AU597748B2 - Novel cephalosporin derivatives with improved pharmacokinetics, process for their preparation and pharmaceutical compositions in which they are present - Google Patents

Abstract

Derivatives corresponding to the general formula: <IMAGE> in which: - R1, R2 and R3 denote H, or R1 and R2 denote H or CH3 and R3 = COOH or else <IMAGE> form a cyclobutyl group and R3 = COOH; - A and B are different and occupy the meta and para positions of the benzene nucleus; one denotes a hydroxyl group and the other is chosen from the groups <IMAGE> and the pharmaceutically acceptable salts and esters of the said derivatives. Use in therapeutics.

Description

COMMONWEALTH OF AUSTRALIA Patents Act 1952 C O M P L E T E SP E. C I T I O N

(ORIGINAL)

Application Number Lodged Complete Specification Lodged Accepted his ocument contains the amendments made und,,r Section 49 and is correct for rin tg.

0 4 9u Published Priority 20 November 1986 Related Art Name of Applicant Address of Applicant Actual Inventor/s

SANOFI

:40, avenue George V, 75008 Paris France :Dominique OLLIERO; Bernard LABEEUW; Gilles ROCHE; Ali SALHI F.B. RICE CO., Patent Attorneys, 28A Montague Street, Balmain NS.W. 2041 Address for Service Complete Specification for the invention entitled: Novel cephalosporin derivatives with improved pharmacokinetics, process for their preparation and pharmaceutical compositions in which they are present The following statement is a full description of this invention including the best method of performing it known to us:- Sia The present invention relates to novel cephalosporin derivatives, a process for their preparation and pharmaceutical compositions in which they are present as the active principle.

In French Patent Application No. 84 14 878 published on 28/03/86 under number 2 570 702, the 10 Applicanj Company described a family of cephalosporin derivatives possessing a broad activity against both Gram-negative germs and Gram-positive germs.

These derivatives include, in particular, compounds substituted in the 3-position by a group: 15 -CH"2OC S- NH-CO-Alk-NH 2 g in which Alk represents an optionally substituted lower alkyl group and in which the substituent NH-CO-Alk- 0

SNH

2 is located in the 3-position or 4-position.

According to the present invention, it has 20 been found that, surprisingly, by slightly modifying the nature of the substituent in the 3-position, compounds are obtained which preserve the good activity of the compounds described in the prior art but additionally have very greatly improved pharmacokinetic properties and, in particular, very high plasma concentrations which persist for a very long time.

Modification of the pharmacokinetic parameters in this way is important insofar as it makes it possible to envisage reducing the dosage and the number of administrations of the product for the same therapeutic effect.

2 The compounds according to the invention correspond to the general formula: HN S 2 O (I) 2 S N C- C NH- N N R N CH 2

OCO

O-C-R B 1 2

COOH

R3 i3 10 in which: R1, R 2 and R3 each denote a hydrogen atom, or R 1 and R each denote a hydrogen atom or a methyl group 2 and R 3 denotes a carboxyl group, or R 1 and R2, taken 3 1 2 S* together with the carbon atom to which they are bonded, 15 form a cyclobutyl ring and R 3 denotes a carboxyl group; and A and B are different and occupy the meta and pa.ra positions of the benzene ring, one representing an OH group and the other denoting either: 20

R

S.14 a group -NH-CO-CH-NH-R in which R 4 represents hydrogen, a methyl group, a hydroxymethyl group or a group (CH 2 n-NH 2 in which n is an integer between 1 and 4, and R5 denotes hydrogen, 5 or if R 4 represents hydrogen, R 5 can also represent a lower alkyl group containing from 1 to 4 carbon atoms;.

or: a group -NH-CO-CH2CH2NH2; or: a group -NH-CO

NH

AdvantageousLy, the hydroxyl group (A or B) is in the meta position bf the benzene ring.

As a consequencie b:f the presence o'f an oxime I 3 group in their formula, the compounds can exist in 2 isomeric forms: syn and anti. The syn isomers, which have the superior therapeutic activity, are the preferred compounds.

If one of the substituents A or B represents a group NHCO-CH-NHR5 in which R 4 is other than hydrogen,

RA

the carbon atom carrying R 4 is an asymmetric carbon.

The compounds can therefore exist in the form of 4 diastereoisomers (arising from the D or L forms of 10 the amino acid) or in the form of a mixture of the 2 *00* diastereoisomers (arising from the DL form of the amino acid). The invention encompasses all these forms.

Advantageously, the substituents A and B of the compounds according to the invention represent respectively, the hydroxyl group in the meta position, the -NHCOCH NH 2 group in the para position of the benzene ring.

It is understood that the compounds (I) indicated below can exist: either in the form indicated Lin rormula 2 or in the tautomeric form HN 0 c 0 S H- C NH 0 A 0 I 2 0-C-R 0

B

-I 2

COOH

R3 in which R 1

R

2

R

3 A and B are as defined above.

The salts of the compounds of formula I (or form an integral part of tle invention.

These include salts with pharmaceutically acceptable acids which can be formed with the amino groups of the molecule, as well as salts with alkali metals or alkaline earth metals or salts with amino acids or amines, such as triethylamine or ethanolamines, which are capable of being formed with the carboxyl 4 group in the 4-position of the compound or, if it exists, with the carboxyl group present in the substituent of the oxime, or with both these carboxyl groups.

