WO2002032891A1 - Flavan compounds, their preparation and their use in therapy - Google Patents

Flavan compounds, their preparation and their use in therapy Download PDF

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Publication number
WO2002032891A1
WO2002032891A1 PCT/GB2001/004641 GB0104641W WO0232891A1 WO 2002032891 A1 WO2002032891 A1 WO 2002032891A1 GB 0104641 W GB0104641 W GB 0104641W WO 0232891 A1 WO0232891 A1 WO 0232891A1
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alkyl
same
different
aryl
alkenyl
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PCT/GB2001/004641
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French (fr)
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Mohammad Michel Afshar
Stephen David Morley
Alastair Iain Hamilton Murchie
Martin James Drysdale
Andrew John Potter
Justin Fairfield Bower
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Ribotargets Limited
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Priority claimed from GB0025581A external-priority patent/GB0025581D0/en
Priority claimed from GB0100423A external-priority patent/GB0100423D0/en
Application filed by Ribotargets Limited filed Critical Ribotargets Limited
Priority to AU2002210686A priority Critical patent/AU2002210686A1/en
Publication of WO2002032891A1 publication Critical patent/WO2002032891A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
    • C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
    • C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
    • C07D231/12—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
    • C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
    • C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
    • C07D311/96—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings spiro-condensed with carbocyclic rings or ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/10—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a carbon chain containing aromatic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention relates to a class of flavan (2,3-dihydro-2-phenylbenzopyran) chemical compounds, their preparation and their use in therapy, particularly in the treatment of bacterial infection.
  • Flavan derivatives have previously been reported as the condensation products of phenols and ketones.
  • Baker et al. J. Chem. Soc, 1957, 3060-3062 identify the condensation product of m-cresol and cyclohexanone as cyclohexanespiro-4-(2 , -hydroxy-7, 4'-dimethyl- 2, 3-tetramethyleneflavan.
  • Livant et al. J. Org.
  • the present inventors have identified a class of flavan derivatives, which are useful in the treatment of bacterial infection.
  • each R " ,R (l and R 7 is the same or different and is selected from H. C1-1 2 alkyl, C 2 . 12 alkenyl, C 2 -12 alkynyl, C 3 -12 aryl, C 4 - 1 X aralkyl; provided that at least one of A and B is OR " , and pharmaceutically acceptable derivatives thereof.
  • C ⁇ - 12 alkoyl, C 3 -. ⁇ 2 aryloyl and C is aralkyl, and each R ⁇ R 6 and R 7 is the same or different and is selected from H. C 1 - 12 alkyl. C2-12 alkenyl. C 2 -. 2 alkynyl. C 3 -12 aryl. C4. 1S aralkyl; provided that at least one of A and B is OR ⁇ and pharmaceutically acceptable derivatives thereof, provided that at least one of A. B. Y 1 . Y " and Y 3 is not H, CH;,. C;H ? or OH ( preferably at least one of A. B, Y 1 . Y ⁇ and Y ⁇ is not H. OH.
  • alkyl means an optionally substituted branched or unbranched, cyclic or acyclic, hydrocarbyl radical.
  • the alkyl group is preferably a C 1 - 12 , more preferably C]-rumble more preferably C1- 4 chain.
  • the alkyl group is preferably a C 3 -i 2 , more preferably Cs-i and more preferably comprises a C 5 , C 6 or C ⁇ ring.
  • the term "alkenyl” means an optionally substituted branched or unbranched, cyclic or acyclic, hydrocarbyl radical comprising at least one double bond.
  • the alkenyl group is preferably a C 2 -1 2 - more preferably C 2 - 6 , more preferably C1-4 chain.
  • the alkenyl group is preferably a C 3 -1 2 , more preferably C 5 - 10 and more preferably comprises a C 5 , C 6 or C 7 ring.
  • alkynyl means an optionally substituted, branched or unbranched, cyclic or acyclic, hydrocarbyl radical comprising at least one triple bond.
  • the alkynyl group is preferably a C 2 - 12 , more preferably C 2 - 4 chain.
  • the alkynyl group is preferably a C4-1 2 , more preferably C 6 - ⁇ o ring.
  • aryl means an optionally substituted C 3 -12, aromatic group, such as phenyl or naphthyl, or a heteroaromatic group containing one or more, preferably one, heteroatom, such as pyridyl, pyrrolyl, furanyl, thienyl.
  • aromatic heterocyclic groups include monocyclic groups such as anovanyl, azetyl, azoninyl, imidazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridinyl, pyri idinyl, pyrrolyl, sym ⁇ triazinyl, triazolyl, furanyl, oxoninyl, pyryliumyl, thioninyl, thiophenyl, thiopyryliumyl, selenophenyl, fiirazanyl, isothiazolyl, isoxazolyl, methylthiabenzenyl, oxazinyl, oxazolyl, thiazinyl, thiazolyl; bicyclic groups such as benzimidazolyl, cinnolinyl, indazolyl, indolyl, indolinyl, indolizin
  • quinolinyl quinolinyl, quinolizinyl, quinoliziniumyl, quinoxalinyl, benzofliranyl, benzopyryliumyl, chromanyl, chromenyl. chromonyl, coumarinyl. isobenzofuranyl, isochromanyl. isochromenyl, benzothiophenyl, arsindolyl, arsinolinyl, isoarsindolyl, isoarsinolinyl. isophosphindolyl, isophosphinolinyl. phosphindolyl. phosphinolinyl; polycyclic groups such as acridinyl, carbolinyl. carbazolyl.
  • aralkyl means an optionally substituted branched or unbranched cyclic or acylic C 4 _i 8 group comprising an alkyl group and an aryl group (for example, benzyl). An aralkyl group may be bonded via the alkyl or aryl group.
  • heterocyclic group means an optionally substituted C 3 - 12 cyclic group containing at least one, preferably one to three, heteroatoms.
  • examples include monocyclic groups such as anovanyl, azetyl, azoninyl, imidazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, sym-triazinyl, triazolyl, furanyl, oxoninyl, pyryliumyl, thioninyl, thiophenyl, thiopyryliumyl, selenophenyl, furazanyl, isothiazolyl, isoxazolyl, methylthiabenzenyl, oxazinyl, oxazolyl, thiazinyl, thiazolyl; bicyclic groups such as
  • non-aromatic heterocyclic groups include monocyclic groups such as azaundecinyl, azepinyl, azetidinyl, aziridinyl, azocinyl, imidazolidinyl, imidazolinyl, piperazinyl, piperidinyl, pyrazolidinyl, pyrazolinyl, pyiTolidinyl, pyrrolinyl, quinuclidinyl, dihydropyranyl, dioxanyl, ethylene oxidyl, oxepinyl, oxetanyl, oxocinyl. pyranyl, pyronyl, tetrahydrofuranyl.
  • alkoyl means alkyl-CO- w herein alkyl is defined above.
  • arvlovl means arvl-CO- wherein arvl is defined above.
  • the alkyl, alkenyl, alkynyl, aryl, aralkyl and heterocyclic groups may be substituted or unsubstituted. Where substituted, there are preferably one to three substituents, more preferably one substituent.
  • Substituents may include halogen atoms and halogen containing groups such as haloalkyl (e.g. trifluoromethyl); oxygen containing groups such as alcohols (e.g.
  • hydroxy, hydroxyalkyl, aryl(hydroxy)alkyl ethers (e.g. alkoxy, alkoxyalkyl, aryloxyalkyl), aldehydes (e.g. carboxaldehyde), ketones (e.g. alkylcarbonyl, alkylcarbonylalkyl, arylcarbonyl, arylalkylcarbonyl, arylcarbonylalkyl), acids (e.g. carboxy, carboxyalkyl), acid derivatives such as esters (e.g.
  • cyano, cyanoalkyl nitro
  • sulfur containing groups such as thiols, thioethers, sulfoxides, and sulfones (e.g. alkylthio, alkylsulfinyl, alkylsulfonyl, alkylthioalkyl, alkylsulfmylalkyl, alkylsulfonylalkyl, arylthio, arylsulfinvl, arylsulfonyl, arylthioalkyl, arylsulfmvlalkyl, arylsulfonylalkyl); carbon containing groups (e.g.
  • alkyl, alkenyl, alkynyl, aryl and aralkyl ); and heterocyclic groups containing one or more, preferably one, heteroatom (e.g. thienyl, furanyl. pynolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, oxadiazolyl, thiadiazolyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, tetrahydro furanyl, pyranyl, pyronyl, pyridyl, pyrazinyl, pyridazinyl.
  • heteroatom e.g. thienyl, furanyl. pynolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl,
  • aromatic heterocyclic groups include monocyclic groups such as uazonyl. uzetyl. azon ⁇ n l. imidazolyl. pyrazinyl. razolyl. pyridazinyl. py ⁇ dinyi. py ⁇ m ⁇ dinyi. pyrrolyl. sym-triazinyl. triazolyl. furanyl. oxon ⁇ n l. pyryliumyl. thioninyl. thiophenyl.
  • thiopyiyhumyl selenophenyl fuiazanyl isothiazolyl, isoxazolyl methylthiabcnzenvl oxazinyl, oxazolyl thiazinyl thiazolyl, bicychc gioups such as benzimidazolyl, cmnolmyl indazolyl, indolyl indolinyl, mdohzinyl, isoindolyl, lsomdohnyl, l tetrahydroisoquinolmy, naphthyiidmyl, phthalazinyl, pteiidmyl, memeinyl, pynohzinyl, quinazolinyl, quinolinyl qumolizmyl, qumoliziniumyl, qumoxalinyl, benzoturanyl, benzopyiyhumyl, chrome
  • halogen means a fluorine, chlorine, biomme oi iodine tadical preferably a fluorine chlorine or bromine radical, preferably a fluonne or chlorine tadical
  • dialkyl ammo gioups ' include cyclic amines of the to ⁇ nul ⁇ -NRR wheie R and R are linked to form a cyclic group pieteiablv containing 5 to 7 moie pt efeiably 6 nng atoms and piefeiablv containing one to thiee heteioatoms
  • Examples of such cvchc g' oup include pipe ⁇ dinvl morphohn ⁇ l and piperaz v l groups
  • ⁇ phaimaLeuticallv acceptable e anv ph ⁇ naceuticallv acceptable salt addition compound oi anv othe- compound w hich upon admmisti ation to ⁇ leupient capable of id g idiiectlv oi -n ⁇ i , ⁇ compound of the ⁇ e->en ⁇ e .-ion oi ⁇ pharmaceutically acceptable metabolite.
  • 'pharmaceutically acceptable metabolite is meant a metabolite or residue of a compound of the present invention which gives rise to a biological activity exhibited by the present compounds.
  • the compounds of the present invention may exist in a number of diastereomeric and enantiomeric forms.
  • the present invention encompasses pure diastereomers and enantiomers as well as mixtures (including racemic mixtures) of diastereomers and enantiomers.
  • T p represents a divalent group comprising groups T where p is selected from the integers 4, 5 and 6, such that the ring comprising -Tp- is a 5-, 6- or 7-membered ring, respectively.
  • p is 4 or 5.
  • Tq represents a divalent group comprising groups T where q is selected from the integers 3, 4 or 5, such that the ring comprising — T t] -is a 5-, 6- or 7-membered ring, respectively.
  • q is 3 or 4.
  • T p one T is X and the remaining T are -CR ;
  • T q one T is X and the remaining T are -CR 1 ;-
  • the compounds of the present invention are of the formula
  • each U may be the same or different and is selected from
  • U when U is not symmetrical (i.e. when U is -X-CR 2 - or -CR 2 -X- CR'i-CR 1 ; it may be bonded to the molecule in two orientations (namely -X-CR' 2 - or -CR' ⁇ -X- and -CR ⁇ -X-CR ⁇ -CR ⁇ - or -CR ⁇ -X-CR ⁇ -CR 1 ;). Both orientations are encompassed by the present invention.
  • U is -CR -X-CR 1 -.
  • each X is selected from 0, S, NR' where each R' may be the same or different and is selected from H. C t .i, alkyl. C;. fl alkenyl. C;-i.> aryl, C4-1; aralkyl. C(0)R “ . SO;R “ and C(0)R “ ; wherein R " may be same or different and is selected from . C t .., alkvl. C;.,, alkenyl. C . ;- ⁇ , > arvl. C-i-i; aralkyl.
  • each X is -CR 1 ;-.
  • Each group R is selected independently from H, C M ; alkyl, C2- 12 alkenyl, C 3 -12 aryl and C4- 1 aralkyl.
  • any two R 1 in a geminal relationship i.e. each R group bonded 5 to the same carbon atom) are the same.
  • each R is hydrogen.
  • each R' is hydrogen or C
  • each R" is C ⁇ - 6 alkyl.
  • A, B, Y 1 , Y 2 , Y J , Z 1 , Z 2 , Z 3 and Z may be the same or different and are selected from H, halogen, Ci-i; alkyl, C 2 -12 alkenyl, C 2 -12 alkynyl, C 3 -12 aryl, C4-1S aralkyl, SR ⁇ , OR J , NR R " ,
  • R 2 , R J and R are the same or different and are selected from H, C ⁇ -12 preferably C ⁇ - 6 alkyl, C 2 -1 2 preferably C;-6 alkenyl, C2- 12 preferably C2- 6 alkynyl, C 3 -1 2 preferably C 3 -1 0 aryl, Ci-1 preferably Ci-r, alkoyl, C 3 -12 preferably C 3 _ ⁇ o aryloyl and C 4 - 18 preferably C 2 aralkyl; and each R", R 6 and R 7 is the same or different and is selected from H, C1- 12 preferably C ⁇ - 6 alkyl, C;-i2 preferably C2- 6 alkenyl, C2-12 preferably
  • A, B, Y , Y " , Y J , Z 1 , Z ⁇ , Z J and Z 4 are selected from C 2 -12 alkenyl, C 3 -1 2 aryl and C4-18 aralkyl groups,
  • substituents are selected from halogen atoms.
