AU660311B2 - Amino acid azetidinyl substituted pyridone derivatives, their preparation and their application as medications - Google Patents

Description

AUSTRALIA

Patents Act 660311 COMPLETE SPECIFICATION

(ORIGINAL)

Class Int. Class Application Number: Lodged: Complete Specification Lodged: Accepted: Published: Priority Related Art: Name of Applicant: Laboratorios Del Dr. Esteve, S.A.

Actual Inventor(s): Jordi Frigola Constansa Juan Pares Corominas Jordi Corbera-Arjona Address for Service: PHILLIPS ORMONDE FITZPATRICK Patent and Trade Mark Attorneys 367 Collins Street Melbourne 3000 AUSTRALIA Invention Title: AMINO ACID AZETIDINYL SUBSTITUTED PYRIDONE DERIVATIVES, THEIR PREPARATION AND THEIR APPLICATION AS MEDICATIONS Our Ref 288673 POF Code: 2975/113338 The following statement is a full description of this invention, including the best method of performing it known to applicant(s): 6006 -1h) The present invention describes new azetidine derivatives of pyridonecarboxylic acids such as 1,4dihydro-4-oxoquinoline-3-carboxylic, 1, 8-naphthyridine- 4-oxo-3-carboxylic and 2,3-dihydro-7-oxo-7H-pyrido( 1,2,3de]-1,4-benzoxazine-6-carboxylic acids, and azetidine derivatives of isothiazolopyridone, such as 2,3,4,9tetrahydroisothiazolo([5,4-b]naphthyridine-3,4-dione, 1,2,8,9-tetrahydro-7H-isothiazolo[4',5':5,6]-pyrido- [1,2-,3-de]-benzoxazine-7,8-dione, therapeutically acceptable salts of these compounds, process for their preparation and their application as medications.

European Patent Application 88403352.3, where azetidi derivatives of quinolones and of pyridobenzoxazin are described. It is also a continuation of Europ Patent Application 90400684.8, where various azet' ine derivatives of naphthyridines, of quinolones d of pyridobenzoxazines are described. It is al a continuation of European Patent Application 9 01036.0, where various azetidine derivatives of othiazolonaphthyridines, of isothiazoloquinolones and of isothiazolopyridobenzoxazines are cribed, all being compounds which have a very goo antimicrobial activity but which nevertheless hav a very low solubility at a physiological pH, which r uces their absorption by oral route and which als have a low solubility in water, which makes them ifficult to use injectable formulations for intravenous, The present invention describes new azetidine derivatives of quinolones, of naphthyridines, of pyridobenzoxazines, of isothiazoloquinolones, of iso*hiazolonaphthyridines and of isothiazolopyridobenzoxazines, in which the azetidine ring is 3-A-aminoazetididine or else 3-A-aminomethylazetidine, in which A is a solubilising group. The solubilising group A is an amino acid residue or a polypeptide chain, preferably a di-, tri- or tetrapeptide, or ele3 these same amino acids or polypeptide chains in which the functional groups are suitably 2 protected by groups such as a benzyloxycarbonyl, a tertbutoxycarbonyl, ethers such as, for example, a benzyl ether or a tert-butyl ether, N-trifluoroacetyl or other well-known protective groups for the synthesis of peptides, as well as their therapeutically acceptable salts.

The compounds which are the subject matter of the present inventio correspond to the general formula I: R4 o o A-N- FI \j I in which X denotes a nitrogen atom or else a carbon atom bonded to a hy~f~ ogen atom or else a carbon atom bonded to a halogn atom in which Y denotes a fluorine, chlorine', bromine atom, or else a carbon atom bonded to a hydroxyl radical (C-OH) or alkoxy radical (C-0-alkyl), or else a carbon atom bonded to an alkyl radical (C-alkyl), or else a carbon atom bonded to a halogenated alkyl radical (C-CF 3 or else a carbon atom i)nded to an alkylamino radical (C-NHalkyl); R, denotes a lower alkyl or cycloalkyl radical, a lower haloalkyl radical, a hydroxyalkyl radical, a vinyl radical, an aryl radical or an aryl radical substituted by one or more'Aluorine atoms, or an alkylamino radical (NHalkyl);

R

2 denotes a hydrogen atom or a lower alkyl radical;

R

3 denotes a hydroxyl radical or a CI-C 4 lower alkoxy radical; 3

R

4 denotes a hydrogen atom, a fluorine atom, a lower alkyl radical, a nitro radical, an amino radical or a sbstituted amino radical; Rs, R, and R, denote a hydrogen atom or a lower alkyl radical. Depending on the nature and the relative position, the azetidine substituents may have up to two chiral centres, each of them with an or "S" configuration; R, denotes a hydrogen atom or a lower alkyl radical; Ii n is a number which may be 0 or 1; X and RI may together form a bond represented by a group C-CHz-CHz-CH-R 9 or C-O-CH 2

-CH-R

9 or C-S-CHz-CH-R, in which R, denotes a hydrogen atom, a lower alkyl radical or a lower haloalkyl radical and which in the tw, latter cases have one chiral centre with an or configuration; RI and R 2 may together form a bond represented by a group -CHRio-S- in which R 10 denotes a hydrogen atom or a lower alkyl radical or a lower haloalkyl radical and in the two latter cases have one chiral centre with an "R" or configuration;

R

2 and R 3 may together form a bond represented by an -S-NH- group; A is a solubilising group which is represented by a residual amino acid or a polypeptide chain consisting of two to four or more amino acid residues which are bonded covalently one to another via peptide bonds, and thOir pharmaceutically acceptable salts.

The amino acid residues of the present invention include the 20 natural amino acids, denoted by means of the three letters symbolising them, as well as other amino acids such as norvaline (Nva), homoserine, 4-hydroxyproline, hydroxylysine, ornithine, and the like.

The conventional symbolic designations of the amino acids are the following: 'Alanine Ala Arginine Arg -4 Asparagine Asn Aspartic acid Asp Glutamic acid Glu Cysteine Cys Glutamine Glu Glycine Gly Histidine His Isoleucine Ile Leucine Leu Lysine Lys Methionine Met Phenylalanine Phe Proline Pro Serine Ser Threonine Thr Tryptophan Trp Tyrosine Tyr Valine Val It must be mentioned that the 3-amino or 3-aminomethyl group of the azetidine ring forms a bond with the carboxylic group of amino acid residue A. In the case of the polypeptide chains the amino acid residues are bonded one to another via a conventional peptide bond; thus the alpha-amino group of the first bonded amino acid residue is bonded to the carboxylic group of the second alpha-amino acid residue, and so on.

The stereochemical configuration of the amino acids of the present invention may be of the D- or Lform or a mixture of the two, the use of amino acid residues in L-form being nevertheless preferable.

The compounds forming the subject matter of the present invention are particularly active against bacterial microorganisms and consequently are useful in human or veterinary medicine for prophylaxis and chemotherapy of local and systemic infections produced by aerobic or anaerobic pathogens, both Gram-positive and Gram-negative. In addition to exhibiting a high antibacterial activity, the compounds forming the subject matter 5 of the present invention show an Increase In solubility both at physiological pH and in water that it permits their use by a parenteral route, as well as an excellent bloavailab llty. Futhermore, the compounds forming the subject matter of the present Invention can act as proxy drugs for previously known azetidinylquinolones, azetidinylnaphthyridines, azetidinylpyridobenzoxazines, azetidinylisothiazoloquinolones, azetidinylisothiazolonaphthyridines or azetidinylisothiazolopyridobenzoxazines, given that the solubilising group A, when A is an amino acid residue or a polypeptide chain, can undergo an enzyme hydrolysis which produces the release of the previously known azetidinylquinolones, azetidinylnaphthyridines, azetidinylpyridobenzoxazines, azetidinylisothiazoloquinolones, and azetidinylisothia-.

zolopyridobenzoxazines.

The compounds o, general formula I and their physiologically acceptable salts, such as the salts of inorganic acids like, for example, hydrochlorides, and the salts of organic acids like, for example, toluenesulphonates or methylsuphonates, are preferably administered in this form in the pharmaceutical compositions.

SYNTHESIS OF THE COMPOUNDS The compounds of general formula II, which have a 3-amino-l-azetidinyl group or a 3-aminomethyl-l-azetidinyl group, both variously substituted, and which are bonded to position 7 of various naphthyridines and quinolones, or which are bonded to position 10 of pyridobenzoxazines or which are bonded to position 7 of isothiazolonaphthyridines and isothiazoloquinolones, or which are bonded to position 11 of isothiazolopyridobenzoxazines, which are used for preparing the heterocyclic compounds of the general formula I, can be prepared b known methods such as those described in, ur an 3 A^pp ications A0o0684.0 and D 0,33iO and The heterocyclic compounds of general formula IV, which can be employed as starting materials for preparing the compounds of general formula II, can be prepared by 6 known methods in the bibliography Koga, A. Itoh, S. Murayama, S.Suzne and T. Irikura, J. Med. Chem., 1980, 23, .1358; H. awa, T. Miyamoto, A. Minamida, Y. Nishimura, H. Okada, H. Uno and J. Matsumoto, J. Med. Chem., 1984, 27, 1543; D.T.W. Chu, P.B. Fernandes, A.K. Claiborne, L. Shen and A.G. Pernet, Drugs Exptl. Chim. Res., 14 379, (1988).

The compounds of general formula VIII, which can be employed as starting materials for preparing the compounds of general formula I, can be prepared by known methods such as those described in European Patent Application 90401860.3.

A general scheme for the preparation of the compounds of the present invention of general formula I.

by means of amino acid coupling techniques is shown in the following diagram and detailed in Examples 2aA, 3aA, 4aA, 2aL, 3aL, 4aL, 2bA, 3bA, 4bA, 2bL, 3bL, 4bL, 2ca, 3ca, 4ca, 3da and 4da.

7- 0 0 P1.-A.P2 or P1 -A

R

PN 11

I

I

H-N--

R

6 PI-A >tY6nN-H P1ANI(H) M, V

A-N-

P1-A AsViI P 1 -A-P2.

P1-A H -N (eli -H

A-N-

1-.qit 8 Diagram 1 shows various synthesis sequences for the preparation of compounds of general formula I in which-A denotes an amino acid residue.

SCHEME I Method A A compound of general formula III is obtained, by reaction of a compound of general formula II in which R 1

R

2

R

3

R

4 RS, Re, R 7

R

8

R

9 R10, X and n have the abovementioned meanings, with a compound of general formula P1-A-P2, in which A has the abovementioned meanings and PI denotes any one of the protective groups for functional groups employed in the synthesis of peptides Bodansky, Principles of Peptide Synthesis (Reactivity and Structure: Concepts in Organic Chemistry, V.16), publ..

Springer Verlag, Berlin Heidelberg 1984. M. Bodansky'and A. Bodansky, The Practice of Peptide Synthesis (Reactivity and Structure: Concepts in Organic Chemistry, V.21), (publ. Springer Verlag, Berlin Heidelberg 1984), such as protectors for the amine functional group, such as, for example, benzyloxycarbonyl, tert-butoxycarbonyl, biphenylisoptopoxycarbonyl, 9-fluorenylmethoxycarbonyl, and other well-known groups; P1 also denotes protective groups for the chain, such as ethers like, for example, benzyl ether, tert-butyl ether, N-trifluoroacetyl, or protectors for the carboxylic group, such as esters like, for example, benzyl esters or tert-butyl esters, as well as other well-known protective groups (Theodora W.