The same applies to the readily hydrolyzable or metabolically labile esters derived from one or other or both of the carboxyl groups.which may be present in the molecule. Among these esters, the following may be mentioned in particular: 10 the phthalidyl esters: I 0 0 the l-acetoxyethyl esters: -CH-OCOCH 3 kI 3 the l-ethoxycarbonyloxyethyl esters: -CH-OCO-OC H and the (4-methyl-2-oxodioxol-4-en-5-yl)methyl esters: -CH-C C-CH 3 S2 3 0 Q 15 C The invention also relates to a process for the preparation of the compounds of formula I (or represented by the following reaction scheme:

H

SCOOtBu N C C NH 2 II 11 0-C-R' T 1 3' COOtBu

R

2 5 A'

B'

(or one of its reactive S derivatives)

COOH

H

Tr-N S N R N CH 2

-OCO

~I 1 0 0-C-R' B' S. I 3 COOtBu R2 0. 3 p tIn these formulae, Tr represents a group 1 protecting the amine group, preferably the trityl 5 group, tBu represents the tert.-butyl group and R' denotes hydrogen or a readily labile ester group, preferably a COOtBu group.

A' and which occupy the meta and para positions of the benzene ring, are different, one of them representing an OH group and the other representing a group derived from the group B by the blocking of the amino group or groups contained therein with a labile group.

Finally, R 1

R

2

R

3 A and B have the meanings defined above.

The iodine compound 1 is reacted with the acid 2, in which the amine group or groups have been protected beforehand, according to a known method, by a group such as tert.-butoxycarbonyl or trichloroethoxycarbonyl.

i

I

I t 6 The reaction generally takes place in solution in a suitable solvent, preferably dimethylformamide, in the presence of potassium bicarbonate or a tertiary amine of low nucleophilicity, such as diisopropylethylamine.

The reaction is carried out at a low temperature of 0 to 20 0

C.

The resulting protected compound 3 is used to prepare the compounds by removal of the protecting 10 groups carried by the amine and carboxyl groups, according to a known process, in particular by hydrolysis in an acid medium using, for example, trifluoroacetic acid or a formic acid/hydrochloric acid mixture.

15 Under these conditions; the compound is isolated directly in the form of the salt of the amino group with the strong acid used for deprotection,'i.e.

r in the form of the trifluoroacetate or hydrochloride.

If appropriate, these salts can be converted 20 to other salts of strong acids -assing a solution of the hydrochloride or trifluo, ,etate over a basic ion exchange resin in the form of the salt of a weak acid (for example formate or acetate).

The solution thus obtained is treated with the 25 strong acid whose salt it is desired to obtain, and the resulting salt is isolated, for example by lyophilization.

The iodine derivatives 1 used as starting materials are known or can be prepared by a known process, especially in the manner indicated in German Patent Application No. 3 311 300.

The protected amino acids 2 are prepared from the corresponding hydroxyamino acids according to the equation: 7- HO 0CHOO H o VCOOH R 6 COOH COOH

SR

6 CONH- H N 6 2 4 2 in which R 6 represents the groups corresponding to:

-CH-NH-R

5

-(CH

2 2

NH

2 or

J--NH

S* in which the amine group or groups have been protected 5 and in which R 4 and R 5 are as defined above.

The reaction to form the amide is generally I carried out not with the acid R 6 COOH but with an activated ester thereof, such as the ester of Nhydroxysuccinimide or the ester of N-hydroxybicyclo[2.

2.1]hept-5-ene-2,3-dicarboxylic acid imide.

The reaction is carried out in solution in *dimethylformamide by heating at between 30 and 80 0

C.

0 O* The carboxylic acid esters and salts of the compounds of the invention are obtained from the 15 compounds by reactions known per se.

Thus, the inorganic salts are obtained by reacting the compounds with an equimolecular amount of an inorganic base such as sodium hydroxide, S' potassium hydroxide or sodium bicarbonate; the salification reaction is carried out in a solvent such as water or ethanol, and the salt obtained is isolated by evaporation of the solution.

The salts of organic bases are obtained by reacting a solution of the acid I with an equimolecular amount of the organic base in a suitable solvent or mixture of solvents. The salt is isolated by precipitation with ether. The esters are obtained by the known esterification processes; for example, a halogen I r 8 derivative will advantageously be reacted with a salt of the acid, such as the sodium salt; the reaction will preferably be carried out in a solvent which is capable of dissolving the starting acid derivative, for example in dimethylformamide.

The syn and anti isomeric forms are obtained by appropriately choosing the reactants.

The examples which follow will provide a clearer understanding of the scope of the invention S. 10 without however limiting it.

As is usual in this family of compounds, the Sproducts according to the invention do not have sharp melting points, but only decomposition points which do J not enable them to be characterized.

15 The products will therefore be characterized by their nuclear magnetic resonance spectra. Unless indicated otherwise, they are run at 250 MHz, the internal standard being hexamethyldisiloxane.

The spectra are run in deuterated dimethyl sulfoxide: 10 mg in 0.5 ml.

The chemical shifts are measured to ±0.01 ppm and the coupling constants to ±0.5 Hz.

The following abbreviations will be used: S singlet S* 25 D doublet D of D doublet of doublets b.s. broadened singlet SM multiplet Q quadruplet T triplet AB AB system J coupling constant Also, the elemental microanalyses were performed in each case and are in agreement wit: the formulae indicated.

r .:i.rr 'I 9 EXAMPLE 1 Bis-trifluoroacetate of 7-[2-(2-aminothiazol-4-yl)-2- (2-carboxyprop-2-yloxyimino)acetamido]-3-[[4-(2-aminoacetyl)amino-3-hydroxybenzoyl]oxymethyl]-3-cephem-4carboxylic acid, syn isomer (SR 43753) :i

R

1

R

2

-CH

3

R

3 -COOH, A -OH B -NH-CO-CH2-NH 2 (4)

S

S

S

j

S

S.

A) 3-Hydroxy-4-[(2-tert.-butoxycarbonylaminoacetyl)amino]benzoic acid A solution of 9.75 g of 4-amino-3-hydroxybenzoic acid and 9.75 ml of triethylamine in 90 ml of dimethylformamide is heated to 80 0 C. 20 g of the Nhydroxysuccinimide ester of N-tert.-butoxycarbonylglycine are added and the reaction mixture is kept at 80 0 C for 4 hours.

