4H, ra,> CH2 x 2), 5.10 (IK, s, - ^> CH), 5.91 (1H, d, ^ CH), 6.88-7.20 (2H, ra, 2µ : HwH | -? JTjJ), 7.37-7.50 (1H, ra, JT H), 7.90 (1H, s, ^ y M), 9.58-9.74 (2H, ra,> NH x 2).

Example IV

25 g of 10- / - D (-) - a- (4-ethyl-2,3-dioxo-1-piperazine carboxamido) -a- (2-thienyl) were dissolved in 10 ml of an aqueous solution containing 0.27 g of sodium hydrogen carbonate. acetamido-7a-methoxy-3-acetoxymethyl-A3-cefera-4-carboxylic acid and 0.32 g of the sodium salt of 1- (2-dimethylamino thy1) -5-raercapto-1,2,3,4-tetrazole . While maintaining the pH at 6.2-6.5, raen reacted the solution at 60 ° C for 16 hours. The reaction mixture was purified by color chromatography (stationary phase: 220 g Amberliat XAD; elution with 100 ml water and then just 100 ml 50% aqueous solution of methanol). The eluate was evaporated to dryness to give 0.32 g of sodium 70V D (-> - 35 ot (4-ethyl-2,3-dioxo-1-piperazinecarboxaraido) -O (2-thienyl) acetamido). s * «/ '.i * ** -38- 7ci-methoxy-3- {5 - / - 1- (2-dimethylaminoethyl) -1.2.3.4-tetrazolyl / thiomethyl} -A ^ -cephem-4-carboxylate in the form of a white powder, mp 166 ° - 175 ° C (dec.), yield 26.17%.

IR (KBr) cm "1: \> C = 0 1760, 1705, 1670, 1610 5 NMR (DMSO - dJ: 5 values: 6 1.10 (3H, t, -CH3), 2.20 (611, m, - CH3 x 2), 2.68 (2H, t, ^> CH2), 3.20-3.70 (6H, m,> CH2 x 3), 3.40 (3H, s, CH30-), 3, 72-4.30 (6H, m,> CH 2 x 3), 5.05 (1H, s, ^ CH), 5.91 (1H, d, ^ CH), 6.90-7.30 (2H, m * 10 'Ks 7'40-7'51 (iH'o H), 9.6-9.75 (2H, m,> MH x 2).

3 3

The following compounds were obtained in an analogous manner.

Sodium-7S- / D (-) - a- (4-ethyl-2,3-dioxo-1-pipera -zinecarboxamido) -a- (2-thienyl) acetamido _ / - 7a-methoxy-3- {5 - / _ 2 - (2-dimethyl-15 aminoethyl) -1.2.3.4-tetrazolyl / thiomethyl} -A'-cephem-4-carboxylate; m.p.

175 ° - 183 ° C (dec.).

IR (KBr) cm -1: υ C = 0 1760, 1705, 1670, 1615.

NMR (DMSO - dj: 6 values: 6 1.11 (311, t, -CH), 2.24 (6H, m, -CH3 X 2), 2.83 (2H, t, 20 J> CH2), 3, 20-3.70 (6H, m,> CII., X 3), 3.42 (3H, s, CH3-0-), 3.70-4.20 - (4H, ra,> CH_ x 2) , 4.60 (2H, t,> CH 3), 5.0 (1H, s, C 1 Cl), 5.92 (1H, d, C 1 Cl), 6.90-7.30 (2H, ra, Η H f-1-i), 7.37-7.48 (1H, ra, YY, yY-H), 9.6-9.75 (2H, ra, NH * 2).

25 3

Sodium-78- / D (-) - a- (4-ethyl-2,3-dioxo-1-pipera -zinecarboxamido) -a- (2-thienyl) acetamido _ / - 7a-methoxy-3- {5- / 1- (2-aminoethyl) -1,2,3,4-tetrazolyl / thiomethyl] -A-1-cephem-4-carboxylate; m.p.

180 ° -187 ° C (dec.).

30 IR KBr) cm -1: v C = O 1765, 1705, 1670, 1600.

NMR (DMSO - d-): 6 values: 6 1.10 (3H, t, -CH 3), 3.23-3.74 (6H, m,> CH x 3), 3.45 (3H, s, -CH-j), 3.77-4.87 (6H, ra,> CH 2 x 3), 4.81 (2H, t,> 0 ^), 5.07 (1H, s, ^ -CH), 5.45 (III, d,> CH), 6.95 - 7.25 (2H, m, 35 Γ,: ι \ 'λ o * 5 ^ v; ij £ 0 - 39 - Η Η

