WO2017100849A1 - Inhibiteurs des 6-oxopurine phosphoribosyltransférases - Google Patents

Inhibiteurs des 6-oxopurine phosphoribosyltransférases Download PDF

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WO2017100849A1
WO2017100849A1 PCT/AU2016/051238 AU2016051238W WO2017100849A1 WO 2017100849 A1 WO2017100849 A1 WO 2017100849A1 AU 2016051238 W AU2016051238 W AU 2016051238W WO 2017100849 A1 WO2017100849 A1 WO 2017100849A1
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alkyl
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spp
mmol
dmtr
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John De Jersey
Luke William GUDDAT
Dana Hockova
Dianne Therese KEOUGH
Radek Pohl
Dominik Rejman
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University of Queensland UQ
Institute of Organic Chemistry and Biochemistry CAS
Institute of Organic Chemistry and Biochemistry of ASCR vvi
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University of Queensland UQ
Institute of Organic Chemistry and Biochemistry CAS
Institute of Organic Chemistry and Biochemistry of ASCR vvi
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/683—Diesters of a phosphorus acid with two hydroxy compounds, e.g. phosphatidylinositols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • 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
    • A61P31/06—Antibacterial agents for tuberculosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P33/00—Antiparasitic agents
    • A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
    • A61P33/06—Antimalarials
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings
    • C07F9/65616—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings containing the ring system having three or more than three double bonds between ring members or between ring members and non-ring members, e.g. purine or analogs
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention relates to compounds which are useful as inhibitors of 6-oxopurine phosphoribosyltransferases such as hypoxanthine-guanine-(xanthine) phosphoribosyltransferase (HG(X)PRT).
  • Inhibitors of these enzymes may be used in the prevention or treatment of microbial infections, including infections caused by protozoal and bacterial species.
  • the compounds of the invention are particularly suited to the prevention or treatment of microbial infections caused by Plasmodium spp., which is responsible for malaria, and infections caused by Mycobacterium tuberculosis.
  • the 6-oxopurine phosphoribosyltransferases (EC:2.4.2.8 HG(X)PRT and/or XGPRT EC:2.4.22) are necessary for both survival and reproduction of many microorganisms (including protozoa and certain bacteria) because, unlike mammalian cells, such microorganisms are auxotrophic for the purine ring. Thus, these microorganisms depend on these enzymes for the synthesis of the 6-oxopurine nucleoside monophosphates required for RNA/DNA production. On the other hand, humans possess two metabolic pathways to synthesise nucleoside monophosphates: de novo and salvage. Partial inhibition of the human enzyme should not have any serious side-effects. This is based on the fact that humans with an inherited genetic defect which results in only 3% of normal activity of this enzyme lead normal lives.
  • 6-oxopurine PRTases represent a target with therapeutic potential.
  • the reactions catalysed by these enzymes are shown in Scheme 1.
  • 6-Oxopurine phosphoribosyltransferase is the generic name for enzymes which add a phosphoribosyl group from PRib-PP onto the N9 atom of a 6-oxopurine to form a nucleoside monophosphate.
  • the 6-oxopurine PRTases found in nature vary in their specificities for the three naturally occurring 6-oxopurines, hypoxanthine, guanine and xanthine. The specific names given to 6-oxopurine PRTases denote this specificity.
  • the human enzyme is called hypoxanthine guanine PRTase (HGPRT) because it can efficiently use both hypoxanthine and guanine as substrates.
  • the Plasmodium falciparum enzyme is called HGXPRT because it can use xanthine in addition to hypoxanthine and guanine as substrates.
  • Some organisms (including human, other mammals and Plasmodium species) encode and synthesize only one 6-oxopurine PRTase.
  • Other organisms encode and synthesize two 6- oxopurine PRTases.
  • Escherichia coli XGPRT and HPRT are Escherichia coli XGPRT and HPRT (Guddat, L.W.
  • 6-oxopurine PRTases are members of purine salvage pathways present in all or virtually all species. These pathways contain a variety of enzymes. Their function is to make all of the required purine nucleotides (for RNA and DNA synthesis and for other purposes) using preformed purines.
  • Organisms which lack the de novo pathway and depend absolutely on the activity of one 6-oxopurine PRTase for the synthesis of purine nucleotides and which are human or animal pathogens are therefore the prime targets for 6-oxopurine PRTase inhibitors.
  • Such organisms include several protozoan parasites including the Plasmodium species responsible for human malaria and Helicobacter pylori, the causative organisms of gastric ulcers.
  • inhibitors of the 6-oxopurine PRTases are potential leads for the development of novel antibiotics. It is also proposed that combination therapy which includes the co-administration with inhibitors of the de novo pathway (such as azaserine) will be a successful therapy.
  • Acyclic nucleoside phosphonates are reverse transcriptase inhibitors and several ANP-based drugs are in current clinical use for the treatment of serious viral infections (e.g. Viread ® , Vistide ® , Hepsera ® ). These compounds consist of a nucleobase, either 6- aminopurine or pyrimidine, linked to a phosphonate group by an acyclic linker.
  • 2- (Phosphonoethoxy)ethyl guanine (PEEG) and 2-(phosphonoethoxy)ethyl hypoxanthine (PEEHx) are good inhibitors of both human HGPRT and Plasmodium falciparum HGXPRT (P HGXPRT):
  • PEEG and PEEHx have IQ values for human HGPRT of 1 ⁇ and 3.6 ⁇ respectively, and 0.1 ⁇ and 0.3 ⁇ for HGXPRT.
  • ANPs are believed to be metabolically stable due to the presence of a phosphonate P-C linkage instead of a P-0 phosphoester, making them resistant towards phosphomonoesterases and nucleotidases.
  • ANPs are also believed to be stable because the bond between the purine and the linker is stable, in contrast to the bond between the purine and the ribose in purine nucleotides.
  • Plasmodium falciparum Pf
  • Plasmodium vivax Pv
  • Drugs such as artemisinin and combination therapies, quinine, chloroquine, primaquine, are the only known treatments for malaria but, because of increasing resistance to these drugs as well as cost-effectiveness, there is an urgent need for the discovery of new targets and therapeutic leads for the development of potent antimalarials.
  • ANPs are potent inhibitors of 6-oxopurine phosphoribosyltransferases such as hypoxanthine-guanine-(xanthine) phosphoribosyltransferase (HG(X)PRT).