The solvent is evaporated off in vacuo and the residue is taken up in the minimum amount of water and poured into 2 liters of sulfate buffer pH 2.

The solid which separates out is filtered off, washed with water and dissolved in 1.2 liters of ethyl acetate. The organic solution is washed 3 times with 500 ml of water and then dried over magnesium sulfate.

The solvent is evaporated off to dryness and the solid residue is taken up in 300 ml of methylene chloride, with vigorous stirring.

The crystals are filtered off, washed with methylene chloride and dried at 100 0 C to give 16.5 g of the expected'product. M.p. 250 0

C.

NMR SPECTRUM run at 60 MHz in solution in deuterated dimethyl sulfoxide 1H at 9.13 ppm ArNHCO) 1H at 8.20 ppm J 8 Hz, aromatic H meta COOH) 2H at 7.47 ppm (M, aromatic protons ortho COOH) 1H at 7.38 ppm J 10 7 Hz, -NH-Boc) 2H at 3.77 ppm J 7 Hz, 2 9H at 1.30 ppm -C(CH 3 3 B) Tert.-butyl 7-[2-(2-tritylaminothiazol-4-yl)-2- (2-tert.-butoxycarbonylprop-2-yloxyimino)acetamido]- 3-[[4-(2-tert.-butoxycarbonylaminoacetyl)amino-3hydroxybenzoyl]oxymethyl]-3-cephem-4-carboxylate, syn isomer g of the protected acid prepared in A and 10.9 ml of diisopropylethylamine are dissolved in S. 10 100 ml of anhydrous dimethylformamide.

The solution is cooled to 4°C and 36 g of tert.butyl 7-[2-(2-tritylaminothiazol-4-yl)-2-(2-tert.- *butoxycarbonylprop-2-yloxyimino)acetamido]-3-iodomethyl- 3-cephem-4-carboxylate, syn isomer, are added. The 15 reaction mixture is stirred at '4C for 5 hours and then poured into an iced solution of sulfate buffer pH 2. The resulting mixture is stirred vigorously for 10 minutes and the solid is then filtered off and washed with water. The solid is taken up in ethyl acetate and the organic solution is washed with sulfate buffer pH 2 followed by water, a saturated solution of sodium bicarbonate and finally water.

The solution is dried over magnesium sulfate and the solvent is then evaporated off to dryness.

25 The product obtained is chromatographed on a column of silica H. Elution with a 90/10 (vol/vol) methylene chloride/ethyl acetate mixture gives 25 g of the expected product, which is used as such for the next step.

NMR SPECTRUM 1H at 10.5 ppm OH) 1H at 9.35 ppm J 9 Hz, 1H at 9.10 ppm ArNHCO-) 1H at 8.80 ppm NH-trityl) 1H at 8.15 ppm J 8 Hz, aromatic H meta CO) 18H at 7.25 ppm trityl aromatic protons, aromatic protons ortho CO and -NH-Boc) 11 1H at 6.64 ppm II thiazole) 1H at 5.73 ppm (D of D, J 9 Hz, J 4 Hz, H 7 2H at 5.10 ppm H and CH 2 1H at 4.81 ppm J 13 Hz,

CH

2 OC=0) 2H1 at 3.72 ppm J 7 Hz, C0-CH 2 2H at 3.57 ppm (AB, JAB 17 Hz, CH 2 S) 33H at 1.30 ppm (3S, H-Boc and -C-(CH3) 2 C) SR 43753 g of the protected product obtained in B are added in small portions to a mixture of 250 ml of 10 trifluoroacetic acid and 25 ml of anisole.

When the addition is complete, the mixture is left at 25 0 C for 1 hour and then evaporated to dryness in vacuo. 500 ml of anhydrous ether are added to the residue and the resulting mixture is stirred vigorously 15 for 15 minutes. The solid is filtered off, washed 3 times with anhydrous ether and then dried in vacuo.

18 g of the expected product are obtained: NMR SPECTRUM 1H at 10.60 ppm OH) 1H at 9.91 ppm ArNHCO) 1H at 9.42 ppm J 9 Hz, -CO-NH-) 4H at 8.10 ppm NH glycine and aromatic H meta C=O) 4H at 7.40 ppm aromatic protons ortho CO and NH 2 thiazole) 1H at 6.71 ppm H thiazole) 1H at 5.82 ppm (D of D, J 9 Hz, J2 4 Hz, H7) 2H at 5.20 ppm H 6 25 and CH OCO) 1H at 4.84 ppm J 13 Hz, CHO2CO) 2H at 3.85 ppm CU-CH 2 2H at 3.64 ppm (AB, JAB 17 Hz, CH 2 S) 6H at 1.40 ppm (2S, 3 2 EXAMPLES 2 TO A) By following the same procedure as in Example 1-A, but varying the products reacted, the protected acids collated in Table 1 are obtained in the same manner.

12 TABLE 1

A

HOOC

-B

NMR spectrum I no A B ormeting: .9...,point :7i.p.: sos,-NHCOCH 2NH-Boc OH 1 :.O2 -NHCOCH 2 CH NHI-Boc 2 -OF -NH-CO-CH-NH-Boc 3 so CH 3 -OHf CO-CHi-NHBoc 4 0 so. -OH-NH-COCH 9999 2O 2 -OH -NHf-C :m.p.:210-2 0

C:

:O Bo c so:: -OH -N}{-CO-CH-(CH 2 3 NHf-13oc 6 NH-Boc NMR SPECTRUM No. 1 (60 MHz, DMSO) 1H at 12.60 ppm-(b.s., GOOH) at 10.40 ppm OH) lI- at 9.05 ppm ArNH-) 1Hl at 8.60 ppm (D, J 3 Hz, aromatic H ortho NH) 1Hl at 7.57 ppm (D of D, J 8 Hz, J 2 =3 Hz, aromatic H para NH) 1B at 7.20 ppm J 7 Hz, -NHBoc) 1H at 6.90 ppm (D,