Greene, Protective Groups in Organic Chemistry, John Wiley and Sons, 1981); P2 denotes any one of the activating groups for the carboxylic functional group which are employed in the synthesis of peptides, such as, for example, acid chloride, a symmetric anhydride, a mixed anhydride such as isovaleric or pivalic, and the mixed anhydride of ethyl or isobutyl carbonate, an active fster such as N-hydroxysuccinimide, N-hydroxyphthalin\ide, cyanomethyl or o-nitrophenyl ester, as well as the use of other well-known activating groups for the carboxylic functional group.

9 The compound of general formula III can also be formed by reaction of II with a compound of general formula P1-A and with the aid of coupling reactants such as DCC (dicyclohexylcarbodiimide), DCC in the presence of 1-hydroxybenzotriazole, or DCC in the presence of N-.hydroxysuccinimide, or by using IIDQ (1-isobutyloxycarbonyl-2-isobutyloxy-1,2-dihydroquinoline) or EDC (N-ethyl-N'-3-dimethylaminopropylcarbodiimide hydrochloride), or of other well-known reactants for the formation of peptide bonds.

The reaction is performed in the presence of an appropriate solvent, for example dimethylformamide, dimethyl sulphoxide, an alcohol, an ether like, for example, ethyl ether, dioxane, 1,2-dimethoxyethane or. tetrahydrofuran, ethyl acetate, dichloromethane, chloroform, benzene, toluene, pyridine, as well as in an organo-aqueous medium like pyridine-water.

SThe reaction takes place in the presence of a base such as N-ethylmorpholine, N-methylmorpholine, sodium hydroxide, pyridine or triethylamine.

The reaction temperature varies between -15 0 C and /that of reflux of the solvent, and the reaction time is between ten minutes and 24 hours.

Method B The preparatidn of the compound of general formula I is obtained by reaction of a compound of general formula III, in which R R 2 i3, R 4

R

5

R

6

R

7

R

a

R

9 Ro 0 X, A, P1 and n Aave the abovementioned meanings, with reactants which are useful for the removal of protective groups for functional groups present in the amino acids and peptides of the compound of general formula III.

This removal of the protective groups can be carried out by means of a catalytic hydrogenation, reduction with sodium in liquid ammonia, hydrazinolysis, acidolysis, for example by employing hydrobromic acid in acetic acid, hydrobromic acid in trifluoroacetic acid, hydrochloric acid in acetic acid or by employing 10 trifluordacetic acid, as well as by means of a hydrolysis with an alkali, an enzyme-catalysed hydrolysis or by employing other methods which are well known in the synthesis of peptides.

The reaction is performed in the absence of other solvents or else in a solvent such as an alcohol like, for example, ethanol, methanol or isopropanol, dimethylformamide, dimethyl sulphoxide, acetic acid, trifluoroacetic acid or water.

The most appropriate temperatures vary between and that of reflux of the solvent, and the reaction time is between ten minutes and 24 hours.

Method C A compound of general formula VII is obtained by reaction of a compound of general formula VIII in which

R

5 Ra, Ry, R 8 (arsdn have the abovementioned meanings and in which RI i-a diphenylmethyl radical, compounds already described in European Patent Application 90401860.3, with a compound of general formula P1-A-P2, or a compound of general formula P1-A in which Pl, A and P2 have the abovementioned meanings, and by employing the techniques for forming peptide bonds, mentioned in Method A.

Compounds of general formula VI are obtained by removal of the protective groups R 11 and P1 from a compound of general formula VII by means of procedures already referred to in the section Method B.

The compounds of general formula I are obtained by- reaction of a compound of general formula IV in which

R

1

R

3

R

4 Ro and X have the abovementioned meanings and Z denotes a halogen, preferably fluorine or chlorine, atom, with an azetidine of general formula VI.

The reaction is performed in the presence of an appropriate solvent, like, for example dimethylformamide, dimethyl sulphoxide, pyridine, trialkylamines such as triethylamine, dichloromethane, chloroform, and ethers such as tetrahydrofuran or dioxane, or a mixture of these solvents. The most appropriate temperatures vary between 11 room temperature and th reflux temperature of the solvent, and the reaction time is between 1 hour and 24 hours.

Method D By hydrogenolysis of a compound of general formula VII, in which R 5

R

6

R

8 A, PI, n and R, have the abovementioned meanings, by means of the use of a palladium catalyst, preferably Pd(OH),/.C under a hydrogen atmosphere and in an appropriate solvent such as alcohol like, for example, ethanol or methanol, and at a hydrogen pressure of between 1 atm and 20 atm and the appropriate temperatures vary between 200C and 7 0 a compound of formula V Is obtained.

The compounds of general formula III are obtained by reaction of a compound of general formula IV, in which..

R

1

R

2 R3, R 4

R

9

R

10 and X have the abovementioned meanings and Z denotes a halogen, preferably fluorine or chlorine, atom,, with an azetidine of general formula V.

The reaction is performed in the presence of an appropriate solvent, for example dimethylformamide, dimethyl sulphoxide, pyridine, trialkylamines such as triethylaminie, dichloromethane, chloroform and ethers such as tetrahydrofuran or dioxane, or a mixture of these solvents. The most appropriate temperatures vary between room temperature and the reflux temperature of the solvent, and the reaction time is between 1 hour and 24 hours.

Method E The corresponding salt is obtained by reaction of a -compound of general formula I with an inorganic or organic acid in an appropriate solvent such as ethanol or methanol.

The preparation of new derivatives according to the invention will be shown in the examples which follow.

Some typical forms of use will also be described in the case of the various fields of application.

-The examples below, given merely by way of illustration, mnust not, however, limit the scope of the invention in any way.

12 Example 2aA. Preparation of 7-E(2S,3R)-2methyl-3 -N-CBZ-Ala-amino-1-azetidinyl ]-1-cyclopropyl-6fluoro-1, 4-dihydro-4-oxo-,1, 8-naphthyridine-3-carboxylic acid.

The ester N-CBZ-kla-N-hydroxysuccinimide (742 mg, 2. 31 mmol) is added 'to a solution of 7- 3R)-2-methyl- 3 -amino- 1-azetidinylj]-1l-cyclopropyl-6-f luoro-1, 4-dihydro- 4-oxo-1,8-naphthyridine-3-.carbo ylic acid (la) (700 mg,/ 2.11 mmol) and N4-methylmorpholine (213 mg, 2.13, mmol) iry, dry dimethylformamide (30 ml) cooled to 080,, and is kept at this temperature for 1 hour and then at r~oom temperature for 8 hours. The resulting solution is added to a solution of hydrochloric acid (200 ml, 0.5N) and a precipitate is f ormed, which is filtered off. washed with.

water, dried over P 2 0 5 and 7-((2S,3R)-2-metbyl,-3-N-CBZ- Ala-amino-1-azetidinyl]- 1-cyclopropyl-6-fluoro-1, 4dihydro-4-oxo-1,,8-naphthyridine-3-carboxylic acid (2aA) (1.1 g, 2.04 nimol) with .a melting point of 211-3 0 C is obtained.

0' :27.7 (c 0.78, DMSO) gf~trosdopic data 1H 1MR(100 MAHz) (DMSO-d 5 1.11 (m,4H); 1.21 (dJ 7.0 Hz, 3H); 1.59 (dJ 6.1 Hz,3H);. 3.60 (in, 1H); 3.85,-4.70 5Hi); 5.01 7.30 7.97 (dJ 11.5 H~z, 1H1); 8.56 1H).

IR (KBr): 3325, 1720, 1680, 1632, 1509, 1449, 1328 cm--.

Example -3aA.- Preparation of 7-[(2S,3E)-2methyl-3-Ala-amnino-1-azetidinyl]J-1-cyclopropyl-6-fluoro- 1, 4-dihyciro-4-oxo-1, 8-naphthyridine-3-carboxylic acid.

mg of 10%-Pd/C ar 'e added to a solutioni of cyclopropyl-6-f luoro-1, 4-dihydro-4-oxo-1, 8-naphthyridine- 3-carboxylic acid (2a.A) (960 mg, 1.78 nimol) in 80 ml' of dimethylformamide, and the mixture is kept under hydrogen atmosphere, for 24 hours. The catalyst is filtered off and washed well with dcimethylformamide. The solvent is evaporated off at reduced pressure and the resulting 13 solid crystallises from an' ethanol-water mixture and propyl-6-fluoro-1, 4-dihydro-4-oxo-1, 8-naphthyridine-3carboxylic acid (3a.A) (500 mg, 1.23 mmol) is obtained M4.P. 220-222*C DcJ 16.9 (c 0.75, DMSO) 'H NI4R (100 MHz) (DMSO-d 6 /TFA): 1.16 1.39 (d,J =7.00K-Hz, 3H), 1.65 (d,J 6.2 Hz, 3H), 3.55-4.00 (in, 2H), 4.00-4.80 4H), 8.02 (dJ 11.6, 1H), 8.15 3H), 8.6 1H), 8.95 (i,1H).

IR (KBr): 3630-2420 1630, 1510, 1500, 1450, 1362, 1320 cmf'.

Exal Preparation of the hydrochloride of (2S, 3R)-2-methyl-3-Ala-amino--azetidinyl]--cyco-.- 16- propyl-6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3carboxylic acid.

0.35 g (0.86 inmol) of 3R)-2-methyl-3-Alaamino-1-azetidinyl ]-1-cyclopropyl-6-fluoro-1, 4-dihydro- 4-oxo-1, 8-naphthyridine-3-carboxylic acid (3aA) are treated with a solution of EtOH-HC1, the solvent is evaporated of f at reduced pressure and 0. 37 g 84 mmiol) of the hydrochloride of 7- 3R) -2-inethyl-3;-Ala-anino- 1-azetidinyl] -1-cyclopropyl-6-f luoro-1, 4-dihydro-4-oxo- 1, 8-naphthyridine-3-carboxylic acid (4a.A) are obtained.

M.p. 190-192-C 20 [a1ID -16.2 (c 0.88, DMSO) 'H NMR (100 MHz) (DMSO-dr/TFA): 1.10 (in,4H), 1.38 (d,J 7.A Hz, 3H), 1.63 (d,J =6.2 Hz, 3H), 3.50-4.00 4.00-4.80 4H), 8.00 J 11.5 Hz, 1H), 8.16 3H), 8.58 1H 9.13 (mn, 1H).

IR (KBr) 3620-2400 1718, 1686, 3,63!L, 1561, 1490, 1449, 1328.

Example 2aL. Preparation of 7-[(2S,3R)-2iethyl-3-N-t-BOC-Leu-amino-1-azettidinyl) -1-cyclopropyl- 6-f luoro-1, 4-dihyciro-4-oxo-1, 8-naphthyridine-3-carboxylic acid.

I4 N-t-BOC-Leu N-hydroxysuccinimide ester (433 mg, 1.32 mmol) is added to a solution of 7-[(2S,3R)-2-methyl- 3-amino-1-azetidinyl]-1-cyclopropyl-6-fluoro-1,4-dihydro- 4-oxo-1,8-naphthyridine-3-carboxylic acid (la) (400 mg, 1.20 mmol) and of N-methylmorpholine (121 mg, 1.20 mmol) in dry dimethylformamide (20 ml) cooled to 0 0 C and this temperature is maintained for one hour and room temperature overnight. The resulting solution is poured onto a solution of hydrochloric acid (200 ml, 0.5N) and a precipitate appears which is filtered off, washed with water and dried in a desiccator with P 2 0, and 7-[(2S,3R)- 2-methyl-3-N-t-BOC-Leu-amino-1-azetidinyl]-1-cyclopropyl- 6-fluoro-1,4-dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid (2aL), (600 mg, 1.10 mmol) is obtained.

M.p. 117-120 0

C

17.0 (c 0.71, DMSO).

1 H NMR (100 MHz) '(DMSO-ds/TFA-dl): 0.85 (d,J 5.9 Hz, 6H), 1.14 4H), 1.35 (s,12H), 1.59 (d,J 5.9 Hz, 3H), 3.55-4.70 6H), 7.97 J 11.5 Hz, 1H), 8.57 1H).

