WO2010074586A1 - Pyrazolo[1,5-a]pyridine and imidazo[1,2-a]pyridine derivatives and their use in cancer therapy - Google Patents

Pyrazolo[1,5-a]pyridine and imidazo[1,2-a]pyridine derivatives and their use in cancer therapy Download PDF

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WO2010074586A1
WO2010074586A1 PCT/NZ2009/000298 NZ2009000298W WO2010074586A1 WO 2010074586 A1 WO2010074586 A1 WO 2010074586A1 NZ 2009000298 W NZ2009000298 W NZ 2009000298W WO 2010074586 A1 WO2010074586 A1 WO 2010074586A1
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pyridin
methyl
nitrobenzenesulfonohydrazide
ethylidene
amino
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PCT/NZ2009/000298
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French (fr)
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Jackie Diane Kendall
Anna Claire Giddens
Kit Yee Tsang
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Pathway Therapeutics Limited
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • 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
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis

Definitions

  • the present invention relates to pyrazolo[1 ,5-a]pyridine and/or -imidazo[1 ,2-a]pyridine derivatives, to their preparation, to their use as agents or drugs for cancer prevention or therapy for treating cancer, either alone or in combination with radiation and/or anticancer drugs.
  • Phosphoinositide-3-kinases are a group of lipid kinases which phosphorylate the 3- hydroxyl of phosphoinositides. They are split into three classes (Class I, Il and III) and play an important role in cellular signalling [Stephens et al., Curr. Opin. Pharmacol. 2005, 5, 357].
  • the Class I enzymes are further split into Class Ia and Ib based on their mechanism of activation; the Class Ia PI3Ks are heterodimeric structures consisting of a catalytic subunit (p110 ⁇ , p110 ⁇ or p110 ⁇ ) in complex with a regulatory p85 subunit, while the Class Ib PI3K (p110 ⁇ ) is structurally similar but lacks a regulatory subunit, and instead is activated by ⁇ y subunits of heterotrimeric G-proteins [Walker et al,. MoI. Cell., 2000, 6, 909].
  • PI3Ks play a variety of roles in normal tissue physiology [Foukas & Shepherd, Biochem. Soc. Trans., 2004, 32, 330; Shepherd, Acta Physiol. Scand., 2005, 183, 3], with p110 ⁇ having a specific role in cancer growth, p110 ⁇ in thrombus formation mediated by integrin ⁇ n ⁇ 3 [Jackson et al., Nat. Med., 2005, 11 , 507], and p110 ⁇ in inflammation, rheumatoid arthritis [Camps et al., Nat. Med., 2005, 11 , 936] and other chronic inflammation states [Barber et al., Nat. Med., 2005, 11 , 933].
  • the PI3K enzymes produce phosphoinositide 3,4,5-triphosphate (PIP3) from the corresponding diphosphate (PIP2), thus recruiting AKT (protein kinase B) through its PH domain, to the plasma membrane. Once bound, AKT is phosphorylated and activated by other membrane bound kinases, and is central to a cascade of events that lead to inhibition of apoptosis [Berrie, Exp. Opin. Invest. Drugs, 2001 , 10, 1085].
  • the p110 ⁇ isoform is selectively amplified and activated in a number of cancer types [Stephens et al., Curr. Opin. Pharmacol., 2005, 5, 357; Stauffer et al., Curr. Med. Chem. - Anti- Cancer Agents, 2005, 5, 449].
  • pan-PI3K inhibitor LY294002 While PI3K isoenzymes play important roles in many cellular processes, published experimental studies in mice with human tumour xenografts show that the pan-PI3K inhibitor LY294002 is well-tolerated, reduces signalling through the PI3K pathway, causes reduction of tumour volume, and is more active in cell lines over-expressing mutant forms of p110 ⁇ than parental control cells [Semba et al., Clin. Cancer Res., 2002, 8, 1957; Hu et al., Cancer Res., 2002, 62, 1087].
  • PI3K and especially the p110 ⁇ isoform
  • LY294002 non- selective
  • PI103 lightly ⁇ -selective
  • ZSTK474 non-selective
  • Het represents pyrazolo[1 ,5-a]pyridine or imidazo[1 ,2-a]pyridine (below);
  • X may represent up to two of halogen, R 1 , OR 1 , OCOR 1 , CONR 1 2 , CO 2 R 1 , SO 2 R 1 , SO 2 NR 1 2 , CN, NO 2 , NR 1 2 or NR 1 COR 1 , placed at any of the available positions A-, 5-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the available positions 5-, 6-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine; Y is H or Me;
  • D is phenyl, naphthyl, or 5- or 6-membered heterocycle or benzoheterocycle, where the heterocyclic ring contains up to two of the atoms S, O, N and is optionally substituted at any available position with up to two of halogen, R 1 , OR 1 , C0NR 1 2 , CO 2 R 1 , SO 2 R 1 , SO 2 NR 1 2 , CN, CF 31 OCF 31 NO 21 NR 1 Z1 NR 1 COR 1 ;
  • R 1 is H or C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R 3 , OR 3 , NR 3 2 , OCOR 3 , CONR 3 2 , CO 2 R 3 , CN 1 SO 2 R 3 , SO 2 NR 3 2 , NR 3 COR 3 or optionally substituted aryl or. heteroaryl, or in the case where R 1 forms part of NR 1 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR 4 ;
  • R 2 is C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R 3 , OR 3 , NR 3 2 , OCOR 3 , CONR 3 2 , CO 2 R 3 , CN 1 SO 2 R 3 , SO 2 NR 3 2l NR 3 COR 3 or optionally substituted aryl or heteroaryl;
  • R 3 is H 1 C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R 4 , OR 4 , NR 4 2 , OCOR 4 , CONR 4 2 , CO 2 R 4 , CN, SO 2 R 4 , SO 2 NR 4 2 , NR 4 COR 4 ; or optionally substituted aryl or heteroaryl, or in the case where R 3 forms part of NR 3 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR 4 ;
  • R 4 is H,
  • X may represent one or two of halogen, R 1 , OR 1 , OCOR 1 , CONR 1 2 , CO 2 R 1 , SO 2 R 1 , SO 2 NR 1 2 , CN, NO 2 , NR 1 2 or NR 1 COR 1 , placed at any of the available positions 4-, 5-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the available positions 5-, 6-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine, where R 1 is defined as above.