I

13 0 6

S

0

S

so 00 00 *00 6* 0@ S 0 @0500 0 @0 0 0S J 8 Hz, aromatic H meta NH) 2H at 3.72 ppm (D, J 7 Hz, -C(=O)-CH 2 9H at 1.35 ppm Boc) NMR SPECTRUM No. 2 1H at 12.50 ppm COOH 1H at 10.43 ppm OH) 1H at 9.24 ppm Ar-NH-) 1H at 8.00 ppm (D, J 8 Hz, aromatic H ortho NH) 1H at 7.40 ppm (S, aromatic H ,)rtho OH) 1H at 7.34 ppm J 8 Hz, aromatic H para OH) 1H at 6.81 ppm J 7 Hz, NH-Boc) 2H at 3.18 ppm J 7 Hz, -CH NH-Boc) 10 2H at 2.52 ppm J 7 Hz, CH2CH2NHBoc) 9H at 1.33 ppm Boc) NMR SPECTRUM No. 3 (60 MHz, DMSO) 1H at 12.40 ppm COOH) 1H at 10.40 ppm OH) 1H at 9.10 ppm ArNH) 1H at 8.10 ppm (D, 15 J 8 Hz, aromatic H ortho NH) 3H at 7.40 ppm (M, -NH-Boc 2 aromatic H) 1H at 4.12 ppm -CO-CH-N) I 3 9H at 1.34 ppm Boc) 3H at 1.27 ppm J 7 Hz,

-CH-N)

CH

-3 NMR SPECTRUM No. 4 20 1H at 12.60 ppm COOH) 1H at 10.45 ppm Ar-OH) 1H at 9.22 ppm ArNHCO) 1H at 8.18 ppm J 8 Hz, aromatic H ortho NH) 2H at 7.40 ppm (aromatic H meta NH) 1H at 7.15 ppm J 7 Hz, NHBoc) 1H at 5.06 ppm CH2OH) 1H at 4.17 ppm COCH-N) 2H at 3.57 ppm CO-CH-N) 9H at 1.34

I-

CH OH CH 2

OH

ppm Boc) NMR SPECTRUM No. 5 (60 MHz, DMSO) 1H at 12.6 ppm COOH) 1H at 10.47 ppm OH) 1H at 9.21 ppm ArNH-CO) 1H at 8.10 ppm (D, J 8 Hz, aromatic H para OH) 2H at 7.43 ppm (M, aromatic protons) 2H at 4.00 ppm COCH2N) 3H at 3.80 ppm N-CH 3 9H at 1.32 ppm Boc) Nor 14 NMR SPECTRUM No. 6 1H at 12.66 ppm COOH) 1H at 10.44 ppm OH) 1H at 9.11 ppm ArNHCO) 1H at 8.12 ppm (D, J 8 Hz, aromatic H ortho NH) 3H at 7.40 ppm (M, aromatic protons -CH-CH2) 1H at 6.75 ppm J NHBoc 7 Hz, CH 2 NHBoc) 1H at 4.08 ppm -CH-CH 2H at

N

2.81 ppm CH -N 4H between 1.4 and 1.7 ppm (M, -2 CH -CH 18H at 1.33 ppm (2S, Boc) 2 2 NMR SPECTRUM No. 7 10 1H at 12.55 ppm COOH) 1H at 10.47 ppm OH) 1H at 9.15 ppm ArNHCO) 1H at 8.13 ppm (D, SJ 8 Hz, aromatic H ortho NH) 3H at 7.40 ppm (M, aromatic protons -CH-CH 2 IH at 6.78 ppm J

I

NHBoc 7 Hz, CH 2 NHBoc) 1H at 4.13 ppm CH-CH 2 2H at 2 2 S:o N 15 2.98 ppm -CH2NHBoc) 1H at 1.92 ppm and 1H at 1.70 ppm -CH-CH 2 18H at 1.33 ppm (2S, Boc) 3 N f A\ B) Reaction of these protected acids with different S iodine compounds according to Example 1-B gives the .corresponding protected cephalosporins, which, when 20 deprotected according to the method of Example 1-C, lead to the different compounds isolated in the syn form as the trifluoroacetates; these are collated in Table 2.

151 15 TABLE 2 Example SR code:

A

no no R R 2 R :NMR

'I-B

j: 2 43851: H H H OH 8 NHCOCH NH 133852 H H H a HNHCOCH NH 29 2 2 9

OH

4 43853 H H H

OH

5 43903 CH3 CH3 COOH OH 11N2 3 A( NCC NH *2 2 6 43904 :CH :CH :COOH: NHCOCH CH NH 1

OH

7 43955 CH 3 CH3 COOH :N-OHNH-CH- 13 2 CH (DL OH 3 2 16 439 83 43984

S

0O

S

S@ OS U 0 5 0 0

S.

S S

S.

S.

S

S..