Xf), 7.40-7.52 (1Κ, π, JT ”1 ^ Η}, 9.51-9.80 (2Η, ra,> NHx 2). Example V

(1) To an ice-cooled suspension of 2.55 g of DL-α-amino-2-furylacetic acid in 35 ml of methylene chloride was added 4.81 ml of trimethylchlorosilan. 5.04 ml of triethylamine were added dropwise to the resulting mixture over a 10 minute period

5-10 ° C. The reaction was left at 15-20 ° C for 1 hour

10 expired. To the reaction mixture were added 7.01 g of a mixture of 4-ethyl-2,3-dioxo-1-piperazine carbonyl chloride and triethylaraine hydrochloride over a period of 15 min at 5-10 ° C, which mixture was 58.02% of the former compound, added "The reaction was allowed to proceed at 10-15 ° C for 1 hour. After adding 20 ml of water, the reaction mixture was stirred at room temperature for 1 hour and then at 5-7 ° C for 30 min. The precipitated crystals were collected by filtration, washed with 5 ml of methylene chloride and 5 ml of water in this sequence and then dried, yielding 4.75 g of DL-α- (4-ethyl-2,3-dioxo-1-piperazine carboxamido) -2-furylacetic acid; m.p. 179 ° - 181 ° C (dec.); yield 85.6 l, IR (KBr) cm v C = O 1735, 1709, 1650.

NMR (DMSO-d,): <5 values: o 1.09 (3H, t, -CH3), 3.18-3.71 (4H, ra,> CK2 x 2), 3.78-4.09 (2H, m,> CH2), 5.53 (1H, d, ^ CH), 6.44 (2H, m,

25 Η H

ji "jj), 7.61 (1H, s, JV, jL-H), 9.69 (1H, d, ^ NH).

(2) 0.94 g of DL-α-β-ethyl-2,3-dioxo-1-piperazine carboxamido) -2-30 furylacetic acid were suspended in 15 ml of anhydrous methylene chloride. 0.35% N-methylmorpholine was added to the suspension to form a homogeneous solution. To the solution, cooled to -20 ° C, was added dropwise over a period of 5 min. A solution of 0.32 ml of ethyl chlorocarbonate in 2 rI anhydrous methylene chloride. The mixture was stirred at -15 ° C for 60 min. To the mixture, cooled again to -20 ° C, a solution of 1.50 g of diphenylmethyl-7-amino-3- / ~ 5- (1- 1) was added dropwise over a period of 10 min. methyl 1,2,3,4-tetrazolyl) thioraethyl-A3-cephem-4-carboxylate in 15 ml of anhydrous methylene chloride. The mixture was further stirred at -10 ° C for 90 min. After the temperature of the reaction mixture was raised to room temperature, the reaction mixture was poured into 10 ml of water and the organic layer was separated and 10 ml of fresh water was added thereto, and the pH thereof was adjusted to 6.5 with an aqueous solution of sodium hydrogen carbonate. The organic layer was separated, washed with 10 ml of water and 10 ml of saturated sodium chloride solution in this order and dried over magnesium sulfate. After removal of the solvent by distillation under reduced pressure, the residue was treated with diethyl ether to obtain 2.0 g of a white powder. By thin layer chromatography developer: a benzene-ethyl acetate mixture (1: 3} / was found to consist of two components (Rf: 0.20 and 0.24). The white powder was subjected to a elution by column chromatography (stationary phase: silica gel - eluent: a benzene-ethyl acetate mixture (2: 1)) to obtain the component corresponding to the component in the above in thin layer chromatography, and the component corresponding to the component in isolating the lower zone in the thin layer chromatography, the amount of the two isolated components totaling 1.1 g.

The upper zone component (probably 25 diphenylmethyl-7V D-α- (4-ethyl: 1-2,3-dioxo-1-piperazine carboxamido) -a- (2-furyl) acetamido _ / - 3 - / _ 5- (1- methyl 1,2,3,4-tetrazolyl) thiomethyl (43-cephem-4-carboxylate); m.p. 428 ° - 138 ° C (dec.), IR (KBr) cm -1: V c = O 1800, 1705, 1680.

NMR (CDCl 3): <5 values: 1.11 (3H, t, -CH 3), 3.30-4.30 (1011, m, J> CH 2 x 5), 3.76

OH, s, -CH3), 4.91 (1H, d,> CH), 5.72-5.93 (2H, ra, ^ CH x 2), 6.25 (2H, m, Η H

), 6.82 (1H, s,> CH), 7.20-7.53 (12H, ra, -CgH5 x 2, X> <35 NH), 9.80 (1H, d, ^ 1IH) ,

• "V

... ». 'I.V

f * - 41 -

The lower zone component (probably diphenylmethyl-7- / L-cc- (4-ethyl-2,3-dioxo-1-piperazine carboxamido) -a- (2-furyl) acetamido _ / - 3- / 5- (1-methyl- 1.2.3.4-Tetrazolyl) thiomethyl (- (A, - (cephem-4-carboxylate)); m.p. 128 ° - 136 ° C (dec.).