  • Z , Z ⁇ , Z J and Z are selected from C1- 1 ; alkyl groups, preferred substituents are selected from halogen atoms, SR' ' , OR 1 and NR 1 R ⁇ , wherein . R . R and R are the same or different and are selected from H and Ci-., alkyl.
  • Z , Z " , Z" and Z are selected from
  • I S aralkyl groups one to three substituents is selected from -COR . CO;lV" and CO " N ; R 1 R wherein R 1 ' " . R J ⁇ R "' and R ⁇ : are selected from I I. C ; alkyl. C;- t ; alkenyl. C;- ⁇ ; alkynyl.
  • aryl and C4- 1 aralkyl groups which may be substituted as defined for alkyl, alkenyl, alkynyl, aryl and aralkyl groups above.
  • preferred substituents are selected from amino, alkylamino, dialkylamino and heterocyclic groups, preferably, a group of the formula -NR , 0 R H where R 1 and R 1 1 are selected from H, C M ; alkyl, C2.
  • alkenyl, C 3 - 12 aryl and C 4 - 18 aralkyl which may be unsubstituted or substituted as defined for alkyl, alkenyl, aryl and aralkyl groups above; preferably R and R are linked to form a cyclic group, preferably containing 5 to 7, more preferably six, ring atoms and preferably containing one to three heteroatoms, which may be unsubstituted or substituted as defined for heterocyclic groups above.
  • Y , and Y J are selected from H, OR J , C ; alkyl and halogen. More preferably, Y 1 and Y J are selected from H, OFI and halogen, more preferably H and halogen.
  • Y 1 or Y J is C 1 -1 2 alkyl which may be unsubstituted or substituted as defined for alkyl groups above.
  • Y or Y J is C 1 -1 2 alkyl substituted with a group selected from amino, alkylamino, dialkylamino and heterocyclic groups.
  • Y 1 or Y J is a group of the formula
  • R 10 and R 11 are selected from H. C ; alkyl, C 2 - 1 ; alkenyl, C3-1; aryl and C4-1 8 aralkyl, which may be unsubstituted or substituted as defined for alkyl, alkenyl, aryl and aralkyl groups above.
  • R ⁇ and R 1 are linked to form a cyclic group, preferably containing 5 to 7. more preferably six, ring atoms and preferably containing one to three heteroatoms.
  • Such heterocyclic groups may be unsubstituted or substituted as defined for heterocyclic groups above. Examples of such heterocyclic groups include piperidinyl. piperazinyl and morpholinyl.
  • Y or Y" is CO;R" w herein A is selected from H. C M; alkyl. C;- ⁇ ; alkenyl. C;.j; alkynyl. C.M; aryl. C.j-.,s ' aralkyl an heterocyclic group w hich may be unsubstituted or substituted as defined for alkyl, alkenyl, alkynyl, aryl, aralkyl and heterocyclic groups above.
  • Y or Y" is CONR ; wherein each R is selected from H, C M ; alkyl. C;. ⁇ ; alkenyl, C;- ⁇ ; alkynyl, C;_ ⁇ ; aryl and C M S aralkyl, which may be unsubstituted or substituted as defined for alkyl, alkenyl, alkynyl, aryl and aralkyl groups above.
  • two R groups may be linked to form a heterocyclic group, preferably containing 5 to 7, more preferably 6 ring atoms, and preferably containing one to three heteroatoms. Such heterocyclic groups may be unsubstituted or substituted as defined for heterocyclic groups above.
  • heterocyclic groups include piperidinyl, piperazinyl, morpholinyl, 1,2,3,4-tetrahydroisoquinolinyl and 1 ,2,3,4- tetrahydro-6,7-isoquinolinediolyl.
  • Y 2 is selected from H, C M alkyl, OR 3 , CO;R 6 and CONR 7 ;. More preferably Y 2 is OR "1 in which R J is preferably H, C M alkyl, C M ; alkoyl or C4-1 8 aralkyl, more preferably FI, C ⁇ - 6 alkyl or Ci-g alkoyl.
  • Z , Z J and Z are selected from H, OR J and halogen, more preferably H, OH and halogen.
  • Z 2 is selected from H, C 2 alkyl, OR 3 , CO;R ⁇ and CONR 7 ;. More preferably Z ⁇ is OR J , in which R J is preferably H, C ; alkyl, C M ; alkoyl or C4-18 aralkyl, more preferably H, C ⁇ - 0 alkyl or C ⁇ -(, alkoyl.
  • At least one of A and B is OR" 3 .
  • one of A and B is OR" and one of A and B is
  • a or B is OR “* where R "" is preferably H.
  • a or B is OR where R J is selected from C M alkyl, C M ; alkenyl, C M; alkynyl, C 3 -1; aryl, C M ; alkoyl, C 3 -1; aryloyl and C 4 - 18 aralkyl, each of which may be substituted or unsubstituted as defined for alkyl, alkenyl, alkynyl, aryl and aralkyl groups above.
  • a or B is OR J where R is C;- ⁇ ; alkenyl which may be substituted as defined for alkenyl groups above.
  • a or B is OR J where R 3 is a group of the forcnula
  • T is H, C ; alkyl, C 3 - 12 aryl or C4-1 2 aralkyl; preferably H or C 3 - 12 aryl (including unsubstituted C 3 -1 2 aryl such as phenyl, benzothiazolyl and quinolinyl, and substituted C 3 -1; aryl such as p-fluorophenoxyphenyl, methylbenzothiazolyl and methylquinolinyl).
  • a or B is OR J where R J is C ; alkyl which may be unsubstituted or substituted as defined for alkyl groups above.
  • a or B is OR 3 where RJ is C e, alkyl substituted with a group selected from -COR 12 , CO;R and CONR'V 5 where R 12 , R 13 , R 14 and R 15 are selected from H, C 2 alkyl, C;- ⁇ ; alkenyl, C 2 - 1 ; alkynyl, C 3 - 1 ; aryl and C4- 1 S aralkyl, which may be substituted as defined for alkyl, alkenyl, alkynyl, aryl and aralkyl groups above.
  • a or B is OR "1 where R J is a group of the formula
  • R 14 and R are as defined above.
  • R 14 is H.
  • R 14 and R may be linked to form a cyclic group preferably 5 to 7, more preferably six ring atoms and containing one to three heteroatoms.
  • Such heterocyclic groups may be unsubstituted or substituted as defined for heterocyclic groups above.
  • Preferred heterocyclic groups include pipe ⁇ dinyl, piperaziny and morpholinyl,
  • a preferred group of compounds of the present invention comprise compounds of the formula
  • m and n may be the same or different and are selected from 1, 2 or 3; each R 1 and R ⁇ may be the same or different and are selected from H, C M alkyl. C M ; alkenyl, C 3 -1; aryl and C4-] 8 aralkyl; A, B, Y , Y ⁇ , Y J , Z , Z " , Z J and Z may be the same or different and are selected from H, C M ; alkyl, C M ; alkenyl, C 3 - 1 ; aryl, Q-is aralkyl, OR 3 and NR 4 R ⁇ ; wherein R J , R and R" are the same or different and are selected from H and C ⁇ - 6 alkyl; provided that at least one of A and B is OR J , and pharmaceutically acceptable derivatives thereof.
  • the compounds of the present invention may be used in the treatment of bacterial infection of humans, other animals or plants.
  • a compound according to the present invention for use in a method of treatment, preferably in the treatment of bactenal infection.
  • use of a compound according to the present invention in the manufacture of a medicament for the treatment of bacterial infection is provided.
  • a method of treatment of bacterial infection comprising administration to a subject in need of such treatment an effective dose of a compound according to the present invention.
  • Bacterial infections include bacterial infections that can afflict humans, other animals or plants. Bacterial infections include infections by Gram Positive Bacteria including Bacillus cereus, Bacillus anthracis, Clostridium botulinum, Clostridium difficile, Clostridium tetani, Clostridial perfringens, Corynebacteria diphtheriae, Enterococcus (Streptococcus D), Listeria Monocytogenes, Pneumoccoccal Infections (Streptococcus pneumoniae), Staphylococcal Infections and Streptococcal Infections; Gram Negative Bacteria including Bactcroides, Bordetella pertussis, Bmcella, Campylobacter Infections, Enterohemmorrhagic Escherichia coli (EFIEC/E.coli 0157-H7), Enteroinvasive Escherichia coli (EIEC), Enterotoxigenic Escherichia coli (ETEC),
  • Acid Fast Bacteria including Mycobacterium tuberculosis, Mycobacterium avium-intracellulare, Mycobacterium leprae, Atypical Bacteria, Chlamydia, Mycoplasma, Rickettsia, Spirochetes, Treponema pallidum, Bonelia recunentis, Borrelia burgdorfii and Leptospira icterohemorrhagiae; and other miscellaneous bacteria including Actinomyces and Nocardia.
  • Bacterial infections also include bacteria of the following genera of bacteria, which are significant phytopathogens affecting fruit, vegetable and ornamental plant production: Pseudomonas. Xa ⁇ tli ⁇ m ⁇ nas, Envinia, Strept ⁇ myces. Clostridium and ⁇ grobacrerium.
  • the term "'treatment” includes prophylactic treatment. It is a featui e of the compounds of the present invention that they inhibit the binding of compounds to RNA Foi example the compounds of the present inv ention inhibit the binding of proteins to RNA Accoidinglv the piesent invention furthei piovides use of a compound of the piesent invention to inhibit the binding of a compound, prefeiablv a piotem, to RNA
  • the piesent invention furthei provides use of a compound of the piesent invention to inhibit the translation of pioteins, preferably bacterial proteins
  • composition comprising a compound of the present invention m combination with a pharmaceutically acceptable excipient
  • a method of preparing a pharmaceutical composition comprising the step of combining a compound of the present invention with a pharmaceutically acceptable excipient
  • theie is prov ided a method of inhibiting a hgand fiom binding to a cognate RNA, comprising contacting undei binding conditions said hgand, a cognate RNA to which said hgand binds undei binding conditions, and a compound accoiding to the present invention, so as to permit inhibition of binding of said hgand to said NA by said compound
  • theie is piovided a piocess for the prepaiation of the compounds of the present invention
  • the compounds may be piepaied according to the following reaction schemes General Reaction Scheme
  • Reagents (i) ketone, dioxan, 10% HCI, 95°C; (ii) ketone, HCl(g), neat; ketone, CH A, p- TSA, 40°C; (iii) ketone, CHCl 3 , -TSA,
  • the compounds may also be prepared according to the following reaction schemes.
  • Reagents (i) ketone, dioxan, 10% HCI, 95"C; (ii) ketone, I-ICl(g), neat; (iii) ketone, CHC1 , -TSA, 40"C; (iv) ketone, FI;0, c.HCl, 55°C.
  • Reagents (i) Acetic anhydride, pyridine, rt; (ii) sodium hydride, R X, THF; (iii) Potassium carbonate, methanol, rt.
  • Reagents (i) ArBr, P(o-tolvl) 3 , Pd(OAc);, NEt 3 , MeCN, 90 "C, N : ; (ii) NaOMe. MeOH, RT.
  • A, B, Y 1 , Y 2 , Y J , Z 1 , Z 2 , Z J and Z 4 may be the same or different and are selected from H, halogen, C 1 - 1 ; alkyl, C;. ⁇ ; alkenyl, C 3 - 1 ; aryl, CM S aralkyl, SR " , OR J ,
  • each R 2 , R 3 and R 4 is the same or different and is selected from H, C f .r > alkyl, C;. f) alkenyl, C 3 .1t) aryl, C ⁇ - 6 alkoyl, C 3 -. 1 i) aryloyl and C4-1; aralkyl, and each R ⁇ R° and R is the same or different and is selected from I , Ci-s, alkyl, C;.(, alkenyl.
  • the medicament employed in the present invention can be administered by oral or p ⁇ renteral routes, including intravenous, intramuscular, intraper ⁇ neal. .subcutaneous. transde ⁇ nal, airway (aerosol), rectal, vaginal and topical (including buccal and sublingual) administration.
  • oral or p ⁇ renteral routes including intravenous, intramuscular, intraper ⁇ neal. .subcutaneous. transde ⁇ nal, airway (aerosol), rectal, vaginal and topical (including buccal and sublingual) administration.
  • the compounds of the invention will generally be provided in the form of tablets or capsules, as a powder or granules, or as an aqueous solution or suspension.
  • Tablets for oral use may include the active ingredients mixed with pham- aceutically acceptable excipients such as inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavouring agents, colouring agents and preservatives.
  • Suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while com starch and alginic acid are suitable disintegrating agents.
  • Binding agents may include starch and gelatin, while the lubricating agent, if present, will generally be magnesium stearate, stearic acid or talc.
  • the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate, to delay absorption in the gastrointestinal tract.
  • Capsules for oral use include hard gelatin capsules in which the active ingredient is mixed with a solid diluent, and soft gelatin capsules wherein the active ingredients is mixed with water or an oil such as peanut oil, liquid paraffin or olive oil.