*5 S S

S*

S

S

S

505055

S

SS

S

S

10 43985 H H CH CH3 CH 3 CH3 CH3 CH3

C

H: NHCO-CH-NH 2 OH ~CH 2OH OHO2

COOH

COOH

COOH

SNUCOCH 2NHCH3

OH

NHCO

OH (DL) N NHCOCH(CH 3

NH:

OH H 2

(L)

16 17 18 14: 11 44049 44128 12 13 44130 :CH 3 :CH3 COGH \NHCOCH(CH 2 2 NH 2 19 OH NH 2

(DL)

14 :44218: H :H :H 44219 :H :H :H NHCOCH(CH9 3 NH: 21: i NH (L OH 2

(L

17 NMR SPECTRUM No. 8 IH at 11.13 ppm Ofl) in at 9.80 ppm (S, ArNHCO) iH at 9.57 ppm J =9 Hz, CONH) IH at 8.52 ppm aromatic H ortho NH) -3H at 8.02 ppm CH NH) in at 7.61 ppm J 8 Hz, aromatic H para NH) 211 at 7.2,5 ppm NH 2 thiazole) 1H at 6.98 ppm J 8 Hz, aromatic H meta NH) 1H at 6.70 ppm H thiazole) -iH at 5.77 ppm (D of D, J =9Hz, J 4 Hz, H iH at 5.20 ppm J 13 Hz, 1 H 7 10 CH 2 OCO) 1H at 5.16 ppm J =4 Hz, H 6 1 H at 4.85 ppm J 13 Hz, CH 2OCO) -3H at 3.77 ppm (S,

NOCH

3 2H at 3.58 ppm (AB, JA 17 Hz, CH 2

S)

SNMR SPECTRUM No. 9 11- at 10.57 ppm OH) IH at 9.90 ppm ArNHCO)- 15 1H at 9.61 ppm J =9 Hz, CON-) -4H at 8.10 ppm aromatic H ortho NH and CH NH) 4H at 7.40 ppm (aromatic protons meta NH and NH 2 thiazole) iH at 6.72 ppm H thiazole) 1HI- at 5.81 ppm (D of D, 0""0 zJ 4H, 7 2H at 5.10 ppm H 6 an~ CH 2 OCO) in at 4.90 ppm J 13 Hz, CH 2 OCO) 2H atc 3.85 ppm CH 2

NH

2 3H it 3.77'ppm N-OCH 3 2at3.65 pm(AB, J AB 17 Hz, CH 2S) NMR SPECTRUM No. dO*in at 10.47 ppm OH) -2H at 9.54 ppm CONH and ArNHCO) hi at 8.07 ppm J 8 Hz, aromatic H ortho NH) 3H at 7.80 ppm CH 2 NH. 2H at 7.45 ppm V(M, aromatic protons meta NH) 2H at 7.20 ppm

NH

2 thiazole) in at 6.70 ppm H thiazole) in at 5.78 ppm (D of D, J 9 Hz, J 2 4 Hz, H 2H at 5.13 ppm H 6 and CH 2 OCO) in at 4.87 ppm J 13 Hz, CH 2 OCO) 31H at 3.80 ppm N-OCH 3 2H at 3.60 ppm (AB, J AB 17 Hz, CH 2 S) 211 at 3.05 ppm (M, CH NH+) 211 at 2.80 ppm CH CH NH) 18 NMR SPECTRUM No. 11 1H at 11.20 ppm OH) 1H at 9.80 ppm ArNHCO)- 1H at 9.56 ppm J 9 Hz, CONH) 1H at 8.52 ppm aromatic H ortho NH) 3H at 8.06 ppm CH NH) lH at 7.60 ppm J =8 Hz, aromatic H para NH) 2H1 at 7.30 ppm NH 2thiazole) H at 6.98 ppm (ID, J 8 Hz, aromatic H meta NH) -1H at 6.71 ppm H thiazole) 1H at 5.82 ppm (ID of D, J 9 Hiz, J 2 4 Hz, 1 7 2H1 at 5.15 ppm. H16 and CH 2

OCO)-

1H at 4.81 ppm J 13 Hz, CH OCO) -2H at 3.84 ppm CH 2 NH) 3 2H at 3.60 ppm (AB, J AB =17 Hz, CH 2

S)-

6H at 1.35 ppm (2S, -C-(CH 3 2 NMR SPECTRUM No. 12 1H at 10.48 ppm OH) 1H at 9.54 ppm ArNHCO)- 1H at 9.45 ppm (ID, J 9 Hz, CONH) 1H at 8.10 ppm J =8 Hz, aromatic H ortho NH) 3H at 7.70 ppm CH NH+) 48 at 7.40 ppm NH 2 thiazole,.

2-3 2 aromatic protons meta NH) 18 at 6.70 ppm H ***thiazole) 1H at 5.80 ppm (D of D, J 1 9 Hz, J 2 4 Hz, H 7 2H at 5.15 ppm 8 6 and CH 2 OCO) 1H at 4.82 ppm J 13 Hz, CH OCO) 28 *at 3.62 ppm (AB, 4 AB =17 Hz, CH S) 2H at 3.05 ppm CH NH)- 2H at 2.80 ppm CH 2 CH 2 NH) 3 6H at 1.34 ppm (2S, NMR SPECTRUM No. 13 1H at 10.60 ppm s. OH) 18 at 9.95 ppm (S, ArNHCO) 18 at 9.47 ppm J 9 Hz, CONH) 48 at 8.20 ppm -ClI1-NH and aromatic protons ortho NH)- CH 3 4H at 7.40 ppm NH 2 thiazole and aromatic protons meta NH) IH at 6.71 ppm H thiazole) -1H at 5.88 ppm (D of D, ~J1 9 Hz, J2= 4 Hz, H 7 2H1 at 5.20 ppm H16 and CH 2 000) 111 at 4.84 ppm. J= 13 Hz, CH OCO) 1H at 4.20 ppm -CH--NH 28 at -2 I- 3 CH 3 19 3.62 ppm (AB, J AB =17 Hz, CH 2 S) -9H at 1.35 ppm

-C-(CH

3 2 and -CHNH C H -3 NMR SPECTRUM No. 14 1H at 10.62 ppm, OH) -1H at 9.83 ppm (S, ArNHC0) 1H1 at 9.59 ppm J 9 Hz, CONH) 4H at 8.30 ppm, CH-NH~ and aromatic H ortho NH) -211 at 7.43 1 3