  • the present invention provides a compound of formula (la) or (lb):
  • R 1 is alkyl, aryl, heteroaryl, alkenyl, alkynyl, arylalkyl, heteroarylalkyl, NR 6 2 , halogen, OR 6 , H or NH 2 ;
  • R 2 is H, -C(0)L 1 P(0)R 7 R 8 or -C(0)L 2 P(0)R 7 R 8 ; the or each R 3 is independently selected from H, -I ⁇ OH, -I ⁇ OR 6 , -CH 2 OC(0)L 1 P(0)R 7 R 8 , -CH 2 C(0)L 1 P(0)R 7 R 8 , -OC(0)L 1 P(0)R 7 R 8 , and -OL 1 P(0)R 7 R 8 ;
  • R 4 is -OC(0)L 2 P(0)R 7 R 8 , -OL 1 P(0)R 7 R 8 or -NHC(0)L 1 P(0)R 7 R 8 ;
  • R 4 is -C(0)L 1 P(0)R 7 R 8 when R 2 is -C(0)L 2 P(0)R 7 R 8 ;
  • R 4 is H when R 3 is other than H
  • R 5 is H, alkyl, aryl, heteroaryl, NR 6 2 , halogen or OR 6 ;
  • R 7 and R 8 are independently selected from OH, OR 6 , SR 6 , NHR 6 , NR 6 2 , and R p ; wherein R p is a prodrug substituent and wherein the or each R 6 , where present, is independently selected from alkyl, alkenyl, alkynyl, aryl, acyl and arylalkyl optionally containing one or more heteroatoms;
  • L 1 is Ci- 4 alkylene, C 2 - 4 alkenylene or C 2 - 4 alkynylene;
  • L is C 2 - 4 alkylene, C 2 - 4 alkenylene or C 2 - 4 alkynylene;
  • L 1 and L 2 are optionally substituted with one or more OH, Ci_ 4 alkyl, C 2 - 4 alkenyl or C 2 - 4 alkynyl;
  • X 1 is N or CH
  • X 2 is C or X 2 -R 5 forms N;
  • X 3 is N or CH
  • X 4 is C or X 4 -R l forms N;
  • Y 1 where present is S or O
  • n 1 or 2;
  • the present invention provides a compound of formula (la) represented by formula (II):
  • A is selected from one of the following: and wherein:
  • R 1 is alkyl, aryl, heteroaryl, alkenyl, alkynyl, arylalkyl, heteroarylalkyl, NR 6 2 , halogen, OR 6 , H or NH 2 ;
  • R 2 is H, -C(0)L 1 P(0)R 7 R 8 or -C(0)L 2 P(0)R 7 R 8 ;
  • the or each R 3 is independently selected from H, -I ⁇ OH, -I ⁇ OR 6 , -CH 2 OC(0)L 1 P(0)R 7 R 8 , -CH 2 C(0)L 1 P(0)R 7 R 8 , -OC(0)L 1 P(0)R 7 R 8 , and -OL 1 P(0)R 7 R 8 ;
  • R 4 is-OC(0)L 2 P(0)R 7 R 8 , -OL 1 P(0)R 7 R 8 or -NHC(0)L 1 P(0)R 7 R 8 ;
  • R 4 is -C(0)L 1 P(0)R 7 R 8 when R 2 is -C(0)L 2 P(0)R 7 R 8 ;
  • R 4 is H when R 3 is other than H
  • R 5 is H, alkyl, aryl, heteroaryl, NR 6 2 , halogen or OR 6 ;
  • R 7 and R 8 are independently selected from OH, OR 6 , SR 6 , NHR 6 , NR 6 2 , and R p ; wherein R p is a prodrug substituent and wherein the or each R 6 , where present, is independently selected from alkyl, alkenyl, alkynyl, aryl, acyl and arylalkyl optionally containing one or more heteroatoms;
  • L 1 is Ci- 4 alkylene, C 2 - 4 alkenylene or C 2 - 4 alkynylene;
  • L is C 2 - 4 alkylene, C 2 - 4 alkenylene or C 2 - 4 alkynylene;
  • L 1 and L 2 are optionally substituted with one or more OH, Ci_ 4 alkyl, C 2 -
  • n 1 or 2;
  • the invention provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the inhibition of a 6-oxopurine phosphoribosyltransferase enzyme such as hypoxanthine-guanine-(xanthine) phosphoribosyltransferase (HG(X)PRT).
  • HG(X)PRT hypoxanthine-guanine-(xanthine) phosphoribosyltransferase
  • the invention provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the prevention or treatment of a microbial infection.
  • the invention provides a method of preventing or treating a microbial infection comprising administering a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, to a subject in need thereof.
  • the invention provides a compound according to the invention, or a pharmaceutically acceptable salt thereof, for use in the prevention or treatment of a microbial infection.
  • the invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable carrier or diluent.
  • the invention also provides a combination comprising a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, together with at least one other therapeutic agent.
  • the compounds of the invention are selective inhibitors for HGXPRT and/or vHGPRT over human HGPRT.
  • the new series of compounds described herein differ most significantly from other ANPs in that they comprise a nitrogen containing heterocyclic group such as pyrrolidine or piperidine which replaces the oxygen atom in the acyclic portion of the earlier molecules.
  • this heterocyclic group provides a branching point in the acyclic portion of the molecule such that the compound may bind at three closely spaced sub- sites of the target. This is believed to increase the binding potential of the compound which leads to a significant improvement in the affinity and/or selectivity for 6- oxopurine phosphoribosyltransferases such as hypoxanthine-guanine-(xanthine) phosphoribosyltransferase (HG(X)PRT).
  • the heterocyclic group provides rigidity between the purine base and the phosphonate groups which, when combined with the relative stereochemistry of the ring substituents, is believed improve orientation of the purine base and the phosphonate groups into the active site of the enzyme.
  • the heterocyclic group also enables groups to be appended in a modular fashion, enabling the construction of a diverse range of molecular architectures with a diverse range of chemical groups, using a range of readily accessible synthetic techniques.
  • the compounds of the invention are particularly well suited to preventing or treating infections caused by those organisms that are auxotrophic for the purine ring present in nucleosides, and thus rely, or substantially rely, on the salvage pathway to provide the purine moiety for reproduction. In some embodiments such microorganisms possess no de novo, or substantially no de novo, pathway for purine synthesis.
  • R p is taken to be a prodrug substituent.
  • the term prodrug refers to a masked form of a compound of the invention such that drug absorption and/or drug delivery into the target organism is typically enhanced.
  • the highly polar phosphonate group (wherein R 2 and R 3 are OH), for example, may be masked with a more lipophilic group to enhance transport across cell membranes.
  • the prodrug may be unmasked by cellular enzymes (including lipases, esterases, reductases, oxidases, nucleases or amidases) or by chemical cleavage such as hydrolysis to release the compound of the invention after the prodrug has entered a cell in the target of interest.
  • prodrugs of the compounds of the invention may provide certain advantages over the correspondingly unmasked compounds of the invention, such as improved levels of uptake into the microorganism (prodrugs are not inhibitors of the enzymes activity or are weakly inhibitory).
  • substituents which may be used to form prodrugs according to the invention include those substituents that may be cleaved in vivo to provide the compound of the invention with a phosphonate residue.