  • X may represent one or two of halogen, R 1 , OR 1 , OCOR 1 , CONR 1 2 , CO 2 R 1 , SO 2 R 1 , SO 2 NR 1 2 , CN, NO 2 , NR 1 2 or NR 1 COR 1 , where one of X is placed at position 5- where Het represents pyrazolo[1 ,5-a]pyridine or at position 6- where Het represents imidazo[1 , 2- ajpyridine, and the other X, if present, is placed at any of the remaining available positions 4-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the remaining available positions 5-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine, where R 1 is defined as above.
  • D is phenyl and is optionally substituted at any available position with up to two of halogen, R 1 , OR 1 , C0NR 1 2 , CO 2 R 1 , SO 2 R 1 , SO 2 NR 1 2 , CN, CF 3 , OCF 3 , NO 2 , NR 1 2 , or NR 1 COR 1 , where R 1 is defined as above.
  • Het represents pyrazolo[1 ,5-a]pyridine (below);
  • R 4 is H 1 C1-6 saturated or unsaturated alkyl.
  • the compound of Formula I as defined above is selected from: 5 ⁇ (1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide
  • the prodrugs are selected from phosphate or carboxylic acid or aminoacid ester prodrugs.
  • the compounds of Formula I may be defined wherein: Het represents pyrazolo[1 ,5-a]pyridine (below);
  • X may represent one or two of halogen, R 1 , OR 1 , OCOR 1 , CONR 1 2 , CO 2 R 1 , SO 2 R 1 , SO 2 NR 1 2 ,
  • Y is H or Me;
  • D is phenyl and is optionally substituted at any available position with up to two of halogen, R 1 , OR 1 , CONR 1 2 , CO 2 R 1 , SO 2 R 1 , SO 2 NR 1 2 , CN, CF 3 , OCF 3 , NO 2 , NR 1 2 , NR 1 COR 1 ;
  • R 1 is H or C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R 3 , OR 3 , NR 3 2 , OCOR 3 , CONR 3 2 , CO 2 R 3 , CN, SO 2 R 3 , SO 2 NR 3 2 , NR 3 COR 3 or optionally substituted aryl or heteroaryl, or in the case where R 1 forms part of NR 1 2 this may form an optionally substituted 4-7 membered saturated
  • R 2 is C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R 3 , OR 3 , NR 3 2 , OCOR 3 , CONR 3 2 , CO 2 R 3 , CN, SO 2 R 3 , SO 2 NR 3 2 , NR 3 COR 3 or optionally substituted aryl or heteroaryl;
  • R 3 is H, C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R 4 , OR 4 , NR 4 2 , OCOR 4 , CONR 4 2 , CO 2 R 4 , CN, SO 2 R 4 , SO 2 NR 4 2 , NR 4 COR 4 ; or optionally substituted aryl or heteroaryl, or in the case where R 3 forms part of NR 3 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR 4 ;
  • the prodrugs are selected from phosphate or carboxylic acid or aminoacid ester prodrugs.
  • a method of cancer prevention or therapy for treating cancers including the step of administering a compound of Formula I as defined in the first or second aspects.
  • the method further includes administering one or more chemotherapeutic agents and/or therapies.
  • agents and/or therapies are selected from:
  • Antitumour antibiotics Antitumour vegetable alkaloids
  • the method further includes the step of administering one or more chemotherapeutic agents to the subject before, during or after the administration of the compound of Formula I to the subject.
  • a compound of Formula I is administered to a human or other primate.
  • Formula I is administered to a farm, sports, or pet animal.
  • a pharmaceutical composition including a compound of Formula I as defined in the first or second aspects, and a pharmaceutically acceptable excipient, adjuvant, carrier, buffer or stabiliser.
  • the pharmaceutical composition is adapted for oral or parenteral administration.
  • the pharmaceutical composition is adapted for cutaneous, subcutaneous, or intravenous injection.
  • the pharmaceutical composition is in a tablet, capsule, powder, or liquid form.
  • the pharmaceutical composition further includes one or more chemotherapeutic agents.
  • chemotherapeutic agents are selected from any one or more of:
  • Antitumour immunological agents In a fifth aspect there is provided the use of a compound of Formula I as defined in the first or second aspects, in the manufacture of a ⁇ medicament for cancer prevention or therapy for the treatment of cancer.
  • the medicament is in tablet, capsule, powder or liquid form.
  • the medicament is suitable for oral or parenteral administration.
  • a seventh aspect there is provided a method of making a compound of Formula I as defined in the first or second aspect, the method including the step of modifying a pyrazolo[1 ,5- a]pyridine-3-carbonyl compound of Formula Il or an imidazo[1 ,2-a]pyridine-3-carbonyl compound of Formula III
  • variables X, Y and R 2 are defined above as for Formula I.
  • a eighth aspect there is provided a method of making a compound of Formula I as defined in the first or second aspects, according to any one or more of the steps as substantially described in Schemes 1 to 7.