CH

2

O

ppm aromatic protons meta NH) 2H at 7.25 ppm

NH

2 thiazole) Ill at 6.69 ppm H thiazole) IIH at 5.79 ppm (D of D, J 9 Hz, J 4 Hz, H 2H 1 217) 10 at 5.15 ppm H 6 and CH 2 OCO) Ill at 4.86 ppm (D, J 13 Hz, CH 2 OCO) Ill at 4.24 ppm CH-NH)- 3H CHI OH 2 at 3.81 ppm N0CH 3 2H at 3.76 ppm J =6 Hz, CH-NH)- 2H at 3.63 ppm (AB, JA 17 Hz, ClH 2

S)

3 B CH 2OH 4 NMR SPECTRUM No. Ill at 10.60 ppm OH) 1H at 9.90 ppm (S, ArNHC0) Ill at 9.42 ppm (ID, J =9 Hz, CONH) 4H at 8.20 ppm CH-NH and aromatic H ortho NH) 2H at *,7.45 ppm aromatic protons meta NH) 2H at 7.30 ppm NH 2 thiazole) IlH at 6.69 ppm H thiazole) IlH at 5.84 ppm (D of D, J 1 9 Hz, J 2 4 Hz, H 7 2H at 5.17 ppm. H 6 and CH 2 000) 1H at 4.84 ppm J 13 Hz, CH 2 OCO) IIIH at 4.25 ppm Tn-NH+) 2H at 3.75 ppm (ID, J =6 Hz, CH-NH) CH OH CH OH 2 -2 2H at 3.68 ppm (AB, J AB 17 Hz, CH 2 S) 6H at 1.35 ppm C(CHl 3 2 NMR SPECTRUM No. 16 1H at 10.57 ppm OH) IlH at 9.98 ppm (S, ArNHCO) IlH at 9.40 ppm J 9 Hz, GONH) 2H1 at 8.80 ppm CH NH+-CH )-1H1 at 8.08 ppm J 2 2 3 8 Hz, aromatic H1 ortho NH) -2H at 7.45 ppm (M, aromatic protons meta NH-) -2H at 7.25 ppm NH 2 thiazole) 1H at 6.68 ppm H thiazole) 1H at 5.82 ppm (D of D, J =9 Hiz, J -4 Hiz, H 7 2H 127 at 5.18 ppm 11 6 and C 11 2 OCO) 1H1 at 4.83 ppm (D, J= 13 Hz, CH OCO) 2H at 4.00 pm(M, CH NHiCH) pp 22 3 2H at 3.61 ppm (AB, J =17 Hiz, CH 2 S) 3H at 2.57 AB2 ppmi CH 2 NH 2

CH

3 6H at 1.37 ppm (2S, C(CH 3 2 NMR SPECTRUM No. 17 ArNHCO) -1H at 9.43 ppm J 9 Hz, CONH) 2H at 8.5t0. ppm OH 1H at 9.5 ppm (,J 8Haromatic H ortho NH) -4H at 7.40 ppm NH 2 thaol n aromatic protons meta NH) 111 at 6.69 ppm H thiazole) -1HI at 5.84 ppm (D of D, J 1 =9Hz 0 *~goB 4 Hz, H 7 2H at 5.20 ppm. H 6 andC 2

O)-

)f 1H at 4.82 ppm J 13 Hz, CH 2 OCO) 2H at 3.66 ppm (AB, J AB= 17 Hz, CH 2 S) 5H between 2.5 and 3.5 ppm CO-CH and CH 2 N) 4H between 1.45 and 2.05 ppm -2 (CH 2of the ring meta and para. to N )-6H at 1.40 ~*ppm (2S, C(G11 3 2 NMR SPECTRUM No. 18 IIH at 10.80 ppm OH) -1H at 10.00 ppm (S, ArNHCO) 1H at 9.44 ppm. J 9 Hz, CONH) -3H at 8.20 ppm NH) 3 1H at 8.05 ppm J 8 Hz, aromatic H ortho NH) -3H at 7.70 ppm N 4H at 7.30 ppm (NH 2 thiazole and aromatic protons meta NH) 1H at 6.66 ppm H1 thiazole) 1H at 5.82 ppm (D of D, J 1 =9 Hz, J2=4 Hz, H 7 2H at 5.20 ppm H 6 and CH 2 OCO) 1H at 4.84 ppm J 13 Hz,

CH

2 OCO) -1H at 4.42 ppm -CjH-NH 2 2H at 3.61 ppm (CH 2 3 NH 2 (AB, J AB =17 Hz, CH 2 S) 2H at 2.83 ppm CH-NH CH 2

-CH

2 CH 2 Nil 21 2H1 at 1.80 ppm -011-N 211 at 1.60 ppm UlH 2 CH 2 C 1-12 NH3 (MI -OH-N 611 at 1.39 ppm (2S, CC

CH

2

CH

2

CH

2

NH~

NMR SPECTRUM No. 19 1H1 at 10.72 ppm OH) 1H at 10.00 ppm (S, ArNHCO) 111 at 9.36 ppm J =9 Hz, CONH) -3H at

S+

**8.40 ppm NH) 3 1H at 8.11 ppm J 8 Hz, aromatic H ortho NH) -311 at 7.90 ppm NH 411 at 7.30 ppm aromatic protons meta NH and NH 2 o thiazole) 11H at 6.66 ppm H thiazole) -11H at 5.81 ppm (D of D, J 1=9 Hz, J 2- 4 HH7 Ha 05.15 ppm 116 and CH 2 OCO) 1H1 at 4.82 ppm J 13 Hz, CH12000) 111 at 4.33 ppm CH-NH+) 211 at (C (12)2 NH 3 a eke 3.64 ppm (AB, J AB 17 Hz, CH 2 S) 21H at 2.90 ppm (M, go CH-NH -211 at 2.15 ppm CH-N11+ 611 at 1. -39 ppm CHCHN CH 2 H 2NH 3 2 CC 2 N1 3 -2 3 15 (2S, 0(011)2 NMR SPECTRUM No. so. 1H at 10.80 ppm OH) 111 at 10.05 ppm ArNHCO)- 1H1 at 9.62 ppm J =9 Hz, 0ONH) -311 at 8.40 ppm NH+) 1H1 at 8.10 ppm J =8 Hz, aromatic H1 para 0OH) 311 at 7.85 ppm NH) 211 at 7.45 ppm aromatic protons) 211 at 7.20 ppm NH 2 thiazole) 1H1 at 6.66 ppm H thiazole) 111 at 5.76 ppm (D of D, J I 9 Hiz, J2= 4 Hz, 117) 211 at 5.14.