IR (KBr): 3318, 2962, 1719, 1631, 1509, 1447, 1368, 1331 cm-.

Example 3aL. Preparation of 7-[(2S,3R)-2methyl-3-Leu-amino-1-azetidinyl]-1-cyclopropyl-6-fluoro- 1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid.

0.54 g (0.99 mmol) of 7-((2,3)-2-methyl-3-N-t- BOC-Leu-amino-1-azetidinyl]-1-cyclopropyl-6-fluoro-1,4dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid (2aL and 15 ml of trifluoroacetic acid are mixed cold and kept at r6om temperature for an hour. Ether is added and the precipitate which appears is filtered off and is washed with ether. The salt thus formed is dissolved in water and adjusted to a pH of approximately 7.6 with NH 3 and a precipitate appears which is filtered with water, is washed and is dried in a desiccator with P 2 0 5 and 0.25 g (0.56 nmmol) of 7- (2S,3R) -2-methyl-3-Leu-amino-1-azetidinyl]-1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carbpxylic acid (3aL) are obtained.

15 M.P. =216-.218*C D]0 +9.7 (c 0.76, DMSO) 1 H NMR (100 M4Hz) (DMSO-d 5 /TFA): 0.92 J 8 Hz, 6H) 1. 13 4H) 1. 65 J 5. 9 Hz, 6H), 3.68 (in, 2H) 4.05-4.80 (in, 4H), 8. 03 J 11. 7 Hz, 1H), 8.15 (b 3H), 8.60 1H), 9.09 (mn, 1H).

IR (KBr) 3331, 2962, 1724, 1636, 1571, 1509, 1449 cm Ex~unple 4aL. Preparation. of the hydrochloride of (2S,3E)-2-inethyl-3-Leu-amino-1-azetidinyl]-1-cyclopropyl-6-fi~uoro-1,4-dihydro-4-oxo-1, 8-naphthyridine-3carboxylic acid.

0.20 g (0.45 inmol) of 7-[(2S.,3R)-2-methy1-3-Leuamino- 1-azetidinyl 1-cyclopropyl-6.-f luoro-1, 4-dihydro-.

4-oxo-1,8-naphthyridine-3-carboxylic acid (3a1 8 -are treated with a solution,,of EtOH-HC1, the solvent is evaporated off at reduced pressure and 0.21 g (0.44 mmol) of hydrochloride of (2S, 3R) -2-methyl-3-Leu-amino-1azetidinyl]-1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-1,8naphthyridine.-3-carboxylic acid (4aL) are obtained.

M.P.18 1-184 0

C

[cx]f +23.8 (c 0.75, DMSO) 1H NMR (100 (;DSO-d 6 /TFA): 089 (d,J= 5.2 Hz,6H),, 1.11 1.63 (d,J =5.5 Hz, 6H), 3.43 (mn, 2H), 4.05-4.80 4H), 8.01 J 11.4 Hz, 1H), 8.15 3H) 8'.58 111), 9.19 (mn, 1H).

IR (KBr): 3600-2400 1718, 1687, 1630, 1562, 1512, 1449, 1325 cmf'.

Example 2bA. Preparation of 7-(3-N-CBZ-Alaamino-1-azetidinyl)-1-(2,4.-difluorophenyl)-6-fluoro-1,4dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid.

The N-CBZ-Ala N-hydr'oxysuccinimide ester (704 mng.

2.2 inmol) is added to a so'lution of 7 -amino- I-azetidinyl) ,4-difluorophenyl)-6-fluoro-1,4-dihydro-4-oxo- 1,8-naphthyridine-3-carboxylid acid (1b) (78,0 mg, 2 inmol) and of N-rnethylmorpholine (202 ing, 2 inmol) in dry DMF ml) cooled to 0 0 C and the mixture is kept at this temperature for one hour and at room temperature 16 overnight. The resultiz~g solution is poured into a solution of HCl (200 ml, 0.5N) and a precipitate appears which -is filtered off, and washed with H 2 and which is dried over P 2 0 5 1.06 g (1.78 nimol) of .7-(3-N-CBZ-Alaamino-1-azetidinyl)-1-(2,4-difluorophenyl)-6-fluoro-1,4dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid (2bA) is obtained.

M.p. =205-208 0

C

=a3 +11.9 (c 0.63, DHSO) 1H NMR (100 Hz) (DMSO-d6/TFA-dl): 1.18 (d,J Hz, 3H), 3.70-4.70 6H), 4.98 7.05-7.80 8H, (6 7.27, 7.97 (d,J 11.3 Hz, 1HI), 8.71 1H).

IR (KBr): 1449, 1509, 1631, 1656, 1719, 33,18 cnf'.

Example 3bA. Preparation of 7-(.3-Ala-axnino-1azetidinyl) ,4-difluorophenyl) -6-f luoro-1, 4-dihydro- 4-oxo-1 ,8-naphthyridine-3-carboxylic acid.

0.8 g (1.34 inmol), of 7-(3-N-CBZ-Ala-amino-1azetidinyl)-1-(2,4-difluorophenyl)-6-fluoro-1, 4-dihydro- 4-oxo-1,8-naphthyridine-3-carboxylic acid (2bA) are dissolved in 2 ml of acetic acid. To this' solution are added 10 ml of a solution of acetic acid saturated with hydrobromic acid. The mixture is kept at room temperature for one hour, ether is added and a precipitate appears which is filtered off a~nd which is washed with ether. The salt is dissolved in water and adjusted to 7H 7.6 and a solid precipitates and is filtered off, washed with water and dried in a desiccator with P 2 0 5 and 0.43 g (0.93 mmol) of 7-(3-Ala-amino-1-azetidinyl)-1-(2,4difluorophenyl)-6-fluoro-1,4-dihydro-4-oxco-1,8naphthyridine-3-carboxylic acid (3bA) are obtained.

M.p. 1 56~-152 0

C

Dc]o +7.80 (c G.591 DMSO) 'H NMR- (100 MHz) (d.-DMSO/TFA) (T 310 1.33 J 7. 0 1 311), 3.70-4.15 (mn, 3H1), 4.15-4.70 (mn, 3H1), 7.10-7.J80 3H1), 8,01 J =11.2 Hiz, 1H1), 8.1. 311) 8.7 1R1), 9.10 (in, 1H1) 17 IR (KBr) 3418, 2262, 3075, 1636, 1561, 1510, 1 459 cmC'.

S Example 4bA. Preparation of the hydrochloride of7-(3-Ala-amino-1-a2zetidinyl)-1-(2,4-difluorophenyl)- 6-f luoro-1 ,4-dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid.

0.33 -g (0.71---nmo1) of 7-(3-Ala-amino-1-azetidinyl) ,4-difluorophenyl) -S-fluoro-1, 4-dihyd.ro-4-oxo- 1,8-naphthyridine-3-carboxylic acid (3bA) are treated with a solution of EtOH-HCl, the solvent is evaporated off at reduced pressure and 0.35 g (0.70 mrmol) of hydrochloride,, of 3-Ala-aini-l-azetidinyl) ,4difluorophenyl)-6-fluoro-1,4-dihydro-4-oxo-1,8naphthlyridine-3-carboxylic acid (4bA) are obtained.

=204-207 0

C

D +5.1 (c 0.84, DMSO).

'H NNR (100 MHz) (DMSO-dr/TFA): 1.31 (d,J= Hz, 3H), 3.70-4.70 (in, 6H), 7.10-7.80 3H), 7.85-'8.15 8.71 1H), 8.94 (in, 1H).

IR (KBr): 3600-2700 1725, 1687, 1632, 1492, 1450.

Exampl1e 2bL. Preparation of 7-(3-N-:t-BOC-Leuamino-1-azetidinyl) 4-dif luorophenyl) -6-f luoro-1, 4dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid.

N-t-BOC-Leu N-hycdroxysuccinimide ester (721 mg, 2.2 minol) is added to a solution of 7- (3 -amino- 1-azetidinyl) 4-dif luorophenyl) -6-f luoro-1, 4-dihydro-4-oxo- 1,8-naphthyridine-3-carboxylic acid (1b) (780 mg, 2 minol) and of N-inethylmorpholine (202 ing. 2 mmiol) in dry DMF (30 ml) cooled to 0 0 C and the mixture is kept at this temperature for one hour and at room temperature overnight. The resulting solution is added to a solution of HCl (200 ml, 0.5N) and a precipitate appears which is filtered off and which is wqas hed with water and which is dried in a desiccator with P 2 0 5 1. 11 g 82 inmol) of 7- (3-N-t-BOC.-Leu-anino--azetidinyl) (2,4-dif luorophenyl) "6-f luoro-lf 4-dihydro-4-oxo-1, 8-naphthyridine-3ceiboxylic acid (2bL) are obtained.

18 M.P. =123-1217*C 8.2 0.75, DMSO).

1H NMR (100 M4Hz) (DMSO-d 6 /TFA-dl): 0.87 J= 5.6 Hz, 1.38 12H), 3.80-4.70 (in, 6H), 7.15-7.90 (in, 3H), 8.0 (d1 11.3 Hz, IH), 8.78 1H).

IR (K 3331, 2956, 1719, 1633, 1509, 1448 cm-'.

Example \,3bL. Preparation of 7-(3-Leu-aminoazetidin-yl)-=( 2, 4-dif luorophenyl) -6-f luoro-1,4-dihydro- 4-oxo-1, 8-naphthyridine-3-carboxylic acid.

0.8 g (1.32 mmol) of 7-(3-N-t-BOC-Leu-amino-1azetidinyl) ,4-difluorophenyl) -6-fluoro-1,4-dihydro- 4-oxo-1,8-.naphthyridine-3-carboxylic acid (2bL) and 10 ml of trifluoroacetic acid are mixed cold and kept for one hour at room temperature. Ether is added and the precipitate which appears is filtered off and is washed with ether. The salt thus formed is dissolved in water and is stirred at PH 7.6 with NH 3 and a precipitate appears, which is filtered off, washed with water and dried in a desiccator with P 2 0 5 and 0.56 g (1.11 mmoi) of 7-(3-Leuamino-1-azetidinyl) 4-difluorophenyl)-6-fluoro-1, 4dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid (3bL) are obtained.

M.p. =214-216 0

C

C a12D =0.55, DHSO).

'H NMR (100 MHz) (DMsO-d./TFA) (T =320 K) 0. 89 (d,J =5.3 Hz, 6H), 1.57 (in, 3.67 (mn, 1H), 4.02 (in, 2H), 4.42 (mn, 3H),7.19-7.86 (mn, 3H), 8.03 J= 11.2 Hz, IH), 8.16 3H), 8.74 1H), 8.99 (in, 1H).

IR (KBr): 3600-2400 (bj, 1725, 1637, 1561, 1509, 1460,, 1357 cm-'.

Example 4bL. Preparation of the hydrochloride of 7-(3-Leu-ainn-1-azetidinyl)-1-(2,4-.difluorophenyl)- 6-f luoro-1, 4-dihyciro-4-oxo-1, e-naphthyridine-3-carboxcylic acid.

0.45 q (0.89 nimol) of 7-(3-Leu-.aiino-1-azetidinyl)-1-(2 ,4-difluorophenyl)-6-fluoro-1,4-dihydro-4-oxo- 1,8-naphthyridine-3-carboxylic acid (3bL) are treated with a solution of EtOH-HCl, the solvent is evaporated 19 off at reduced pressure and a solid is obtained, which is crystalli.sed with EtOH-ether and 0.43 g (0.79 inmol) of the hydrochloride of 7-(3-Lfu-amino-1-azetidinyl)-1-(2,4difluorophenyl) -6-f luoro-1, 4-dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid (4bL) are obtained.