  • a phosphoramidate moiety formed by condensing the amine group of an amino acid with a phosphonate (or activated form thereof) may undergo hydrolysis in vivo to reform the phosphonate group.
  • R p may be an amino acid residue (including an ester derivative of an amino acid residue) or R p may be an optionally substituted alkoxy group.
  • Suitable prodrug substituents include, but are not limited to: proteins; antibiotics (and antibiotic fragments); amino acids (D- and/or L-) including derivatives thereof (such as esters and amides), attached to the -P(O)- moiety in the compounds of the invention via a nitrogen atom or an oxygen atom; peptides (up to 10 amino acids) attached to the -P(O)- moiety in the compounds of the invention via a nitrogen atom or an oxygen atom; drug moieties attached to the -P(O)- moiety in the compounds of the invention via a nitrogen atom or an oxygen atom; steroids; cholesterols; folic acids; vitamins; polyamines; carbohydrates; polyethylene glycols (PEGs); cyclosaligenyls; substituted 4 to 8-membered rings, with or without heteroatom substitutions, with 1,3-phosphodiester, 1,3- phosphoramidate/phosphoester or 1,3-phosphoramidate attachments; acylthioethoxy, (
  • the prodrug is a group attached to the -P(O)- moiety in the compounds of the invention via a nitrogen atom or an oxygen atom.
  • Prodrug substituents can include a residue of an amino acid, including derivatives thereof (such as esters and amides).
  • Especially preferred prodrugs of the compounds of the invention are phosphoramidates formed from an alkyl ester of phenylalanine attached via a nitrogen atom to the -P(O)- moiety (such as the isopropyl or ethyl ester of phenylalanine, particularly (L)- phenylalanine) and phosphoesters formed from an alkyl group such as RCOSCH 2 CH 2 0- (eg SATE), lipophilic esters (eg Hostetler esters) and pivaloyloxymethyl esters (POM).
  • RCOSCH 2 CH 2 0- eg SATE
  • lipophilic esters eg Hostetler esters
  • POM pivaloyloxymethyl esters
  • alkyl includes saturated aliphatic groups, including straight-chain alkyl groups (e.g., methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, etc.) and branched-chain alkyl groups (isopropyl, tert-butyl, isobutyl, etc.).
  • alkyl refers to straight chained alkyl.
  • C x _ y alkyl wherein x is 1-5 and y is 2-10 indicates an alkyl group (straight- or branched-chain) containing the specified number of carbon atoms.
  • Ci_ 4 alkyl includes, but is not limited to, methyl, ethyl, propyl, butyl, isopropyl, tert-butyl and isobutyl.
  • alkylene refers to a divalent alkyl group.
  • alkenyl includes unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one double bond.
  • alkenyl includes straight-chain alkenyl groups (e.g., ethenyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, etc.) and branched-chain alkenyl groups.
  • alkenyl refers to straight chained alkenyl.
  • a straight chain or branched chain alkenyl group has 6 or fewer carbon atoms in its backbone (e.g., C 2 -C 6 for straight chain, C3-C 6 for branched chain).
  • cycloalkenyl groups may have from 3-8 carbon atoms in their ring structure, and more preferably have 5 or 6 carbons in the ring structure.
  • C 2 -C 6 includes alkenyl groups containing 2 to 6 carbon atoms.
  • alkenylene refers to a divalent alkenyl group.
  • alkynyl includes unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but which contain at least one triple bond.
  • alkynyl includes straight-chain alkynyl groups (e.g., ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, etc.) and branched-chain alkynyl groups.
  • alkynyl refers to straight chained alkynyl.
  • a straight chain or branched chain alkynyl group has 6 or fewer carbon atoms in its backbone (e.g., C 2 -C6 for straight chain, Ci-C 6 for branched chain).
  • C 2 -C 6 includes alkynyl groups containing 2 to 6 carbon atoms.
  • alkynylene refers to a divalent alkynyl group.
  • aryl refers to aromatic monocyclic (eg phenyl) or polycyclic groups (e.g., tricyclic, bicyclic, e.g., naphthalene, anthryl, phenanthryl).
  • Aryl groups can also be fused or bridged with alicyclic or heterocyclic rings which are not aromatic so as to form a polycycle (e.g., tetralin, methylenedioxyphenyl).
  • heteroaryl represents a monocyclic or bicyclic ring, typically of up to 7 atoms in each ring, wherein at least one ring is aromatic and contains from 1 to 4 heteroatoms selected from the group consisting of O, N and S.
  • Heteroaryl groups within the scope of this definition include but are not limited to: benzimidazole (otherwise known as benzoimadazole), acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolinyl, isoquinolinyl, oxazolyl, isoxazolyl, indoiyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, tetrahydroquinoline.
  • benzimidazole otherwise known as benzoimadazole
  • acridinyl carbazolyl
  • cinnolinyl quinoxalinyl
  • pyrrazolyl indolyl
  • benzotriazolyl furanyl
  • heteroaryl is also understood to include the N-oxide derivative of any nitrogen-containing heteroaryl.
  • heteroaryl substituent is bicyclic and one ring is non-aromatic or contains no heteroatoms, it is understood that attachment is via the aromatic ring or via the heteroatom containing ring, respectively.
  • acyl includes compounds and moieties which contain the acyl radical (CH 3 CO-) or a carbonyl group such as CH 3 CH 2 CH 2 CO-.
  • alkoxy includes substituted and unsubstituted alkyl, alkenyl, and alkynyl groups covalently linked to an oxygen atom.
  • alkoxy groups include methoxy, ethoxy, isopropyloxy (isopropoxy), propoxy, butoxy, and pentoxy groups and may include cyclic groups such as cyclopentoxy.
  • esteer includes compounds and moieties that contain a carbon or a heteroatom bound to an oxygen atom that is bonded to the carbon of a carbonyl group.
  • esteer includes alkoxycarboxy groups such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, etc.
  • the alkyl, alkenyl, or alkynyl groups are as defined above.
  • halogen includes fluorine, chlorine, bromine and iodine. In some embodiments halogen refers to fluorine or chlorine.
  • heteroatom includes atoms of any element other than carbon or hydrogen.
  • Preferred heteroatoms are nitrogen, oxygen, sulfur and phosphorus.
  • Particularly preferred heteroatoms are nitrogen and oxygen.
  • optionally substituted typically refers to where a hydrogen atom on a group has been substituted with a non-hydrogen group. Any optionally substituted group may bear one, two, three or more optional substituents.
  • any number of the listed functional groups and molecules may be combined to create a larger molecular architecture.
  • the terms aryl and alkyl can be combined to form an arylalkyl group, an example of which is a phenylmethyl group, otherwise known as a benzylic group. It is to be understood that when combining functional groups and molecules to create a larger molecular architecture, hydrogens can be removed or added, as required to satisfy the valence of each atom.