IR (KBr) Cia-1: \ C = 1800, 1705, 1680.

NMR (CDCl 3): 5 values: 1.08 (3H, t, -CH 3), 3.20-4.30 (10H, m,> CH 2 x 5), 3.86 C3H, s, -CH 3), 5.00 (1H, d, ^ CH), 5.61-5.98 (2H, m, - ^ CHx2), 6.31 (211, m, Η H _ 10 \ j), 6.83 (1H , ξ, ΟΙΙ ΟΙΙ), 7.29 (11H, m, -CgI15 x 2 J7 TL-H) / 7/94 (III, d,> NH), 9.88 (1H, d,> NH).

(3) In a mixture of 14 ml of anhydrous chloroform and 3.5 ml of anhydrous tetrahydrofuran, 0.86 g of the 15 component in the upper zone, obtained under (2), were dissolved. To the solution, cooled to -75 ° C, was added 1.97 ml of a solution of lithium methoxide (1.948 mmol / ml) in methanol and the resulting mixture was stirred at -75 ° C for 3 min. After adding 0.20 ml of tert-butyl hypochlorite, the mixture was stirred at -70 to -65 ° C for 15 min.

20 Ma addition of 0.20 ml acetic acid, the temperature of the reaction system was raised to room temperature. The reaction mixture was freed from solvent by distillation under reduced pressure and the residue was dissolved in a mixture of 10 ml ethyl acetate and 10 ml water. The organic layer was separated and 10 ml of water was added thereto, after which its pH was adjusted to 2.5 with 2N hydrochloric acid, acidic. The organic layer was separated, washed twice with 10 ml portions of saturated sodium chloride solution, dried over magnesium sulfate and freed from solvent by distillation under reduced pressure. The residue was purified by column chromatography (stationary phase: 15 g of silica gel; eluent: a benzene-ethyl acetate mixture (2: 1)) to obtain 0.30 g of a white powder.

The white powder thus obtained was dissolved in 3.0 ml of anisole and 3.0 g of trifluoroacetic acid were added to the solution under ice cooling. The resulting mixture was stirred at the same temperature for 30 min. After removal of the solvent by distillation under reduced pressure, 5 ml of ethyl acetate and 5 ml of water were added to the residue, and a saturated sodium hydrogen carbonate solution was then added thereto with stirring to adjust the pH to 6.5, after which the residue was dissolved, the aqueous layer 5 was separated and 10 ml of methyl acetate were added thereto, the pH was adjusted to 2.0 with 2N hydrochloric acid and the organic layer was separated. The aqueous layer was further extracted with 10 ml of methyl acetate and the extract was combined with the organic layer obtained above. The combined organic layer 10 was washed twice with 10 ml portions of saturated sodium chloride solution, dried over magnesium sulfate and freed from solvent by distillation under reduced pressure. The residue was treated with diethyl ether to give 0.17 g of 7B-V Da- (4-ethyl-2,3-dioxo-1-piperazinecarboxamido) -a- {2-furyl) acetamido - / - 7a-methoxy-3- / 5- (1-Raethyl-1,2,3,4-tetrazolyl) thiomethyl-A-1-cephem-4-carboxylic acid in white powder form m.p. 130 ° - 138 ° C (dec.); yield 70.8%.

IR (KBr) cm -1: C = O 1770, 1700, 1670.

NMR (DMSO - d.): <$ Values: 6 1.07 (3H, t, -CH3) 3.28-4.01 (8H, m,> CH2 x 4), 3.38 (3H, s , -CH3), 3.88 (3H, s, -CH3), 4.10-4.45 (2H, m, ^ CI ^), 5.03 (1H, s, ^ CH), 5.73 (1H, d, ^ CH), 6.39 (2H, m,

Η H

X? ), 7.54 (1H, bs,), 9.51-9.70 (2H, m,> NH x 2).

The following compounds were obtained in an analogous manner.

78- / Da- (4-ethyl-2,3-dioxo-1-piperazinecarbox-amido) -a- (2furvl) -acetamido _ / - 7a-methoxy-3 - [_ 5- (1-carbamoylmethyl-1,2,4.4) -tetrazolyl) thiomethyl-alpha-alpha-cephem-4-carboxylic acid.

30 7β - / _ D / -a- (4-ethyl-2,3-dioxo-1-piperazinecarboxamido) -a- (2-furyl) -acetamido _ / ^ - 7a-methoxy-3- {5 - / _ l - {2-hydroxyethyl) -1,2,3,4-tetrazolyl / thiomethyl} -A'-cepfera-4-carboxylic acid.