  • Fonnulations for rectal administration may be presented as a suppository with a suitable base comprising for example cocoa butter or a salicylate.
  • Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray fonnulations containing in addition to the active ingredient such earners as are known in the art to be appropriate.
  • the compounds of the invention will generally be provided in sterile aqueous solutions or suspensions. buffered to an appropriate pH and isotonicity. Suitable aqueous vehicles include Ringer ' s solution and isotonic sodium chloride. Aqueous suspensions according to the invention i
  • suspending agents such as cellulose derivatives, sodium alginate, polyvinyl- pyrrolidone and gum tragacanth, and a wetting agent such as lecithin.
  • Suitable preservatives for aqueous suspensions include ethyl and n-propyl p-hydroxybenzoate.
  • the compounds of the invention may also be presented as liposome formulations.
  • a suitable dose will be in the range of 0.01 to 100 mg per kilogram body weight of the recipient per day, preferably in the range of 0.2 to 10 mg per kilogram body weight per day.
  • the desired dose is preferably presented once daily, but may be dosed as two, three, four, five or six or more sub-doses administered at appropriate intervals throughout the day. These sub-doses may be administered in unit dosage forms, for example, containing 10 to 1500 mg, preferably 20 to 1000 mg, and most preferably 50 to 700 mg of active ingredient per unit dosage form.
  • HPLC HP 1 100 Column: ABZ-. 3.3em*4,6mmD
  • Solvents A - Water - 0, 1 " -, Formic acid - I Ommoi ammonium acetate B - 95% Acetonitrile/water + 0.05" o formic acid
  • the bis-CBZ compound was hydrogenated in monomethoxyethyleneglycol/formic acid over 10%>Pd/C for 19h at RT.
  • the reaction mixhire was filtered through silica gel and concentrated in vacuo. Purification was by semi-preparative HPLC gave the desired product.
  • Compounds of the present invention showed inhibitory activities in in vitro translation assays utilizing E, coli extracts.
  • the plasmid pBestLuc which contains the gene for firefly luciferase downstream of an E. coli promoter and a ribosome binding site is used as a template.
  • the activity of the firefly luciferase enzyme results in a strong luminescent signal.
  • the luminescence generated is a direct measurement of protein expression and of translation efficiency.
  • Translation reactions in the presence of compound are started by mixing a translation premix that contains Mg" , plasmid template, amino acids, nucleotidetriphosphates, phosphocreatine, creatine phosphokinase and folinic acid with an S30 extract that contains RNA polymerase, ribosomes and translation factors (prepared from E.coli MR ⁇ 600 cells) followed by incubation at 37°C.
  • the activity of the translated luciferase protein is measured by adding an aliquot of the translation reaction to the non-luminescent substrate luciferin and the luminescence measured. The luminescence is quantified in a luminescence plate reader (Wallac Victor). Compounds were assayed 3-5 times over a range of concentrations and an IC 0 calculated.
  • Example 3 70%
  • Example 4 - 48%
  • the compounds of the present invention therefore show good in vitro biological activity.
  • the in vivo therapeutic efficacy of the compounds of the invention is measured by intramuscular injection to mice experimentally infected with a pathogenic gram positive or gram negative bacterium (e.g. methicillin-resistant Staphylococcus aureus (MRSA). Clostridium difficile. Klebsiella pneumoniae. Eschericia coli. l laemophilus ⁇ nfiuenzue. etc. ).
  • MRSA strain A2 " 223 can be used.
  • MRSA strain A27223 is prepared for experimental infection by growth on two large Brain Heart Infusion Agar plates. On each plate, 0.5 ml of frozen stock culture is plated out. Plates are then incubated for 18 hours at 30°C.
  • mice The next day each plate is washed with 20 ml of Brain Heart Infusion Broth and then pooled together. A microscopic direct count of microorganisms is done using a 1 : 1000 dilution of plate wash. After a direct count is obtained, the number of organisms per milliliter is calculated. The count is adjusted to the desired amount of inoculum by diluting in 4% hog mucin.
  • the desired challenge (amount of organisms given to mice) is 2.4 x 10 8 cfu/0.5 ml/mouse for MRSA strain A27223.
  • the mice are infected intraperitoneally with 0.5 ml of challenge. Ten non-treated infected mice are used as controls. Mice used are adult male ICR mice. The average weight of the animals should range from 20 to about 26 grams.
  • a PDjo (protective dose-50, the dose of drug given, which protects 50%> mice from mortality) runs for 5 days. During this time, mortality of mice is checked every day and deaths are recorded. The cumulative mortality at each dose level is used to calculate a PD 50 value for each compound. Surviving mice are sacrificed at the end of day 5 by CO 3 inhalation. The actual calculation of PD 5 0 is perfo ⁇ ned with a computer program using the Spearman- Karber procedure.
  • a compound according to the invention is effective for the treatment of bacterial infection if it has a PD-n of about 100 mg kg or less.
  • Topical administration assav Compounds according to the invention can be used for the treatment of dermatological infections. It is often desirable to treat cerost ⁇ cal infections with topically applied antibiotics because such application allows direct delivery of the antibiotic agent to the site of infection, and because it permits delivery in higher concentrations at the affected site than would, for example, systemic administration.
  • mice with experimentally induced skin infections are used. Any recognised experimental model of skin infection is acceptable.
  • One of the most commonly used models is Staphyloccocus aureus infection of mice. Infection may be induced by intracutaneous injection of S. aureus bacteria or topical application of bacteria to experimentally wounded or inflamed skin of immunosuppressed mice. Immunosuppression is commonly achieved by, for example, cyclophosphamide treatment of the animals (usually 2 mg/mouse for 5 days prior to infection).
  • Experimental dermatological infection of mice is described in, for example Abe et al., 1992, J. Dermatol. Sci. 4: 42-48; Akiyama et al., 1996, J. Dermatol, Sci. 11 : 234-238; and Molne & Tarkowski, 2000, J. Invest. Dermatol. 1 14: 1 120- 1 125, which are incorporated herein by reference.
  • a compound of the invention is assayed by topical application of the compound in a preparation , e.g. an ointment, appropriate for such topical application.
  • a preparation e.g. an ointment
  • suitable vehicles for topical administration can be selected from suitable vehicles for topical administration.
  • a range of differing concentrations of the compound being assayed is applied to the infected area of different infected mice.
  • concentration of the compound to use in such assays or in treatment of infection in general can be estimated by considering the amount of the compound effective to kill or inhibit the pathogen in culture.
  • concentration of compound in a topical preparation will range from about 0.0001% to about 10%. (wwv) of the compound being assayed, preferably about 0.001 % to about 2%. about 0.0 1% to about IV about 0.001% to about 0. 1%. or 0.0 ( 11% to 0-01 ",,-.
  • the preparation can also be applied with varying frequency, ranging from example, from once an hour to once a day.
  • the effectiveness of the topical treatment can be evaluated in several ways. For example, visual inspection of the infected area can be used. If the area of infection, as evidenced by inflammation or skin lesions, either does not increase in size or decreases in size relative to its size at the time of the first treatment, the agent can be judged effective. Alternatively, or in addition, microscopic examination of samples taken from infected skin can be conducted, and the degree of inflammation or infiltration of the infection into the tissue can be evaluated by a professional skilled in the interpretation of histological specimens, e.g. a pathologist, dermatologist or medical technologist.
  • Differences are determined by examining samples taken from untreated, infected control animals.
  • two areas of infection can be established on, for example, opposite sides of the back of the same mouse. In that instance, only one side is treated with the compound being assayed and the untreated site serves as a control. If the inflammation (judged by the presence of inflammatory cells) or infiltration of the pathogen does not increase following treatment, the agent is judged effective as a bacteriostatic agent effective against that pathogen in vivo. If the inflammation or infiltration decrease (e.g.
  • Additional controls can include skin samples taken from infected animals treated with other drugs of known effectiveness against the experimental pathogen (e.g. vancomycin for S. aureus). Such controls can provide a measure of the effectiveness of the compound relative to that of standard antibiotics.

Abstract

The present invention provides compounds, their preparation and their use in therapy, particularly the treatment of bacterial infection, of the formula (I) wherein p is selected from 4, 5 and 6; q is selected from 3, 4 and 5; each T is the same or different and is selected from -X- and -CR1 2-; each R1 may be the same or different and is selected from H, C¿1-12? alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl and C4-18 aralkyl; each X may be the same or different and is selected from -CR?1 ¿2-, O, S, and NR' where each R' may be the same or different and is selected from H, C1-6 alkyl, C2-6 alkenyl, C2-12 alkynyl, C3-10 aryl, C4-12 aralkyl, C(O)R'', SO2R'' and C(O)NR''2 wherein R'' is selected from C1-6 alkyl, C2-6 alkenyl, C3-10 aryl, C4-12 aralkyl; A, B, Y?1, Y2, Z1, Z2, Z3 and Z4¿ may be the same or different and are selected from H, halogen, C¿1-12? alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl, C4-18 aralkyl, SR?2, OR3, NR4R5, CO¿2R6 and CONR7 2 wherein each R?2, R3 and R4¿ is the same or different and is selected from H, C¿1-12? alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl, C1-12 alkoyl, C3-12 aryloyl and C4-18 aralkyl, and each R?5, R6 and R7¿ is the same or different and is selected from H, C¿1-12? alkyl, C2-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl, C4-18 aralkyl; provided that at least one of A and B is OR?3¿, and pharmaceutically acceptable derivatives thereof.

Description

FLA VAN COMPOUNDS, THEIR PREPARATION AND THEIR USE IN THERAPY
The present invention relates to a class of flavan (2,3-dihydro-2-phenylbenzopyran) chemical compounds, their preparation and their use in therapy, particularly in the treatment of bacterial infection.
Although many pharmaceutical compounds and compositions are available for the treatment of bacterial infections, there remains a continuing need for improved treatments.
Flavan derivatives have previously been reported as the condensation products of phenols and ketones. Baker et al. (J. Chem. Soc, 1957, 3060-3062) identify the condensation product of m-cresol and cyclohexanone as cyclohexanespiro-4-(2,-hydroxy-7, 4'-dimethyl- 2, 3-tetramethyleneflavan. Livant et al. (J. Org. Chem., 1997, 62, 737-742) report analogous condensation products arising from the condensation of resorcinol and cyclopentanone (to give 2',4'J-trihydroxy-233-propanoflavan-4-spirocyclopentane) and the condensation of resorcinol and cyclohexanone (to give 2',4,J-trihydroxy-2,3-butano- flavan-4-spirocyclohexane).
2,3-Butano-flavan-4-spirocyclohexane derivatives have also been reported by Sumitomo as useful in radiation-sensitive positive resist compositions (EP-A-0458988 and EP-A- 0525185).
The present inventors have identified a class of flavan derivatives, which are useful in the treatment of bacterial infection.
According to the present invention there is provided a compound for use in therapy, wherein the compound is of formula
Figure imgf000003_0001
wherein p is selected from 4, 5 and 6; q is selected from 3, 4 and 5; each T is the same or different and is selected from -X- and -CR 2-; each R1 may be the same or different and is selected from H, Cι_ι alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl and C4-18 aralkyl; each X may be the same or different and is selected from -CR , O, S and NR' where each R' may be the same or different and is selected from H, Ci-g all y 1, C2-6 alkenyl, C2-12 alkynyl, C3-10 aryl, C4-12 aralkyl, C(O)R", SO2R" and C(0)NR"2 wherein R" is selected from Cι-6 alkyl , C -6 alkenyl, C3-10 aryl, C4-12 aralkyl; A, B, Y1 , Y2, YJ, Z 7?, ZJ and Z4 may be the same or different and are selected from H, halogen, C1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl, C4-1;; aralkyl, SR2, OR3, NR R5, CO2R6 and CONR7 2 wherein each R2, R3 and R is the same or different and is selected from H, C ι-ι alkyl, C2-12 alkenyl, .12 alkynyl,
Figure imgf000003_0002
aryl,
C ι-12 alkoyl, €3-12 aryloyl and C4-18 aralkyl, and each R",R(l and R7 is the same or different and is selected from H. C1-12 alkyl, C2.12 alkenyl, C2-12 alkynyl, C3-12 aryl, C4-1 X aralkyl; provided that at least one of A and B is OR", and pharmaceutically acceptable derivatives thereof.