  • the structures of some of the compounds of the invention include asymmetric carbon atoms. It is to be understood accordingly that the isomers arising from such asymmetry (e.g., all enantiomers, stereoisomers, rotamers, tautomers, diastereomers, or racemates) are included within the scope of this invention.
  • the present invention includes within its scope all of these stereoisomeric forms either isolated (in, for example, enantiomeric isolation), or in combination (including racemic mixtures and diastereomic mixtures).
  • the invention thus also relates to compounds in substantially pure stereoisomeric form with respect to the asymmetric centres of the amino acid residues, e.g., greater than about 90% de, such as about 95% to 97% de, or greater than 99% de, as well as mixtures, including racemic mixtures, thereof.
  • the skilled person will appreciate that there are a range of techniques available to produce achiral compounds of the invention in racemic, enantioenriched or enantiopure forms.
  • enantioenriched or enantiopure forms of the compounds may be produced through stereoselective synthesis and/or through the use of chromatographic or selective recrystallisation techniques.
  • the compounds of the invention may be prepared by appending a natural or unnatural amino acid to the phosphonate group to form a prodrug. Accordingly, a racemic mixture of amino acids may be used to prepare a racemic mixture of a compound of the invention, an enantioenriched amino acid may be used to prepare an enantioenriched mixture of a compound of the invention and an enantiopure amino acid may be used to prepare an enantiopure compound of the invention.
  • Y 1 where present is selected from S and O
  • Y 2 where present is selected from halogen.
  • the compound of the invention has the following formula where Y 1 is selected from S and O:
  • Y 1 is O.
  • R 1 is alkyl, aryl, heteroaryl, alkenyl, alkynyl, arylalkyl, heteroarylalkyl, NR 6 2 , halogen, OR 6 , H or NH 2 .
  • R 1 is H.
  • R 1 is NH 2 .
  • R 2 is H.
  • R 2 is -C(0)L 1 P(0)R 7 R 8 .
  • R 2 is -C(0)L 2 P(0)R 7 R 8 .
  • each R 3 is independently selected from H, -I ⁇ OH, -I ⁇ OR 6 , -CH 2 OC(0)L 1 P(0)R 7 R 8 , -CH 2 C(0)L 1 P(0)R 7 R 8 , -OC(0)L 1 P(0)R 7 R 8 , and -OL 1 P(0)R 7 R 8 .
  • n 2 wherein one R 3 is H and the other R 3 is H, -I ⁇ OH, -I ⁇ OR 6 , -CH 2 OC(0)L 1 P(0)R 7 R 8 or -CH 2 C(0)L 1 P(0)R 7 R 8 .
  • R 4 is-OC(0)L 2 P(0)R 7 R 8 , -OL 1 P(0)R 7 R 8 or -NHC(0)L 1 P(0)R 7 R 8 .
  • R 4 is -C(0)L 1 P(0)R 7 R 8 when R 2 is -C(0)L 2 P(0)R 7 R 8 .
  • R 4 is H when R 3 is other than H.
  • R 5 is H, alkyl, aryl, heteroaryl, NR 6 2 , halogen or OR 6 , t) R 5 is H.
  • R 7 and R 8 are independently selected from OH, OR 6 , SR 6 , NHR 6 , NR 6 2 , and R p .
  • the or each R 6 is independently selected from alkyl, alkenyl, alkynyl, aryl, acyl and arylalkyl optionally containing one or more heteroatoms.
  • R 7 and R 8 are independently selected from OH, OR 6 and R p , wherein R 6 is Ci_ 3 alkyl.
  • R 7 and R 8 are OH.
  • R 7 and R 8 are R p .
  • R p is an amino acid residue, or derivative or ester thereof, or an optionally substituted alkoxy group.
  • R p is a Ci_ 3 alkyl ester derivative of phenylalanine or -OCH 2 CH 2 SC(0)R (SATE) wherein R is Ci_ 4 alkyl.
  • L 1 is Ci_ 4 alkylene.
  • L is C 2 - 4 alkylene, C 2 _ 4 alkenylene or C 2 _ 4 alkynylene optionally substituted with one or more OH, Ci_ 4 alkyl, C 2 _ 4 alkenyl or C 2 _ 4 alkynyl.
  • L is C 2 _ 4 alkylene.
  • L 1 is methylene or ethylene and L 2 is ethylene.
  • IInn aa pprreeffeerrrreedd eemmbbooddiimmeenntt AA iiss R 5 is H.
  • the present invention provides compounds of the formula (II) represented by formula (III):
  • R 1 is H or NH 2 ;
  • R 2 is H, -C(0)L 1 P(0)R 7 R 8 or -C(0)L 2 P(0)R 7 R 8 ;
  • R 3 is H
  • R 4 is -OC(0)L 2 P(0)R 7 R 8 , -OL 1 P(0)R 7 R 8 or -NHC(0)L 1 P(0)R 7 R 8 ;
  • R 4 is -C(0)L 1 P(0)R 7 R 8 when R 2 is -C(0)L 2 P(0)R 7 R 8 ;
  • R 7 and R 8 are independently selected from OH, OR 6 , SR 6 , NHR 6 , NR 6 2 , and R p ;
  • R p is a prodrug substituent;
  • R 6 is independently selected from alkyl, alkenyl, alkynyl, aryl, acyl and arylalkyl optionally containing one or more heteroatoms;
  • L 1 is Ci_ 4 alkylene;
  • L is C 2 - 4 alkylene
  • the compound of formula (la) is selected from:
  • treatment may include alleviating or ameliorating the symptoms, diseases or conditions associated with the microbial infection being treated, including reducing the severity and/or frequency of the microbial infection.
  • prevention may include preventing or delaying the onset of, inhibiting the progression of, or halting or reversing altogether the onset or progression of the particular symptoms, disease or condition associated with a microbial infection.
  • microorganism includes any microscopic organism or taxonomically related macroscopic organism within the categories algae, bacteria, fungi and protozoa or the like.
  • the microorganism is a bacteria or protozoa, most preferably a protozoa.
  • the present invention is predicated, in part, on the susceptibility of microorganisms to inhibition of 6-oxopurine phosphoribosyltransferases such as the hypoxanthine-guanine-(xanthine) phosphoribosyltransferase (HG(X)PRT) enzyme involved in the purine salvage pathway.
  • 6-oxopurine phosphoribosyltransferases such as the hypoxanthine-guanine-(xanthine) phosphoribosyltransferase (HG(X)PRT) enzyme involved in the purine salvage pathway.
  • the compounds of the invention are particularly well suited to those organisms that are auxotrophic for the purine ring present in nucleosides, and thus rely, or substantially rely, on the salvage pathway to provide the purine moiety for reproduction.
