Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
  • A61K31/445—Non condensed piperidines, e.g. piperocaine
  • A61K31/4453—Non condensed piperidines, e.g. piperocaine only substituted in position 1, e.g. propipocaine, diperodon
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/16—Amides, e.g. hydroxamic acids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/16—Amides, e.g. hydroxamic acids
  • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/16—Amides, e.g. hydroxamic acids
  • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
  • A61K31/167—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide having the nitrogen of a carboxamide group directly attached to the aromatic ring, e.g. lidocaine, paracetamol
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
  • A61K31/19—Carboxylic acids, e.g. valproic acid
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
  • A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
  • A61K31/22—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids of acyclic acids, e.g. pravastatin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
  • A61K31/425—Thiazoles
  • A61K31/426—1,3-Thiazoles
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
  • A61K31/4402—Non condensed pyridines; Hydrogenated derivatives thereof only substituted in position 2, e.g. pheniramine, bisacodyl
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
  • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
  • A61K31/445—Non condensed piperidines, e.g. piperocaine
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/04—Antibacterial agents

Definitions

• This invention relates to the use of hydroxamic acid derivatives as antibacterial agents.
• bacterial pathogens are classified as either Gram-positive or Gram- negative. Many antibacterial agents (including antibiotics) are specific against one or other Gram-class of pathogens. Antibacterial agents effective against both Gram- positive and Gram-negative pathogens are therefore generally regarded as having broad spectrum activity.
• antibacterial agents including the penicillins and cephalosporins, tetracyclines, sulfonamides, monobactams, fluoroquinolones and quinolones, aminoglycosides, glycopeptides, macrolides, polymyxins, lincosamides, trimethophm and chloramphenicol.
• penicillins and cephalosporins including the penicillins and cephalosporins, tetracyclines, sulfonamides, monobactams, fluoroquinolones and quinolones, aminoglycosides, glycopeptides, macrolides, polymyxins, lincosamides, trimethophm and chloramphenicol.
• penicillins and cephalosporins tetracyclines
• sulfonamides monobactams
• fluoroquinolones and quinolones aminoglycosides
• glycopeptides
• MRSA methicillin resistant Staphylococcus aureus
• MRCNS methicillin resistant coagulase negative Staphylococci
• Penicillin resistant Streptococcus pneumoniae and multiply resistant Enterococcus faecium.
• Pathogenic bacteria are often resistant to the aminoglycoside, ⁇ -lactam (penicillins and cephalosporins), and chloramphenicol types of antibiotic. This resistance involves the enzymatic inactivation of the antibiotic by hydrolysis or by formation of inactive derivatives.
• the ⁇ -lactam (penicillin and cephalosporin) family of antibiotics are characterised by the presence of a ⁇ -lactam ring structure. Resistance to this family of antibiotics in clinical isolates is most commonly due to the production of a "penicillinase" ( ⁇ -lactamase) enzyme by the resistant bacterium which hydrolyses the ⁇ -lactam ring thus eliminating its antibacterial activity.
• Vancomycin-resistant enterococci are particularly hazardous in that they are frequent causes of hospital based infections and are inherently resistant to most antibiotics. Vancomycin works by binding to the terminal D-Ala-D-Ala residues of the cell wall peptidioglycan precursor.
• the high-level resistance to vancomycin is known as VanA and is conferred by a genes located on a transposable element which alter the terminal residues to D-Ala-D-lac thus reducing the affinity for vancomycin.
• This invention is based on the finding that certain hydroxamic acid derivatives have antibacterial activity, and makes available a new class of antibacterial agents.
• the inventors have found that the compounds with which this invention is concerned are antibacterial with respect to a range of Gram-positive and Gram-negative organisms.
• Bacterial polypeptide deformylases (EC 3.5.1.31), are a conserved family of metalloenzymes (Reviewed: Meinnel T, Lazennec C, Villoing S, Blanquet S, 1997, Journal of Molecular Biology 267, 749-761) which are essential for bacterial viability, their function being to remove the formyl group from the N-terminal methionine residue of ribosome-synthesised proteins in eubacteria. Mazel et al. (EMBO J. 13(4):914-923, 1994) have recently cloned and characterised an E. coli PDF.