M.P. =190-195*C +15.6 (c 0.76, DMSO) 1H NMR (100 MHz) (DMSO-d./TFA): 0.87 J= 5.1 Hz, 6H) 1.54 (in, 3H), 3.60-4.70 6H), 7.15-7.80 (in, 3H), 8.03 J =11.2 Hz, 1H), 8.16 3H), 8.75 1H), 9.35 1H).

IR 3600-2400 1718, 1687, 1631, 1512, 1450 cm- 1 Example 2ca. Preparation of 7-(3-methyl-3-N:-..

CBZ-D-Ala-ainino-1-azetidinyl) -1-cyclopropyl-6, 8-difluoro- 1,4-iy r--oo3qioincroyi aci4.

4' solution of N-CBZ-D-alanine (319 mg, 1.43 iriiol) is neutralised in 7 ml of dry tetrahyc~rofuran and cooled to -15 0 C with N-me-Zhyliorpholine (145 ing, 1.43 minol) and isobutyl chioroformate (195 mg, 1.43 minol) is added. Five minutes later a suspension of ,7-(3-niethyl-3-anino.-lazetidinyl) -cyclopropyl-6 ,8-difluoro-1, 4-dihydro-4-oxco- 3-quinolinecarboxylic acid (1c) (500 mg, 1,-13 nunol) in ml of dimethylfornaride is added, the mixture is kept 2 5 at -15 0 C for 15 minutes and is left at room temperature with vigourous stirring for 8 hourts. The mixture is then added to a cold solution of HCl (100 ml, 0.5N) and a precipitate forms, which is filtered off, washed with water and dried with P 2 0, and 570 mng (1.03 mmiol) of 7-(3-iethyl-3-N-CBZ-D-Ala-amino-1-azetidinyl)-l-cycllopropyl-6,8-difluoro-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid (2ca), with a melting point of 138-1410C, are obtained.

Spectroscopic data-, 'H NMR (100 MHz) (DMSO-d 6 /TFA-dj):0.85-1.28 1.54 3H), 3.79-4.60 6H), 5.00 2H), 7.30 51), 7.64 J 13.1 Hz, 1H), 8.55 1H).

20 IR (KBr) 3320, 1725, 1680, 1627, 1528, 1459, 1413, 1327 cm-'.

Example 3ca. Preparation of 7-(3-methyl-3-D- AJa- amino- 1-azet iinyl) -1-cyclopropyl-6, 8-dif luoro-1, 4dihydro-4-oxo-3-quinolinecarboxylic acid.

mg of 10% Pd/C are added to a solution of 7- (3-methyl-3-N-CBZ-D-Ala-amino-1-azetidinyl).1-cyclopropyl-6, 8-dif luoro-l, 4-dihydro-4-oxoquinolinecarboxylic acid (2ca) (196 mng, 0.35 mmol) in 15 ml of dimethylformamide and the mixture is kept under a hydrogen atmosphere for 24 hours. The catalyst is filtered of f and is washed well with dimethylformamide. The solvent is evaporated off at reduced pressure a nd the resulting solid crystallises from an ethanolA iater mixture and.

7-(3-methyl-3-D-Ala-amino-l-azetidinyl)-l-cyclopropyl- 6, 8-difluo-ro-1, 4-dihydro-4-oxo-quinolinecarboxylic acid (3ca) (63-mg, 0.15 mmol) is obtained.

M.P. 154-160 0

C,

1 H NMR (100 MHz) (DMSO-ds/TFA): 1.15 (in, 4H), 1.36 J 6.5 Hz, 31H), 1.57 3H), 3.65-4.70 6H), 7.61 J 11.4 Hiz, 1H), 8.05 3H), 8.56 1H), 8.90 (in, 1H).

IR (KBr): 3600-2400 1718, 1675, 1628, 1528, ,1467, 1326 cm-.

Example 4ca. Preparation of the hydrochloride of 7- (3-methyl-3-j-Ala-amino-l-azetidinyl) -1-cyclopropyl- 6, 8-difluoro-i, 4-dihydro-4-oxo-3-quinolinecarboxylic acid.

44 mg (0.11 mmol) of 7-(3-methyl-3-D-Ala-amino- 1-azetidinyl) -1-cyclopropyl-6 ,8-difluoro-1,4-dihydro-4oxo-quinolinecarboxylic acid (3cvzA) are treated with a solution of EtOH-HCl, the solvent is evaporated off at reduced pressure and 42 mng (0.09 minol) of the hydrochloride of 3-methyl-3,-D-Ala-amino-1-azetidinyl) -1cyclopropyl-6, 8-difluoro-1, 4-dihydro-4-oxo-quinolinecarboxylic acid (4ca) are obtained.

M.P. =181-185 0

C

21 'H NMR (100 M.Hz) (DMISO-d6/TFA): 1.12 (in, 411), 1.34 J 5.5 Hz, 311), 1.55,(s, 311), 3.70-4.70 611), 7.68 J 12.7 Hz, 1H1), 8.01 3H), 8.55 LH), 8.79 (in, 1H).

IR (KBr): 3600-2400 1718, 1688, 1627, 1528, 1465, 1328 cmf'.

Examprle 3da. Preparation of 7-(3-methyl-3-pa-amino--azetidinyl) -1-cyclopropyl-6-fluoro-8-chloro- 1, 4-diydro-4-oxo-3-quinolinecarboxylic acid.

\A solution of N-CBZ-D-alanine (1.35 g, 6 nimol) in ml zf dry tetrahydrofuran cooled to -150 0 C is neutralis4 d with N-methylmorpholine (0.61 g 6 nimol) and isobutyl chioroformate (0.82 g, 6 inmol) is added minutes later, a solution of 3-amino-3-inethyl-1-.benzhydrylazetidine (1,52 g, 6 inmol) in 20 ml, of tetrahydrofuran is added, the mixture is kept at -10 0

C

for 15 minutes. The solution is allowed to return to room tempei~ture, the precipitate which forms is filtered off and the'filtrate is evaporated to dryness and, after one crystallisation from AcOEt-hexane, 3-methyl-3-N-CBZ-D- Ala.-amino-1-benzhydrylazetidine (1,83 g, 4 mmol), with a melting point of 90-98*C, is obtained.

1.68 g (3.6 inmol) of 3-methyl-3-N-CBZ-D-Alaamino-1-benzhydrylazetidine aro treated with ethanol saturated with hydrochloric acid, the solvent .,Is evaporated off at reduced pressure and 1.69 g nmol) of 3-methyl-3-N-CBZ-D-Ala-amino--bnzhydyazetid .ne hydrxochloride, with a melting point of 93-970C, are obtained.

160 mg of Pd(OH) 2 /C are added to a solution of -3-iethyl-3-N-CBZ-D-Ala-amino-1,-benzhydrylazetidine,-' hydrochloride (1.63 g, 3.3 mmol) in 150 ml of methanol and the mixture is left overnight under a hydrogen atmosphere (16 atm). The catalyst is filtered off, the solvent is evaporated off at reduced pressure and 3-methvl-3-D,-Ala-aminoazetidine hydrochloride (600 mg, 3.1 nimol) is obtained.

100 mg (0.33 inmol) of 1-cyclopropyl-6,7-difluoro-.

8-chloro-1, 4-dihydro-4-ox-o-3-quinolineearboxylic acid, 22 129 mg (0.6o6 inmol) of 3-methyl-3-D-Ala-aininoazetidine hydrochloride and 290 mg (2.8 mmol) of triethylamine are mixed -in 8 ml of anhydrous pyridine. The solution is kept at 70 0 C far 15 hours. The solvent is evaporated of f at reduced preasure and the resulting crude product crystallises, from anl e'thanol1-water mixture and 66 mg 15 mmol) of 1- (3-methyl-3-D-Ala-amino-1-azetidinyl) -1-cyclopropyl- 6-f luoro-8-chloro-Y, 4-d-thydro-4-oxo-3-quinolinecarboxylic acid (3da) are obtain~d.

M4.P. 167-171'C 'H NMR (100 MAHz) (DMSO-d 6 /TFA): 0. 90-1.20 (in, 4H),r 1.37 J 7.0 Hz, 3H), 1.57 3H), 3.81 111), 4.10-4.70 5H), 7.75 J =13.7 Hz, 1H), 8.1 (in, 3H), 8.72 1H), 9.08 (in, 1H1).

IR (KBr): 3600-2400,-(b) 1718, 1668, 1618, 1543, 1443, 1405, 1312 cm-'.

Example 4da. Preparation of the hydrochloride of 7- 3-iethyl-3-D-Ala-amino-1-azetidinyl) -1-cyclopropyl- 6-f luoro-8-chloro-1, 4-dihydro-4-oxo-3-quinolinecarboxylic acid.

mng (0.08 inmol) of 7-(3-methyl-3-D-Ala-amino- 1-azetidinyl)-1-cyclopropyl-6-fluoro-8-chloro-1, 4-dihy o-4-oxo-3-quinolin carboxylic acid (3da,) are treated 4 )th a solution of tOH-NC1, the solvent is evaporated off at reduced pressure and 36 mg, minol) of the hydrochloride of 7- (3-methyl-3,-Q-Ala-ainino-1-azetidinyl) 1-cyclopropyl-6-fluoro-8-chloro-114-dihydro-4-oxo-3quinolinecarboxyli, acid are obtained.

176-180 0

C.

'H NMR (1.00 MHz) (DMSO-do/TFA) 0. 90-1.20 (mn, 4H),I 1.36 J -a 6.9 Hz, 3H) 1,56 3M), 3.80 (me, IH)F 4.10-4.70 SM) 7.75 J 13.3 Hz, 1H) 8.00 (in, 3H), 8.70 iM), 9.00 (in, 11).

IR (KBr): 3600-2400 1718, 1681, 1625, 1450, 1406, 1318 cm'.

TABLE I TABLZ I (CoaL.)

A-

Example Salt Stersochamistry M.P. (0 C)

(DMSO)

I 4. i I- IR (cma 7 (Kbr) 36JO-24 20 16.30, 1510I, 1!300,_ 14!W, 1362, 132-- 3620-2400 1718, Ib~b, 1b31, L561, 1490, 1449, 1328 Al a (2S. 31A) 220-222 -f1 t, 9 (C-0,.75) 1~ 1- 4 &k 12S, 3R) 190-192 +16.2 0. as) 2&LC13. N -t-S0CLout(2S* 3R) 117-120 +17,0 3318, 2962, 1719, 1631, 1509, 1447, (c-0.71) 1368, 1331 UaL I CH H H 14 Lou (2S,3R) 216-218 +9.7 3331, 2962, 1724, 163h, itb71, 1509, 3 c-0,76) 1449 4&L 1 CH 3 I H N LOU Q2S,3 R) HC1 181-184 *23.8 3600-2400 4 b 1 1718, 168-7, 1630, 1562, I 3(c-0.75) 1512, 1449, 1325 it Hi II N -CLIZ-Ad 1 205-208 +11.9 3318, 1719, 1656, 1631, 1509, 1449 63)j IfIf If H U Ala- 150-152 aZ, 34111, 32b2, 30'1!A. 163., 1561b, 1510o 1459 TAaL. I (Cat.)