  • variables X, Y and R 2 are defined according to the first or second aspect, the method including any one of the following steps: (i) condensation with a sulfonohydrazide; or
  • a compound of Formula I according to the first or second aspects when produced by a method according to the seventh, eighth and/or ninth aspects.
  • the present invention broadly relates to a new class of compounds for use as agents or drugs for cancer prevention, therapy and related methods.
  • PI3K inhibitors are thought to be valuable for the treatment of cell proliferation disorders and particularly as anti tumour agents.
  • Het represents pyrazolo[1 ,5-a]pyridine or imidazo[1 ,2-a]pyridine (below);
  • R 1 is H or C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R 3 , OR 3 , NR 3 2 , OCOR 3 , CONR 3 2 , CO 2 R 3 , CN, SO 2 R 3 , SO 2 NR 3 2 , NR 3 COR 3 or optionally substituted aryl or heteroaryl, or in the case where R 1 forms part of NR 1 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR 4 ;
  • R 2 is C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R 3 , OR 3 , NR 3 2 , OCOR 3 , CONR 3 2 , CO 2 R 3 , CN, SO 2 R 3 , SO 2 NR 3 2 , NR 3 COR 3 or optionally substituted aryl or heteroaryl;
  • R 3 is H, C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R 4 , OR 4 , NR 4 2 , OCOR 4 , CONR 4 2 , CO 2 R 4 , CN, SO 2 R 4 , SO 2 NR 4 2 , NR 4 COR 4 ; or optionally substituted aryl or heteroaryl, or in the case where R 3 forms part of NR 3 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR 4 ;
  • R 4 is H
  • X represents one or two of halogen, R 1 , OR 1 , OCOR 1 ,
  • X may represent one or two of halogen, R 1 , OR 1 , OCOR 1 , CONR 1 2 , CO 2 R 1 , SO 2 R 1 , SO 2 NR 1 2 , CN, NO 2 , NR 1 2 or NR 1 COR 1 , where one of X is placed at position 5- where Het represents pyrazolo[1 ,5-a]pyridine or at position 6- where Het represents imidazo[1 ,2- a]pyridine, and the other X if present is placed at any of the remaining available positions 4-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the remaining available positions 5-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine, where R 1 is defined as above.
  • D is phenyl and is optionally substituted at any available position with up to two of halogen, R 1 , OR 1 , CONR 1 2 , CO 2 R 1 , SO 2 R 1 , SO 2 NR 1 2 , CN, CF 3 , OCF 3 , NO 2 , NR 1 2 , or NR 1 COR 1 , where R 1 is defined as above.
  • Het represents pyrazolo[1 ,5-a]pyridine (below);
  • R 4 is H, C1-6 saturated or unsaturated alkyl.
  • the compound of Formula I as defined above is selected from: /V-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide
  • physiologically acceptable salt used throughout the specification is to be taken as meaning any suitable acid or base derived salt and, in particular, those formed from hydrochloric, sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic, maleic, methanesulfonic, isoethonic acids and the like and potassium carbonate, sodium or potassium hydroxide, ammonia, triethylamine, triethanolamine and the like.
  • prodrug means any compound which releases an active parent drug according to formula (I) in vivo when such prodrug is administered to a subject.
  • Prodrugs of a compound of formula (I) are prepared by modifying functional groups present in the compound of formula (I) in such a way that the modifications may be cleaved in vivo to release the parent compound.
  • prodrugs include phosphate prodrugs of phenols or alcohols, or carboxylic acid ester or amino acid ester prodrugs.
  • Such prodrugs may be made by any number of standard methods recognised in the art.
  • phosphate prodrugs may be prepared by methods similar to those described by G. S. Gill et a/., Org. Prep. Proc. Int. 2006, 38(6), 604, and amino acid ester prodrugs may be prepared by methods similar to those described by L. Ribeiro et a/., Arch. Pharm. 2007, 340, 32.
  • Suitable prodrugs include phosphate or carboxylic acid or aminoacid ester derivatives of Formula I.
  • halo or halogen group used throughout the specification is to be taken as meaning a fluoro, chloro, bromo or iodo group.
  • Suitable substitution for the optionally substituted aryl, heteroaryl and 4-7 membered saturated rings include alkyl, unsaturated alkyl, hydroxyalkyl, alkoxyalkyl, O-alkyl, C(O)-alkyl, C(O)NH 2 , C(O)NH(alkyl), C(O)N(alkyl) 2 , CO 2 H, CO 2 -alkyl, S-alkyl, S(O)-alkyl, SO 2 -alkyl, SO 2 NH 2 , SO 2 NH(alkyl), SO 2 N(alkyl) 2 , OH, CN, CF 3 , OCF 3 , NO 2 , NH 2 , NH(alkyl), N(alkyl) 2 , N(alkyl)C(O)(alkyl), halo.
  • the compounds of Formula I in addition to the biological activity as PI3K inhibitors, have been found to have a level of metabolic stability that is unexpected.
  • Metabolic stability of a drug molecule is a desirable property, since extensive metabolism reduces the systemic exposure and decreases the half-life of the compound. The rate of metabolism can be assessed using a liver microsomal assay. Greater stability of a drug molecule in liver microsomes often means that the drug will have improved metabolic stability in vivo [Li, Curr. Top. Med. Chem. 2004, 4, 701].
  • a method of cancer prevention or therapy for treating cancers including the step of administering a compound of Formula I to a subject in need thereof.
  • the method includes administration of a compound of Formula I together with administering one or more suitable chemotherapeutic agents and/or therapies to a patient in need thereof.