• PDF Bacterial polypeptide deformylases
• the natural antibiotic actinonin (see for example J.C.S Perkin I, 1975, 819) is a hydroxamic acid derivative of Structure (A):
• Hydroxamic acid derivatives are also known in the field of matrix metalloproteinase (MMP) inhibition. Many examples of the class have been synthesised and their MMP inhibitory properties reported. A smaller number have been reported to be active in animal models of diseases mediated by MMPs, for example various cancers and rheumatoid arthritis.
• MMP matrix metalloproteinase
• Ri represents hydrogen, or C 1 -C 6 alkyl, C r C 6 alkyl substituted by one or more halogen atoms, amino, hydroxy, or C,-C 6 alkoxy;
• R 2 represents a group R 10 -(X) n -(ALK) m - wherein
• R 10 represents hydrogen, or a alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, cycloalkyl, aryl, or heterocyclyl group, any of which may be unsubstituted or substituted by (C C 6 )alkyl, (C 1 -C 6 )alkoxy, hydroxy, mercapto, (C 1 -C 6 )alkylthio, amino, halo (including fluoro, chloro, bromo and iodo), trifluoromethyl, cyano, nitro, -COOH, -CONH 2 -COOR A , -NHCOR A , -CONHR A , -NHR A , -NR A R B , or - CONR A R B wherein R A and R B are independently a (C r C 6 )alkyl group, and ALK represents a straight or branched divalent alkyiene, C 2 -C 6 al
• X represents -NH-, -O- or -S-
• n and n are independently 0 or 1 ;
• A represents (i) a group of formula (IA), (IB), (IC) or (ID)
• R 3 represents hydrogen or C,-C 6 alkyl and R 4 represents the side chain of a natural or non-natural alpha amino acid or R 3 and R 4 when taken together with the nitrogen and carbon atoms to which they are respectively attached form an optionally substituted saturated heterocyclic ring of 5 to 8 atoms which ring is optionally fused to a carbocyclic or second heterocyclic ring,
• R 5 and R 6 independently represent hydrogen, or optionally substituted C r C 8 alkyl, cycloalkyl, aryl, aryl(C 1 -C 6 alkyl), heterocyclic, or heterocyclic(C 1 -C 6 alkyl), or R 5 and R 6 when taken together with the nitrogen atom to which they are attached form an optionally substituted saturated heterocyclic ring of 3 to 8 atoms which ring is optionally fused to a carbocyclic or second heterocyclic ring, and R 7 represents hydrogen, C r C 6 alkyl, or an acyl group.
• R 5 and R 6 taken together with the nitrogen atom to which they are attached do not form an optionally substituted saturated heterocyclic ring of 3 to 8 atoms when R and R 3 are hydrogen, R 2 is hydrogen, alkyl, phenyl, benzyl, 4-chlorophenylmethyl, 4-nitrophenylmethyl, or 4-aminophenylmethyl and R 3 is hydrogen, methyl, isopropyl, isobutyl or benzyl; and (b) R 5 is not 2-pyridyl or 2- thiazolyl when R., R 3 and R 6 are hydrogen, R 2 is n-pentyl and R 4 is isopropyl; and (c) R 5 and R 6 are not both ethyl when R and R 3 are hydrogen, R 2 is n-pentyl and R 4 is methyl or isopropyl.
• the invention provides a method for the treatment of bacterial infections in humans and non-human mammals, which comprises administering to a subject suffering such infection an antibacterially effective dose of a compound of formula (I) as defined above.
• the compounds of formula (I) as defined above may be used as component(s) of antibacterial cleaning or disinfecting materials.
• the various aspects of the invention can be applied against vancomycin-, quinolone- and " ⁇ -lactam”-resistant bacteria and the infections they cause.