According to a further aspect of the present invention there is provided a compound of the formula
Figure imgf000004_0001
wherein p is selected from 4, 5 and 6; q is selected from 3, 4 and 5; each T is the same or different and is selected from -X- and -CR'2-; each R1 may be the same or different and is selected from H, Cι-i2 alkyl, C _ι alkenyl, C2-12 alkynyl, C3-12 aryl and C4-18 aralkyl; each X may be the same or different and is selected from -CR'2-, O, S and NR' where each R' may be the same or different and is selected from H, -g alkyl, C2-6 alkenyl, C2-12 alkynyl, C3-H) aryl, C4-12 aralkyl, C(0)R' \ SO2R" and C(0)NR"2 wherein R" is selected from Cι-6 alkyl, C2-6 alkenyl, C3-10 aryl, C4-12 aralkyl; A, B, Y , Y", Y"\ Z1, Z~, ZJ and Z may be the same or different and are selected from H, halogen, C 1-1 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl, C4-18 aralkyl, SR:, OR3, NR R5, CO2R6 and CONR7 2 wherein each R2, R3 and R4 is the same or different and is selected from H. C1-1 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl,
Cι-12 alkoyl, C3-.ι2 aryloyl and C is aralkyl, and each R\R6 and R7 is the same or different and is selected from H. C1-12 alkyl. C2-12 alkenyl. C2-.2 alkynyl. C3-12 aryl. C4.1S aralkyl; provided that at least one of A and B is OR\ and pharmaceutically acceptable derivatives thereof, provided that at least one of A. B. Y1. Y" and Y3 is not H, CH;,. C;H? or OH ( preferably at least one of A. B, Y 1. Y~ and Y~ is not H. OH. halogen, unsubstituted C 1-4 alkyl or halogen ≤ubstimred C1.4 alkyH; and 'or provided that both Tp is not -(CH;);- and Tq is not -( CH;)4- tind the compound is not 2'.4'J-trihydro\y-2.3-piOpanofla\-an-4-spιiOcyclopentane.. As used herein, the term "alkyl" means an optionally substituted branched or unbranched, cyclic or acyclic, hydrocarbyl radical. Where acyclic, the alkyl group is preferably a C1-12, more preferably C]-r„ more preferably C1-4 chain. Where cyclic, the alkyl group is preferably a C3-i2, more preferably Cs-i and more preferably comprises a C5, C6 or Cγ ring. As used herein, the term "alkenyl" means an optionally substituted branched or unbranched, cyclic or acyclic, hydrocarbyl radical comprising at least one double bond. Where acyclic, the alkenyl group is preferably a C2-12 - more preferably C2-6, more preferably C1-4 chain. Where cyclic, the alkenyl group is preferably a C3-12, more preferably C5-10 and more preferably comprises a C5, C6 or C7 ring.
As used herein, the term "alkynyl" means an optionally substituted, branched or unbranched, cyclic or acyclic, hydrocarbyl radical comprising at least one triple bond. Where acyclic, the alkynyl group is preferably a C2-12, more preferably C2-4 chain. Where cyclic, the alkynyl group is preferably a C4-12, more preferably C6-ιo ring.
As used herein, the term "aryl" means an optionally substituted C3-12, aromatic group, such as phenyl or naphthyl, or a heteroaromatic group containing one or more, preferably one, heteroatom, such as pyridyl, pyrrolyl, furanyl, thienyl. Further examples of aromatic heterocyclic groups include monocyclic groups such as azecinyl, azetyl, azoninyl, imidazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridinyl, pyri idinyl, pyrrolyl, sym~ triazinyl, triazolyl, furanyl, oxoninyl, pyryliumyl, thioninyl, thiophenyl, thiopyryliumyl, selenophenyl, fiirazanyl, isothiazolyl, isoxazolyl, methylthiabenzenyl, oxazinyl, oxazolyl, thiazinyl, thiazolyl; bicyclic groups such as benzimidazolyl, cinnolinyl, indazolyl, indolyl, indolinyl, indolizinyl, isoindolyl, isoindolinyl, isoquinolinyl, naphthyridinyl, phthalazinyl, pteridinvl, purinvl, pyrrolizinyl, quinazolinyl. quinolinyl, quinolizinyl, quinoliziniumyl, quinoxalinyl, benzofliranyl, benzopyryliumyl, chromanyl, chromenyl. chromonyl, coumarinyl. isobenzofuranyl, isochromanyl. isochromenyl, benzothiophenyl, arsindolyl, arsinolinyl, isoarsindolyl, isoarsinolinyl. isophosphindolyl, isophosphinolinyl. phosphindolyl. phosphinolinyl; polycyclic groups such as acridinyl, carbolinyl. carbazolyl. phenanthridinyl. phenazinyl. periniidinvl. phenanthrolinvl. xanthenyl. naphthothiophenyl. phenoxathiiin l. thianthrenyl. acridarsinyl. arsanthπdinyl. phenarsazinyl. phenothiazinyl. phenoxazinyl. As used herein, the term "aralkyl" means an optionally substituted branched or unbranched cyclic or acylic C4_i8 group comprising an alkyl group and an aryl group (for example, benzyl). An aralkyl group may be bonded via the alkyl or aryl group.
As used herein, the term "heterocyclic group" means an optionally substituted C3-12 cyclic group containing at least one, preferably one to three, heteroatoms. Examples include monocyclic groups such as azecinyl, azetyl, azoninyl, imidazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, sym-triazinyl, triazolyl, furanyl, oxoninyl, pyryliumyl, thioninyl, thiophenyl, thiopyryliumyl, selenophenyl, furazanyl, isothiazolyl, isoxazolyl, methylthiabenzenyl, oxazinyl, oxazolyl, thiazinyl, thiazolyl; bicyclic groups such as benzimidazolyl, cinnolinyl, indazolyl, indolyl, indolinyl, indolizinyl, isoindolyl, isoindolinyl, isoquinolinyl, naphthyridinyl, phthalazinyl, pteridinyl, purinyl, pyrrolizinyl, quinazolinyl, quinolinyl, quinolizinyl, quinoliziniumyl, quinoxalinyl, benzofuranyl, benzopyryliumyl, chromanyl, chromenyl, chromonyl, coumarinyl, isobenzofliranyl, isochromanyl, isochromenyl, benzothiophenyl, arsindolyl, arsinolinyl, isoarsindolyl, isoarsinolinyl, isophosphindolyl, isophosphinolinyl, phosphindolyl, phosphinolinyl; polycyclic groups such as acridinyl, carbolinyl, carbazolyl, phenanthridinyl, phenazinyl, perimidinyl, phenanthrolinyl, xanthenyl, naphthothiophenyl, phenoxathiinyl, thianthrenyl, acridarsinyl, arsanthridinyl, phenarsazinyl, phenothiazinyl, phenoxazinyl. Further examples of non-aromatic heterocyclic groups include monocyclic groups such as azaundecinyl, azepinyl, azetidinyl, aziridinyl, azocinyl, imidazolidinyl, imidazolinyl, piperazinyl, piperidinyl, pyrazolidinyl, pyrazolinyl, pyiTolidinyl, pyrrolinyl, quinuclidinyl, dihydropyranyl, dioxanyl, ethylene oxidyl, oxepinyl, oxetanyl, oxocinyl. pyranyl, pyronyl, tetrahydrofuranyl. ethylene sulphidyl. thiepinyl, thietanyl. thiocinyl, moipholinyl, oxazolidinedionyl. oxazolidonyl. propane soltonyl, thiazolidinyl, thiomorpholinyl. thiozanyl.
As used herein the tern, alkoyl means alkyl-CO- w herein alkyl is defined above. The term arvlovl means arvl-CO- wherein arvl is defined above. The alkyl, alkenyl, alkynyl, aryl, aralkyl and heterocyclic groups may be substituted or unsubstituted. Where substituted, there are preferably one to three substituents, more preferably one substituent. Substituents may include halogen atoms and halogen containing groups such as haloalkyl (e.g. trifluoromethyl); oxygen containing groups such as alcohols (e.g. hydroxy, hydroxyalkyl, aryl(hydroxy)alkyl), ethers (e.g. alkoxy, alkoxyalkyl, aryloxyalkyl), aldehydes (e.g. carboxaldehyde), ketones (e.g. alkylcarbonyl, alkylcarbonylalkyl, arylcarbonyl, arylalkylcarbonyl, arylcarbonylalkyl), acids (e.g. carboxy, carboxyalkyl), acid derivatives such as esters (e.g. alkoxycarbonyl, aryloxycarbonyl, aralkyloxycarbonyl, alkoxycarbonylalkyl, alkylcarbonylyoxy, alkylcarbonylyoxyalkyl) and amides (e.g. aminocarboαyl, mono- or dialkylaminocarbonyl, aminocarbonylalkyl, mono- or dialkylaminocarbonylalkyl, arylaminocarbonyl); and carbamates (e.g. alkoxycarbonylamino, aryloxycarbonylamino, aralkyloxycarbonylamino, amino-carbonyloxy, mono- or dϊalkylaminocarbonyloxy, arylaminocarbonyloxy), and ureas (e.g. mono- or dialkylaminocarbonylamino or arylaminocarbonylamino); nitrogen containing groups such as amines (e.g. amino, mono- or dialkylamino, amino substituted with one or two groups selected from alkyl, alkenyl, aryl ad aralkyl groups, aminoalkyl, mono- or dialkylaminoalkyl), azides, nitriles (e.g. cyano, cyanoalkyl), nitro; sulfur containing groups such as thiols, thioethers, sulfoxides, and sulfones (e.g. alkylthio, alkylsulfinyl, alkylsulfonyl, alkylthioalkyl, alkylsulfmylalkyl, alkylsulfonylalkyl, arylthio, arylsulfinvl, arylsulfonyl, arylthioalkyl, arylsulfmvlalkyl, arylsulfonylalkyl); carbon containing groups (e.g. alkyl, alkenyl, alkynyl, aryl and aralkyl); and heterocyclic groups containing one or more, preferably one, heteroatom (e.g. thienyl, furanyl. pynolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, oxadiazolyl, thiadiazolyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, tetrahydro furanyl, pyranyl, pyronyl, pyridyl, pyrazinyl, pyridazinyl. piperidyl, piperazinyl, moφholinyl, thianaphthyl, benzofuranyl, isobenzofliranyl. indolyl. oxyindolyl, isoindolyl. indazolyl, indolinyl. 7-azaindolyl, benzopyranyl, coumarinyl. isocoumarinyl. quinolinyl. isoquinolinyl. naphthridinyl. cinnolinyl, quinazolinyl. pyridopyridyl. benzoxaziny , quinυ alinyl. chromenyl. chromanyl. isochromanyl, phthalazinyl and carbolinyl.). Further examples of aromatic heterocyclic groups include monocyclic groups such as uzecinyl. uzetyl. azonϊn l. imidazolyl. pyrazinyl. razolyl. pyridazinyl. pyπdinyi. pyπmϊdinyi. pyrrolyl. sym-triazinyl. triazolyl. furanyl. oxonπn l. pyryliumyl. thioninyl. thiophenyl. thiopyiyhumyl, selenophenyl fuiazanyl isothiazolyl, isoxazolyl methylthiabcnzenvl oxazinyl, oxazolyl thiazinyl thiazolyl, bicychc gioups such as benzimidazolyl, cmnolmyl indazolyl, indolyl indolinyl, mdohzinyl, isoindolyl, lsomdohnyl,
Figure imgf000008_0001
l tetrahydroisoquinolmy, naphthyiidmyl, phthalazinyl, pteiidmyl, puiinyl, pynohzinyl, quinazolinyl, quinolinyl qumolizmyl, qumoliziniumyl, qumoxalinyl, benzoturanyl, benzopyiyhumyl, chromanyl, chromenyl, cluomonyl, coumaπnyl, isobenzofuianyl, lsochromanyl, isochromenyl, benzothiophenyl, arsindolyl, aisinolinyl, isoarsindolyl, lsoarsmolinyl, isophosphindolyl, lsophosphinohnyl, phosphindolyl, phosphinolmyl, polycychc groups such as acπdinyl, caibohnyl, carbazolyl, phenanthπdmyl, phenazinyl, peiimidinyl, phenanthiolinyl, xanthenyl, naphthothiophenyl, phenoxathimyl, thianthienyl, acndarsmyl, aisanthπdmyl, phenaisazmyl, phenothiazmyl, phenoxazmyl Further examples of non-aromatic heterocyclic groups include monocyclic gioups such as azaundecinyl azepmyl, azetidinyl, azind yl, azocinyl, imidazohdinyl, lmidazohnyl, pipeiazinyl, pipe dmyl, pyrazohdinyl, pyrazolinyl, pyirohdmyl, pyiTolmyl, qumuchdinyl, dihydropyranyl, dioxanyl, ethylene oxidyl, oxepinyl, oxetanyl, oxocmyl, pvranyl, pyronyl, tetrahydrofuranyl, ethylene sulphidyl, thiepmyl, thietanyl, thiocmyl, moφhohnyl oxazohdmedionyl, oxazohdonyl, piopane soltonyl, thiazohdmyl, thiomorphohnyl, thiozanyl A group may contain more than one substituent Preferably the overall size of a gioup including substituents is less than 30, preferably less than 24, preferably less than 18, pteferably less than 12 carbon atoms
As used herein the term "halogen" means a fluorine, chlorine, biomme oi iodine tadical preferably a fluorine chlorine or bromine radical, preferably a fluonne or chlorine tadical
As used herein, "dialkyl ammo gioups ' include cyclic amines of the toπnulα -NRR wheie R and R are linked to form a cyclic group pieteiablv containing 5 to 7 moie pt efeiably 6 nng atoms and piefeiablv containing one to thiee heteioatoms Examples of such cvchc g' oup include pipeπdinvl morphohn\ l and piperaz v l
Figure imgf000008_0002
groups
α phaimaLeuticallv acceptable e is meant anv phαπnaceuticallv acceptable salt addition compound oi anv othe- compound w hich upon admmisti ation to α leupient capable of
Figure imgf000008_0003
id g idiiectlv oi -nϋιi , α compound of the ρι e->enι e .-ion oi α pharmaceutically acceptable metabolite. By "'pharmaceutically acceptable metabolite" is meant a metabolite or residue of a compound of the present invention which gives rise to a biological activity exhibited by the present compounds.
It will be appreciated that the compounds of the present invention may exist in a number of diastereomeric and enantiomeric forms. The present invention encompasses pure diastereomers and enantiomers as well as mixtures (including racemic mixtures) of diastereomers and enantiomers.