7β- / Da- (4-ethyl-2,3-dioxo-1-piperazinecarbox-amido) -a- {2-f-uryl) acetamido ^ -7a-methoxy-3V 5- (1-vinyl-1,2.3.4 -tetrazo-35-lyl) thiomethyl / -A-1-cephem-4-carboxylic acid.

V Ij * - 43 -

Example VI

(1) To a suspension of 2.5 g of D (-) - α-amino-1,4-cyclohexadi-enylacetic acid in 20 ml of water, 2.3 ml of triethylamine were added dropwise with ice-cooling to form a solution. To the solution at 5-10 ° C were added dropwise 6.3 g of 4-ethyl-2,3-dioxo-1-piperazine carbonyl chloride (purity 58.02%) over a period of 30 min, keeping the pH at 6.5- 8.0 was adjusted with 2.5 ml of triethylamine. The mixture was stirred at 10-15 ° C for an additional 30 min. The reaction mixture was washed with 20 ml of ethyl acetate and 30 ml of fresh ethyl acetate were added thereto, and the pH thereof was adjusted to 2.2 with 6N hydrochloric acid. The organic layer was separated and the aqueous layer was further extracted with 15 ml of ethyl acetate. The extract was combined with the above organic layer, washed 2 times with 20 ml portions of water and then 15 with 20 ml of a saturated aqueous sodium chloride solution. The washed organic layer was dried over magnesium sulfate and freed from solvent by distillation under reduced pressure to obtain 4.2 g of D {-) - α - (4-ethyl-2,3-dioxo-1-piperazine carboxamido) -1,4-cyclohexadienylacetic acid in white powder, m.p. 104 ° - 108 ° C; yield-20 yields 80.0%.

IR (KBr) cm -1: v C = 0 1705, 1670.

MMR (DMSO-d.} 5 values: 6 1.10 (3H, t, -CH3), 2.61 (4H, m,> CH2 x 2), 3.18-3.69 (4H, m, ^ > CH2 x 2), 3.70-4.13 (3H, m, J> CH2, ^ CH), 4.56 (1H, d, ^ CH), 5.60 (2H, m, ^> CH) x 2), 5.70 (1H, s, ^ CH), 9.49 (1H, d,> NH), (2) To a solution of 2.3 g of D (-) - a- (4-ethyl - 2,3-dioxo-1-piperazinecarboxamido) - 1,4-cyclohexadienylacetic acid in 20 ml anhydrous methylene chloride were added at -20 ° C 0.79 ml N-methylmoropholine, 0.75 ml ethyl-30 chlorocarbonate was added dropwise to the mixture. over a period of 5 min. at -20 to -15 ° C.

To the mixture, which was stirred for an additional 60 min at -20 to -15 ° C, a solution of 3.0 g of diphenylmethyl-7-amino-3- was added dropwise over a period of 15 min. At -20 ° C. 5- (1-methyl-1,2,3,4-tetrazolyl) thio-3 methyl-1-A-cephem-4-carboxylate in 30 ml of anhydrous methylene chloride. The mixture was stirred at -10 ° C for an additional 120 min. It Λ 6.

The reaction mixture was poured into 50 ml of water and the organic layer was separated. 30 ml of water were added to the organic layer and the pH thereof was adjusted to 7.0 with saturated aqueous sodium hydrogen carbonate solution. The organic layer was then separated, washed with 30 ml of water and 30 ml of saturated aqueous sodium chloride solution in this order, dried over magnesium sulfate and freed from solvent by distillation under reduced pressure. 30 ml of benzene were added to the residue and the precipitated crystals were collected by filtration to obtain 3.9 g of diphenylmethyl 7 3- / D (-) - a- (4-ethyl-2,3-dioxo-1- niperazine carboxamido) -et- (1,4-cyclohexadienyl) acetamido / -3- - - .3 / - 5- (1-methyl-1,2,3,4-tetrazolyl) thioraethyl / / - Z1 -cephem-4-carboxylate, m.p. 151 ° -155 ° C; yield 80.7%, IR (KBr) cm 2 C = 0 1770, 1700, 1665.

NMR (CDCl3) δ values: 15 1.15 (3H, t, -CH3), 2.62 (4H, m,> -CH2 x 2), 3.23-3.70 (6H, m,> CH2 x 3), 3.71 (3H, s, -C! ^), 3.70-4.35 (4H, ra,> CH2 x 2), 4.92-5.28 (2H, n, x 2) , 5.57 (211, m, ^ KZH x 2), 5.75-5.97 (211, ra, ^ CHx 2), 6.78 (lil, s, ^ - CH), 7.75 (ΠΗ , m, -CgH5 x 2,> ΝΗ), 9.57 (1H, d,> NH).