  • agents and therapies can be of any suitable type as would be well known to a skilled person. However, a non-limiting list would include agents and therapies selected from:
  • Alkylation agents for example cisplatin, carboplatin
  • Antimetabolites for example methotrexate, 5-FU
  • Antitumour antibiotics for example adriamymycin, bleomycin
  • Antitumour vegetable alkaloids for example taxol, etoposide
  • Antitumour hormones for example dexamethasone, tamoxifen
  • Antitumour immunological agents for example interferon ⁇ , ⁇ , y
  • the method can further include the step of administering one or more chemotherapeutic agents to the subject before, during or after the administration of the compound of Formula I to the subject.
  • While these compounds of Formula I will typically be used in cancer prevention or cancer therapy of human subjects, they can be used to target cancer cells in other warm blooded animal subjects such as other primates, farm animals such as cattle, and sports animals and pets such as horses, dogs, and cats.
  • reference to cancer prevention or cancer therapy is not intended to be a reference to a cure for cancer or to absolute prevention.
  • the reference is intended to include reference to a reduction in the likelihood of contraction of cancer or a mitigation of development, or like outcome.
  • a pharmaceutical composition including a compound of Formula I as defined above, and a pharmaceutically acceptable excipient, adjuvant, carrier, buffer or stabiliser.
  • the pharmaceutical composition will preferably take the form of a tablet, capsule, powder, or liquid form. It is further preferred that the composition will be suitable for oral or parenteral administration.
  • the pharmaceutically acceptable excipient, adjuvant, carrier, buffer or stabiliser can be of any known type and should be non-toxic and should not interfere with the efficacy of the active ingredient.
  • the precise nature of the carrier or other material will depend on the route of administration, which may be oral, or by injection, such as cutaneous, subcutaneous, or intravenous injection.
  • compositions of the invention formulated for oral administration may be in tablet, capsule, powder or liquid form.
  • a tablet may comprise a solid carrier or an adjuvant.
  • Liquid pharmaceutical compositions generally comprise a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil. Physiological saline solution, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol may be included.
  • a capsule may comprise a solid carrier such as gelatin.
  • compositions may be formulated for intravenous, cutaneous or subcutaneous injection
  • the active ingredient will be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has a suitable pH, isotonicity and stability.
  • a parenterally acceptable aqueous solution which is pyrogen-free and has a suitable pH, isotonicity and stability.
  • isotonic vehicles such as Sodium Chloride injection, Ringer's injection, Lactated Ringer's injection.
  • Preservatives, stabilisers, buffers antioxidants and/or other additives as are known to be suitable for such use may be included as required.
  • compositions preferably also include one or more chemotherapeutic agents as defined above.
  • the medicament is preferably in the form of a tablet, capsule, powder or liquid form.
  • the medicament will preferably be suitable for oral or parenteral administration.
  • the medicament will be formulated as described above.
  • a method of making a compound of Formula I as defined above including the step of modifying a pyrazolo[1 ,5-a]- pyridine-3-carbonyl compound of Formula Il or an imidazo[1 ,2-a]pyridine-3-carbonyl compound of Formula III
  • variables X, Y and R 2 are defined above as for Formula I.
  • the compounds of Formula I can preferably be made according to any one or more of the steps as described in Schemes 1 to 7 in the section titled "Methods for preparing compounds".
  • the method of making a compound of Formula I from compound of Formula Il or Formula III involves one of the following steps:
  • ketone 3 can be made by ⁇ /-amination of pyridine 1 using a suitable O-substituted hydroxyl- amine such as O-(mesitylsulfonyl)hydroxylamine or O-(2,4-dinitrophenyl)hydroxylamine to form ⁇ /-aminopyridinium 2.
  • a suitable O-substituted hydroxyl- amine such as O-(mesitylsulfonyl)hydroxylamine or O-(2,4-dinitrophenyl)hydroxylamine to form ⁇ /-aminopyridinium 2.
  • Cyclisation under basic conditions with a suitable alkyne forms substituted pyrazolo[1 ,5-a]pyridine 3.
  • carboxylic acid derivative 4 (where LG is a suitable leaving group) can be reacted with bis(trimethylsilyl)acetylene to form ketone 5. Then reaction with N- aminopyridinium 2 under basic conditions with the addition of a fluoride source gives pyrazolo[1 ,5-a]pyridine 6.
  • halogen-containing compounds 9 The synthesis of halogen-containing compounds 9 is depicted in Scheme 3.
  • Protected amine 7 can be deprotected with acid to form amine 8, which can then undergo a Sandmeyer reaction to introduce the halogen atom.
  • Compound 12 can be made, as shown in Scheme 4, either by condensation of ketone 10 with sulfonohydrazide 11 , or by condensation with hydrazine followed by sulfonylation with a suitably substituted sulfonyl chloride 13.
  • fluorobenzene 14 can be substituted by a primary or secondary amine to form amine 15, or by an alcohol with NaH to form ether 16.
  • Amine 18 can be made according to Scheme 6. Hydroxyalkylpiperazine 17 can be reacted with a minimum of two molar equivalents of a sulfonyl chloride, and then with a primary amine to form secondary amine 18.
  • ⁇ /-Methylation of 19 can be carried out using CH 2 N 2 to afford 20, as shown in Scheme 7.
  • Table 1 gives examples of compounds representative of the invention, and preparable by the methods outlined in Schemes 1-7.
  • Example 1 /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-methyl-5-nitro- benzenesulfonohydrazide (E1).