  • there are known techniques such as genomic sequencing or gene deletion experiments
  • the compounds of the invention are particularly well suited to preventing or treating a microbial infection caused by a microorganism that cannot sustain itself in the absence of the salvage pathway.
  • such microorganisms possess no de novo, or substantially no de novo, pathway for purine synthesis.
  • the bacterial infection may be caused by one or more species selected from one or more of the Gram-negative bacterial genera: Acinetobacter; Actinobacillus; Bartonella; Bordetella; Brucella; Burkholderia; Campylobacter; Cyanobacteria; Enterobacter; Erwinia; Escherichia; Francisella; Helicobacter; Hemophilus; Klebsiella; Legionella; Moraxella; Morganella; Mycobacterium; Neisseria; Pasteur ella; Proteus; Providencia; Pseudomonas; Salmonella; Serratia; Shigella; Stenotrophomonas; Treponema; Vibrio; and Yersinia. Specific examples include, but are not limited to, infections caused by Helicobacter pylori and uropathogenic Escherichia coli.
  • the bacterial infection may be caused by one or more species selected from one or more of the Gram-positive bacterial genera: Actinobacteria; Bacillus; Clostridium; Corynebacterium; Enterococcus; Listeria; Nocardia; Staphylococcus; and Streptococcus.
  • Protozoal infections include, but are not limited to, infections caused by Leishmania, Toxoplasma, Plasmodia (which are understood to be the causative agent(s) of malarial infection), Theileria, Anaplasma, Giardia, Tritrichomonas, Trypanosoma, Schistosoma, Coccidia, and Babesia. Specific examples include Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium knowlesi, Plasmodium ovale and Giardia lamblia.
  • the compounds of the invention are particularly well suited to the prevention or treatment of infections caused by Plasmodia spp. (especially Plasmodium falciparum and Plasmodium vivax), Giardia spp. (especially Giardia lamblia), Trypanosoma spp., Schistosoma spp., Helicobacter spp. (such as Helicobacter pylori), Mycobacterium tuberculosis, and certain uropathogenic Escherichia coli.
  • Plasmodia spp. especially Plasmodium falciparum and Plasmodium vivax
  • Giardia spp. especially Giardia lamblia
  • Trypanosoma spp. Schistosoma spp.
  • Helicobacter spp. such as Helicobacter pylori
  • Mycobacterium tuberculosis and certain uropathogenic Escherichia coli.
  • nucleosides may modulate the activity of one or more kinases, including certain nucleosides bearing saccharide and/or polyphosphate moieties (for example diphosphate or triphosphate moieties).
  • polyphosphate moieties for example diphosphate or triphosphate moieties.
  • microbial infections include bacterial or fungal wound infections, mucosal infections, enteric infections, septic conditions, pneumonia, trachoma, ornithosis, trichomoniasis, fungal infections and salmonellosis, such as in veterinary practice.
  • the compounds of the invention may also be used for the treatment of resistant microbial species or in various fields where antiseptic treatment or disinfection of materials is required, for example, surface disinfection.
  • the term "subject" is intended to include organisms such as mammals, e.g. humans, dogs, cows, horses, pigs, sheep, goats, cats, mice, rabbits, rats, and transgenic non-human animals.
  • the subject is a human, e.g. a human suffering from, at risk of suffering from, or potentially capable of suffering from a microbial infection.
  • the subject is a cell.
  • the compounds of the invention may be in crystalline form or as solvates (e.g. hydrates) and it is intended that both forms are within the scope of the present invention.
  • solvate is a complex of variable stoichiometry formed by a solute (in this invention, a compound of the invention) and a solvent. Such solvents should preferably not interfere with the biological activity of the solute. Solvents may be, by way of example, water, acetone, ethanol or acetic acid. Methods of solvation are generally known within the art.
  • the present invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of a compound as hereinbefore defined, or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable carrier or diluent.
  • Pharmaceu tic ally acceptable acid addition salts may be prepared from inorganic and organic acids. Examples of inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like.
  • organic acids include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like.
  • the nitrogen atom in the acyclic portion of the compounds of the invention may undergo reaction with an acid to form the acid addition salt.
  • Pharmaceutically acceptable base addition salts may be prepared from inorganic and organic bases.
  • Corresponding counterions derived from inorganic bases include the sodium, potassium, lithium, ammonium, calcium and magnesium salts.
  • Organic bases include primary, secondary and tertiary amines, substituted amines including naturally- occurring substituted amines, and cyclic amines, including isopropylamine, trimethyl amine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2- dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobromine, purines, piperazine, piperidine, and N-ethylpiperidine.
  • the compound of the invention possesses a phosphonate group the compound may undergo reaction with a base to form the base addition salt.
  • Acid/base addition salts tend to be more soluble in aqueous solvents than the corresponding free acid/base forms.
  • composition is intended to include the formulation of an active ingredient with encapsulating material as carrier, to give a capsule in which the active ingredient (with or without other carrier) is surrounded by carriers.
  • the compounds as hereinbefore described, or pharmaceutically acceptable salt thereof may be the sole active ingredient administered to the subject, the administration of other active ingredient(s) with the compound is within the scope of the invention.
  • the compound could be administered with one or more therapeutic agents in combination.
  • the combination may allow for separate, sequential or simultaneous administration of the compound as hereinbefore described with the other active ingredient(s).
  • the combination may be provided in the form of a pharmaceutical composition.
  • the route of administration and the nature of the pharmaceutically acceptable carrier will depend on the nature of the condition and the mammal to be treated. It is believed that the choice of a particular carrier or delivery system, and route of administration could be readily determined by a person skilled in the art. In the preparation of any formulation containing the compound care should be taken to ensure that the activity of the compound is not destroyed in the process and that the compound is able to reach its site of action without being destroyed. In some circumstances it may be necessary to protect the compound by means known in the art, such as, for example, micro encapsulation or coating (such as the use of enteric coating). Similarly the route of administration chosen should be such that the compound reaches its site of action.
  • Those skilled in the art may readily determine appropriate formulations for the compounds of the present invention using conventional approaches. Identification of preferred pH ranges and suitable excipients, for example antioxidants, is routine in the art. Buffer systems are routinely used to provide pH values of a desired range and include carboxylic acid buffers for example acetate, citrate, lactate and succinate. A variety of antioxidants are available for such formulations including phenolic compounds such as BHT or vitamin E, reducing agents such as methionine or sulphite, and metal chelators such as EDTA.
  • phenolic compounds such as BHT or vitamin E
  • reducing agents such as methionine or sulphite
  • metal chelators such as EDTA.
  • the compounds as hereinbefore described, or pharmaceutically acceptable salt thereof may be prepared in parenteral dosage forms, including those suitable for intravenous, intrathecal, and intracerebral or epidural delivery.