• the compounds (I) act by inhibition of intracellular PDF, the most potent antibacterial effect may be achieved by using compounds which efficiently pass through the bacterial cell wall.
• compounds which are highly active as inhibitors of PDF in vitro and which penetrate bacterial cells are preferred for use in accordance with the invention.
• the antibacterial potency of compounds which are potent inhibitors of the PDF enzyme in vitro, but are poorly cell penetrant may be improved by their use in the form of a prodrug, ie a structurally modified analogue which is converted to the parent molecule of formula (I), for example by enzymic action, after it has passed through the bacterial cell wall.
• (C 1 -C 6 )alkyl means a straight or branched chain alkyl moiety having from 1 to 6 carbon atoms, including for example, methyl, ethyl, n- propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl and n-hexyl.
• divalent (C 1 -C 6 )alkylene radical means a saturated hydrocarbon chain having from 1 to 6 carbon atoms and two unsatisfied valencies.
• (C 2 -C 6 )alkenyl means a straight or branched chain alkenyl moiety having from 2 to 6 carbon atoms having at least one double bond of either E or Z stereochemistry where applicable.
• the term includes, for example, vinyl, allyl, 1- and 2-butenyl and 2-methyl-2-propenyl.
• divalent (C 2 -C 6 )alkenylene radical means a hydrocarbon chain having from 2 to 6 carbon atoms, at least one double bond, and two unsatisfied valencies.
• C 2 -C 6 alkynyl refers to straight chain or branched chain hydrocarbon groups having from two to six carbon atoms and having in addition one triple bond. This term would include for example, ethynyl, 1-propynyl, 1- and 2- butynyl, 2-methyl-2-propynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 2-hexynyl, 3- hexynyl, 4-hexynyl and 5-hexynyl.
• divalent (C 2 -C 6 )alkynylene radical means a hydrocarbon chain having from 2 to 6 carbon atoms, at least one triple bond, and two unsatisfied valencies.
• cycloalkyl means a saturated alicyclic moiety having from 3-8 carbon atoms and includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.
• cycloalkenyl means an unsaturated alicyclic moiety having from 3-8 carbon atoms and includes, for example, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl. In the case of cycloalkenyl rings of from 5-8 carbon atoms, the ring may contain more than one double bond.
• aryl refers to a mono-, bi- or tri-cyclic carbocyclic aromatic group, and to groups consisting of two covalently linked monocyclic carbocyclic aromatic groups. Illustrative of such groups are phenyl, biphenyl and napthyl.
• heteroaryl refers to a 5- or 6- membered aromatic ring containing one or more heteroatoms, and optionally fused to a benzyl or pyridyl ring; and to groups consisting of two covalently linked 5- or 6- membered aromatic rings each containing one or more heteroatoms; and to groups consisting of a monocyclic carbocyclic aromatic group covalently linked to a 5- or 6- membered aromatic rings containing one or more heteroatoms;.
• Illustrative of such groups are thienyl, furyl, pyrrolyl, imidazolyl, benzimidazolyl, thiazolyl, pyrazolyl, isoxazolyl, isothiazolyl, triazolyl, thiadiazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, 4-([1 ,2,3]-thiadiazoly-4-yl)phenyl and 5-isoxazol-3-ylthienyl.
• heterocyclyl or “heterocyclic” includes “heteroaryl” as defined above, and in particular means a 5-7 membered aromatic or non-aromatic heterocyclic ring containing one or more heteroatoms selected from S, N and O, and optionally fused to a benzene ring, including for example, pyrrolyl, furyl, thienyl, piperidinyl, imidazolyl, oxazolyl, thiazolyl, thiadiazolyl, pyrazolyl, pyridinyl, pyrrolidinyl, pyrimidinyl, morpholinyl, piperazinyl, indolyl, benzimidazolyl, maleimido, succinimido, phthalimido and 1 ,3-dioxo-1 ,3-dihydro-isoindol-2-yl groups.