(3) 0.80 g of diphenylmethyl-73- / D (-) - a- (4-ethvl-2,3-dioxo-1-piperazinecarboxamido) -a were dissolved in a mixture of 15 ml of anhydrous chloroform and 4 ml of anhydrous tetrahydrofuran. - (1,4-cyclohexadienyl) acetamido-3- / 5- (1-methyl-1,2,3,4-tetrazolyl) thiomethyl-1-A-cephem-4-carboxylate. After adding 1.0 g of "molecular sieve 3A", the solution was cooled to -70 ° C. To the solution were added 1.8 ml of a solution of lithium methoxide (1.948 nmol / ml) in methanol. The mixture was stirred at -70 ° C for 3 min. Then 0.18 ml of tert-butyl hypochlorite were added all at once and the mixture was stirred at -70 to -65 ° C for 15 min and 0.2 ml of acetic acid was added thereto, after which the temperature of the mixture was raised to room temperature. After removal of the molecular sieve by filtration, the reaction mixture was freed from solvent by distillation under reduced pressure. The residue was dissolved by adding 20 ml of ethyl acetate and 10 ml of water and the pH of the solution was adjusted to 6.5 with saturated aqueous sodium hydrogen carbonate. o. _? * - 45 - solution. The organic layer was separated and 10 ml of water was added thereto, and the pH thereof was adjusted to 2.5 with 6N hydrochloric acid. The organic layer was separated again, washed with 10 ml of water and 10 already saturated aqueous sodium chloride solution in this order and freed from solvent by distillation under reduced pressure. The residue was purified by column chromatography (stationary phase: silica gel; eluent: benzene-ethyl acetate mixture (2: 1)}. After removal of the solvent from the eluate by distillation under reduced pressure, the residue was treated with diethyl ether. obtained 0.5 g of diphenylmethyl-7B- / D (-) - a- (4-ethyl-2,3-dioxo-1-piperazine-carboxamido) -a (1,4-cyclohexadienyl) acetamido _ / - 7a-raethoxy-3- / 5- {1-methyl-1,2,3,4-tetrazolyl) thiomethyl-y-A-cephem-4-carboxylate in white powder form; yield 60.3 i.

IR (KBr) cm -1: v C = O 1775, 1710, 1675.

15 NMR (CDCl 3) <5 values: 1.17 (3H, t, -GH3) 7 2.75 (4H, m,)> CH2 x2), 3.29-3.63 (6H, m,> CH2) x 3), 3.47 (3H, s, -CH ^), 3.77 (3H, s, -CH3), 3.80-4.09 (4H, m,> CII2 X 2), 4.99 (1H, s. ^ CI1), 5.61 (2H, m, ^ -CH x 2), 5.95 (1H, s, ^ CI0, 6.87 (1H, s,> CH), 7.23 -7.45 (11H, m, -C 8 H 5 x 2, 20 ^ NH), 9.55 (µl, d, - ^ NH).

(4) Dissolved 4 g of diphenylmethyl-73- / D (-) - ei- (4-ethyl-2,3-dioxo-1-piperazinexarboxamido) -a (1,4-cyclohexadienyi) acetamido_ / -7a-methoxy-3- / 5- (1-methyl-1,2,3,4-tetrazolyl) thiomethyl-alpha-cephem-4-carboxylafc. 5 ml of trifluoroacetic acid were added to the solution under ice cooling and the mixture was stirred at the same temperature for 30 min. After removing the solvent from the reaction mixture, 10 ml of methyl acetate and 10 ml of water were added to the residue and their pH was adjusted to 6.5 by adding a saturated aqueous sodium hydrogen carbonate solution while stirring. The aqueous layer was separated and 5 ml of methyl acetate was added thereto, its pH adjusted to 2.2 with 6N hydrochloric acid and then the mixture was left in a refrigerator overnight. The precipitated crystals were collected by filtration, washed twice with 5 ml portions of water and then with 3 ml of methyl acetate and dried to give v - 46 - 0.2 g of 73- / D (-) - a- (4-ethyl -2,3-dioxo-1-piperazinecarboxamido) -a- (1,4-cyclohexadienyl) acetamido _ / - 7a-methoxy-3 ~. / 5- (1-methyl-1,2,3,4-tetrazolyl) thiomethyl __ / - A ^ -cephem-4-carboxylic acid, mp. 177 ° - 178 ° C (dec.); yield 62.7%.

5 IR (KBr) cm -1: v C = 0 1770, 1710, 1685.

NMR (DMSO - d /) 6 values: 5 1.10 (3H, t, -CH 3), 2.18 (4H, m,> Zl 2 x 2), 3.34-3.73 (6H, ra,> -CH2 x 3), 3.41 <3H, s, -CH ^, 3.79-4.37 (4H, m,> CU2 x 2), 3.93 (3H, s, -CII3), 4, 97 (1H, d, ^ 'cyl), 5.06 (10.s, ^ CH), 5.61 (2H, m, 10 ^ CH x 2), 5.87 (lil, s, JcH), 9 .30 (1H, d,> NH), 9.53 (1H, s,> rNH).