  • Step 1.1 A fresh solution of 0-(mesitylsulfonyl)hydroxylamine in CH 2 CI 2 (30 mL, 0.38 mol L "1 , 11.4 mmol) [T. Eichenberger ef a/., HeIv. Chim. Acta 1986, 69(6), 1521] was added to 4- pyridinecarbonitrile (1.19 g, 11.4 mmol) in CH 2 CI 2 (20 mL) at 0 0 C. After 2 h, the solvent was removed in vacuo to leave crude 1-amino-4-cyanopyridinium 2-mesitylenesulfonate as a yellow solid.
  • Step 1.2 A solution of 3-acetylpyrazolo[1 ,5-a]pyridine-5-carbonitrile (50 mg, 0.27 mmol) and 2- methyl-5-nitrobenzenesulfonohydrazide (69 mg, 0.30 mmol) [I. Kh. Fel'dman et al., Zh. Obshch. Khim., 1963, 33, 38] in MeOH (5 mL) was refluxed for 18 h.
  • Example 2 AT-(I -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)- ⁇ /,2-dimethyl-5-nitro- benzenesulfonohydrazide (E2).
  • Example 3 /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)propylidene)-2-methyl-5-nitro- benzenesulfonohydrazide (E3).
  • Step 3.1 AICI 3 (921 mg, 6.91 mmol) was added to a solution of propionic anhydride (0.74 mL, 5.76 mmol) and bis(trimethylsilyl)acetylene (1.43 mL, 6.31 mmol) in dry CH 2 CI 2 (20 mL) at 0 0 C.
  • Step 3.2 A solution of 1-(trimethylsilyl)pent-1-yn-3-one (520 mg, 3.38 mmol) in DMF (5 mL) was added to a suspension of 1-amino-4-cyanopyridinium 2-mesitylenesulfonate (1.62 g, 5.08 mmol), KF (589 mg, 10.1 mmol) and K 2 CO 3 (1.4Og, 10.1 mmol) in DMF (20 mL) at 0 0 C, and then slowly warmed to room temperature overnight. The reaction mixture was diluted with water and extracted twice with EtOAc. The combined extracts were washed with water, 1 M HCI, water, brine, and then dried (Na 2 SO 4 ) and the solvent removed in vacuo.
  • Step 3.3 Reaction of 3-propionylpyrazolo[1 ,5-a]pyridine-5-carbonitrile (71 mg, 0.36 mmol) using the conditions of Step 1.2 gave ⁇ -( ⁇ ( ⁇ -cyanopyrazoloti . ⁇ -aJpyridin-S-yOpropylidene) ⁇ - methyl-5-nitrobenzenesulfonohydrazide (E3) as a yellow solid (105 mg, 71%).
  • Example 4 /V-(1 -(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)propylidene)- ⁇ /,2-dimethyl-5-nitro- benzenesulfonohydrazide (E4).
  • Example 5 /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)butylidene)-2-methyl-5-nitro- benzenesulfonohydrazide (E5).
  • Step 5.1 Reaction of butyryl chloride (2.00 mL, 19.3 mmol) using the conditions of Step 3.1 gave 1-(trimethylsilyl)hex-1-yn-3-one as a brown oil (2.98 g, 92%).
  • 1 H NMR ⁇ 400 MHz, CDCI 3 ) 2.54 (t, J 7.3 Hz, 2H), 1.70 (m, 2H) 1 0.95 (t, J 7.4 Hz, 3H), 0.24 (s, 9H).
  • Step 5.2 Reaction of 1-(trimethylsilyl)hex-1-yn-3-one (147 mg, 0.88 mmol) using the conditions of Step 3.1 gave 3-butyrylpyrazolo[1 ,5-a]pyhdine-5-carbonitrile as an off-white solid (153 mg, 82%).
  • Step 5.3 Reaction of 3-butyrylpyrazolo[1 ,5-a]pyridine-5-carbonitrile (57 mg, 0.27 mmol) using the conditions of Step 3.1 gave /V-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)butylidene)-2-methyl- 5-nitrobenzenesulfonohydrazide (E5) as a yellow solid (68 mg, 60%).
  • Example 6 W-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)butylidene)-A/,2-dimethyl-5-nitro- benzenesulfonohydrazide (E6).
  • Example 7 /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5- nitrobenzenesulfonohydrazide (E7).
  • Step 7.1 A solution of 3-acetylpyrazolo[1 ,5-a]pyridine-5-carbonitrile (295 mg, 1.59 mmol) and hydrazine monohydrate (402 mg, 8.04 mmol) in MeOH (4 mL) heated at 65 0 C for 3 h. After cooling to room temperature, the residue was concentrated in vacuo to leave 3-(1-hydrazono- ethyl)pyrazolo[1 ,5-a]pyridine-5-carbonitrile as a yellow solid which was used in the next step without purification.
  • Step 7.2 To a solution of 3-(1-hydrazonoethyl)pyrazolo[1 ,5-a]pyridine-5-carbonitrile in CH 2 CI 2 (3 mL) and pyridine (0.57 mL, 7.05 mmol), was added 2-fluoro-5-nitrobenzenesulfonyl chloride (400 mg, 1.67 mmol). Reaction mixture was stirred at room temperature for 1 h. The solvents were removed in vacuo.
  • Example 8 2-((2-Aminoethyl)(methyl)amino)- ⁇ / l -(1 -(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide (E8).
  • Example 9 ⁇ P-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(methyl- amino)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E9).
  • Example 10 /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)- ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E10).
  • Example 11 ⁇ P-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)- ethyl)(methyl)amino)- ⁇ /-methyl-5-nitrobenzenesulfonohydrazide (E11 ).
  • Example 12 ⁇ T-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((3-(dimethylamino)- propyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E12).
  • Example 14 /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)- ethyl)(methyl)amino)- ⁇ /-methyl-5-nitrobenzenesulfonohydrazide (E14).
  • Example 15 /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(piperidin- 1-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E15).