• acyl means a group R 20 C(O)- where R 20 is (C 1 -C 6 )alkyl, (C 2 -C 6 )alkenyl, (C 3 -C 7 )cycloalkyl, phenyl, heterocyclyl, phenyl(C r C 6 )alkyl, heterocyclyl(C r C 6 )alkyl, (C 3 -C 7 )cycloalkyl(C r C 6 )alkyl, phenyl(C 2 -C 6 )alkenyl, heterocyclyl(C 2 -C 6 )alkenyl, (C 3 -C 7 )cycloalkyl(C 2 -C 6 )alkenyl, any of which R 20 groups may be substituted.
• substituted as applied to any moiety herein means substituted with up to four substituents, each of which independently may be (C.,-C 6 )alkyl, benzyl, (C r C 6 )alkoxy, phenoxy, hydroxy, mercapto, (C 1 -C 6 )alkylthio, amino, halo (including fluoro, chloro, bromo and iodo), trifluoromethyl, nitro, -COOH, -CONH 2 -COR A , -COOR A , -NHCOR A , -CONHR A , - NHR A , -NR A R B , or -CONR A R B wherein R A and R B are independently a (C r C 6 )alkyl group.
• substituted means benzyl
• the phenyl ring thereof may itself be substituted with any of
• side chain of a natural alpha-amino acid and “side chain of a non-natural alpha-amino acid” mean the group R in respectively a natural and non-natural amino acid of formula NH 2 -CH(R x )-COOH.
• side chains of natural alpha amino acids include those of alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamic acid, histidine, 5- hydroxylysine, 4-hydroxyproline, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, ⁇ -aminoadipic acid, ⁇ -amino-n-butyric acid, 3,4-dihydroxyphenylalanine, homoserine, ⁇ - methylserine, omithine, pipecolic acid, and thyroxine.
• amides for example as a NHCOC r C 6 alkyl amide
• Salts of the compounds of the invention include physiologically acceptable acid addition salts for example hydrochlo des, hydrobromides, sulphates, methane sulphonates, p-toluenesulphonates, phosphates, acetates, citrates, succinates, lactates, tartrates, fumarates and maleates. Salts may also be formed with bases, for example sodium, potassium, magnesium, and calcium salts.
• R. may be, for example, hydrogen, hydroxy, methoxy, methyl, or trifuoromethyl. Hydrogen is currently preferred.
• R 2 may be, for example: optionally substituted alkyl, C 3 -C 6 alkenyl, C 3 -C 6 alkynyl or cycloalkyl;
• cycloalkyl(C.,-C 6 alkyl)- cycloalkyl(C 3 -C 6 alkenyl)- or cycloalkyl(C 3 -C 6 alkynyl)- optionally substituted in the cycloalkyl ring;
• R 2 groups include methyl, ethyl, n- and iso-propyl, n- and iso-butyl, n-pentyl, iso-pentyl 3-methyl- but-1-yl, n-hexyl, n-heptyl, n-acetyl, n-octyl, methylsulfanylethyl, ethylsulfanylmethyl, 2-methoxyethyl, 2-ethoxyethyl, 2-ethoxymethyl, 3- hydroxypropyl, allyl, 3-phenylprop-3-en-1-yl, prop-2-yn-1-yl, 3-phenylprop-2- yn-1-yl, 3-(2-chlorophenyl)prop-2-yn-1-yl, but-2-yn-1-yl, cyclopentyl, cyclohexyl, cyclopentylmethyl, cyclopentylethyl,
• Presently preferred groups at R 2 are n-propyl, n-butyl, n-pentyl, benzyl and cyclopentylmethyl.
• R 3 may be, for example, hydrogen or methyl, with hydrogen presently preferred.