In the compounds according to the present invention Tp represents a divalent group comprising groups T where p is selected from the integers 4, 5 and 6, such that the ring comprising -Tp- is a 5-, 6- or 7-membered ring, respectively. Preferably, p is 4 or 5.
Tq represents a divalent group comprising groups T where q is selected from the integers 3, 4 or 5, such that the ring comprising — Tt]-is a 5-, 6- or 7-membered ring, respectively. Preferably q is 3 or 4.
Preferably, in Tp one T is X and the remaining T are -CR ;
Preferably, in Tq one T is X and the remaining T are -CR1;-
Preferably, the compounds of the present invention are of the formula
Figure imgf000010_0001
wherein each U may be the same or different and is selected from
-X-CR1 .-
-CRS-X-CR - and
-CR -X-CR1. -CR1
The groups A, B, Y , Y~, YJ, Z , Z", ZJ, Z , X and R are as previously defined.
It will be appreciated that when U is not symmetrical (i.e. when U is -X-CR 2- or -CR 2-X- CR'i-CR1; it may be bonded to the molecule in two orientations (namely -X-CR'2- or -CR'α-X- and -CR^-X-CR^-CR^- or -CR^-X-CR^-CR1;). Both orientations are encompassed by the present invention.
Preferably, U is -CR -X-CR1-.
In one preferred group of compounds each X is selected from 0, S, NR' where each R' may be the same or different and is selected from H. Ct.i, alkyl. C;.fl alkenyl. C;-i.> aryl, C4-1; aralkyl. C(0)R". SO;R" and C(0)R"; wherein R" may be same or different and is selected from . Ct.., alkvl. C;.,, alkenyl. C.;-ι,> arvl. C-i-i; aralkyl. In another preferred group of compounds each X is -CR1;-.
Each group R is selected independently from H, C M; alkyl, C2-12 alkenyl, C3-12 aryl and C4-1 aralkyl. Preferably, any two R1 in a geminal relationship (i.e. each R group bonded 5 to the same carbon atom) are the same.
Preferably, each R is hydrogen.
Preferably, each R' is hydrogen or C|-6 alkyl. 0 Preferably, each R" is Cι-6 alkyl.
A, B, Y1, Y2, YJ, Z1, Z2, Z3 and Z may be the same or different and are selected from H, halogen, Ci-i; alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl, C4-1S aralkyl, SR~, ORJ, NR R",
(5 CO2R6 and CONR7 2; wherein R2, RJ and R are the same or different and are selected from H, Cι-12 preferably Cι-6 alkyl, C2-12 preferably C;-6 alkenyl, C2-12 preferably C2-6 alkynyl, C3-12 preferably C3-10 aryl, Ci-1 preferably Ci-r, alkoyl, C3-12 preferably C3_ιo aryloyl and C4-18 preferably C 2 aralkyl; and each R", R6 and R7 is the same or different and is selected from H, C1-12 preferably Cι-6 alkyl, C;-i2 preferably C2-6 alkenyl, C2-12 preferably
10 C2-6 alkynyl, C3_ι; preferably C3-10 aryl and C4-18 preferably C4-12 aralkyl. When A, B, Y1, Y", Y3, Z1, Z , ZJ and Z4 are selected from C M; alkyl, C2-12 alkenyl, C3-12 aryl and C4- is aralkyl groups the groups may optionally be substituted as defined for alkyl, alkenyl, alkynyl, aryl and aralkyl groups above. In one aspect of the invention, when A, B, Y , Y", YJ, Z1, Z~, ZJ and Z4 are selected from C2-12 alkenyl, C3-12 aryl and C4-18 aralkyl groups,
25 preferred substituents are selected from halogen atoms. In a further aspect of the invention, when A, B, Y , Y~, Y". Z , Z~, ZJ and Z are selected from C1-1; alkyl groups, preferred substituents are selected from halogen atoms, SR'', OR1 and NR1 Rπ, wherein . R . R and R are the same or different and are selected from H and Ci-., alkyl. In a further aspect of the invention, when A. B. Y . Y~. Y". Z , Z", Z" and Z are selected from
30 C M; alkyl. C;-ι; alkenyl C;.ι; alkynyl. C.;-ι; aryl and C4.IS aralkyl groups one to three substituents (preferably one substituent) is selected from -COR . CO;lV" and CO"N;R1 R wherein R 1'". RJ\R "' and Rι: are selected from I I. C ; alkyl. C;-t; alkenyl. C;-ι; alkynyl. C3-1; aryl and C4-1 aralkyl groups which may be substituted as defined for alkyl, alkenyl, alkynyl, aryl and aralkyl groups above. In a further aspect of the invention, when A, B, Y1, YJ, Y", Z , Z", ZJ and Z are selected from C alkyl, preferred substituents are selected from amino, alkylamino, dialkylamino and heterocyclic groups, preferably, a group of the formula -NR, 0RH where R1 and R1 1 are selected from H, C M; alkyl, C2.12 alkenyl, C3-12 aryl and C4-18 aralkyl, which may be unsubstituted or substituted as defined for alkyl, alkenyl, aryl and aralkyl groups above; preferably R and R are linked to form a cyclic group, preferably containing 5 to 7, more preferably six, ring atoms and preferably containing one to three heteroatoms, which may be unsubstituted or substituted as defined for heterocyclic groups above.
In a preferred embodiment , Y , and YJ are selected from H, ORJ, C ; alkyl and halogen. More preferably, Y1 and YJ are selected from H, OFI and halogen, more preferably H and halogen.
In a further preferred embodiment, Y1 or YJ is C1-12 alkyl which may be unsubstituted or substituted as defined for alkyl groups above. Preferably, Y or YJ is C1-12 alkyl substituted with a group selected from amino, alkylamino, dialkylamino and heterocyclic groups. Preferably, Y1 or YJ is a group of the formula
-CH-NRl ϋRπ
where R10 and R11 are selected from H. C ; alkyl, C2-1; alkenyl, C3-1; aryl and C4-18 aralkyl, which may be unsubstituted or substituted as defined for alkyl, alkenyl, aryl and aralkyl groups above. In a preferred embodiment, R } and R1 are linked to form a cyclic group, preferably containing 5 to 7. more preferably six, ring atoms and preferably containing one to three heteroatoms. Such heterocyclic groups may be unsubstituted or substituted as defined for heterocyclic groups above. Examples of such heterocyclic groups include piperidinyl. piperazinyl and morpholinyl.
In a further preferred embodiment Y or Y" is CO;R" w herein A is selected from H. C M; alkyl. C;-ι; alkenyl. C;.j; alkynyl. C.M; aryl. C.j-.,s' aralkyl an heterocyclic group w hich may be unsubstituted or substituted as defined for alkyl, alkenyl, alkynyl, aryl, aralkyl and heterocyclic groups above.
In a further preferred embodiment, Y or Y" is CONR ; wherein each R is selected from H, C M; alkyl. C;.ι; alkenyl, C;-ι; alkynyl, C;_ι; aryl and CM S aralkyl, which may be unsubstituted or substituted as defined for alkyl, alkenyl, alkynyl, aryl and aralkyl groups above. In a preferred embodiment two R groups may be linked to form a heterocyclic group, preferably containing 5 to 7, more preferably 6 ring atoms, and preferably containing one to three heteroatoms. Such heterocyclic groups may be unsubstituted or substituted as defined for heterocyclic groups above. Examples of such heterocyclic groups include piperidinyl, piperazinyl, morpholinyl, 1,2,3,4-tetrahydroisoquinolinyl and 1 ,2,3,4- tetrahydro-6,7-isoquinolinediolyl.
Preferably Y2 is selected from H, C M alkyl, OR3, CO;R6 and CONR7;. More preferably Y2 is OR"1 in which RJ is preferably H, CM alkyl, C M; alkoyl or C4-18 aralkyl, more preferably FI, Cι-6 alkyl or Ci-g alkoyl.
Preferably, Z , ZJ and Z are selected from H, ORJ and halogen, more preferably H, OH and halogen.
Preferably Z2 is selected from H, C 2 alkyl, OR3, CO;Rδ and CONR7;. More preferably Z~ is ORJ, in which RJ is preferably H, C ; alkyl, C M; alkoyl or C4-18 aralkyl, more preferably H, Cι-0 alkyl or Cι-(, alkoyl.
At least one of A and B is OR"3. Preferably, one of A and B is OR" and one of A and B is
It will be appreciated that by free rotation about the bond that joins the phen> 1 group containing A and B to the rest of the molecule, the groups A and B and the groups Y and Y~ are equivalent.
In one embodiment. A or B is OR"* where R"" is preferably H. In a further embodiment, A or B is OR where RJ is selected from C M alkyl, C M; alkenyl, C M; alkynyl, C3-1; aryl, CM; alkoyl, C3-1; aryloyl and C4-18 aralkyl, each of which may be substituted or unsubstituted as defined for alkyl, alkenyl, alkynyl, aryl and aralkyl groups above. In a preferred embodiment A or B is ORJ where R is C;-ι; alkenyl which may be substituted as defined for alkenyl groups above. Preferably, A or B is ORJ where R3 is a group of the forcnula
-CH2CH=CH-T
where T is H, C ; alkyl, C3-12 aryl or C4-12 aralkyl; preferably H or C3-12 aryl (including unsubstituted C3-12 aryl such as phenyl, benzothiazolyl and quinolinyl, and substituted C3-1; aryl such as p-fluorophenoxyphenyl, methylbenzothiazolyl and methylquinolinyl).
In an alternative preferred embodiment A or B is ORJ where RJ is C ; alkyl which may be unsubstituted or substituted as defined for alkyl groups above. Preferably, A or B is OR3 where RJ is C e, alkyl substituted with a group selected from -COR12, CO;R and CONR'V5 where R12, R13, R14 and R15 are selected from H, C 2 alkyl, C;-ι; alkenyl, C2- 1; alkynyl, C3-1; aryl and C4-1 S aralkyl, which may be substituted as defined for alkyl, alkenyl, alkynyl, aryl and aralkyl groups above. Preferably, A or B is OR"1 where RJ is a group of the formula
-CH;-CO-NR14R15
where R14 and R are as defined above. Preferably R14 is H. Alternatively, R14 and R may be linked to form a cyclic group preferably 5 to 7, more preferably six ring atoms and containing one to three heteroatoms. Such heterocyclic groups may be unsubstituted or substituted as defined for heterocyclic groups above. Preferred heterocyclic groups include pipeπdinyl, piperaziny and morpholinyl,
A preferred group of compounds of the present invention comprise compounds of the formula
Figure imgf000015_0001
wherein m and n may be the same or different and are selected from 1, 2 or 3; each R1 and R~ may be the same or different and are selected from H, CM alkyl. CM; alkenyl, C3-1; aryl and C4-]8 aralkyl; A, B, Y , Y~, YJ, Z , Z", ZJ and Z may be the same or different and are selected from H, C M; alkyl, C M; alkenyl, C3-1; aryl, Q-is aralkyl, OR3 and NR4R\; wherein RJ, R and R" are the same or different and are selected from H and Cι-6 alkyl; provided that at least one of A and B is ORJ, and pharmaceutically acceptable derivatives thereof.
In a further aspect of the invention there is provided each of the individual compounds set out in the examples.
The compounds of the present invention may be used in the treatment of bacterial infection of humans, other animals or plants.
According to a further aspect of the present invention there is provided a compound according to the present invention for use in a method of treatment, preferably in the treatment of bactenal infection. According to a further aspect of the present invention there is provided use of a compound according to the present invention in the manufacture of a medicament for the treatment of bacterial infection.
According to a further aspect of the present invention there is provided a method of treatment of bacterial infection comprising administration to a subject in need of such treatment an effective dose of a compound according to the present invention.
Bacterial infections include bacterial infections that can afflict humans, other animals or plants. Bacterial infections include infections by Gram Positive Bacteria including Bacillus cereus, Bacillus anthracis, Clostridium botulinum, Clostridium difficile, Clostridium tetani, Clostridial perfringens, Corynebacteria diphtheriae, Enterococcus (Streptococcus D), Listeria Monocytogenes, Pneumoccoccal Infections (Streptococcus pneumoniae), Staphylococcal Infections and Streptococcal Infections; Gram Negative Bacteria including Bactcroides, Bordetella pertussis, Bmcella, Campylobacter Infections, Enterohemmorrhagic Escherichia coli (EFIEC/E.coli 0157-H7), Enteroinvasive Escherichia coli (EIEC), Enterotoxigenic Escherichia coli (ETEC), Haemophilus influenzae, Helicobacter pylori, Klebsiella pneumonia, Legionella spp., Moraxella catarrhalis, Neisseria gonnorrhoeae, Neissεria meningitidis, Proteus spp., Pseudomonas aeruginosa. Salmonella spp., Shigella spp., Vibrio cholera and Yersinia; Acid Fast Bacteria including Mycobacterium tuberculosis, Mycobacterium avium-intracellulare, Mycobacterium leprae, Atypical Bacteria, Chlamydia, Mycoplasma, Rickettsia, Spirochetes, Treponema pallidum, Bonelia recunentis, Borrelia burgdorfii and Leptospira icterohemorrhagiae; and other miscellaneous bacteria including Actinomyces and Nocardia. Bacterial infections also include bacteria of the following genera of bacteria, which are significant phytopathogens affecting fruit, vegetable and ornamental plant production: Pseudomonas. Xaπtliυmυnas, Envinia, Streptυmyces. Clostridium and Λgrobacrerium.