A-

2 iR (cm 1 (KI' z) II 2a 0 Example R 7 1 XR A Stersochemistry Salt. Mp. (O C) R (cm S(KQ)r) (DMsQ) 411A H H H N Ala I C1 204-207 +5,1 3600-2100 1725, 1687, 1632, 1492, Ic-0.84) 1450 2bL H Hj N N-t-BOC-.eu 123-127 t8.2 3331, 295ti, 1719, 1633, 1509, 1448 (c-0.75) 3bL H If LOU 214-216 +3.8 3600-2400 1725, 1637, 1561, 1509, (c-0,55) 1460, 1357 4bL -H -La HCI 190-195 +15.6 3600-2400 1718, 1687, 1631, 1512, H U(c-0.76) 1450

F

H H 4N N-tBOC-Cly 228-230 3331, 1720, 1659, 1632, 1509, 1445, 1151 3bC H ii N Gly 235-236 3600-2400 1638, 1568, 1512, 141,t, (dec.) 1370 4 if I H N Gly I 274-277 3b00-2400 1710, tb71, lb12, 1b12, (doc.) 1571, 1509, 14b0 TABiLE I lCont.) 1H (cm-)(tr ,1680, 1627, 1528, 1459, 3620-2400 1718, 1675, 1628, 1528, 1467, 1326 3600-2400 1718, 1688, 1627, 1528, 1465, 1328 3325, 1725, 1622, 1448 3600-2400 jb), 1718, 1668, 1618, 1543, 1443, 1405, 1312 3b00-2400 1715i, 1651, 1bi5 1406, 1318 35b0-3z00 17is, 1655, 1505s, 14UU, 1401, 1325 TADLK- I (Coat.) 1= -201 IR(cu) (KHr) Example a RS R-1A Streochamistry Salt m.p. (O C) I0 c1 3ea 1

R

7

B

8 X 0lHI I -A (2S, 3-R) -2l4( 57.9 3 500 1630, 1± 09, 147-S 1420, C I(c-0.43) 1340 4ai C- H I -Ala (2S,3R) HCI 207-211 3600-2400 (bj, 1718, 1687, 1637, 1506, ]3 1462, 1393, 1318 f H-t-BOC-Nva 141-143 t9. 9 (c-0.S1) I i i i 3331, 1-19, 1633, 1509, 1449 3400-2500 1725, 1635, 1569, 1510, 1459, 1368, 1275 F 176-179 +7.3 (c-0.54) 4 I--f-f-I 4 1 4bft i 2aa

F

A

Au- KCi N-CBZ-D-Ala (2S, 3) 196-200 177-182 234-136 +12.1 (c-077) -20 8 (c-0 4 61) 16,7 7 9 3400-2500 1725, 1697, 1639, 1509, 1449 3368, 1720, 1632, 1509, 1448, 1331 1400-200 (b 172b3 I3b, 1ui, fttuo 151, 1!)0 1331 N Cli1II I if D-Ala I J A I rA1U. I (ot.) Exap_ SRj 20 Exra~pIe lR I R 8 X A atereochemistry S4ilt M. P O C) uji IR (cim (Kbr)

(OHSO)

H N -ly A aa 14 (2S-3) HCI 238-240 13.4 3600-2400 1718, iba, 1543, 1443, (c 0. 73) 1331, 2b& H 20C0-.03 -12.6 32-93, 1719, 16!6, 1631, 1543, 1509, (c-0.81) 1449, 1331 3ba H H H N D-kla 155-151 -8.2 3418, 3263, 3075, L635, 1560, 1509, 1460 (c-0,69) dba l D-Ala uC 202-205 -5.9 3600-2400 1725, 1685, 1631, 1492, (c-0,53) 1449 2f& 3ta 4rA

-H

A

III

H-CBZ-D-Ala f-Ala 2-Ala 12BR, 3S) (2R, 3S) 207-210 21U-221 -21. 1 3312 (c-0,B2J 1325 -16,2 33b0 (c-0,64) 1362 1718, 1681, 1632, 1543, 1512, 1237 -2400 lb32, 1509, 1500, 14:1, 1325 -2400 1718, 65, jail., ibL), 1450, 1325 193!-195 lidi 6) ThLE. I (Cont.)

RS

I I~ii t I N 5 Example Stereochemistry m.p. (0 C) 20

(DMSO)

IR (cm (KBr) 2aAA CU 3 H H N N-CI3Z-Ala-Ala (2S-3R) 121-123 3300, 1718, 1680, 1630, 1447 (C-0.77) 3aAA C H H N Ala-Ala (2S,3R) 194-198 12.0 3600-2400, 1632, 1543, 1452, 1368, 1325 4aAA CH H H N Ala-Ala (2S,3R) UCi 188-191 +16,6 3600-2400, 1687, 1668, 1632, 1568, 1475, 3 1325

F

2eA CH H H C-H N-CBZ-Ala (2S,3R) 119-122 -21.8 3325, 1718, 1629, 1506, 1462, 1325 3c-0,7)

F

3eA~ CH 3 1 H H C-H A~a (2S,3R) 152-156 -40,5 3600-23D0, 1630, 1509, 1475, 1387, 1318 (c-0.77) 1 6 f C-11 (2S, 3R) Ts0II 172-175 -21.0 (c-0.9) 3600-2300, 1718, 1687, 1630, 1520, 1466) u TABLE~ o (Coat-) 0 R5mx1 R R X A Stareachamistry Salt M.P. (O C) [a0 IR (cm1 (KBr) 2eN If 11 H -BOC-Nval (2 S-3R) 123-127 3362, 1718, 1629, 1507, 1466, 1318 F_ 3 i 7 1 (c-0,84) 12 3HC H C-H jNval (2S 3R) 205-209 -17.0 3600-2300, 1615, 1630, 1508, 1468, 1325 4e H H UCH vl(2S, 3R) T30H 164-157 -12.3) 3600-2300, 1718, 1681,,'1629, 1507, 1466, 4-]F C 31H I C- 112 -31 TABLE 11 E xainplel Solvent 'H NHR (.100 Hz) 1.11 1.21 (d,J =7.0 Rz, 3H); 1.59 (d,J =6.1 Hz, 3H); 3.60 (m,tH); DMSO-d,/TFA-dj. 3,85-4,70 5K); 5.01 (9,2H); 7.30 7,97 (d,J =11.5 HZ# 1K); 8,56 1H) 1,16 1,39 (d,J 7.00 Hz, 3H); 1,65 (d,J 6.2 Hz, 3H); DMSO-do/TFA 3,55-4.00 4.00-4,80 4H), 8.02 (d,j 11.6 Hz, 1K), 8.15 b, 3H); 8.60 (u4KH); 8.95 (m,1HI)'* 1.10 Cm,4H), 1,38 (d,J -7-.00 Hz, DtSO-do/TFA 3H); 1,63 (d,J 6,2 Hz, 3H); 3P50-4.00 4,00-4.80 8,00 (d,J -11.5 Hz, 1H); 8.16 3H); 8.58 9,13 (m,1H) 0,85 (d,J 5,9 Hz, 6E); 1.14 (m,4H); DMSO-d*/TFA-d% 1,35 Os,I 1.59 (d,J 5.9 Hz, 3K); 3.56'-4,70 c.a. e 6H) 7,.97 (d,J -11.5 Hz, 1K); 8,57 (a,1K) 0.92 (d,J a 4.8 Hz, 6H); 1.13 (m,4H); 1.65 5,9 Hz, 6H); 3.68 (m,2Hi); DMSO-da/TFA 4,05-4,80 8.03 (d,J 11.7 Hi;, 1H); 8.15 3H); 8.60 (s,IK; 9.09 (nOIH) 32 TABLE II (Cont.) Example Solvent H NMR (100 MHz) 0,89 (d,J 5.2 Hz, 6H); 1.11 (m,4H); 1.63 (d,J 5.5 Hz, 6H); 3,43 (m,2H); 4aL DMSO-d/TFA 4A)5-4,80 4H); 801 (d,J 1,4 Hz, 1K); 8,15 b 3H); 8.58 9,19 (mIH) 1,18 (d,J 7.0 Hz, 3H); 3.70-4.70 2bA DMSO-do/TFA-di 6H) 4,98 7.05-7.80 8H (d8- 7.97 (d,J 11,3 Hz, 1H); 8,71 (s,IH) 1.33 (dJ =7,0 Hz, 3H); 3.70-4.L5 3bA DMSO-d/TFA 4.15-4.70 7.10-7,80 3H); 8.01 (dJ 11,2 Hz, 1H) 8.15 3H); 8.72 9.10 (m,1H) 1,31 (dJ a 6,5 Hz, 3H); 3.70-4.70 4bA DMSO-do/TFA 7,10-7.80 3H); 7.85- 8.15 Q:,71 8,94 (m,1H) 0,87 (dJ 5,6 HZ, 6H); 1.38 2bL DMSO-d/TFA-d (s,12 3.80-4.70 6H); 7.15- 7,90 8.05 (dJ 11.3 Hz, 1K); 8.78 33 TABLE. II (Cont.) Example Solvent -H NMR (100 MHz) 0.89 (d,J S.3 Hz, 6H); 1(37 4L 367 4,02 4,42 3bL DMSO-do/TFA 7,19-7,86 8.03 (d,J 11.2 Hz, 1H); 8,16 3H); 8.74 8.99 (m,1H) 0,87 (d,J 5.1 Hz, 6H); 1.54 (m,3M); 3.60-4.70 6H); 7.15-7,80 4bL DMSO-d./TFA 8.03 (dJ 11.2 Hz, 1H); 8.16 3H); 8,75 9.35 (m,1H) 1.33 3.49 3.80-4.70 2bG DMSO-d./T'A-dL 7,10-7.80 7.99 (d,J 10,6 Hz, 1H); 8P73 3,53 (dJ 5.9 Hz, 2H); 3.65-4.70 3bG DMSO-do/TFA 5H); 7,15-7,80 7,80- 8,15 4f (8 -8t03 d,J 11,3 8,77 8.97 (m,1H) 3,51 (d,J 59 Hz, 2H); 3,70-4.80 4bG DMSO-d/TPA (c.a 7,05-7,80 7.80- 8.15 4H (8a 7.98, dJ 11,3 8.70 Cu,1K); 9.00 (mIH) 34 TABLE II (Cont.) Example Solvent 'H NMR (100 MHz) 0,85-1,28 7H); 1,54 (s,3H); 2ca DMSO-d/TA-dz 3,79-4,60 6H); 5,00 (s,2K); 7,30 7.64 (dJ 13.1 Hz, Itl); 8.55 (s,IH) 1.15 1,36 (d,J 6.5 Hz,*3M); 3ca DMSO-do/TFA 1,57 3,65-4.70 6H); 7.67 (dJ 11,4 Hz, AH); 8.05 (0, 3H); 8,56 8,90 (m,1H) 1.12 1.34 (d,J 5,5 Hz, 3H); 4ca DMSO-d./TFA 1,55 3,70-4.70 6H); 7,68 (d,J 12.7 Hz, 1K); 8.01 3H); 8,55 8.79 (m,1H) 0.75-1.34 (ca 1,50 (a,3H); 2da DMSO-do/T'A-d% 3.84-4.64 6H); 4,98 (a,2H); 7,25 7.69 (dJ 12.9 Hz, 1H); 8.68 (s,lli) 0.90-1.20 1,37 (d,J 7.0 Hz, 3H); 1.57 3,81 (mlH); 4.10- 3da DMSQ-d,/TA 4.70 5H); 7,75 (d,J 13,7 Hz, 1H); 8,1 8,72 9.08 Cm,IH) 35 TABLE 11 (Cont.) Example Solvent 'Hi NMR (100 MHz) 0,90-1.20 1.36 (d.j 6.9 Hz, 3H); 1.56 3.80 (rm,MK; 4,10- 4da DMSO-d*/TFA 4.70 5K); 7.75 (d,J =13.,j Hz,' 1H); 8..0 8.70 9,00 (m,1H) 1. 15 a. 6H) 3.60-4.45 c. 4.97 5.74 ,7.3 liz, 2ea DtISO-do/TFA-d, 1H): 7.10-7.80 8H (a 7,27-, 7.87 12.6,1H)f- 8.69 (s,1H) 1.32 6H); 3.60-4.50 5,75 (d 1 .J 7.2 iiz, 1!I)i 7.10- 3ea DMSQ-do/TFA 7,60 3H); 7.86 (a1,j -=12.7 Hz, 1H); 8.10 3H); 8i.-4 8.75 (m,1,H) 1.33 6H); 3,60-4,.50 5,76 (d,J =7,2 Hz, In); 7.10- 4e& DMSO-d./TPX 7.60 3H); 7.87 (d,j 12.7 Hz, 1H); 8.10 3H); 8,65 8.76 0.01 1.00-1.70 13H (du 1.33, a, 3,70-4.70 6H); 2bN DMSO-dq,/TFA-d, 7,10-7,85 Ic-4., 3H); 8.01 (d,J 11,3 Rz, In); 8.75 (s,1H) 36 TABLE. IT (Cont.) Example Solvent 'H NMR (100 MHz) 0.82 1.28 1.61 3.60-4.70 6R); 7.10- 3bN DMSO-d/TFA 7.85 3H); 7.94 (d,J 11.3 Hz; 1H); 8.10 8,70 9,01