  • the pharmaceutical forms suitable for injectable use include sterile injectable solutions or dispersions, and sterile powders for the extemporaneous preparation of sterile injectable solutions. They should be stable under the conditions of manufacture and storage and may be preserved against reduction or oxidation and the contaminating action of microorganisms such as bacteria or fungi.
  • the solvent or dispersion medium for the injectable solution or dispersion may contain any of the conventional solvent or carrier systems for the compound, and may contain, for example, water, ethanol, polyol (for example, glycerol, propylene glycol and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
  • the proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • the prevention of the action of microorganisms can be brought about where necessary by the inclusion of various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal and the like.
  • agents to adjust osmolality for example, sugars or sodium chloride.
  • the formulation for injection will be isotonic with blood.
  • Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin.
  • Pharmaceutical forms suitable for injectable use may be delivered by any appropriate route including intravenous, intramuscular, intracerebral, intrathecal, epidural injection or infusion.
  • Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients such as those enumerated above, as required, followed by filtered sterilization.
  • dispersions are prepared by incorporating the various sterilised active ingredient into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above.
  • preferred methods of preparation are vacuum drying or freeze-drying of a previously sterile-filtered solution of the active ingredient plus any additional desired ingredients.
  • compositions include oral and enteral formulations of the present invention, in which the active compound may be formulated with an inert diluent or with an assimilable edible carrier, or it may be enclosed in hard or soft shell gelatin capsule, or it may be compressed into tablets, or it may be incorporated directly with the food of the diet.
  • the active compound may be incorporated with excipients and used in the form of ingestible tablets, buccal or sublingual tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. The amount of active compound in such therapeutically useful compositions is such that a suitable dosage will be obtained.
  • the tablets, troches, pills, capsules and the like may also contain the components as listed hereafter: a binder such as gum, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such a sucrose, lactose or saccharin may be added or a flavouring agent such as peppermint, oil of wintergreen, or cherry flavouring.
  • a binder such as gum, acacia, corn starch or gelatin
  • excipients such as dicalcium phosphate
  • a disintegrating agent such as corn starch, potato starch, alginic acid and the like
  • a lubricant such as magnesium stearate
  • a sweetening agent such as sucrose, lactose or saccharin may be added or a flavouring agent such as peppermint, oil of winter
  • tablets, pills, or capsules may be coated with shellac, sugar or both.
  • a syrup or elixir may contain the active compound, sucrose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavouring such as cherry or orange flavour.
  • any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts employed.
  • the active compound may be incorporated into sustained-release preparations and formulations, including those that allow specific delivery of the active compound to specific regions of the gut.
  • Liquid formulations may also be administered enterally via a stomach or oesophageal tube.
  • Enteral formulations may be prepared in the form of suppositories by mixing with appropriate bases, such as emulsifying bases or water-soluble bases. It is also possible, but not necessary, for the compounds of the present invention to be administered topically, intranasally, intravaginally, intraocularly and the like.
  • the present invention also extends to any other forms suitable for administration, for example topical application such as creams, lotions and gels, or compositions suitable for inhalation or intranasal delivery, for example solutions, dry powders, suspensions or emulsions.
  • topical application such as creams, lotions and gels
  • compositions suitable for inhalation or intranasal delivery for example solutions, dry powders, suspensions or emulsions.
  • the compounds of the present invention may be administered by inhalation in the form of an aerosol spray from a pressurised dispenser or container, which contains a propellant such as carbon dioxide gas, dichlorodifluoromethane, nitrogen, propane or other suitable gas or combination of gases.
  • a propellant such as carbon dioxide gas, dichlorodifluoromethane, nitrogen, propane or other suitable gas or combination of gases.
  • the compounds may also be administered using a nebuliser.
  • Pharmaceutically acceptable vehicles and/or diluents include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents and the like.
  • the use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, use thereof in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
  • Dosage unit form refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required pharmaceutically acceptable vehicle.
  • the specification for the novel dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding active materials for the treatment of disease in living subjects having a diseased condition in which bodily health is impaired as herein disclosed in detail.
  • the principal active ingredient may be compounded for convenient and effective administration in therapeutically effective amounts with a suitable pharmaceutically acceptable vehicle in dosage unit form.
  • a unit dosage form can, for example, contain the principal active compound in amounts ranging from 0.25 ⁇ g to about 200 mg. Expressed in proportions, the active compound may be present in concentrations ranging from about 0.25 ⁇ g to about 200 mg/mL of carrier. In the case of compositions containing supplementary active ingredients, the dosages are determined by reference to the usual dose and manner of administration of the said ingredients.
  • the term "effective amount" refers to an amount of compound which, when administered according to a desired dosing regimen, provides the desired therapeutic activity. Dosing may occur once, or at intervals of minutes or hours, or continuously over any one of these periods. Suitable dosages may lie within the range of about 0.1 ng per kg of body weight to 1 g per kg of body weight per dosage. A typical dosage is in the range of 1 ⁇ g to 1 g per kg of body weight per dosage, such as is in the range of 1 mg to 1 g per kg of body weight per dosage. In one embodiment, the dosage may be in the range of 1 mg to 500 mg per kg of body weight per dosage. In another embodiment, the dosage may be in the range of 1 mg to 250 mg per kg of body weight per dosage. In yet another embodiment, the dosage may be in the range of 1 mg to 100 mg per kg of body weight per dosage, such as up to 50 mg per body weight per dosage.
  • Preparative column chromatography was carried out on silica gel (40-60 ⁇ ; Fluka) and elution was performed at the flow rate of 40 ml/min.
  • the following solvent systems were used for TLC and preparative chromatography: toluene-ethyl acetate 1: 1 (T); chloroform-ethanol 9: 1 (CI); ethyl acetate-acetone-ethanol-water 6: 1: 1:0.5 (H3); ethyl acetate-acetone-ethanol-water 4: 1: 1: 1 (HI).
  • the concentrations of solvent systems are stated in volume percent (% v/v).
  • Purity of prepared compounds was determined by LC- MS performed on Waters Auto Purification System with 2545 Quaternary Gradient Module and 3100 Single Quadrupole Mass Detector using LUNA C18 column (Phenomenex, 100 x 4.6 mm, 3 ⁇ ) at a flow rate of 1 ml/min. Typical conditions were: mobile phase, A - 50mM NH 4 HC0 3 ; B - 50 mM NH 4 HC0 3 in 50% aq. CH 3 CN; C - CH 3 CN; A ⁇ B/10 min, B ⁇ C/10 min, C/5 min.
  • the column was washed with ethanohwater (1: 1) mixture (100 ml) and crude product was eluted with 3% NH 3 in ethanol: water (1: 1) (-150 ml, UV absorption was monitored).
  • the crude material was purified by rpHPLC using a linear gradient of methanol in water.