As used herein, the term "'treatment" includes prophylactic treatment. It is a featui e of the compounds of the present invention that they inhibit the binding of compounds to RNA Foi example the compounds of the present inv ention inhibit the binding of proteins to RNA Accoidinglv the piesent invention furthei piovides use of a compound of the piesent invention to inhibit the binding of a compound, prefeiablv a piotem, to RNA
It is also a featuie of the compounds of the pi esent invention that they inhibit the tianslation of piotems particularly bacterial proteins Accoidinglv, the piesent invention furthei provides use of a compound of the piesent invention to inhibit the translation of pioteins, preferably bacterial proteins
According to a further aspect of the present invention there is provided a pharmaceutical composition comprising a compound of the present invention m combination with a pharmaceutically acceptable excipient
According to a further aspect of the present invention there is provided a method of preparing a pharmaceutical composition comprising the step of combining a compound of the present invention with a pharmaceutically acceptable excipient
According to a furthei aspect of the piesent invention theie is prov ided a method of inhibiting a hgand fiom binding to a cognate RNA, comprising contacting undei binding conditions said hgand, a cognate RNA to which said hgand binds undei binding conditions, and a compound accoiding to the present invention, so as to permit inhibition of binding of said hgand to said NA by said compound
According to a furthei aspect of the piesent invention theie is piovided a piocess for the prepaiation of the compounds of the present invention The compounds may be piepaied according to the following reaction schemes General Reaction Scheme
Figure imgf000018_0001
Reagents: (i) ketone, dioxan, 10% HCI, 95°C; (ii) ketone, HCl(g), neat; ketone, CH A, p- TSA, 40°C; (iii) ketone, CHCl3, -TSA,
For example :-
Figure imgf000018_0002
The compounds may also be prepared according to the following reaction schemes.
Scheme 1
Figure imgf000018_0003
Reagents: (i) ketone, dioxan, 10% HCI, 95"C; (ii) ketone, I-ICl(g), neat; (iii) ketone, CHC1 , -TSA, 40"C; (iv) ketone, FI;0, c.HCl, 55°C.
Scheme 2
Figure imgf000019_0001
Reagents: (i) N-Bromosuccinimide, DMF, rt; (ii) Benzyl bromide, cesium carbonate, DMF, rt; (iii) X = H, tert-Butly lithium then CO; ,g), -78 °C - rt; (iv) Ethyl chlorotbrmate, N-methyl morpholine, THF, RT, aq. NH;,; (v) N-Cyclohexylcarbodiimide, NH -methyl polystyrene, hydroxybenzotriazole, R' R NH, DCM, rt; (vi) H; ,g). 10% Pd/C.
Scheme 3
Figure imgf000019_0002
Reagents: ( ϊ) Acetic acid, ethanol. foπnaldehvde. R I R. . -~-NNH. SO "C. Scheme 4
Figure imgf000020_0001
Reagents: (i) Acetic anhydride, pyridine, rt; (ii) sodium hydride, R X, THF; (iii) Potassium carbonate, methanol, rt.
Scheme 5
Figure imgf000020_0002
Reagents: (i) ArBr, P(o-tolvl)3, Pd(OAc);, NEt3, MeCN, 90 "C, N:; (ii) NaOMe. MeOH, RT.
According to a further aspect of the present invention there is provided a compound per sc of the foπnula
Figure imgf000021_0001
wherein p is selected from 4, 5 and 6; q is selected from 3, 4 and 5; each T is the same or different and is selected from -X- and -CR1;-; each R1 may be the same or different and is selected from FI, Cj-i; alkyl, C;-ι; alkenyl, C3-1; aryl and C -1 S aralkyl; each X may be the same or different and is selected from -CR ;-, 0, S and NR' where each R' may be the same or different and is selected from H, Cι-6 alkyl, C;-(, alkenyl, C3-10 aryl, C4-1; aralkyl, C(0)R'\ SO;R" and C(0)NR"; wherein R" is selected from Cι-6 alkyl, , C;-e alkenyl, C3-10 aryl, C4-1; aralkyl;
A, B, Y1, Y2, YJ, Z1, Z2, ZJ and Z4 may be the same or different and are selected from H, halogen, C1-1; alkyl, C;.ι; alkenyl, C3-1; aryl, CMS aralkyl, SR", ORJ,
NR4R5, CO;R6 and CONR7; wherein each R2, R3 and R4 is the same or different and is selected from H, Cf.r> alkyl, C;.f) alkenyl, C3.1t) aryl, C ι-6 alkoyl, C3-.1i) aryloyl and C4-1; aralkyl, and each R\R° and R is the same or different and is selected from I , Ci-s, alkyl, C;.(, alkenyl. C3-1U aryl, C4-1; aralkyl; provided that at least one of A and B is OR3, provided that both Tp is not -(CH;) - and Tq is not -( CH; - and provided that the compound is not 2\4\7-trihydiOxy-2.3-propunυflavan-4-spirQeyclopentane,
The medicament employed in the present invention can be administered by oral or pαrenteral routes, including intravenous, intramuscular, intraperπυneal. .subcutaneous. transdeπnal, airway (aerosol), rectal, vaginal and topical (including buccal and sublingual) administration.
For oral administration, the compounds of the invention will generally be provided in the form of tablets or capsules, as a powder or granules, or as an aqueous solution or suspension.
Tablets for oral use may include the active ingredients mixed with pham- aceutically acceptable excipients such as inert diluents, disintegrating agents, binding agents, lubricating agents, sweetening agents, flavouring agents, colouring agents and preservatives. Suitable inert diluents include sodium and calcium carbonate, sodium and calcium phosphate, and lactose, while com starch and alginic acid are suitable disintegrating agents. Binding agents may include starch and gelatin, while the lubricating agent, if present, will generally be magnesium stearate, stearic acid or talc. If desired, the tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate, to delay absorption in the gastrointestinal tract.
Capsules for oral use include hard gelatin capsules in which the active ingredient is mixed with a solid diluent, and soft gelatin capsules wherein the active ingredients is mixed with water or an oil such as peanut oil, liquid paraffin or olive oil.
Fonnulations for rectal administration may be presented as a suppository with a suitable base comprising for example cocoa butter or a salicylate.
Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams or spray fonnulations containing in addition to the active ingredient such earners as are known in the art to be appropriate.
For intramuscular, intraperitoneal. subcutaneous and intravenous use. the compounds of the invention will generally be provided in sterile aqueous solutions or suspensions. buffered to an appropriate pH and isotonicity. Suitable aqueous vehicles include Ringer's solution and isotonic sodium chloride. Aqueous suspensions according to the invention i
may include suspending agents such as cellulose derivatives, sodium alginate, polyvinyl- pyrrolidone and gum tragacanth, and a wetting agent such as lecithin. Suitable preservatives for aqueous suspensions include ethyl and n-propyl p-hydroxybenzoate.
The compounds of the invention may also be presented as liposome formulations.
In general a suitable dose will be in the range of 0.01 to 100 mg per kilogram body weight of the recipient per day, preferably in the range of 0.2 to 10 mg per kilogram body weight per day. The desired dose is preferably presented once daily, but may be dosed as two, three, four, five or six or more sub-doses administered at appropriate intervals throughout the day. These sub-doses may be administered in unit dosage forms, for example, containing 10 to 1500 mg, preferably 20 to 1000 mg, and most preferably 50 to 700 mg of active ingredient per unit dosage form.
The invention will now be described with reference to the following Examples. It will be appreciated that what follows is by way of example only and that modifications to detail may be made whilst still falling within the scope of the invention.
EXPERIMENTAL
Chemical Synthesis
The following general techniques were employed.
LCMS Method
Examples 1 -5
HPLC: HP 1 100 Column: ABZ-. 3.3em*4,6mmD
Temperature: 20"C
Solvents: A - Water - 0, 1 " -, Formic acid - I Ommoi ammonium acetate B - 95% Acetonitrile/water + 0.05" o formic acid
Flow rate: lmL/min Gradient: Total time 8 minutes
- 100% A for 0J minutes ramp up to 100% B over 3.5 minutes
- 100% B for 3.5 minutes ramp down to 0% B over 0.3 minutes
Detection: UN detection at 230nm, 254nm and 270nm Mass spec: HP 1 100 MSD Method: Electrospray, +'ve ion
The following examples were synthesized using the procedures outlined in Reaction Scheme 1.
Example 1
Figure imgf000024_0001
Cvclopentanone (l eq.) and resorcinol (3eq.) were heated at 95°C for 24h in a mixture of dioxane/10% aqueous HCI (5/1 , v/v). After cooling to RT, the solvents were evaporated in vacuo and the residue partitioned between DCM and water. The organic layer was dried over M S04 and concentrated in vacuo. The crude product was purified by column chromatography on silica, eluting with mixtures of 10-30% EtOAc in hexane to yield the desired product as a colourless oil. LC retention time 4.67 minutes. [M~H]~ 353. [M-Νa]1" 375.
Example O 02/32891
24
Figure imgf000025_0001
A mixture of resorcinol (3eq.), cyclohexanone (2eq.) and 100 mg of p-toluenesulfonic acid monohydrate (0.025eq.) in chlorofonn is stiired at 40°C for 24 hours. The mixture cooled and filtered, and filtered solid washed with chlorofonn, light petroleum (40-70). The solid is dissolved in methanol and precipitated when mixed with a 6-fold excess of water. The resulting solid is filtered and dried under vacuo.
LC retention time 5.00 minutes, [M+H]+ 381 , [M+Na]+ 403.
Example 3
Figure imgf000025_0002
3-Methoxyphenol ( leq.) and cyclohexanone ( l eq.) were cooled to 0°C and saturated with ITCKg). The mixture was stirred as efficiently as possible for 24h at RT. The thick mass was partitioned between EtOAc and saturated sodium bicarbonate solution, and the organic solution washed with water and brine and dried over MgS04. The EtOAc was treated with decolourising charcoal, and concentrated in vacuo. The crude product was purified by column chromatography on silica, ciuting with 10% EtOAc in he.xane to yield the desired product as a colourless oil.
LC retention time 5.80 minutes. [M-H]" 37~.
Example
Figure imgf000026_0001
As Example 3 using m-cresol
LC retention time 5.40 minutes, [M+Pl 409, [M+Na 431
Example 5
Figure imgf000026_0002
As Example 2 using pyrogallol.
4-
LC retention time 4.69 minutes, [M+H]" 4I3, [M+Na]"" 435
Example 6
Figure imgf000027_0001
As Example 2 using 4-chlororesorcinol.
LC retention time 5.52 minutes, [M+Na] 471 , 473, 475.
Example 7
Figure imgf000027_0002
Compound from example 6 was dissolved in dichloromethane and treated with triethylamine (5 eq.) and acetyl chloride (3.3 eq.) at 0 C. The mixture was allowed to warm to ambient temperature over 2h. then the reaction was quenched by addition of water. The organic layer was separated and the aqueous layer extracted with dichloromethane. The combined organic fractions were washed with water and brine and dried over anhydrous magnesium sulfate. After concentration in vacuo the residue was purified by column chromatography on silica eluting with mixtures of ethyl acetate and hexane to yield the product as a white solid.
LC retention time 5.52 minutes. |"M 57h. ?8, Example 8
Figure imgf000028_0001
4-Pyrogallol (4eq.) in water/c.FlCl (36%) ( 10: 1) was heated at 55DC, then tetrahydro-4H- thiopyran-4-one (leq.) was added portionwise over 30 min. After end of addition, another volume eq. of c.FICl was added. The mixture was stirred at 55°C for 21h. The reaction mixture was concentrated in vacuo and adsorbed onto silica gel. The crude product was purified by column chromatography on silica, eluting with 10-100% EtOAc in hexane to yield the desired product as a colourless oil.
LC retention time 4.14 minutes, [M+Na] 471.
Example 9
Figure imgf000028_0002
As example 8 using 4-chlororesorcinol
LC retention time 4.93 minutes. Example 10
Figure imgf000029_0001
4-Chlororesorcinol (8eq.) in water/c.HCl (36%) (10: 1) was heated at 55°C, then tetrahydro- 4H-pyran-4-one (leq.) was added dropwise over 30 min. After end of addition, another volume eq. of c.FICl was added. The mixture was stined at 55°C for 21 h. Upon cooling, the mixture is diluted with chloroform and the mixture shaken and the precipitated solid filtered off The solid was dissolved in EtOAc/MeOH and adsorbed onto silica gel. The crude product was purified by column chromatography on silica, eluting with 70-100%> EtOAc in hexane to yield the desired product as a colourless oil.
LC retention time 4.21 minutes.
Example 1 1
Figure imgf000029_0002
As example 10, using resorcinol (4eq. ) and tetrahydro-4H-pyran-4-one ( leq.
LC retention time 3.74 minutes. [M~Hf385J. [M-Na]" 407.:
Example L
Figure imgf000030_0001
As example 10 using 4-bromoresorcinol and cyclohexanone.
LC retention time 5.68 minutes.
Example 13
Figure imgf000030_0002
0)
Figure imgf000030_0003
As example 10 using resorcinol and benzyl-4-oxo-l-pιperidϊnecarboxylate.
(ii)
Figure imgf000031_0001
The bis-CBZ compound was hydrogenated in monomethoxyethyleneglycol/formic acid over 10%>Pd/C for 19h at RT. The reaction mixhire was filtered through silica gel and concentrated in vacuo. Purification was by semi-preparative HPLC gave the desired product.
LC retention time 2.61 minutes, [M+H]+383.3.
Example 14
Figure imgf000031_0002
As example 13 using 4-chlororesorcinol.