ppm 116 and OH 000C) 111 at 4.95 ppm J =13 Hz, 2 11 000C) 111 at 4.35 ppm C-H+ 3 t38 p (CfH 2 )2 NH 13 (S,,NOCH 3 211 at 3.60 ppm CH 2 S) 211 at 2.88 ppm 011-N+ 211 at 2.06 ppm OH-N CH 2 CH2 NH 3CH 2 CH 2 NH 3 22 22 NMR SPECTRUM No. 21 1H at 10.75 ppm OH) 1H at 10.01 ppm (S, ArNHCO) 1H at 9.58 ppm J 9 Hz, CONH) 3H at 8.40 ppm NHI 1H at 8.09 ppm J 8 Hz, aromatic H para OH) 3H at 7.80 ppm NH3) 2H at 7.46 ppm aromatic protons) 2H at 7.20 ppm

NH

2 thiazole) 1H at 6.71 ppm H thiazole) -2 1H at 5.79 ppm (D of D, J1 9 Hz, J 2 4 Hz, H 7 2H at 5.18 ppm H6 and CHO 2 CO) 1H at 4.92 ppm (D, 10 J 13 Hz, CH OCO) II at 4.30 ppm CH-NH 3H at

(CH

2

NH

3.82 ppm NOCH3) 2H at 3.60 ppm (AB, JAB 17 Hz,

'*CH

2 S) 2H at 2.78 ppm CH-NH) -2H at 1.80 ppm 2 CH CH CH NH 3 2 2 -2 3 H-NH) 2H at 1.62 ppm CH-NH) CH CH CH NH CH CH CH NH 2 2 2 3 2 2 2 3 The products according to the invention were studied for their pharmacological properties.

The bacteriostatic action was determined in vitro by the dilution method. The study was carried out on both Gram-positive strains and Gram-negative strains.

The results for various products according to the invention, expressed as minimum inhibitory concentrations (MIC Pg/ml), are collated in Table 3.

23 TABLE 3 PRODUCT SR No STRAIN-- 43753: 43852: 43904: 43853: 44128: 44219: 44130: 44218: @0*

S.

S

Staph. aureus Smith 2 :0.25 4 0, 025 8 0.5 8 0.25 :0.25 Escherichia coli Cl/Cal El Tn3 0. 12 :0.06 0. 25 0.06 12 0. 12 Escherichia coli SOL RL 90: 2 0.25 2 0.25 2 0.12 2 Kiebsiella pneumoniae R30: 1 :4 .1I 4 1 :4 :1 :8 Proteus vulgaris GN 76/C-1 0.25 0.12 12 16 0.12 Providencia 155 :1 Pseudomnonas aeruginosa :2 NCTC 8203 .2 1 1 .2 :2 :1 2 :2 1 mommol.- I F; 24 These results show that the products according to the invention have a broad spectrum of activity and possess a very good intrinsic activity.

Furthermore, the pharmacokinetic behavior of the products according to the invention was studied in baboons after administration at a dose of 20 mg/kg by intramuscular injection.

Blood samples taken at various times after administration are used to determine the plasma cont 10 centration of the product studied by microbiological j analysis.

4 It is thus possible to construct the curve showing the plasma concentration as a function of time, and to determine different pharmacokinetic parameters of the compound studied: the elimination half-life (t 1 p) is calculated by 2 the formula ln 2, in which represents the elimination gradient; the area under the curve (AUC) is determined by the trapezium method.

Furthermore, the protein binding is obtained by comparing two standard scales, one being prepared in baboon plasma and the other in phosphate buffer (pH 7, 0.03 M).

The plasma concentrations obtained with various products of the invention are collated in Table 4.

They are expressed in pg/ml.

For the purpose of comparison, this table includes the results obtained with 2 products of the prior art, corresponding respectively to the formulae: I -W 25 C CO -NH N CH 0-C-CooH f 7 CH 2 OCO-R 2CF 3C CCII COO H 0 000 00 0 S 0 0 00000 0 *0 0 0

S

0*0 0* 5 0 0@ R NHCOCH 2NH2 R NHCOCH 2CH 2NH2 Compound A Compound B

S

*00000 0.000.

S

09 *0 0 00 000000 ft ft 262 TABLE 4 TIME PRODUCT (minutes)-- SR 43753: SR 43904: SR 44128: SR 44130:: A :B *.10 36.8 12.6 62.9 39.1 ::20.8 :20 69.1 ND 115.3 60.9 36.8 :16.4 95.2 52.4 134.8 81.9 ::44.4 :16.9 128.6 73.0 176.6 104.7 ::59.3 17.6 1 39.2 76.1 187.9 121.6 ::60.0 :20.7 146.0 94.1 187.8 143.4 ::55.5 :24.1 120 151.1 76.1 194.6 141.6 ::53.3 :20.7 0180 126.1 59.7 168.8 137.8 ::40.0 17,6 240 120.9 57.3 141.9 3.34.8 ::36.2 :12.5 300 112.6 53.4 123.0 109.1 ::27.2 8.3 *330 9. 399 ND 93.8 ::23.7 6.8 *360 80.9 34.3 104.7 90.4 17.3 4.6 ND not determined 27 The pharmacokinetic parameters of the products according to the invention and the comparison products, determined according to the same experiments, have been collated in Table 5 below.