In an analogous manner, the following compounds were obtained: "7β- / D (-) - a- (4-ethyl-2,3-dioxo-1-piperazinecarbox-amido) -a- (1,4-cyclohexadienyl) acetamido _ / - 7a-methoxy-3 -acetoxymethyl-A -15 cefem-4-carboxylic acid.

7β- / D (-) - a— (4-ethvl-2,3-dioxo-1-piperazinecarbox-amido) -α- (1,4-cyclohexadienyl) acetaraido __ / - 7n-methoxy-3- / i 2- (5-methyl - 1,3,4-thiadiazolyl) thiomethyl-alpha-4-em-4-carboxylic acid.

Example VII

(1) 2.0 g of a- (4-ethyl-2,3-dioxo-1-piperazinecarboxamido) -o (2-trichloroethoxycarbonylamino-4-thiazolyl) -acetic acid were dissolved in 15 r anhydrous methylene chloride. 0.43 ml of N-raethylmorphorline were added to the solution at -20 ° C. To the resulting mixture was added dropwise over a period of 5 min a solution of 0.41 ml of ethyl chlorocarbonate in 2 ml of methylene chloride while maintaining the temperature at -20 to -25 ° C. The reaction was left for 1.5 hours expired at the same temperature. To the reaction mixture was added dropwise at -15 to -20 ° C over a period of 15 min a solution of 2.0 g of diphenylmethyl-7-amino-3- / 5- (1-methyl-1,2,3,4-tetrazolyl) thiomethyl-α-β-cephem-4-carboxylate in 30 ml of water free methylene chloride added. The reaction was effected for 2 hours at -10 to -15 ° C. After the temperature of the reaction mixture was raised to room temperature, the reaction mixture was poured into 30 ml of water and its pH was adjusted to 6.5 with a saturated aqueous sodium hydrogen carbonate solution. The organic V V 1 - 47% t layer was separated and 30 ral water was added again, after which its pH was adjusted to 2.0 with 6N hydrochloric acid. The organic layer was separated, washed with 30 ml of water and 30 ml of saturated aqueous sodium chloride solution in this order, dried over magnesium sulfate and freed from solvent, by distillation under reduced pressure to give 3.3 g of a white foamy residue were obtained. Thin residue chromatography (developer; a 1: 3 mixture of benzene and ethyl acetate) found that this residue consisted of two components.

The white foamy residue was subjected to elution by column-10 chromatography (stationary phase: silica gel; eluent: a 3: 2 mixture of benzene and ethyl acetate), 1.3 g of a white powder having a melting range of 166-179 ° C (dec.), Which corresponded to the component in the upper zone in thin layer chromatography, and 1.4 g

O

of a white powder with a melting point of 180-190 ° C (dec.), which corresponded to the component in the lower zone in the thin layer chromatography.

The compound in the upper zone: probably diphenylmethyl-7- / ~ D-α- (2-trichloroethoxycarbonylamino-4-thiazolyl) -a- (4-ethyl-2,3-dioxo-1-piperazinecarboxamido) acetamido _ / - 3- £ 5- (1-raethyl-1,2,3,4-tetrazolyl) thiomethyl-A-1-cephem-4-carboxylate.

IR (KBr) cm -1: v C = O 1780, 1710, 1685.

MHR (CDCl2) 5 values: 1.12 (3H, t, -CH3), 2.90-3.87 (8H, ra,;> CH2 x 4), 3.47 (3H, s, ^> N- CH3), 4.12, 4.20 (2H, ABq,> CH2), 4.24, 4.90 (2H, ABq, ^ CH2) f 5.04 (1H, d, ^ CH), 5.56 (1H, d, ^ CH), 5.98 (1H, d, d, $ SCH), 6.76 (1H, 25 s, ^ CH), 6.89 (1H, s, ^ = CH), 7 .26 (11H, m, -C8, x2, NH), 8.62 (1H, bs,> NH), 9.70 (1H, d,> NH).

The lower zone component: probably diphenylmethyl-7- / L-α- (2-trichloroethoxycarbonylamino-4-thiazolyl) -a- (4-ethyl-2,3-dioxo-1-piperazinecarboxamido) acetamido _ / - 3- / 5- ( 1-methyl-1,2,3,4-tetrazolyl) thioraethyl-1-alpha-cep em-4-carboxylate.

IR (KBr) cm v C = 0 1780, 1710, 1685.