LC retention time 2.93 minutes, [M+H]+ 45 1.2. 453.2,
Example 15
Figure imgf000032_0001
The bis-CBZ compound from example 13(i) was dissolved in dry THF under an atmosphere of dry nitrogen. A I M sol of LiAlFL in TFIF was added dropwise and the solution stirred and heated at 70UC for 30 min. Upon cooling, water was added and the mixture filtered and concentrated in vacuo. The crude mixture was purified by semi- preparative HPLC to yield the desired product .
LC retention time 2.68 minutes, [M-Hπf " 41 1.3.
Example 16
Figure imgf000032_0002
As example 15 using the bis-CBZ compound from example 14.
LC retention time 2.98 minutes, [M+H]~ 479.2, 481.2.
Example 17
Figure imgf000033_0001
As example 8 using 3,4-Dimethoxyphenol and cyclohexanone
LC retention time 5.03 minutes, [M+Na]J~ 491.
The following examples were synthesized using the procedures outlined in Reaction Scheme 2.
Example 18
Figure imgf000033_0002
Compound from example 2 was dissolved in DMF at ambient temperahire and NBS added ( 1.0 eq,). After stirring for 2 hours and concentration in vacuo the residue was purified by column chromatography on silica eluting with a 1/1 mixture of ethyl acetate and hexane. The mono-bromo compound was eluted with an Rr = 0.3 and the di-bromo compound eluted with an Rt-= 0.6 to yield the products as a white solid.
Mono-bromo 18a compound. LC retention time 5.27 mmures. [M~H]~ 45lL 4(ι0. Di-bromo 18b compound. LC retention time . " minutes. [M~TIj~ 52". 53°. 541. O 02/32891
Example 19
Figure imgf000034_0001
Compound from example 18a was stirred at ambient temperature in DMF with benzyl bromide (4 eq ) and caesium carbonate (4 eq ) overnight The reaction was then diluted with ethyl acetate and the organic fraction was washed with water and brine and dried ovei anhydious magnesium sulfate After concentration in vacuo the residue was purified by column chromatography on silica eluting with mixtutes of ethyl acetate and hexane to yield the product as a white solid
LC retention time 6 39 minutes, [M-rH] 729, 73 1
Example 20
Figure imgf000034_0002
Compound tiom example 19 was dissolved in di v THF and cooled to -78l'C undei an atmosphere of nitrogen t-Burv i lithium (2 5 eq) w as added slow Iv diopwise and the mixtuie sfined αt -~S( C .oi 30 minutes Cαibυn dioxide gas w as then bubbled ihi oiigh the icaction mixtuie lot 1 . minutes follow ed nv \\ ai ming "Ό ambient tempc-atui c undei a carbon dioxide atmosphere. After concentration in vacuo the residue was purified by column chromatography on silica eluting with mixtures of ethyl acetate and hexane to yield the product as a colourless oil.
5 LC retention time 5. 18 minutes.
Example 21
Figure imgf000035_0001
10
To compound 20 in THF at 0°C was added triethylamine (1 eq) and ethyl chloro formate (1 eq) and the mixture stireed for 30 minutes. A solution of ammonia (1: 1 aqueous ammonia:THF) was then added and stirred for 1 hour. The solution was poured into ethyl acetate and washed with water. After drying and concentration in vacuo, the compound was dissolved in methanol: ethyl acetate (1 : 1) and hydrogenated at latm. over 10% Pd/C for 4 hours. The reaction was then filtered through celite and concentrated in vacuo to yield the desired product.
LC retention time 4.86 minutes, [M+H]"r 424,
T 0
Example 22
Figure imgf000036_0001
Compound 20 was dissolved in DCM and shaken gently for 30 minutes with solid- supported DCC (9 eq) and HOBt (0.1 eq) for 30 minutes. Morpholine (1.5 eq) was then added and the mixture shaken gently for 3 days. After filtration, the solution was applied to a pad of silica and the pad washed with ethyl acetate. The intermediate was isolated by elution with mixtures of dichloromethane, methanol and aqueous ammonia and concentration in vacuo. The residues were re-dissolved in a mixture of ethyl acetate and hydrogenated at l atm. over 10% Pd/C for 4 hours. The reaction was then filtered through celite and concentrated in vacuo to yield the desired product.
LC retention time 4.89 minutes, [M+H]+ 493.
Example 23
Figure imgf000036_0002
As example 22 using 3-piperidinemethanol.
LC retention time 4.90 minutes. [M-I-l Example 24
Figure imgf000037_0001
As example 22 using l-(2-hydroxyethl)piperazine.
LC retention time 3.63 minutes, [M+H] 536
Example 25
Figure imgf000037_0002
As example 22 using 3-diethylaminopropylamine.
LC retention time 3.84 minutes, [M÷H]" 536
Example 26
Figure imgf000038_0001
As example 22 using l-(3-aminopropyl)-imidazole.
LC retention time 3J9 minutes, [M+FI] 531
Example 27
Figure imgf000038_0002
As example 22 using butylamine.
LC retention time 5.09 minutes, [M+H] 480
Example 28
Figure imgf000039_0001
As example 22 using 2-methylbutylamine.
LC retention time 5.34 minutes, [M+H] 494
Example 29
Figure imgf000039_0002
As example 22 using 2-(l-cyclohexenyl)ethylamine.
LC retention time 5.79 minutes, [M+H] 534
Example 30
Figure imgf000040_0001
As example 22 using 4-phenylbutylamine.
LC retention time 5.48 minutes, [M+H] 556
Example 31
Figure imgf000040_0002
As example 22 using 3-phenylpropylamine.
LC retention time 5.38 minutes, [M+H]-1" 542
Example 32
Figure imgf000041_0001
As example 22 using tryptamine.
LC retention time 4.050 minutes.
The following examples were synthesized using the procedures outlined in Scheme 3.
Example 33
Figure imgf000041_0002
Compound 2 was dissolved in a mixhire of acetic acid, 33% aqueous ethanol and 37% aqueous formaldehyde (0.9 eq). Diethylamine (0.9 eq) was added and the solution stiiτed at 80°C overnight. The mixhire was concentrated in vacuo, resuspended in dichloromethane and the mixture applied to a pad of silica. The pad washed with ethyl acetate, then the intermediate was isolated by elution with mixtures of dichloromethane. methanol and aqueous ammonia followed by concentration in vacuo to yield the desired product.
LC retention time 3.°(. minutes. [M— H]' Ab. Example 34
Figure imgf000042_0001
As example 33 using l-(2-methoxyphenyl)pϊperazine.
LC retention time 4.27 minutes, [M+H]4" 5
Example 35
Figure imgf000042_0002
As example 33 using 4-methylpiperidine.
LC retention time 4, 1 " minutes. [M-l-1] 492. Example 36
Figure imgf000043_0001
As example 33 using 3-hydroxypiperidine.
LC retention time 3.89 minutes, [M+H] 494.
Example 37
Figure imgf000043_0002
As example 33 using 4-{aminomethyl)piperidine.
LC retention time 3.42 minutes. [M+H]~ 507,
Example 38
Figure imgf000044_0001
As example 33 using N-benzylmethylamine.
LC retention time 4.15 minutes, [M+H] 514.
Example 39
Figure imgf000044_0002
As example 33 using benzyl-N-(2-aminoethyl)carbamate.
LC retention time 4.179 minutes. [M~H]"~ 587
Example 40 O 02/32891
44
Figure imgf000045_0001
As example 33 using 3-piperidinemethanol.
LC retention time 3.87 minutes, [M+H]+ 508.
Example 41
Figure imgf000045_0002
As example 33 using morpboline.
LC retention time 3.92 minutes, [M+H]" 480.
Example 42
Figure imgf000046_0001
As example 33 using l ,2,3,4-Tetrahydro-6J-isoquinolinediol hydrobromide.
LC retention time 3.58 minutes, [M+H]+ 558.
The following examples were synthesized using the procedures outlined in Scheme 4.
Example 42
Figure imgf000046_0002
Compound 2 was dissolved in anhydrous pyridine and acetic anhydride (2 eq) added. After stirring for two hours, the mixture was concentrated to dryness, redissolved in ethyl acetate and washed with concentrated copper sulfate solution. The organic extract was then dried with anhydrous magnesium sulfate. concentrated in vacuo and purified by column chromarography on silica eluting with mixtures of ethvl acetate and hexane to yield vhe product as a white solid. LC retention time 5.06 minutes, [M+NH3]"48;
Example 44
Figure imgf000047_0001
0)
Figure imgf000047_0002
Compound 43 was dissolved in DMF at 0°C and sodium hydride ( 1.2 eq) added. After effervescence had ceased, allyl bromide (1.5 eq) was added and the mixhire stirred for 30 minutes at 0°C then 1 hour at ambient temperahire. The mixure was poured into dichloromethane, washed with water, dried with magnesium sulfate and concentrated to dryness to yield the desired product.
LC retention time 5.37 minutes, [M+NH4]~ 522.
( ii ) The O-allyl compound was re-dissolved in anhydrous methanol containing anhydrous potassium carbonate (0,2 eq) and stined at ambient temperature for two houis. After concentration in vacuo the residue w as purified by column chromatography on silica eluting with mixtuies of ethyl acetate and hexane to yield the product as a colυui less oil. LC retention time 5.44 minutes. [M+H] 42
Example 45
Figure imgf000048_0001
As example 43 using methyl iodide.
LC retention time 5.29 minutes, [M+l-Lp 395.
Example 46
Figure imgf000048_0002
As example 43 using benzyl bromide.
LC retention time 5.64 minutes, [M+H]"" A
The following compounds were svnthesized using the procedure outlined m Sch erne Example 47
Figure imgf000049_0001
(i)
Figure imgf000049_0002
The compound from example 44(i) ( 1 eq), 6-bromoquinaldine (2 eq) and P(o-tolyl)3 (0.3 eq) in a solution of triethylamine (3eq) and acetonitrile was degassed, followed by the addition of Pd(OAc); (0.2 eq). The reaction was heated at 90 °C in a sealed tube for 18 hours. After cooling to RT, the solvent was evaporated in vacuo. The residue was partitioned between DCM and water. The organic layer was dried over Na;S04 and concentrated in vacuo. The crude product was purified by column chromatography on silica, eluting with mixtures of 10-50% EtOAc in hexane to yield the desired bis-acetate.
LC retention time 5.37 minutes, [M-^H] 646.
( ii) The bis-acetate was stirred in a solution of NaOMe (2 eq) and MeOH at RT for 3 hours. The solvent was evaporated in vacuo and the residue partitioned between DCM and water. The organic layer was dried over Na;SO4 and concentrated / vacuo to afford the desired product as an oil. O 02/32891
49
LC retention time 5.56 minutes, [M+H] 56-2
Example 48
Figure imgf000050_0001
As example 47 using 3-bromo-4'-fluorodiphenyl ether.
LC retention time of the bis-acetate, 5.68 minutes, [M+NFL] 708. LC retention time of the di-phenol, 6.13 minutes.
Example 49
Figure imgf000050_0002
As example 47 using 5-bromo-2-methylbenzothiazole.
LC retention time of the bis-acetate. 5.53 minutes. [M-H]" 652. LC retention time of the di-phenol. 5.96 minutes. Biological Results
In vitro Translation assav
Assays were performed as described by Spirin, A.S. ( 1988) Science 242, 1162 and Zubay, G. (1973) Ann. Rev. Genet. 65, 856.
Compounds of the present invention showed inhibitory activities in in vitro translation assays utilizing E, coli extracts. The plasmid pBestLuc, which contains the gene for firefly luciferase downstream of an E. coli promoter and a ribosome binding site is used as a template. The activity of the firefly luciferase enzyme results in a strong luminescent signal. The luminescence generated is a direct measurement of protein expression and of translation efficiency.
Translation reactions in the presence of compound are started by mixing a translation premix that contains Mg" , plasmid template, amino acids, nucleotidetriphosphates, phosphocreatine, creatine phosphokinase and folinic acid with an S30 extract that contains RNA polymerase, ribosomes and translation factors (prepared from E.coli MRΕ600 cells) followed by incubation at 37°C. The activity of the translated luciferase protein is measured by adding an aliquot of the translation reaction to the non-luminescent substrate luciferin and the luminescence measured. The luminescence is quantified in a luminescence plate reader (Wallac Victor). Compounds were assayed 3-5 times over a range of concentrations and an IC 0 calculated.
Figure imgf000051_0001
- 1 00 44
Figure imgf000052_0001
Methylation assay
Compounds where assayed for their ability to bind to position A1067 of rRNA by measuring the inhibition of the Thiostrepton Resistence (JSR) gene product in the presence of 50μM compound. The assay was carried out as described in co-pending United Kingdom Patent Application No. 0009772.5.
Figure imgf000052_0002
Inhibition of EFG dependant GTP hvdrolvsis
Compounds were tested fo their ability to inhibit EFG dependant GTP hydrolysis. Assays were performed as described by Rodnina. .VI. V. and Wϊntermeyer. \V. { 19 "^ 5 ) - P AS, 92. 1 Q4 - 1 Q4Q. Release of free phosphate from γ"CPGTP ( l OOμM) was measured the presence of E. coli riboson.es ( 0.2μλl ). EFG ( OJμλH and Compound ( 50uλl) by TLC. The percentage GTPase activity was compared with nonnal GTPase activity, and the well characterised inhibitor thiostrepton at the 10 minute time point in each case.