(MI'H)

0,86 (t,J 6.9 Hz, 3H); 1.22 (m,2H1f; 1.64 3.60-4,70 6); 4bN DMSO-d./TFA 7,10-7,85 3H); 8.04 (d,J= 11,3 Hz, 1K); 8.12 8.75 (a, IK); 9.11. (m,lK) 1,03 1.19 (dJ 7.2 Hz, 3H); 1,54 (d,J 6.0 Hz, 3H); 3.57 (m,1H); 2aa DMSO-de/TFA-dL 3.85-4,80 (ca 5H); 4.96 (s,2H); 7.23 7.87 (d,J 11.5 Hz, 1H); 8.53 (s,1H) 1.08 (mp4); 1.35 (d,47 6,6 Hz, 3H); 3aa OKSO-d./TFA 1.59 (dJ a 6.1 Hz, 3H); 3.50-4.80 6H); 7.95 (dJ 11.9 Hz, 1H); 8,11 m,3M); 8.55 8,93 (m,1H) 1.07 1.36 (dJ 7.0 Iz,3H); 4aa DMSO-do/TFA 1.58 (d,J 6.,0 Hz, 3H); 3,50-4.80 6H); 7,91 (dJ 11.5 Hz, 1H); 8,04 8,54 8.95 (m,LH) TABLE II (Cont.) Example 1 Solvent 'H NMR (100 MHz) 1.18 (dJ 7,0 Hz, 3H); 3,70-4.70 2ba DMSO-ds/TFA-d, 6H); 4,99 (s,2H; 7.10-7,80 8H, (8 7,28, s, 7.98 (d,J ="11.3 HzlH); 8.71 (s,IH) 1,32 (d,J 7,0 Hz, 3H); 3.70-4,15 ,4,15-4,70 7,10-7,80 3ba DMSO-do/TFA 3H); 8.00 (d,J 11.1 Hz, 1H); 8, 13 3H) 8, 71 H) 9.08 (miH) 1.31 (d,J 6,5 Hz, 3H); 3.70-4.70 4ba DMSO-d*/TFA 7.10-7.80 3H); 7.85- 8,15 8.70 8,93 (m,IR) 1.12 1.22 (dJ 7.0 Hz, 3H); 2fa DMSO-ds/TFA-d 1,60 (d,J 6,1 Hz, 3H); 3.61--(m,IH); 3,85-4,70 (c-a 5H); 5.02 (1 2) 7.31 7.98 (d,J =11~.5 Hz,IH); 8.57 (a,IH) 1.16 1,39 Cd,J 7.0 Hz, 3H); 3fa DMSO-d./TFA 1.65 (dJ 6.2 Hz, 3.55-4.00 4.00-4.80 4H); 8.02 (d,J 11,6 Hz, 1K); 8.15 3H); 8.60 8,95 (m,1H) 1.11 1.39 (dJ 7,0 Hz, 3H); 4fas DMSO-ds/TFA 1,64 (dJ 6,2 Hz, 3H); 3.50-4.00 4,00-4,80 4H); 8.01 (d,J 11.5 Hz, 18); 8.17 3H); 8.59 8.97 (m,1H) 38 TABLE II (Cont.) Example Solvent 'H NMR (100 Mz) 0.90-1.26 (m,13H); 1.56 (d,J 6 Hz, 3H); 3.55 3.98 4,2 2aAA DMSO-de/TFA-dx 4.45 4.58 (rn,liH); (300 MHz) 4,78 and 4,85 (system AB,J 12.0 Hz, 2R); 7.87 (d,J 12< ,Hz, LH); 8.47 (s,1H) 0.9-1,4 (m,13H); 1.60 (d,J 5.74 Hz,I 3H); 3.5-4.8 7.85-8.20 (m,4H 3eAA DMSO-d,/TFA (6 8,00, d,J 11,2 8.56'-- 8.63 (b2H) 0,9-1,5 (m,13 1.61 (d,J 6..2 Hz, 4eAA DMSO-qla/TEA 3H); 3.5-4.7 7.88 (d,J 11.2! Hz, 1K); 8.05 8.4-8.6 (n,3H 16 8.52, s)) 1.20 3.60-4.40 4.97 2eA DMSO-da/TFA-d 5.74 (d,J 6.7 Hz, 1H); 7.10-8.00 (m,9H (6 7.26, s),(6 7.88, dJ 11,56 8.71 H) 1.28 2.40 4.15 (n,3H)i 3eA DMSO-do/TFA 5,74 (d,J 7.0 Hz, 1K); 7.10-8.3 t m,7H (6 7.87, d,J 12.7 Hz)); 8.65 8.75 (m,IH) 4eA DMSO-da/TFA 1,26 (m,61i); 22,h S.71 7.3 Hz, 7.06-8 (m,I11i, (6 and7.51, system ABJ 8.2 7,85, dJ =13.13 8.60 (s,Nl); 8.75 39 TABLE 1 (Cont.) Example Solvent 'H NMR (100 MHz) 0.77 0,95-1.60 (m,16H,(S 1.28, 3.75 (m,2H; 4.1 (m,3H); 2eN DMSO-do/T'A-d. 5.72 (d,J =7.0 Hz, 1K); 7..tO-7,95- (mr,4H,(6 =7.83, d,J 12,.6 Hz)); 8.61 (s,1H) 7.77 (m,3EI); 1.23 1.64 (rn,N&{H 3.59 4.11 5.68 3eN DMvSOdo/TFA 7,0 Hz, 1H); 7,00-8,10 (m,6H,(6= 7.720 d,J 12.0 8.58 (s,IH); 8.74 -(m,1H) 0,86 7,6 Hz, 3H); 1.29 1.63 2.28 3,60-3.80 (rn,2H); 4.00-4.20 4.40 (m,1H) 4eN DMSO-dq% 5. 75 (d,J 7. 0 Hz, 11) 7, 09 and 7.47 (300 MHz) (system AB,J =8,0 H7,- 4H); 7.43 IH) 7.74 7.685-8. 00 2H, (S=7.95, d,J 12,0 8,05 b, 2,H) 8.79 and 8.81 (2 x s, 1H) 8.93 40 Solubility A large number of the compounds of the invention exhibit an increase in solubility in water as well as in solubility at pH 7.4 when compared with the reference compounds. The increase in solubility at physiological pH can promote the absorption of the product and high solubility in water allows the compound to be administered parenterally.

FSolubility Solubility Example A Salt in water pH 7.4 la H -23.0 16,5 4aA Ala HCI >500 >500 4aL Leu 1HCI >500 >500 Ilb H 3.0 3,7 4bA Ala HCl 128,4 66.9 4bG Gly HCI >500 7.6 4bL Leu HCI >500 23.1 4aa 0-Ala HCd 494.9 385.6 4fa 0-Ala HC1 >500 >500 4aAA Ala-Ala HC1 >500 >500 4eA Ala TsOR >500 >500 4eN Nval TsOH >500 >500 41 BIOLOGICAL ACTIVITY The antimicrobial pharmacological activity of these.compounds was studied according to the references set out below.

Antimicrobial .,pharmacological activity Daquet and Y.A. Chabbect, techniques en bact6riolo9j.,r Vol 3, Flammarion M6decine-Sciences, Paris 1972, and W.B. Hugo and A.D. Rusell, Pharmaceutical Microbiolo f, Blackwell Scientific Publications, London, 1977.

Culture medium and solvent: Antibiotics agar, No. 1 (Oxoid CM 327) Triptone-soya broth (Oxoid CM 129) Ringer j physiological solution (Oxoid BR 52) Dextrose agar (BBL 11165) Microorganisms "Bacillus subtills" ATCC 6633 "Citrobacter freundii" ATCC 112606 "Enterobacter aerogenes ATCC 15038 "En terobacter cloacae" ATCC 23355 "Bacillus cereus" ATCC 1178 "EscherLchia coll" ATCC 10799 "Escherichia coll" ATCC 23559 'Klebsiella pneumoniae" ATCC 10031 "Proteus vulgaris" ATCC 8427 "Morg. moganil" ATCC 8019 "Pseudowonas aeruginosa" ArCC 9721 "Pseudomonas aeruginosa" ATCC 10145 "Salmonella typhimurlum" ATCC 14028 "Salmonella typhimuriur" ATCC 6539 "Serratia marcescens" ATCC 13880 "Shlgella flexneril" ATCC 12022 "Staphylococcus epidermis ATCC 155-1 "Staphylococcus aureus ATCC 25178 "Streptococcus faecalis" A2'CC 10541" Preparation of the inoculations Each of the microorganisms is seeded as a strtak in tubes of antibiotics agar no.1 and incubated at 37*C 42 for 20 hours. A culture loop is then taken, seeded in a triptone-soya broth and incubated at 37°C for 20 hours.

The culture obtained is diluted to I with a Ringer physiological solution, so as to obtain a standardised suspension of 10-10 ufc/ml for each organism.

Preparation of the medium containing the derivatives of general formula I: Starting with a solution of 100 ig/ml, each product is diluted in dextrose agar (melted beforehand and kept at 50"C) by successive dilutions so as to obtain the. following concentrations: 64, 32, 16, 4, 2, 1, 0.25 and 0.125 pg of derivative/ml of medium.

Subsequently, each concentration of each product is distributed into Petri dishes 10 cm in diameter, in a..

proportion of 10 ml of medium per dish and as many dishes as microorganisms to be tested.

Once the medium is cool, the dishes are seeded with the inoculations in a proportion of 0.4 ml of inoculation per dish. They are spread with a Driglasky loop and the supernatant is collected. The seeded dishes are incubated at 37°C for 20 hours.

The results obtained are described in the following tables. The concentrations are given in g/ml.