• R 4 may be, for example the characterising group of a natural ⁇ -amino acid, for example isopropyl, benzyl, or 4-hydroxyphenylmethyl, in which any functional group may be protected, any amino group may be acylated and any carboxyl group present may be amidated; or
• Alk is a (C r C 6 )alkylene or (C 2 -C 6 )alkenylene group optionally interrupted by one or more -0-, or -S- atoms or -N(R 12 )- groups [where R 12 is a hydrogen atom or a (C.,-C 6 )alkyl group], n is 0 or 1 , and R 9 is hydrogen or an optionally substituted phenyl, aryl, heterocyclyl, cycloalkyl or cycloalkenyl group or (only when n is 1)
• R 9 may additionally be hydroxy, mercapto, (C r C 6 )alkylthio, amino, halo, trifluoromethyl, nitro, -COOH, - CONH 2 -COOR A , -NHCOR A , -CONHR A , -NHR A , -NR A R B , or -CONR A
• R 8 is hydroxyl, amino, (C 1 -C 6 )alkoxy, phenyl(C 1 -C 6 )alkoxy, (C 1 -C 6 )alkylamino, di((C r C 6 )alkyl)amino, phenyl(C r C 6 )alkylamino; or
• a heterocyclic(C 1 -C 6 )alkyl group either being unsubstituted or mono- or di- substituted in the heterocyclic ring with halo, nitro, carboxy, (C C 6 )alkoxy, cyano, (C r C 6 )alkanoyl, trifluoromethyl (C r C 6 )alkyl, hydroxy, formyl, amino, (C C 6 )alkylamino, di-(C r C 6 )alkylamino, mercapto, (C r C 6 )alkylthio, hydroxy(C C 6 )alkyl, mercapto(C 1 -C 6 )alkyl or (C 1 -C 6 )alkylphenylmethyl; or
• each of R a , R b and R c is independently hydrogen, (C 1 -C 6 )alkyl, (C 2 - C 6 )alkenyl, (C 2 -C 6 )alkynyl, phenyl(C 1 -C 6 )alkyl, (C 3 -C 8 )cycloalkyl; or
• R c is hydrogen and R a and R b are independently phenyl or heteroaryl such as pyridyl; or
• R c is hydrogen, (C r C 6 )alkyl, (C 2 -C 6 )alkenyl, (C 2 -C 6 )alkynyl, pheny C ⁇ C 6 )alkyl, or (C 3 -C 8 )cycloalkyl, and R a and R b together with the carbon atom to which they are attached form a 3- to 8- membered cycloalkyl or a 5- to 6-membered heterocyclic ring; or
• R a , R b and R c together with the carbon atom to which they are attached form a tricyclic ring (for example adamantyl); or
• R a and R b are each independently (C 1 -C 6 )alkyl, (C 2 -C 6 )alkenyl, (C 2 - C 6 )alkynyl, phenyl(C.,-C 6 )alkyl, or a group as defined for R c below other than hydrogen, or R a and R b together with the carbon atom to which they are attached form a cycloalkyl or heterocyclic ring, and R c is hydrogen, -OH, -SH, halogen, -CN, -C0 2 H, (C ⁇ C ⁇ perfluoroalkyl, - CH 2 OH, -C0 2 (C 1 -C 6 )alkyl, -0(C r C 6 )alkyl, -0(C 2 -C 6 )alkenyl, -S(C r C 6 )alkyl, -SO(C r C 6 )alkyl, -S0 2
• R 4 groups include methyl, ethyl, isopropyl, benzyl, 4- chlorobenzyl, 4-hydroxybenzyl, phenyl, cyclohexyl, cyclohexylmethyl, pyridin- 3-ylmethyl, tert-butoxymethyl, naphthylmethyl, iso-butyl, sec-butyl, tert-butyl, 1-benzylthio-1-methylethyl, 1-methylthio-1-methylethyl, 1-mercapto-1- methylethyl, 1-methoxy-1-methylethyl, 1-hydroxy-1-methylethyl, 1-fluoro-1- methylethyl, 4,4-dimethyl-prop-1-en-4-yl, 4,4-dimethyl-prop-4-yl hydroxymethyl, 2-hydroxethyl, 2-carboxyethyl, 2-methylcarbamoylethyl, 2- carbamoylethyl, and
• R 3 and R 4 when taken together with the nitrogen and carbon atoms to which they are respectively attached may form an optionally substituted saturated heterocyclic ring of 5 to 8 atoms.