NMR (CDCl3) 6 values: 1.05 (3H, t, -CH3), 3.12-4.10 (811, m,> CH2 x 4), 3.81 (3il, s, -CH3), 4 , 16, 4.23 (2H, ABq,,> 0 ^), 4.84 (2H, bs,> CH2), 4.97 (1H, d, 35 ^ CH), 5.67 (1H, d, d, ^ CH), 6.12 (1H, d,> CH), 6.75 (1H, s, ^ CH), *** - ·· 4 -: * * y -a '- 48 - t 6 .84 (1H, s, ^ CII), 7.23 (10H, s, -Cg ^ x 2), 7.90-8.30 (2H, m, ^ NH x 2), 9.79 (1H, d, NH).

(2) 1.2 g of the component were dissolved in a mixture of 15 ml anhydrous chloroform and 5 ml anhydrous tetrahydrofuran

5 of the upper zone, obtained under (1), The solution was added at -75 ° C

2.17 ml of a solution of lithium methoxide (1.948 mmol / ral) in methanol was added and the resulting mixture was stirred at -75 ° C for 3 min. To the mixture at -75 ° C, 0.22 ml of tert. butyl hypochlorite was added all at once and the resulting mixture was stirred at -75 ° C for 10 min. After adding a solution of 0.2 ml acetic acid in 2 ml chloroform, the temperature of the reaction system was raised to room temperature. The reaction mixture was freed from solvent by distillation under reduced pressure and the residue was dissolved by adding 20 ml of ethyl acetate and 20 ml of water and the pH was adjusted to 6.5 with saturated aqueous sodium hydrogen carbonate solution. The organic layer was separated and 20 ml of water were added thereto, and the pH thereof was adjusted to 2.0 with 2N hydrochloric acid. The organic layer was separated again, washed with 10 ml of water and 10 ml of saturated sodium chloride solution in this order, dried over magnesium sulfate and freed from solvent by distillation under reduced pressure. The residue was purified by calorachromatography (stationary phase: silica gel; eluent: a 3: 2 mixture of benzene and ethyl acetate) to give 0.45 g of diphenylmethyl-7S-, [D-α- (2-trichloroethoxycarbonylaminoi) -4-thiazolyl) -a- (4-ethyl-2,3-dioxo-1-piperazine carboxamido) acetamido __ / - T7a-methoxy-3 - /> 5- (1-methyl-1,2,3,4-tetrazolyl) thiomethyl __ / -A ^ -cephem-4-carboxylate in white powder form, m.p. 155 ° - 177 ° C (dec.); yield 36.4%, IR (KBr) cm -1: v C-O 1785, 1710, 1685.

NMR (CDCl 3) 6 values: 1.10 (3H, t, -CH 3) 3.20-4.30 (10H, m, J> CH 2 x 5), 3.41 (311, s, -CH 3), 3.73 (3H, s, -CH3), 4ƒ73, 4.79 (2H, ABq,> CH2), 4.92 (1H, s, .CH), 5.74 (1H, d,!> CH) ), 6.74 (1H, s, >> CH), 7.04 (1H, s, £cH), 7.20 (11H, m, -CgHj. X 2 ^ NH), 8.34 (1H, bs,]> NH), 9.78 (1H, d,] NH).

(3) Ir> 7 ml of 85% formic acid were dissolved 0.4 35 g of diphenylmethyl-78- £ D-α- (2-tfichloroethoxycarbonylamino-4-thiazolyl) -a- * - 49 - (4-ethyl- 2,3-dioxo-1-piperazinecarboxamido) acetamido-7-armethoxy-3- / 5- (1-methyl-1,2,3,4-tetrazolyl) thioraethyl-A-1-cephem-4-carboxylate, obtained under (2). To the solution was added 0.4 g of zinc dust under ice-cooling and the mixture was stirred at the same temperature for 3 hours. The insoluble material was removed by filtration and the filtrate was freed from solvent by distillation under reduced pressure. The residue was dissolved by adding 7 ml of ethylene chloride, 3 ml of acetone and 10 ml of water and the pH of the solution was adjusted to 7.0 with a saturated aqueous sodium hydrogen carbonate solution. The organic layer was separated, washed with saturated aqueous sodium chloride solution, dried over magnesium sulfaac and freed from solvent by distillation under reduced pressure to obtain 0.21 g of a white powder. The white powder was dissolved in 2 ml of anisole and 2 ml of trifluoroacetic acid were added thereto under ice cooling, after which the solution was stirred at the same temperature for 30 min. The insoluble material was removed by filtration and the filtrate was freed from solvent by distillation under reduced pressure. The residue was treated with 5 ml of ethyl acetate to obtain 0.19 g of the trifluoroacetic acid salt of 73- / D-ct- (2-amino-4-thia-Z0-zolyl) -a- (4-ethyl-2,3-dioxo -1-piperazinecarooxamido) acetamido _ / - 7-methoxy-3 - / _ 5- (1-methyl-1,2,3,4-tetrazolyl] biomethyl _ / - A ^ -cephem-4-carboxylic acid, mp 175 ° - 190 ° C ( dec.); yield GO, 9%.