43 I EXAMPLE S i MICROORGANISMS Pipemidic 4aA 4aL 4 4bA 4bE acid I faillua asubtiliLs 8 042 0.12 0.3 0 ATCC 6633 Bacillus cereus 16 1 0,12 4 ATCC 11778 Strep, faecalis >64 8 2 8 1 ATCC 10541 1 Staph. aureus 64 0.5 0,25 1 I 4 ATCC 25178 Staph. epidermidis 64 0,5 0,25 4 1 ATCC 155-1 1 Ps. aeruginosa 32 4 2 8 4 ATCC 9721 Ps. aeruginosa 32 8 8 16 8 ATCC 10145 Citr. freundii 4 0,25 0.12 11 1 ATCC 11606 Morg. morganii 8 0.5 0.25 2 2 ATCC 8019 Proteus vulgaris 16 1 0.25 8 i 2 ATCC 8427 lKflb. pneumoniae 2 0.06 0.06 1 0I 2 ATCC !0031 Sal. typhimurium 8 0.5 0,12 2 ATCC 14028 Sal. typhi 4 015 0.12 1 ATCC 6539 Esacherichia coli 16 045 0.12 2 1 ATCC 10799 Eacherichia coli 2 0,12 0.06 1 0,25 ATCC. 23559 Ent. aerogenes 32 .5 0,12 1 ATCC 15038 Ent, cloacae 8 0.12 0,12 1 ATCC 23355 Serr. marcescens 16 1 0,25 8 2 ATCC 13880 Shigella flexnerii 4 0.12 0.06 1 ATCC 12022 -44w

MICROORGANISMSEXMLS

4 1 4ca 4bN 4aa Bacillups'ubtt UIs I. I 0.25 0.25 0. 2- ATCC 6J Bact)uiu cereua, 1 005 0 ATCC 11778 Strep. faecalLs 16 8 4 ATCC 10541 Staph. aureus 1 0.5 4 I 0.25 ATf~c 25178 Staph. epidermi2dis, 2 0.5 I 2 1 ATCC 155-1 Ps. aerug2.nosa 16 44 4 ATCC 9721" Ps. aerutginosa 32 16 168 ATCC 10145 Citr. freundii 0.511., ATCC 11606 4org. morganii 4 1 2 2 ATCC 80191 Proteus vulgaris 82 42 ATCC 842.7 1 leb. pneumoniae 10.25 0.12 0. ATcc 100311 Sal. typhimurium 4 1 1 *ATCC 14028,.

Sal. typhi 1 0.510.

ATCC 6539 Eacherichia coli 2 1 2 I IATCC 1079 Escherichia coli 0.5 0'5 0.25 02 ATcc 23559 Ent. aerogenes I. 1 ATCC 15038 Ent. cloacae 1 0'5 0,5 0.25 ATCC 23355 Serr. marcescens 1.6 2 24 ATCC 13880 Shigella flexnerii 0.5 0,,25 0,12 0.12.

«:"ATCC 12022 MICROORGANISMS, EXAM:LES ~4e Bacillus subtilis) 0.5 1 0.3 0,25 A,TCC 6633.

Bacillus cereus 1 16 1 0.3 ATCC 117787 strep. faecalis 1 j >16 8 4 ATCC 10541 Staph.t aureus 1 4 1 ATCC 25178 Staph. epidermidis 0.25 ]8 16 ATOC 155-1 j Ps. aeruginosa, >16 >16 8 4 ATCC 9721 s aeruginosa. >16 >16 8 16 ATOC 10145 citr. freundii 4 2 2 2 ATCC 116062 m rg. oranii 8 4 44 ATCC 8019 Proteus vulgaris 8 8 16 8 ATCC 8427 tKleb. pneumoniae 8 8 2 0.25 ATCC 10031b Sal. typhimurium 4 2 2 1 ATCC 14028 Sal. typhi 4 2- 2 I 0.3 ATCC 6539 Escherichia coli 4 2 2 0.51 ATCC 1.0799 Escherichia coli 1 2 1 ATCT 235,59 Ent. aerogenes 4 2 2 2 ATCC 15038 Ent. cloacae 4 2 2 1 ATCC 23355 Serr. marcescens 16 8 8 6 ATCC 13880 ShigeIla flexnerii 1 ATCC 12022 46 In human therapeutics the administration dose is, of course, a function of the susceptibility of the infective strain, of the nature of the administered compound and of the route of administration. It will generally be between approximately 0.200 and approximately 300 mg per each kilogram of weight and per day. The derivatives of the invention will be, for example, administered in the form of tablets, solutions or suspensions, or of gelatine capsules.

.0 Two particular galenic forms of the derivatives forming the subject matter of the present invention will be shown below by way of exaaples.

Example of formula per tablet Compound of example 4aL Microcrystalline cell Povicone Wheat starch Colloidal silica dioxide Magnesium stearate Tablet weight 250 mg 69 mg 15 mg 36 mg 2 mg 3 mq 375 mg Example of formula per gel.capsule Compound of example 4aL 250 mg Polyoxyethylenated glyceride 85 mg Glycerine pahenate 15 mq Soft gelatine excipient q.s. 450 mg Example of formula per injectable Compound of example 4aL 100 mg LM HCI q.s. pH 3-4 HO0 for injection q.s. 10 ml

Claims (8)

1. Cmpn corresponding to the general formula I: So 0 F N X N R A-N- (CH)n I P. and their pharmaceutically acceptable saltsA AT-whl 'formuAra- X denotes a nitrogen atom or else a carbon atom. bonded to a hydrogen atom or else a carbon atom bonded to a halogen atom in which Y denotes a fluorine, chlorine or bromine atom, or else a carbon atom bonded to a hydroxyl radical (C-OH) or alkoxy radical (C-O-alkyl), or else a carbon atom bonded to an alkyl radical (C-alkyl), or else a carbon atom bonded to a halogenated alkyl radical or else a carbon atom bonded to an alkylamino radical (C-NH-alkyl); R, denotes a lower alkyl or cycloalkyl radical, a lower haloalkyl radical, a hydroxyalkyl radical, a vinyl radical, an aryl radical or an aryl radi(al substituted by one or more fluorine atoms, or an alkylamino radical Halkyl); R 2 denotes a hydrogen atom or a lower alkyl radical; R 3 denotes a hydroxyl radical or a CI-C 4 lower alkoxy radical; R 4 denotes a hydrogen atom, a fluorine atom, a lower alkyl radical, a nitro radical an amino radical or a substituted amino radical; R R, and R 7 denote a hydrogen atom or a lower allyl radical; depending on the nature and the relative position, the azetidine substituents may have up to two chiral centres, each of them with an or "S" configuration; R 8 denotes a hydrogen atom or a lower alkyl radical; n is a number which may be 0 or 1; X and R 1 may together form a bond represented by a group C-CH 2 -CH 2 -CH-R 9 or C-0-CH 2 -CH-R 9 or C-S-CH 2 -CH-R 9 in which R 9 denotes a hydrogen atom, a lower alkyl radical or a lower haloalkyl radical and which in the two latter cases have one chiral centre with an or "S" configuration; R 1 and R 2 may together form a bond represented by a group -CHR 10 in which R10 denotes a hydrogen atom or a lower alkyl radical or a lower haloalkyl radical and in the two latter cases hfve one chirl ~gitre with an or "S" configuration; R 2 and R 3 may together form a bond represented by an -S-NH- group; A is a solubilising group which is represented by a residual amino acid or a polypeptide chain consisting of two or more amino acid residues which are bonded covalently one to another via peptide bonds and wherein the amino acid or polypeptide chain may be protected by any one of the protective groups for functional groups and wherein the amino acid residues include the 20 natural amino acids, denoted by means of the three letters symbolising them, as well as other amino acids such as norvaline (Nva), homoserine,

4-hydroxyproline, hydroxylysine and ornithine; and whdrein the stereochemical configuration of the ahfino acids may be of the D- or L-form or a mixture of the two. 2. A compound according to Claim 1, selected from the i D group including: (2S, 3R) -2-methyl-3-N-CBZ-Ala-amino-l-azetidinyl] -1- a cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine- 3-carboxylic acid, V. 3R) -2-methyl-3-Ala-amino-l-azetidinyl] -l-cyclo- propyl-6-fluoro-1,4-dihydro-4-oxo-l,8-naphthyridine- -48- 49 3-carboxylic acid, hydrochloride of 7-f (2S,3-R)-2-methyl-3-Ala-amino-1- azetirdiny1 -1-cyclopropyl-6-f luoro-1 I 4-dihydro-4-oxo-1, 8- naphthyridine-3-carboxylic acid,