  • Example 17 AT-(I -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(3- morpholinopropyl)amino)-5-nitrobenzenesulfonohydrazide (E17).
  • Example 18 /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4- (methylsulfonyl)piperazin-i -yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E18).
  • Step 18.1 Methanesulfonyl chloride (2.98 mL, 38.5 mmol) was added dropwise to a solution of 1-(2-hydroxyethyl)piperazine (2.00 g, 15.4 mmol) and NEt 3 (5.4 mL, 38.5 mmol) in CH 2 CI 2 (20 mL) at 0 0 C. After stirring for 10 mins, methylamine (10 mL, 33 wt. % solution in EtOH) was added, and the resulting solution stirred at room temperature for 24 h. The solvents were removed in vacuo.
  • Step 18.2 Reaction of E7 (60 mg, 0.15 mmol) and ⁇ /-methyl-2-(4-(methylsulfonyl)piperazin-1- yl)ethanamine (99 mg, 0.45 mmol) using the conditions of Example 9 gave ⁇ / ⁇ H ⁇ -cyano- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4-(methylsulfonyl)piperazin-1-yl)ethyl)- amino)-5-nitrobenzenesulfonohydrazide (E18) as a yellow solid (47 mg, 52%).
  • Example 19 ⁇ r-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-2-yl- methyl)amino)-5-nitrobenzenesulfonohydrazide (E19).
  • Example 20 ⁇ T-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-/V-methyl-2-(methyl- (pyridin-2-ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E20).
  • Example 21 ⁇ T-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-4- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E21 ).
  • Example 22 ⁇ T-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-2- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E22).
  • Example 23 /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-/V-methyl-2-(methyl(2- (pyridin-2-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E23).
  • Example 24 AT-(I -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-4- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E24).
  • Example 25 2-((2-(1W-lmidazol-1-yl)ethyl)(methyl)amino)- ⁇ r-(1-(5-cyanopyrazolo[1,5-a]- pyridin-3-yl)ethylidene)-5-nitrobenzenesulfonohydrazide (E25).
  • Example 26 2-((2-(1H-lmidazol-4-yl)ethyl)(methyl)amino)-/V-(1-(5-cyanopyrazolo[1,5-a]- pyridin-3-yl)ethylidene)-5-nitrobenzenesulfonohydrazide (E26).
  • Example 27 /V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-methyl-5-nitro- benzenesulfonohydrazide (E27).
  • Step 27.1 A solution of 4-(terf-butoxycarbonylamino)pyridine (4.94 g, 25.4 mmol) and O-(2,4- dinitrophenyl)hydroxylamine (5.06 g, 25.4 mmol) [C. Legault et al., J. Org. Chem. 2003, 68(18), 7119] in MeCN (100 mL) was heated at 40 0 C for 18 h. The solvent was removed in vacuo, and then the residue was taken up in DMSO (150 mL) and cooled to 0 0 C. K 2 CO 3 (7.03 g, 50.9 mmol) followed by 3-butyn-2-one (1.99 mL, 25.4 mmol) were then added.
  • Step 27.2 A solution of tert-butyl 3-acetylpyrazolo[1 ,5-a]pyridin-5-ylcarbamate (81 mg, 0.29 mmol) and trifluoroacetic acid (0.43 ml_, 5.8 mmol) in CH 2 CI 2 (5 mL) was stirred at room temperature for 18h. The solvents were removed in vacuo to leave the trifluoroacetate salt of 1-(5-aminopyrazolo[1 ,5-a]pyridin-3-yl)ethanone as a brown solid (86 mg, 100%).
  • Step 27.3 A solution of NaNO 2 (0.378 g, 5.5 mmol) in H 2 O (4 mL) was added dropwise to a suspension of the trifluoroacetate salt of 1-(5-aminopyrazolo[1 ,5-a]pyridin-3-yl)ethanone (1.7g, 4.2 mmol) in 48% HBr (5 mL) in an ice-salt bath at a rate so that the internal temperature of the reaction was maintained below -5 °C. The reaction was stirred for 10 mins, then a solution of CuBr (1.51 g, 10.5 mmol) in 48% HBr (5 mL) was added dropwise to maintain the internal temperature of the reaction between -5 and 0 °C.
  • Step 27.4 Reaction of 1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethanone (60 mg, 0.25 mmol) using the conditions of Step 1.2 gave / ⁇ (1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2- methyl-5-nitrobenzenesulfonohydrazide (E27) as an orange solid (88 mg, 78%).
  • Example 28 /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-/V,2-dimethyl-5-nitro- benzenesulfonohydrazide (E28).
  • Example 29 /V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5-nitro- benzenesulfonohydrazide (E29).
  • Step 29.1 Reaction of 1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethanone (155 mg, 0.65 mmol) using the conditions of Step 7.1 gave 5-bromo-3-(1-hydrazonoethyl)pyrazolo[1 ,5-a]pyridine as a yellow solid.
  • Step 29.2 Reaction of crude 5-bromo-3-(1-hydrazonoethyl)pyrazolo[1 ,5-a]pyridine using the conditions of Step 7.2 gave /V-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5- nitrobenzenesulfonohydrazide (E29) as a yellow solid (66 mg, 22% over 2 steps).
  • Example 30 /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(methyl- amino)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E30).
  • Example 31 ⁇ f-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2- (dimethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E31).
  • Example 32 ⁇ f-(1 -(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)- ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E32).
  • Example 33 /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)- ethyl)(methyl)amino)- ⁇ /-methyl-5-nitrobenzenesulfonohydrazide (E33).
  • Example 35 /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4- (methylsulfonyl)piperazin-1-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E35).