IR (KBr) cm -1: v C = O 1765, 1700, 1675.

The following compounds were obtained in an analogous manner.

Trifluoroacetic acid salt of 7β - / _ D-α- (2-amino-4-thiazolyl} -a- (4-ethyl-2,3-dioxo-1-piperazine carboxamido) acetamido _ / - 7a-methoxy-3 - / _ 5- (1 -carbamoylmethyl-1,2,3,4-tetrazolyl) thiomethyl-A-1-cephem-4-carboxylic acid.

30 Trifluoroacetic acid salt of 73- / D-α- (2-amino-4-thiazolyl) -a- (4-ethyl-2,3-dioxo-1-piperazine carboxamido) acetamido - / - 7a-methoxy-3-acetoxymethyl-A-cephem -4-carboxylic acid.

Trifluroacetic acid salt of 7S - / _ Da- (2-amino-4-thiazolyl) -a- (4-ethyl-2 »3-dioxo-1-piperazine carboxamido> aeeetamido _ / - 7a-35 methoxy-3- / 2- (5 -methyl-1,3,4-thiadiozolyl) thiomethyl-1-1-cephem-4-carboxylic acid.

-. u »*. - 50 - * (re) 0.16 g of the trifluoroacetic acid salt of 73- / Da- (2-amino-4-thiazolyl) -a- (4-ethyl-2,3-dioxo-) were dissolved in 3 ml of anhydrous acetone. 1-piperazine-carboxamido) acetamido 7-7a-methoxy-3- / 5- (1-methyl-3'-1,2,3,4-tetrazolyl) thiomethyl / -Δ-cephem-4-carboxylic acid, obtained under (3), and 0.0197 g (equimolar amount) of pyridine were added and the solvent solution was freed by distillation under reduced pressure. Water was added to the residue and crystals were collected by filtration to give 73- / D-α- (2-amino-4-thiazolyl) -a- (4-ethyl-2,3-dioxo-1-piperazine carboxamido) acetamido / -7a-methoxy-3- / 5- (1- 3-methyl-1,2,3,4-tetrazolyl) thiomethyl-A-cephem-4-carboxylic acid.

15 y * * ·· »’

Claims (4)

Pharmaceutical preparation which can be used for the treatment of bacterial infections in humans and animals, characterized in that it comprises one or more 7α-thoxycephalosporins of the general formula 1, in which represents a hydrogen atom, an acetoxy group or a optionally substituted 1,2,3,4-tetrazol-5-ylthio or 1,3,4-thiadiazol-2-ylthio group, A represents an alkyl group having 1 to 5 carbon atoms and B represents a thienyl, furyl, aminothiazolyl or cyclohexadienyl group, and / or one or more pharmaceutically acceptable salts thereof. 2. A 7a-methoxycephalosporin or pharmaceutically acceptable salt thereof, characterized in that it is a compound of the general formula 1, wherein R 1, A and B have the meanings set out in claim 1, or a pharmaceutically acceptable salt thereof. 3. Process for the preparation of a 7a-methoxy-cephalosporin or a pharmaceutically acceptable salt thereof, characterized in that a compound of the formula 1 in which R 1, R 1, A and B have the meanings set out in claim 1 , prepared in a manner known per se. 4. Compounds and methods as described in the description and / or examples. 25 * £ Cl o och3 V ~ f i N— N. N-CONH-CH — CONH -, - (r, \ B rf I 1 ^ l · »u COOR, 0. OCHj CHjCH, J'J —CONH— CH - CONH —i- * · \ fi I ru d 1b B JN-vj ^ 9ch3 0 COOH Rt — NH —CH — CONH - | -OOB Y-Nv ^ —CHjR2 y ~ -f 0. A— N N— COOH COOR1a 2 '-' 3 OCHj Ré — nh —j— ° y- (0 J H '~ y ~ CH2R2 A - ^ ^ —CONH-CH — COOH 0 COOR, aui 5 0 0 V “f ς m och3 A — NN — CONH-CH — CONH -i- [Λ 1 +" WBJ Nvj ^ 1—1CH * Rt 7 ° COOR1a 6 Π Π R 2 Mg. Yf ^ Aι, 9 A-N N-CONH -CH-CONH - fy -FY-CH-. R, H 2 N-CH-COOH 0. B (/ T I yy coor1s g b 10 a-n n —conh-ch-co— B «~ A n = τι— Hi-N ^ S ^ HN ^ S ^ Toyama Chemical Co., Ltd., Tokyo, Japan