• R 5 and R 6 may independently be, for example, hydrogen, methyl, ethyl, tert-butyl, n- heptyl, cyclopentyl, cyclohexyl, phenyl, 2-ethoxycarbonyl-eth-2-yl, pyrid-2-yl, 1 ,1 ,3,3- tetramethylbutyl, benzyl, 2,6-dimethyl-4-tert-butyl-phenyl,diphenylmethyl, 4- chlorophenyl-phenylmethyl, 2-fluorophenyl-phenylmethyl, 1 -(4-fluorophenyl)-1 - phenyl-1-amino-methyl, 1 ,1-diphenylprop-3-yl, 3-phenyl-thiazoiyl, or 2-hydroxyethyl; or R 5 and R 6 when taken together with the nitrogen atom to which they are attached may form a saturated 5- to 8-membered monocyclic N
• Examples of such rings are substituted or unsubstituted 1-pyrrolidinyl, piperidin-1-yl, 1-piperazinyl, hexahydro-1-pyridazinyl, morpholin-4-yl, tetrahydro-1 ,4-thiazin-4-yl, tetrahydro-1 ,4-thiazin-4-yl 1 -oxide, tetrahydro-1 ,4- thiazin-4-yl 1 ,1 -dioxide, hexahydroazipino, or octahydroazocino.
• R 5 be methyl or hydrogen
• R 6 be methyl
• R 7 may be, for example, hydrogen, or a group R 20 C(O)- where R 20 is a (C r C 6 )alkyl group such as methyl or ethyl.
• a specific example of a compound having PDF inhibiting and antibacterial activity in accordance with the invention is: N 1 -(1S-dimethylcarbamoyl-2,2-dimethyl-1-propyl)-N 4 -hydroxy-2R-butyl- succinamide
• compositions with which the invention is concerned may be prepared for administration by any route consistent with the pharmacokinetic properties of the active ingredient(s).
• Orally administrable compositions may be in the form of tablets, capsules, powders, granules, lozenges, liquid or gel preparations, such as oral, topical, or sterile parenteral solutions or suspensions.
• Tablets and capsules for oral administration may be in unit dose presentation form, and may contain conventional excipients such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinyl-pyrrolidone; fillers for example lactose, sugar, maize-starch, calcium phosphate, sorbitol or glycine; tabletting lubricant, for example magnesium stearate, talc, polyethylene glycol or silica; disintegrants for example potato starch, or acceptable wetting agents such as sodium lauryl sulphate.
• binding agents for example syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinyl-pyrrolidone
• fillers for example
• Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a dry product for reconstitution with water or other suitable vehicle before use.
• Such liquid preparations may contain conventional additives such as suspending agents, for example sorbitol, syrup, methyl cellulose, glucose syrup, gelatin hydrogenated edible fats; emulsifying agents, for example lecithin, sorbitan monooleate, or acacia; non-aqueous vehicles (which may include edible oils), for example almond oil, fractionated coconut oil, oily esters such as glycerine, propylene glycol, or ethyl alcohol; preservatives, for example methyl or propyl p-hydroxybenzoate or sorbic acid, and if desired conventional flavouring or colouring agents.
• suspending agents for example sorbitol, syrup, methyl cellulose, glucose syrup, gelatin hydrogenated edible fats
• emulsifying agents for example lecithin, sorbitan monooleate, or acacia
• non-aqueous vehicles which may include edible oils
• almond oil fractionated coconut oil
• oily esters such as glycerine, propylene
• the active ingredient(s) may be made up into a cream, lotion or ointment.
• Cream or ointment formulations which may be used for the drug are conventional formulations well known in the art, for example as described in standard textbooks of pharmaceutics such as the British Pharmacopoeia.
• the active ingredient(s) may also be administered parenterally in a sterile medium.