  • reaction mixture was warmed to rt, concentrated in vacuo and final product ii was obtained by column chromatography on silica gel using a linear gradient of ethyl acetate in toluene in 88% yield (9.26 g, 16.97 mmol).
  • the final product was obtained as an ⁇ 5:4 mixture of two rotamers.
  • the final product vi was obtained by column chromatography on silica gel using a linear gradient of ethyl acetate in toluene in 59% (2.28 g, 4.16 mmol) yield as an ⁇ 10:9 mixture of two rotamers.
  • TBDMSCl (1.97 g, 13.1 mmol) was added to a mixture of vii (6 g, 8.75 mmol) and imidazole (0.9 g, 13.1 mmol) in DMF.
  • the reaction mixture was stirred at rt under an atmosphere of argon for 3 days.
  • the reaction mixture was concentrated in vacuo, dissolved in CHC1 3 (150 ml) and washed with sat soln. NaHC0 3 .
  • the organic phase was dried over Na 2 S0 4 , filtered and evaporated.
  • the obtained diastereoisomers were separated by column chromatography on silica gel using gradient of ethyl acetate in toluene.
  • IR v max (CHCl 3 ) 3550 (vw, vbr), 3487 (vw, vbr), 3335 (w, vbr), 2982 (s), 2956 (s), 2932 (s), 2896 (m, sh), 2885 (m), 2860 (m), 1690 (vs), 1473 (m), 1464 (m), 1415 (vs), 1392 (s), 1387 (s), 1375 (m), 1367 (s), 1252 (s), 1172 (s), 1141 (s), 1102 (vs), 1068 (w, sh), 1012 (vs, sh), 1002 (vs), 990 (vs, sh), 939 (w), 887 (m), 839 (s).
  • IR v max (CHCl 3 ) 3610 (w), 3359 (w, vbr), 2983 (s), 2932 (m), 2885 (m), 1688 (vs), 1495 (w, sh), 1478 (m), 1456 (m), 1415 (vs), 1393 (s), 1368 (s), 1245 (s), 1167 (s), 1109 (s), 1056 (vs), 1030 (vs), 965 (s), 461 (vw).
  • IR v max (CHCl 3 ) 3610 (w), 3359 (m, br), 2985 (s), 2932 (m), 1688 (vs), 1478 (m), 1456 (m), 1415 (vs), 1394 (s), 1368 (s), 1245 (s), 1167 (s), 1109 (s), 1055 (vs), 1030 (vs), 964 (s).
  • reaction mixture was concentrated in vacuo and the desired product was obtained by chromatography on silica gel using a linear gradient of ethanol in CHC1 3 in 31% yield (0.82 g, 1.24 mmol) in the form of grey foam as an -1: 1 mixture of two ro tamers.
  • Precursor xiv (0.89 g, 2.42 mmol) was converted via the sequence of general methods C, E, F, and B to Compound 1 in 22% overall yield (0.3 g, 0.54 mmol) in the form of a white amorphous lyophilizate as an ⁇ 6:7 mixture of two ro tamers.
  • Precursor xiii (0.49 g, 1.24 mmol) was converted via the sequence of general methods C, E, F, and B to Compound 2 in 11% overall yield (79.5 mg, 0.14 mmol) in the form of white amorphous lyophilizate as an ⁇ 6:7 mixture of ro tamers A:B:
  • Precursor xiii (0.6 g, 1.63 mmol) was converted via the sequence of general methods D, E, F, and B to title compound 3 in 31% overall yield (291 mg, 0.51 mmol) in the form of white amorphous lyophilizate as an ⁇ 6:7 mixture of ro tamers A:B:
  • IR v max (KBr) 3120 (m, br), 2775 (m, br), 2365 (w, vbr), 1691 (vs), 1632 (vs), 1605 (s, sh), 1570 (m, sh), 1534 (w), 1478 (m), 1415 (w), 1169 (m), 1148 (m, sh), 1067 (m), 1052 (m, sh), 979 (w), 898 (m), 782 (w), 640 (w).
  • Precursor xvii (60 mg, 0.092 mmol) was converted using general method B to title compound 4 in 63% yield (32.8 mg, 0.058 mmol) in the form of white amorphous lyophilizate as an ⁇ 6:7 mixture of rotamers A:B:.
  • Precursor ixS (0.39 g, 0.72 mmol) was converted via the sequence of general methods D, E, F, and B to title compound 5 in 10% overall yield (42.3 mg, 72.4 ⁇ ) in the form of white amorphous lyophilizate as an ⁇ 1: 1 mixture of ro tamers A:B:
  • Precursor ixF (0.31 g, 0.59 mmol) was converted via the sequence of general methods C, E, F, and B to title compound 6 in 16% overall yield (56.9 mg, 97 ⁇ ) in the form of white amorphous lyophilizate as an ⁇ 6:7 mixture of ro tamers A:B.
  • IR v max (KBr) 2360 (w, vbr), 1690 (s), 1633 (vs), 1590 (m, sh), 1551 (w), 1516 (w), 1459 (w), 1420 (w), 1342 9we, 1218 (m), 1175 (m), 1141 (m), 1070 (m), 1053 (m), 972 (w), 895 (w), 792 (w), 647 (w).
  • Precursor xxx (1.16 g, 2.2 mmol) was converted via the sequence of general methods C, E, F, and B to title compound 9 in 12% overall yield (154 mg, 260 ⁇ ) in the form of white amorphous lyophilizate as an ⁇ 1: 1: 1: 1 mixture of four isomers (two epimeric rotamers A and two epimeric rot
  • IR v max (KBr) 3129 (s, br), 2770 (m, vbr, sh), 2370 (w, vbr), 1691 (vs), 1633 (vs, br), 1605 (s, br, sh), 1571 (m, sh), 1534 (m), 1478 (m, sh), 1415 (w), 1171 (s), 1145 (m, sh), 1065 (s, br), 975 (w), 900 (m, br), 782 (w), 642 (w).
  • Human HGPRT was stored in 0.1 M Tris-HCl, 0.01 M MgCl 2 , pH 7.4, 200 ⁇ PRib-PP, 1 mM dithiothreitol (DTT), -80°C.
  • P/HGXPRT was stored in 0.01 M phosphate, 60 ⁇ hypoxanthine, 200 ⁇ PRib-PP, pH 7.2, 1 mM DTT as previously described (Keough, D. T., et al., Mol. Biochem. Parasitol. 1999, 98 (1), 29-41). This difference is because the Pf enzyme is completely inactive under the conditions used to store the human enzyme.
  • the buffer was 0.1 M Tris-HCl, 0.01 M MgCl 2 , pH 7.4.
  • the R values were calculated by Hanes' plots at a fixed concentration of guanine (60 ⁇ ) and at variable concentrations of PRib-PP (14 - 1000 ⁇ ) depending on the K m(app) in the presence of the inhibitor.