7-f (2S,3R)-2-iethyl-3-N-t-BOC-Leu-amino-1-azetidinyl]- 1-cyclopropyl-5-fluoro-1 ,4-dihydro-4-oxo-1, 8-naphth- yridine-3-carboxylic acid, 7-f (2S,3R)-2-methyl-3-Leu-amino-1-azetidinylJ-l-cyclo- propyl-6-fluoro-1,4-dihydro-4-oxo-1,8-naphthyridine-3- 16 -carboxylic acid, -hydrochloride of 7-f (2S,3R)-2-methyl-3-Leu-amino-1- azetidinyl -1-cyclopropyl-6-f luoro-1 ,4-dihydro-4-oxo-1, 8- naphthyridine-3-carboxyiic acid, 7-(3-N-CBZ-Ala-amino-l-azetidinyl)-1-(2,4-difluoro-. phenyl) -6-f luoro-1 4-dihydro-4-oxo-1, 8-naphthyridine-3- carboxylic acid, 7-(3-Ala-amino-1-azetidinyl)-l-(2,4-difluorophenyl)-6- fluoro-1: 4-dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid, hydrochloride of 7-(3-Ala.'amino--azetidinyl)-1-(2t4- difluorophenyl)-6-fluoro-114-dihydro-4-oxo-1,8-naph- thyridine-3-carboxylic acid, 7-(3-N-t-BOC-Leu-amino -1-azetidinyl)-l-(2 ,4-difluoro- phenyl) -6-f luoro-1, 4-dihydro-4-oxo-1, 8-naphthyridine-3- carboxylic acid, 7-(3-Leu-aah~no-1-azetidinyl)-1-(2,4-difluorophenyl)-6- fluoro-1, hydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid, hydixochloridel of 3-Leu-amino-1-azetidinyl) ,4- difluorophenyl)-6-fluoro-1,4-dihydro-4-oxo-1,8-naph- thyrdine3-crboxlicacid, 3-N-li-BOC-Gly-amino-1-azetidiny1)-l-(2,4-dif1uoro- phenyl) -6-f luoro-1, 4-dihydro,-4-oxo-1, 8-naphthyridine-3- carboxylic acid, 3-Gly-aznino-2-azetidinyl)-l-(2 ,4-difluorophenyl) -6- fluoro-1, 4-dihydro-4-oxo-1, 8-naphthyridine-3-carboxylic acid, 50 hydrochloride of 3-Gly-amino-1-azetidinyl) 2,4- difluoropheny -6-f luoro-1, 4-dihydro-4-oxo-l, 8-naph- thyridine-3-carboxylic acid,, 3-methyl-3-N-CI3Z-D-Ala-amino-1-azetidinyl) -l-cyclo- propyl-6,8-difluoro-1,4-dihydro-4-oxo-3-quinolinecar- boxylic acid, 7- (3-methyl-D-3-A.la-amino-1-azetidinyl) -3-cyclopropyl- 6"1 8-dif luoro-1, 4-dihydro-4-oxo-3-quinolinecarboxylic acid, -hydrochloride of 7-(3-methy1--3-D-Ala-amino-1-azeti- dinyl) -l-cyclopropyl-6, 8-difluoro-1, 4-dihydro-4-oxo-3- quinolinecarboxylic acid, 3-methyl-3-NI-CBZ-D-Ala-amino-l-azetidinyl) -1-cyclo- pr ,pyl-6- fluoro-8-chloro-1 ,4-dihydro-4-oxo-3-quinoline-. carboxylic acid, 3-methyl-3-D-Ala-amino-1-azetidiniyl) -1-cyclopropyl- 6-f luoro-8-chloro-1 ,4-dihydro-4-oxo-3-quinolinecarboxylic acid, hvd.kochloride of 7- (3-methyl-3-D-Ala-amino-1-azeti- dinyl) -1-cyc~lopropyl-6-fluoro-8-chloro-1, 4-dihydro-4-oxo- 3-quin,.Li necarbobcylic acid, (2g,3R) -,-methyl-3-N-CBZ-Q-Aa-amino--azetidiny1]- 1- 4-difluorophenyl) -6-f luoro-1 ,4-dihydro-4-oxo-3- quinolinecarboxylic acid, (2S,3R)-2-zethyl-3-D-Ala-amino-1-azetidiny13-1-(214- dif luorophenyl) 6- f luoro-114 4-dihyciro-4 -oxo- 3-quinoline- carboxylic acid, hydrochl~oride of -methyl-3-D-Ala-amino-l- azetidinyl] 4-difluorophenyl luoro-1, 4-dihydro- 4-oxoh.3-quinoJlinecarboxylic acid, 7-(3-N-t-BOC-Nva-amino-1-azetidilyl)-l-(2 ,4-difluoro- phenyl) -6-f luoro-l, 4-dihydro-4-oxo-1, 8-naphthyridine-3- carboxylic acid, 7-(3-Nva-amino-l-azetidinyl)-1-(214-difluorophenyl)-6- f luoro- 114-dihye~zo- J-oxo- 118 -naphthyridine- 3-c arboxy ic acid, hydrochloride of 3-Nva-amino-l-azetidinypl)-1-(2, 4- difluorophenyl)-6-fluoro-1,4-dihydro-4-oxo-1, 8-naph- 51 thyridine-3-carboxylic acid, 3-R)-2-methyl-3-N-CBZ-D--Ala-aimino-1-azetidinyl]- 1-cyc-lopropyl-6-f luoro-1, 4-dihydro-4-oxo-1, 8-naph- thyridine-3-carbo acid, 7-C (2S,3R)-2-methyi.-3-D-Ala-amino-1-azetidiny1]-l- cyclopropyl-6-f luoro-1, 4-dihydro-4-oxo-1 8-naphthyridine- 3-carboxylic acid, hydrochloride of (2S.,3R) -2-methy1-3-D-Ala-amino-l- azetidinyl) -l-cyc2.opropyl-6-fluoro-1, 4-dihydzo-4-oxo-1, B- naphthyridine-3-carboxylic acid, 3-N-CBZ-D-Ala-anlino-1-azetidinyl)-1-(2 ,4-difluoro- phenyl) -6-f luoro-1, 4-dihydro-4-oxo-1, 8-naphthyridine-3- carboxylic acid, 3-D-Ala-amino-l-azetidinyl)-l-(2, 4-difluorophenyl) 6-f luoro-1, /gdihycro-4-oxo-1, 8-naphthyridine-3-carboxylic acid, hydro chloride of 7- (3-flAla-amino-1-azetidinyl) (214- dif luor ophenyl) -6-f luoro-l 4-dihydro-4-oxo-1, 8-naph- thyridine-3-carboxylic acid, (2S,3R)-2-methyl-3-N-CBZ-Ala-Ala-axnino-l-azeti- dinyl]-1-cyclopropyl-6-fluoro-1,4-dihydro-4-oxo-1,8- naphthyridine-3-carboxylic acid, (2.a,3R)-2-methyl-3-Ala-Ala-amino-l-azetidil-1- cyclopropy1-6-f 1L"ro-1, 4-dihydro-4-oxo-1, 8-naphthyridine- 3-carboxylic acid, hydrochloride of 7- 3B) -2-methyl-3-Ala-Ala-amino- 1-azetidinyl] -1-cyclopropyl-6-fluoro-1, 4-dihydro-4-oxo- 1, 8-naphthyridine-3-carboxylic acid, (2.a,3R)-2-methyl-3-N-CBZ-Ala-amino-l-azetidinyl]-1- (2,4-difluoropheny1)-6-fluoro-1,4-dihydro-4-oxo-3-quilo- linecarbo~ylic acid, (2a,3B)-2-methyl-3-Ala-anino-1-azetidifl-1-(2,4- difluoropheny1)-6-fluoro-1,4-dihydro-4-oxo-3-qilo- linecarboxylic acid, p-toluenesulphonate of (2$1 3R) -2-methyl-3-Ala-amino- 1-azetidinylJ-1-(2,4-difluorophenyl)-6-fluioro-1,4-di- hydro0-4-oxo-3-quinolinecarboxylic acid, 7- 3R) -2-methyl-3-N-BOC-Nval-amino-l- s-zetidinyl] l-(2,4d-.difluorophenyl)-6-fluoro-,4-dihydro-4-nco-3- quinolinecarboxylic acid, -7-[(25_,3R)-2-methyl-3-Nval-amino-l-azetidinyl)-l-(2,4- dif.1uorophenyl)-6-fluoro-1,4-dihydru-A4-oxo-3-quino- linecarboxylic acid, -p-toluenesulphonate of (2.,3R)-2-methyl-3-Nval- amino-l-azetidinyl]-l-(2,4-,difluorophenyl)-6-fiuoro-l,4- dihydro-4-oxo-3-quinolinecarboxylic acid. 3. A process for the preparation of a compound according to either one of Claims 1 and 2, including: Mi if the compound is of the general formula III 0 o 4 F 6 R 5 R 3 R3 R _N x N PI-A-N-(CH 2 )n RI in which R 1 R 2 1 R 3 0 R 4 R 5 1 R 6 R 7 RV R 9 R0,X, n and A have the abovementioned meanings, and wherein P1 denotes any one of the 'protective groups~ for functional groups employed in the synthesis of peptides; then by reaction of a compound of general formula II F4 R N N R 2 in which Rio R 2 R 3 1 R 4 1 R 5 0, ~6 R 7 RV R 9 Rl$X and n have the abovementioned meanings; with a 39 compound of general formula Pl-A-P2, in which A and -52- and P1 have the abovementioned meanings and P2 denotes any one of the activating groups for the carboxylic functional group which are employed in the synthesis of peptides, or with a compound of general formula Pl-A and with the aid of well-known coupling reactants for the formation of peptide bonds; (ii) if the compound is of general formula I, in which R 1 R 2 R 3 R 4 R 5 R 6 R7' R 8 R 9 X, A and n have the abovementioned meanings; then by reaction of a compound of general formula III, in which R 1 R2, R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 1 0 X, A, P1 and n have the abovementioned meanings, with reactants which are useful for the removal of protective groups for functional groups present in the amino acids and peptides of the compound of general formula III; (iii) if the compound is of general formula I, then by reaction of a compound of general formula IV R4 0 0 R 3 2 R 1 IV in which R 1 R 2 R 3 R 4 R, R10 and X have the S abovementioned meanings and Z denotes a halogen atom, with a compoun' of geseral formula VI 2 R6 R R N-H A-N-(CH2) n 1 2 r RB VI in which R, R, 6 R 7 R 8 A and n have the above- mentioned meanings and wherein the compound of general formula r V I is obtained by removal of the protective groups R 11 and W -53- from a compound of general formula VII Pl-A-N-(CH2) n R 8 VII in which R 5 R 6 R 7 R 8 A, P1 and n have the abovementioned meanings, and R11 denotes a diphenylmethyl radical and wherein the compound of general formula VII is obtained by reaction of a compound of general formula VIII R k N-R 11 H-N-(CH 2 R 8 VIII in which Rg, R 6 R 7 R 8 R 1 1 and n have the abovementioned meanings, with a compound of general formula P1-A-P2, or a compound of general formula P1-A in which PI, A and P2 have the abovementioned meanings and by employing the techniques for forming peptide bonds. (iv) if the compound is of general formula III, then by .3t reaction of a compound of general formula IV, in which R R 2 R 3 R 4 R 9 R 10 and X and Z have the S abovementioned meanings with a compound of general formula V; R R 5 N- H P-A-N-(CH 2 )n -54- in which R 5 R 6 R 7 R 8 P and n have the abovementioned meanings, an_ wherein the compound of general formula V is obtained by hydrogenolysis of a compound of general formula VII, in which R 5 R 6 R 7 R A, PI, n and R11 have the abovementioned meanings, by means of the use of a palladium catalyst under a hydrogen atmosphere and in an appropriate solvent and at a hydrogen pF assure of between 1 atm and 20 atm and the appropriate temperatures vary between C and 70 C; and wherein the corresponding salt of a compound of general formula I is obtained by reaction of a compound of general formula I with an inorganic or organic acid in an appropriate solvent. 4. A process according to claim 3 wherein Z is fluorine or chlorine. A process according to e her one of claims 3 or 4 wherein the palladium catalyst is PB(0H) 2 /C. 6. A process according to any one of claims 4 to 6 wherein the appropriate solvent is ethanol or methanol. 7. A pharmaceutical composition including a compound according to either one of claims 1 or 2 and a pharmaceutically acceptable carrier.

8. A method of treatment of microbial infection in a patient requiring treatment including administration of a compound according to either one of claims 1 and 2 or a S pharmaceutical composition according to claim 7.

9. A method of treatment according to claim 8 including 3'G the prophylaxis and chemotherapy of local and systemic infections produced by aerobic or anaerobic pathogens, both Gram-positive and Gram-negative. A method of treatment according to either one of claims 8 or 9 including administration parenterally.

11. A method of administering azetidinylquinolones, azetidinylnaphthyridines, azetidinylpyridobenzoxazines, azetidinylisothiazoloquinolones, azetidinylisothiazolo- naphthyridines or azetidinylisothiazolopyridobenzoxazines S including administering to a patient requiring treatment a compound according to either one of claims 1 or 2 wherein the compound undergoes enzyme hydrolysis to produce the azetidinylquinolones, azetidinylnaphthyridines, azetidinylpyridobenzoxazines, azetidinylisothiazolo- quinolones, azetidinylisothiazolonaphthyridines or azetidinylisothiazolopyridobenzoxazines in situ to the patient.

12. A compound according to claim 1 substantially as describe, herein with reference to any one of the non- comparatiie examples.

13. A process for the preparation of a compound according to claim 1 substantially as described herein with reference to any one of the non-comparative example-. DATED: 13 January 1995 PHILLIPS ORMONDE FITZPATRICK Attorneys for: LyMORATORIOS DEL DR ESTEVE, S.A. .3t S IIe -56- 57 PATENT "Amino acid azetidinyl substituted pyridone derivatives, their (preparation and their application as medications" Applicant: LABORATORIOS DEL Dr ESTEVE S.A. ABSTRACT The present invention relates to the compounds S corresponding to the general formula I I. 0 0 and their pharmaceutically acceptable salts, in which formula X denotes a nitrogen atom or else a carbon atom bonded to a hydrogen atom, bonded to a halogen atom, a hydroxyl or alkoxy radical, an alkyl radical, a halo- genated alkyl radial or an alkylamino radical; RI denotes a lower alkyl or cycloalkyl radical, a lower haloalkyl radical, a hydroxyalkyl radical, a vinyl radical, an aryl radical or an aryl radical substituted by one or more fluorine atoms, or an alkylamino radical; R 2 denotes a hydrogen atom or a lower alkyl radical; R 3 denotes a hydroxyl radical or a C 1 -C lower alkoxy radical; R 4 denotes a hydrogen atom, a fluorine atom, a lower alkyl radical, a nitro radical, an amino radical or a substituted amino radical; 58 Rs, R 6 and R 7 denote a hydrogen atom or a lower alkyl radical; n is a number which may be 0 or 1; X and RI may together form a bond represented by a group C-CHz-CHz-CH-Rg or C-O-CH-CHz-CH-R, or C-S-CH 2 -CH-Rg, in which R, denotes a hydrogen atom, a lower alkyl radical or a lower haloalkyl radical; R I 'and R 2 may together form a bond represented by a group -CHRi 0 in which R 1 0 denotes a hydrogen atom or a lower alkyl radical or a lower haloalkyl radical; R 2 and R 3 may together form a bond represented by a group -S-NH-; A is a solubilising group which is represented by a residual amino acid or a polypeptide chain consisting of two to four or more amino acid residues which are bonded covalently one to another by peptide bonds, and process for their preparation.