  • Example 36 ⁇ P-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-2- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E36).
  • Example 38 /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-3- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E38).
  • Example 39 /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-4- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E39).
  • Example 41 /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-/V-methyl-2-(methyl(2- (pyridin-2-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E41).
  • Example 42 ⁇ f-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin- 4-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E42).
  • Example 44 2-((2-(1 W-lmidazol-4-yl)ethyl)(methyl)amino)-/V-(1 -(5-bromopyrazolo[1 ,5-a]- pyridin-3-yl)ethylidene)-5-nitrobenzenesulfonohydrazide (E44).
  • Example 45 /V-(1-(6-Bromoimidazo[1,2-a]pyridin-3-yl)ethylidene)-2-fluoro-5-nitro- benzenesulfonohydrazide (E45).
  • Incubation mixtures were prepared by combining 32 ⁇ L of the test compound solution (50 ⁇ L of 0.5 mM stock solution diluted to 5 mL with MiIIi Q water), 32 ⁇ L of MgCI 2 - EDTA solution (24 mM MgCI 2 .6H 2 O and 5 mM EDTA tetra sodium salt in MiIIi Q water), 64 ⁇ L microsome solution (1 part pooled male CD1 mouse liver microsomes supplied by Gentest: 1 part MiIIi Q water: 6 parts 400 mM pH 7.4 phosphate buffer), and 32 ⁇ L of 5 mM NADPH in MiIIi Q water.
  • the resulting supernatants were diluted with water 1:1 v/v (100 ⁇ L aliquots of each) prior to LCMS- QQQ analysis (Agilent Zorbax SB-C18 2.1x50mm ⁇ micron column using a gradient of 20% of solution A (5 mM NH 4 CO 2 H and 0.1% HCO 2 H in water) and 80% of solution B (5 mM NH 4 CO 2 H and 0.1% HCO 2 H in methanol) to 100% of solution B over 5 mins, at a flow rate of 0.4 mL/min).
  • solution A 5 mM NH 4 CO 2 H and 0.1% HCO 2 H in water
  • solution B 5 mM NH 4 CO 2 H and 0.1% HCO 2 H in methanol
  • IC 50 values are an average of two or more determinations, except for those in italics which are a single determination.
  • the compounds described in Table 1 are all inhibitors of Pl 3-kinase. In particular, they inhibit at least one of the Pl 3-kinase isoforms with IC 50 ⁇ 1 ⁇ M. Some of these examples have IC 50 ⁇ 10 nM. In addition, they show inhibition of cellular proliferation in at least one of the two cell lines described above with IC 50 ⁇ 20 ⁇ M.

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Abstract

The invention relates to pyrazolo[1,5-a]pyridine and/or -imidazo[1,2-a]pyridine derivatives, to their preparation, to their use as agents or drugs for cancer prevention or therapy for treating cancer, either alone or in combination with radiation and/or anticancer drugs.

Description

PYRAZOLO[1, 5-a] PYRIDIN E AND I MIDAZO[1, 2-a] PYRI DIN E DERIVATIVES AND THEIR USE IN CANCER THERAPY
TECHNICAL FIELD The present invention relates to pyrazolo[1 ,5-a]pyridine and/or -imidazo[1 ,2-a]pyridine derivatives, to their preparation, to their use as agents or drugs for cancer prevention or therapy for treating cancer, either alone or in combination with radiation and/or anticancer drugs.
BACKGROUND TO THE INVENTION
Phosphoinositide-3-kinases (PI3Ks) are a group of lipid kinases which phosphorylate the 3- hydroxyl of phosphoinositides. They are split into three classes (Class I, Il and III) and play an important role in cellular signalling [Stephens et al., Curr. Opin. Pharmacol. 2005, 5, 357]. The Class I enzymes are further split into Class Ia and Ib based on their mechanism of activation; the Class Ia PI3Ks are heterodimeric structures consisting of a catalytic subunit (p110α, p110β or p110δ) in complex with a regulatory p85 subunit, while the Class Ib PI3K (p110γ) is structurally similar but lacks a regulatory subunit, and instead is activated by βy subunits of heterotrimeric G-proteins [Walker et al,. MoI. Cell., 2000, 6, 909].
PI3Ks play a variety of roles in normal tissue physiology [Foukas & Shepherd, Biochem. Soc. Trans., 2004, 32, 330; Shepherd, Acta Physiol. Scand., 2005, 183, 3], with p110α having a specific role in cancer growth, p110β in thrombus formation mediated by integrin αnβ3 [Jackson et al., Nat. Med., 2005, 11 , 507], and p110γ in inflammation, rheumatoid arthritis [Camps et al., Nat. Med., 2005, 11 , 936] and other chronic inflammation states [Barber et al., Nat. Med., 2005, 11 , 933]. The PI3K enzymes produce phosphoinositide 3,4,5-triphosphate (PIP3) from the corresponding diphosphate (PIP2), thus recruiting AKT (protein kinase B) through its PH domain, to the plasma membrane. Once bound, AKT is phosphorylated and activated by other membrane bound kinases, and is central to a cascade of events that lead to inhibition of apoptosis [Berrie, Exp. Opin. Invest. Drugs, 2001 , 10, 1085].