• the drug can either be suspended or dissolved in the vehicle.
• adjuvants such as a local anaesthetic, preservative and buffering agents can be dissolved in the vehicle.
• Safe and effective dosages for different classes of patient and for different disease states will be determined by clinical trial as is required in the art. It will be understood that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed, the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease undergoing therapy.
• Example 4 By methods described in the literature analogous to those used for Example 1-3 above, the following compounds of formula (I) above, wherein A is a group of formula (IA) were prepared:
• the title compound was prepared by Ugi reaction of the appropriate homochiral succinate ester with trimethylacetaldehyde and ammonia, followed by conversion to the desired hydroxamic acid, as described in GB-2298423-A.
• the starting succinates were prepared by the method described in WO 92/13831.
• the title compound was prepared by a method analogous to that used for the preparation of compounds of Examples 1-4, except that pipe ⁇ dine was used in the coupling reaction in place of the amino acid derivative.
• the E. coli PDF gene was cloned in pET24a(+) (designated pET24-PDF) and was used to transform BL21 DE3 cells from Novagen Inc, (Madison, Wisconsin). Clones were selected at 37°C on YT agar plates (8g/l typtone, 5g/yeast extract, NaCI 5g/l, agar 15g/l) supplemented with 30 ⁇ g/ml kanamycin.
• the cells from a 1 litre expression were resuspeneded in 2x 25ml of ice cold phosphate buffered saline.
• the cell suspension was sonicated on ice using an MSE Soniprep 150 fitted with a medium probe and at an amplitude of 20-25 microns in 6x20 second pluses.
• the resulting suspension was then cleared by centrifugation at 20,000 xg for 15 minutes. The supernatant was then used for further purification of the enzyme.
• E. coli lysate from a 11 culture in phosphate buffered saline (PBS) were adjusted to 2M ammonium sulphate.
• a 15ml phenyl sepharose column was equilibrated with PBS/2M ammonium sulphate at 4°C.
• the lysate was loaded on the column and washed with equilibration buffer.
• the column was eluted by reducing the ammonium sulphate concentration from 2M to 0M over 10 column volumes.
• 5ml fractions were collected and analysed by SDS-PAGE.
• the fractions containing the majority of the 20kDa PDF were pooled.
• the pooled fractions were concentrated using a 3kDa cutoff membrane to a volume of 5ml.
• the assay was performed in a single 96 well plate in a final volume of 10O ⁇ l containing:
• the assay was incubated at 37°C for 30 minutes.
• the free amino group of the deformyiated (Met-Ala-Ser) product was detected using fluorescamine, by the following additions:
• Standard control reactions are a no inhibitor reaction which provides the zero inhibition figure and a no enzyme and no inhibitor reaction which provides the 100% inhibition figure.
• the data was analysed by conversion of the fluorescence units to % inhibition and the inhibitor concentration plotted against % inhibition.
• the IC 50 represents the concentration of inhibitor (nM) required to decrease enzyme activity by 50%.
• test compounds were found to inhibit bacterial PDF in vitro.
• MIC Minimal inhibitory concentrations (MIC) of the test compounds against E. coli strain DH5 ⁇ (Genotype; F- ⁇ 80d/acZ ⁇ M15 ⁇ (/acZYA-argF)U169 eoR recA1 endA ?sdR17(r k ' ,m k + )pt ⁇ oA supE44 ⁇ ' thiX gytA96 re/A1) obtained from GibcoBRL Life Technologies, were determined as follows. Stock solutions of test compound were prepared by dissolution of each compound in dimethylsulfoxide at 10mM.
• 2xYT broth typtone 16g/l, yeast extract 10g/l, sodium chloride 5g/l obtained from BIO 101 Inc, 1070 Joshua Way, Vista, CA92083, USA
• Microtiter plates were incubated at 37°C for 18 hours in a humidified incubator. The test compounds had MIC's of 200 ⁇ M or less against one or both of the test organisms.

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