  • Pf D6 Sud-Leone laboratory line, sensitive to most antimalarial drugs and W2 (Indochina) line, resistant to chloroquine and pyrimethamine, were maintained in RPMI- 1640-LPLF complete medium, containing 10% human plasma, at 4% hematocrit and 1% to 8% parasitemia as previously described (Trager, W.; Jensen, J. B., Science 1976, 193 (4254), 673-675). Cultures were routinely synchronised using D-sorbitol. To evaluate the anti-malarial activity of the compounds, the previously described [ H] -hypoxanthine growth inhibition assay was utilized (Desjardins, R. E., et al. Antimicrob.
  • the results of the study are shown in Table 1 below.
  • the IQ values provided in the Table correspond to the value obtained for the parent compound of the prodrugs listed in the Table.
  • the parent compounds were either synthesized and tested or the prodrug compounds were converted to the parent compound by hydrolysis of the prodrug attachments.
  • the prodrug attachments greatly enhance transport of the parent compound across the cellular membrane to act against the D6 and W2 malaria strains in the cell based assay.
  • the compounds of the invention are effective inhibitors against both strains of malaria and some of the compounds display selectivity for parasitical enzyme HG(X)PRT over human enzyme HGPRT.
  • the values for Mycobacterium tuberculosis HGPRT were determined in 0.1 M Tris- HC1, 12 mM MgCl 2 , pH 7.4 at 25 °C.
  • the concentration of guanine was fixed at 67.5 ⁇ and the concentration of the variable substrate, PRib-PP, was in the range 20-1500 ⁇ , depending on the value for ⁇ m(app) at that concentration of inhibitor.
  • the concentration of inhibitor in the assay ranged from 1.4 ⁇ (for the best inhibitor) to 56 ⁇ (for the weakest inhibitor).
  • the reaction was initiated by the addition of enzyme to give a final concentration of 200 nM.
  • the reaction was followed at 257.5 nm for 60 s with a total change in absorbance of 0.05 units.
  • the ⁇ m(app) , V max and values were calculated using Prism6 (GraphPad Software, Inc., La Jolla, CA). Cell based assays under normal growth conditions
  • M. tuberculosis H37Rv/a (ATCC 25177) was grown in Middlebrook 7H9 broth medium supplemented with OADC (oleic acid dextrose catalase), 0.5% glycerol and 0.02% tyloxapol. Freshly seeded cultures were grown at 37°C, for approximately 14 days, to mid-exponential phase (OD600 0.4-0.8) for use in inhibition assays. The potency of the inhibitors was measured by a resazurin reduction microplate assay, as previously described (Taneja, N. K. and Tyagi, J. S., J. Antimicrob. Chemother., 2007, 60 (2), 288-293; West, N. P., et al., Chem.
  • M. tuberculosis grown to mid-exponential phase (OD600 0.4-0.8), was diluted to OD 6 oo 0.001 in 7H9S media (Middlebrook 7H9 with OADC, 0.5% glycerol, 0.75% tween-80, 1% tryptone) containing 0.5% DMSO.
  • 7H9S media Middlebrook 7H9 with OADC, 0.5% glycerol, 0.75% tween-80, 1% tryptone
  • DMSO DMSO
  • 96-well microtitre plates were set up with 100 ⁇ of inhibitors, serially diluted into 7H9S media. 100 ⁇ of diluted M. tuberculosis, representing ⁇ 2xl0 4 CFU/mL was added to each well.
  • hypoxic assay is identical to the normoxic REMA with the following modifications:
  • PMA phorbol 12-myristate 13-acetate
  • THP-1 cells 5xl0 5 cells/ml
  • PMA is capable of differentiating the monocytic THP-1 cells into macrophages.
  • 200 ⁇ of induced THP-1 cells were added to a 96-well plate and incubated for 3 days at 37°C/5% C0 2 . This facilitated differentiation into macrophages.
  • Candidate compounds were added to the infected macrophages and incubated for 4 days at 37°C/5% C0 2 . Fluorescence was then recorded. Media for THP-1 cells RPMI, L-glutamine (2mM final), 10% FBS, ⁇ -mercaptoethanol (0.05mM final). Made fresh and kept for 1-2 weeks at 4°C

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Abstract

L'invention concerne des composés qui sont utiles en tant qu'inhibiteurs des 6-oxopurine phosphoribosyltransférases telles que l'hypoxanthine-guanine- (xanthine) phosphoribosyltransférase (HG(X)PRT).
PCT/AU2016/051238 2015-12-15 2016-12-15 Inhibiteurs des 6-oxopurine phosphoribosyltransférases Ceased WO2017100849A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111018858A (zh) * 2018-10-10 2020-04-17 复旦大学 杂环类衍生物及其制备方法和用途

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
KASPERSEN, F.M. ET AL.: "Unconventional nucleotide analogues. Part XIII. (2S,4S)-2- Hydroxymethyl- and 2-Carboxy-4-(purin-9-yl)pyrrolidines", JOURNAL OF THE CHEMICAL SOCIETY, PERKIN TRANSACTIONS 1: ORGANIC AND BIO-ORGANIC CHEMISTRY, no. 16, 1975, pages 1617 - 1622, XP008089127 *
KEOUGH, D.T. ET AL.: "Inhibition of the Escherichia coli 6-Oxopurine Phosphoribosyltransferases by Nucleoside Phosphonates: Potential for New Antibacterial Agents", JOURNAL OF MEDICINAL CHEMISTRY, vol. 56, no. 17, 2013, pages 6967 - 6984, XP055391560 *
LI, Y. ET AL.: "Synthesis of 9-(1-amino-4-fluoromethyl-3-pyrrolidinyl)-2,6-dichloropurine from trans-4-hydroxy-D-proline", CHINESE JOURNAL OF APPLIED CHEMISTRY, vol. 25, no. 3, March 2008 (2008-03-01), pages 345 - 349, XP055391559 *
PETERSON, M.L. ET AL.: "Synthesis and biological evaluation of 4-purinylpyrrolidine nucleosides", JOURNAL OF MEDICINAL CHEMISTRY, vol. 34, no. 9, 1991, pages 2787 - 2797, XP002418258 *
POHL, R. ET AL.: "Synthesis, conformational studies, and biological properties of phosphonomethoxyethyl derivatives of nucleobases with a locked conformation via a pyrrolidine ring", ORGANIC & BIOMOLECULAR CHEMISTRY, vol. 13, no. 16, 2015, pages 4693 - 4705, XP055391557 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111018858A (zh) * 2018-10-10 2020-04-17 复旦大学 杂环类衍生物及其制备方法和用途
CN111018858B (zh) * 2018-10-10 2022-11-18 复旦大学 杂环类衍生物及其制备方法和用途

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