TABLE

C

C.

e g.

C

C.

C S

S

C.

C

C

*0 CS C

C

PARAMETERS PRODUCTS SR 43753 SR 43904 SR 44128 SR 44130 A B Max plasma concentration (pg/ml) 151.1 94.1 194.6 143.4 69.5 24.1 T max (min) 120 90 120 90 45 Plasma concentration at 6h (pg/ml) 80.9 34.3 104.5 90.4 17.3 4.8 tj (min) NC 265 NC NC 195 99 AUC 0-6h (pg.

ml- 1 .min) 42178 20859 53890 42403 17033 5276 Excretion in the urine dose, 6h) 18 17 72 37 23 18 Protein binding 84 71 84 92 45 48 NC: not calculable 28 The results given in Tables 4 and 5 show that the plasma concentrations of products of the invention are extremely high and long-lived.

If a comparison is made with the reference products A and B, the 6-hour plasma concentrations obtained for their hydroxylated homologs are respectively 4.6 and 7 times higher. Likewise, if the area e" under the curve is considered, the increase is res- *pectively 2.5 and 4 times, compared with the reference 10 products.

The compounds according to the invention c therefore have very advantageous pharmacokinetic parameters which make it possible substantially to reduce the amount of active principle used and the number of 15 daily administrations which are necessary for a given therapeutic effect.

Finally, the toxicity of the products according to the invention is sufficiently low for them to be used in therapy.

The products of the invention can therefore be employed as antibiotics in human or veterinary medicine.

They have a broad spectrum and can be used for all bacterial infections caused by sensitive germ.s.

The products can be administered by a general route (parenteral, oral, rectal) or by the topical route.

The pharmaceutical compositions are prepared from the compounds in a soluble form obtained by salification of at least one of the acid groups or amine gcoups present therein.

The pharmaceutical compositions containing the antibiotic according to the invention as the active ingredient, in combination with a pharmaceutically acceptable vehicle, can be solid or liquid and cin take the form of, for example, injectable preparations, 29 tablets, gelatine capsules, granules, ointments, creams, gels or suppositories. They contain, for exampte, from 50 to 1000 mg of active principle. The dosage can vary within wide limits, in particular depending on the type and severity of the infection to be treated and depending on the method of administration.

Most frequently, the adult dosage for administra-

S.

tion by injection is between 0.250 g and 4 g per day.

As an example of a pharmaceutical preparation, 10 it is possible to prepare an injectable solution contai-' ning the following in each ampoule SR 43753 1 g Water for injectable preparations 5 ml 15 Sodium carbonate q.s. for pH 6.3 0 *r

Claims (8)

1. Cephalosporin derivatives corresponding to the general formula: H2N S 0 11 s 0-C-R 0 O 2 COOH R 3 *3 in which: R1, R 2 and R 3 each denote a hydrogen atom, or R 1 and R2 each denote a hydrogen atom or a methyl group and R 3 denotes a carboxyl group, or R1 and R 2 taken together with the carbon atom to which they are bonded, form a cyclobutyl ring and R 3 denotes a carboxyl group; and A and B are different and occupy the meta and para positions of the benzene ring, one representing a S* hydroxyl group and the other denoting: a group -NH-CO-CH-NH-R S R 4 in which R 4 represents hydrogen, a methyl group, a hydroxymethyl group or a group (CH) n-NH2, in which n is an integer between 1 and 4, and R 5 denotes hydrogen, or if R 4 represents hydrogen, R 5 can also represent a lower alkyl group containing from 1 to 4 carbon atoms; a group -NHCOCH 2 CH 2 NH 2 or a group -NHCO NH and also the pharmaceutically acceptable salts and esters of the said derivatives.

2. Compounds as claimed in claim 1 in which the i; 31 hydroxyl group is in the meta position of the benzene ring.

3. Cephalosporin derivatives as claimed in one of claims 1 or 2, corresponding to the general formula in which the oxime is in the syn form.

4. Compounds as claimed in claims 1, 2 or 3 in which A represents OH in the meta position and B denotes -NHCOCH 2 NH2 in the para position.

5. A process for the preparation of the derivatives as claimed in claim 1, which consists in e reacting, in solution in a polar aprotic solvent, a compound of the formula: H S Tr-N 0 I I S C C NH C RCHI 0 C COOtBu in which R 1 and R 2 are as defined in claim 1, R' 1 2 3 denotes hydrogen or a readily labile ester group and Tr represents a group protecting the amine group, with an acid of the formula: A' COOH (or one of its reactive derivatives) B' in which A' and which occupy the meta and para positions of the benzene ring, are different, one of them representing a hydroxyl group and the other representing a group corresponding to the groups: -NHCOCH CH2NH2 or NHCO pen n S32 in which R 4 and R 5 are as defined in claim 1 and in which the amino group or groups have been protected by a labile group; removing the protecting groups present on the amine and carboxyl groups by hydrolysis in a strong acid medium; isolating the resulting compound in the form of a salt of the amino groups present in the molecule; if appropriate, converting the said salt to another salt of the amino groups; and if appropriate, isolating, from an amine salt, the corresponding compound I in the form of the base and, if appropriate, converting it, by known processes, to a salt e or ester derived from one or both of the carboxyl groups present in the molecule. es S.

6. Pharmaceutical compositions containing at least one of the compounds of formula as claimed in claim 1 as the active principal in combination with a pharmaceutically acceptable vehicle.

7. A pharmaceutical composition as claimed in claim 6, in the form of dosage units.

8. A pharmaceutical composition as claimed in one of claims 6 or 7, containing 50 to 1000 mg of active principle. DATED this 13th day of March 1990 SANOFI Applicant: F.B. RICE CO.