GTPase activity (%) in lOmin Normal control (no compound)- 70%
Thiostrepton - 25%
Example 1 - 10%
Example 2-
Example 3 - 70% Example 4 - 48%
Example 6 - 5%
Antibacterial activity
Compounds were tested for their ability to inhibit bacterial growth as follows. Incubation volumes were 70μl per well in Mueller-FIϊnton Broth Isosensitest Broth (Oxoid, Basingstoke, UK) and the inoculum 10-4 (10,000 ) cfu/well. Compound was dissolved in DMSO and was added at defined concentrations resulting in 5% DMSO per well. 96 well plates were incubated aerobically at 37°C with shaking in a Molecular Devices Spectra Max Plus 384 micro-plate reader with continuous monitoring of culture absorbance at 600nm.
Example 1
Figure imgf000053_0001
Example 2 Di
Figure imgf000054_0001
Example 6
Figure imgf000054_0002
The compounds of the present invention therefore show good in vitro biological activity.
In vivo antibacterial assay
The in vivo therapeutic efficacy of the compounds of the invention is measured by intramuscular injection to mice experimentally infected with a pathogenic gram positive or gram negative bacterium ( e.g. methicillin-resistant Staphylococcus aureus (MRSA). Clostridium difficile. Klebsiella pneumoniae. Eschericia coli. l laemophilus ϊnfiuenzue. etc. ). As an example. MRSA strain A2"223 can be used. MRSA strain A27223 is prepared for experimental infection by growth on two large Brain Heart Infusion Agar plates. On each plate, 0.5 ml of frozen stock culture is plated out. Plates are then incubated for 18 hours at 30°C. The next day each plate is washed with 20 ml of Brain Heart Infusion Broth and then pooled together. A microscopic direct count of microorganisms is done using a 1 : 1000 dilution of plate wash. After a direct count is obtained, the number of organisms per milliliter is calculated. The count is adjusted to the desired amount of inoculum by diluting in 4% hog mucin. The desired challenge (amount of organisms given to mice) is 2.4 x 108 cfu/0.5 ml/mouse for MRSA strain A27223. The mice are infected intraperitoneally with 0.5 ml of challenge. Ten non-treated infected mice are used as controls. Mice used are adult male ICR mice. The average weight of the animals should range from 20 to about 26 grams.
Compounds are generally tested at 4 dose levels (e.g. 25, 6.25, 1.56 and 0.39 mg/kg) and prepared in 5% cremophor, unless otherwise specified. When MRSA A27223 is the challenging microorganism, vancomycin is used as the control compound, and is dosed at 6.25, 1.56, 0.39 and 0.098 mg/kg. It is prepared in 0.1M phosphate buffer. There are generally five infected mice per dose level, and they are treated with 0.2 ml of the test compound, preferably by intramuscular injection. Treatment begins 15 minutes and 2 hours post-infection.
A PDjo (protective dose-50, the dose of drug given, which protects 50%> mice from mortality) runs for 5 days. During this time, mortality of mice is checked every day and deaths are recorded. The cumulative mortality at each dose level is used to calculate a PD50 value for each compound. Surviving mice are sacrificed at the end of day 5 by CO3 inhalation. The actual calculation of PD50 is perfoπned with a computer program using the Spearman- Karber procedure.
A compound according to the invention is effective for the treatment of bacterial infection if it has a PD-n of about 100 mg kg or less. Topical administration assav Compounds according to the invention can be used for the treatment of dermatological infections. It is often desirable to treat dennatologϊcal infections with topically applied antibiotics because such application allows direct delivery of the antibiotic agent to the site of infection, and because it permits delivery in higher concentrations at the affected site than would, for example, systemic administration.
In order to assay the effect of a compound according to the invention when applied topically, animals with experimentally induced skin infections are used. Any recognised experimental model of skin infection is acceptable. One of the most commonly used models is Staphyloccocus aureus infection of mice. Infection may be induced by intracutaneous injection of S. aureus bacteria or topical application of bacteria to experimentally wounded or inflamed skin of immunosuppressed mice. Immunosuppression is commonly achieved by, for example, cyclophosphamide treatment of the animals (usually 2 mg/mouse for 5 days prior to infection). Experimental dermatological infection of mice is described in, for example Abe et al., 1992, J. Dermatol. Sci. 4: 42-48; Akiyama et al., 1996, J. Dermatol, Sci. 11 : 234-238; and Molne & Tarkowski, 2000, J. Invest. Dermatol. 1 14: 1 120- 1 125, which are incorporated herein by reference.
Once an experimental infection is established, the effectiveness of a compound of the invention is assayed by topical application of the compound in a preparation , e.g. an ointment, appropriate for such topical application. One skilled in the art can selected suitable vehicles for topical administration.
A range of differing concentrations of the compound being assayed is applied to the infected area of different infected mice. The concentration of the compound to use in such assays or in treatment of infection in general can be estimated by considering the amount of the compound effective to kill or inhibit the pathogen in culture. Generally, the concentration of compound in a topical preparation will range from about 0.0001% to about 10%. (wwv) of the compound being assayed, preferably about 0.001 % to about 2%. about 0.0 1% to about IV about 0.001% to about 0. 1%. or 0.0(11% to 0-01 ",,-. The preparation can also be applied with varying frequency, ranging from example, from once an hour to once a day.
The effectiveness of the topical treatment can be evaluated in several ways. For example, visual inspection of the infected area can be used. If the area of infection, as evidenced by inflammation or skin lesions, either does not increase in size or decreases in size relative to its size at the time of the first treatment, the agent can be judged effective. Alternatively, or in addition, microscopic examination of samples taken from infected skin can be conducted, and the degree of inflammation or infiltration of the infection into the tissue can be evaluated by a professional skilled in the interpretation of histological specimens, e.g. a pathologist, dermatologist or medical technologist.
Differences are determined by examining samples taken from untreated, infected control animals. Alternatively, two areas of infection can be established on, for example, opposite sides of the back of the same mouse. In that instance, only one side is treated with the compound being assayed and the untreated site serves as a control. If the inflammation (judged by the presence of inflammatory cells) or infiltration of the pathogen does not increase following treatment, the agent is judged effective as a bacteriostatic agent effective against that pathogen in vivo. If the inflammation or infiltration decrease (e.g. by 10%) or more) the agent is judged to be effective as a bacteriocidal agent effective against the pathogen in vivo, Additional controls can include skin samples taken from infected animals treated with other drugs of known effectiveness against the experimental pathogen (e.g. vancomycin for S. aureus). Such controls can provide a measure of the effectiveness of the compound relative to that of standard antibiotics.

Claims

CLAIMS:
A compound for use in therapy, wherein the compound is of the formula
Figure imgf000058_0001
wherein p is selected from 4, 5 and 6; q is selected from 3, 4 and 5; each T is the same or different and is selected from -X- and -CR'2-; each R1 may be the same or different and is selected from H, Cι.n> alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl and C4.18 aralkyl; each X may be the same or different and is selected from -CR1:-, 0, S and NR' where each R' may be the same or different and is selected from H, Cι-6 alkyl, C2.6 alkenyl, C2-12 alkynyl, C3-10 aryl, C4-12 aralkyl, C(0)R\ S02R" and C(O)NRV wherein R is selected from C |-6 alkyl, C2-6 alkenyl, C3-10 aryl, C4-12 aralkyl;
A, B, Y1, Y2, YJ, Z1,
Figure imgf000058_0002
A and Z may be the same or different and are selected from H, halogen, C 1-12 alkyl, C2-12 alkenyl, C2-12 alkynyl, C3-12 aryl. C4.ιs aralkyl, SR", OR3, NR4R5, CO2R6 and C0NR+ wherein each R2, R3 and R is the same or different and is selected from H. C M: alkyl, C -12 alkenyl. C2-12 alkynyl. C3-12 aryl, C1.12 alkoyl. C _ι2 aryloyl and C4-1X aralkyl. and each R" ,Rf' and R is the same or different and is selected from H, C1-12 alkyl. Cι-\ι alkenyl. C2-.1 alk nyl. C3.12 aryl, Cj-is aralkyl: provided that at least one of A and B is OR" , and pharmaceutically acceptable derivatives thereof. A compound of the formula
Figure imgf000059_0001
wherein p is selected fiom 4, 5 and 6, q is selected from 3, 4 and 5, each T is the same or different and is selected fiom -X- and -CRV, each R1 may be the same or different and is selected from H, C\ 12 alkyl, C2-1 alkenyl, C2-12 alkynyl, C3-12 aryl and C4 ι8 aialkyl, each X may be the same or different and is selected fiom -CR , O, S and NR' where each R' may be the same or different and is selected fiom H C|.6 alkyl, C2 6 alkenyl, C2 12 alkynyl, C3-10 aryl, C4 12 aralkyl, C(0)R\ SO2R" and C(0)NRV wheiein R" is selected from Cι-6 alkyl, C2 & alkenyl, C3 10 aryl, C4-12 aralkyl, A, B, Y , Y~, Y", Z , Z", ZJ and Z may be the same or different and are selected from H, halogen, C1-12 alkyl, C2 12 alkenyl, C2 12 alkynyl, C,,-i2 aryl, C4 t aralkyl, SR2, OR3, NR4R\ C02Rή and CONR7 2 wherein each R2, R3 and R4 is the same or different and is selected from H, Cι-i2 alkyl, C2-12 alkenyl, C 12 alkynyl, C. 12 aryl. C M; alkoyl. C; 12 aiv lov l and C is aralkyl and each R" Rfi and R' is the same 01 different and is selected horn H Ci 12 αlkv l C2 12 alkenv l, C2.Ϊ -llkynv l, C. 12 ai yl
C4 is αialkvl. piov ided that at least one of λ and B is OR" and phαmtαceuticallv acceptable dem αtiv es theieot piov iueu that at least one ot A B l \ ' and " is not H CI 1. C;LL ι OI I
3. A compound according to claim 1 or 2 wherein p is 4 or 5 and q is 3 or 4.
4. A compound according to claim 1 or 2, wherein the compound is of the fonnula
Figure imgf000060_0001
wherein each U may be the same or different and is selected from -X-CR1,-
-CR'-X-CR'- and
-CR 2-X-CRVCR 2-
5. A compound according to claim 4 wherein each U is -CR 2-X-CR 2~.
6. A compound according to any preceding claim wherein any two vicinal R groups are the same.
7. A compound according to any preceding claim wherein X is the same or different and is selected from 0, S and NR' where each R' may be the same or different and is selected from H. C|.„ alkyl. C;_,, alkenyl, C..-1.1 aryl. CM; aralkyl. C(0)R". SO2R " and C(O)NR' *; wherein each R " may be the same or different and is selected from C M, alkyl. C;.ι, alkenvl. C3-.1t. arvl and C4-1; aralkvt.
8. A compound according to any one of claims 1 to 6, wherein the compound is of formula
Figure imgf000061_0001
wherein m and n may be the same or different and are selected from 1, 2 or 3; each R1 and R~ may be the same or different and are selected from H, C1-1 alkyl, Cι-12 alkenyl, C3-12 aryl and C4-13 aralkyl;
A, B, Y1, Y~, Y~, Z , Z2, ZJ and Z4 may be the same or different and are selected from H, C1-12 alkyl, Cι-12 alkenyl, C3-12 aryl, C -ιs aralkyl, OR3 and NR4R5; wherein RJ, R and R" are the same or different and are selected from H and Cι-6 alkyl; provided that at least one of A and B is OR"', and phaiTnaceutically acceptable derivatives thereof.
9. A compound according to claim 8 wherein at least one of A and B is OH.
10, A compound according to claim 8 or 9 wherein m and n are selected from 1 and 2.
1 1 , A compound according to claim 8, 9 or 10 wherein m and n are the same.
L A compound according to any preceding claim wherein each R is H.
13. A compound according to any preceding claim wherein each Y1 and Y~ is selected from H, ORJ, Cι-ι2 alkyl and halogen.
14. A compound according to any preceding claim wherein Y1 or YJ is Cι-ι2 alkyl substihited with a group selected from amino, alkylamino, dialkylamino and heterocyclic groups.
15. A compound according to any one of claims 1 to 12 wherein Y1 or YJ is CONR7 2
16. A compound according to any preceding claim wherein each Z , ZJ and Z is selected from H and ORJ and halogen
17. A compound according to any preceding claim wherein Y~ is OR .
18. A compound according to any preceding claim wherein each Z" is OR\
19. A compound according to any preceding claim wherein one of A and B is OH and one of A and B is FI.
20. A compound according to any one of claims 1 to 18 wherein A or B is OR where RJ is C2-12 alkenyl.
21. A compound according to claim 20 wherein A or B is ORJ where J is a group of the formula
-CH2CH=CH-T
where T is H or C...12 aryl.
22. A compound per se as defined in any one of claims 1 to 2 1 provided that both Tp is not -t CH;)5- and Tq is not -(CFL - and provided that the compound is not 2\4"J- tnhydroxy-2.3-propanυtlavan-4-spirocyclopentane.
23. Use of a compound according to any one of claims 1 to 22 in the manufacture of a medicament for use in the treatment of bacterial infection.
24. A method of treating bacterial infection comprising administering to a subject in need of such treatment an effective dose of a compound as defined in any one of claims 1 to 22.
25. A phamtaceufical composition comprising a compound as defined in any one of claims 1 to 22 in combination with a pharmaceutically acceptable excipient.
26. Use of a compound as defined in any one of claims 1 to 22 to inhibit the binding of a compound to RNA.
27. Use of a compound as defined in any one of claims 1 to 22 to inhibit bacterial protein translation.
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