The p110α isoform is selectively amplified and activated in a number of cancer types [Stephens et al., Curr. Opin. Pharmacol., 2005, 5, 357; Stauffer et al., Curr. Med. Chem. - Anti- Cancer Agents, 2005, 5, 449]. In addition there is a high frequency of non- random mutations in specific sites (primarily in the C2 domain and or the activation loop) of the kinase in several human cancer cell lines, including colon, brain, breast and stomach [Samuels et al., Science,
2004, 304, 554]. This results in a constitutively active enzyme [Ikenoue et al., Cancer Res.,
2005, 65, 4562; Kang et al., Proc. Natl. Acad. ScL USA, 2005, 102, 802], making p110α one of the most highly mutated oncogenes found in human tumours. Structural studies have shown that many of the mutations occur at residues lying at the interfaces between p110a and p85α or between the kinase domain of p110α and other domains within the catalytic subunit [Miled et al., Science, 2007, 317, 239; Huang et al., Science, 2007, 318, 1744].
While PI3K isoenzymes play important roles in many cellular processes, published experimental studies in mice with human tumour xenografts show that the pan-PI3K inhibitor LY294002 is well-tolerated, reduces signalling through the PI3K pathway, causes reduction of tumour volume, and is more active in cell lines over-expressing mutant forms of p110α than parental control cells [Semba et al., Clin. Cancer Res., 2002, 8, 1957; Hu et al., Cancer Res., 2002, 62, 1087].
Thus PI3K, and especially the p110α isoform, is an interesting target for drug intervention. Several classes of compounds have been identified as inhibitors; for example LY294002 (non- selective) [Walker et al., MoI .Cell., 2000, 6, 909], PI103 (slightly α-selective) [Knight et al., Cell, 2006, 125, 733; Hayakawa et al., Bioorg. Med. Chem. Lett., 2007, 17, 2438; Raynaud et al, Cancer Res. 2007, 67, 5840], ZSTK474 (non-selective) [Yaguchi et al., J. Natl. Cancer Inst, 2006, 98, 545; Kong et a I., Cancer Sci. 2007, 98, 1639], TGX221 (β-selective) [Jackson et al., Nat. Med., 2005, 11 , 507], oxazines (γ-selective) [Lanni et al., Bioorg. Med. Chem. Lett, 2007, 17, 756], IC87114 (δ-selective) [Sadhu et al., PCT Int Appl. WO 2001/81346, Nov 2001 ; Billottet et al, Oncogene, 2006, 25, 6648], AS605240 (γ-selective) [Camps et al., Nat. Med., 2005, 11 , 936], the imidazo[1 ,2-a]pyridines (α-selective) [Hayakawa et al., Bioorg. Med. Chem., 2007, 15, 403; Hayakawa et al., Bioorg. Med. Chem., 2007, 15, 5837], [WO 2001083481], aminothiazole (non-selective) [Knight et al., Ce//, 2006, 125, 733], imidazo[4,5- c]quinoline NVP-BEZ235 [Maira et al., MoI. Cancer Then, 2008, 7, 1851 ; Garcia-Echeverria, et al., PCT Int. Appl. WO 2006/122806, Nov 2006], GDC-0941 (α, δ-selective) [Folkes et al., J. Med. Chem., 2008, 51 , 5522]. PCT/NZ2008/000164 (our previous application) describes Pyrazolo[1 ,5-a]pyridines and their use in cancer therapy.
Figure imgf000004_0001
Figure imgf000004_0002
oxazines IC87114 AS605240 lmidazo[1 ,2-a]py ridine
Figure imgf000004_0003
Aminothiazole NVP-BEZ235
OBJECT OF THE INVENTION
It is an object of the present invention to provide a novel class of pyrazolo[1 ,5-a]pyridine or imidazo[1 ,2-a]pyridine derivatives and/or uses thereof in methods of cancer prevention or therapy and/or pharmaceutical compositions thereof and/or uses thereof in the manufacture of a medicament and/or methods of making the novel class of pyrazolo[1 ,5-a]pyridine or imidazo[1 ,2-a]pyridine derivatives or to at least provide the public with a useful alternative.
SUMMARY OF THE INVENTION
In a first aspect of the invention there is provided compounds of Formula I,
Figure imgf000004_0004
- A - wherein Het represents pyrazolo[1 ,5-a]pyridine or imidazo[1 ,2-a]pyridine (below);
Figure imgf000005_0001
pyrazolo[1 ,5-a]pyridine imidazo[1 ,2-a]pyridiπe
X may represent up to two of halogen, R1, OR1, OCOR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, NO2, NR1 2 or NR1COR1, placed at any of the available positions A-, 5-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the available positions 5-, 6-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine; Y is H or Me;
Z is CO, CR1 2, S(O)x, N(R1)CO, N(R1)CR1 2, N(R1JS(O)x where x = O, 1 , 2; D is phenyl, naphthyl, or 5- or 6-membered heterocycle or benzoheterocycle, where the heterocyclic ring contains up to two of the atoms S, O, N and is optionally substituted at any available position with up to two of halogen, R1, OR1, C0NR1 2, CO2R1, SO2R1, SO2NR1 2, CN, CF31 OCF31 NO21 NR1 Z1 NR1COR1 ;
R1 is H or C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R3, OR3, NR3 2, OCOR3, CONR3 2, CO2R3, CN1 SO2R3, SO2NR3 2, NR3COR3 or optionally substituted aryl or. heteroaryl, or in the case where R1 forms part of NR1 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR4;
R2 is C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R3, OR3, NR3 2, OCOR3, CONR3 2, CO2R3, CN1 SO2R3, SO2NR3 2l NR3COR3 or optionally substituted aryl or heteroaryl; R3 is H1 C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R4, OR4, NR4 2, OCOR4, CONR4 2, CO2R4, CN, SO2R4, SO2NR4 2, NR4COR4; or optionally substituted aryl or heteroaryl, or in the case where R3 forms part of NR3 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR4; R4 is H, C1-6 saturated or unsaturated alkyl, C(O)-alkyl, CO2-alkyl, S(O)y-alkyl where y = O, 1 , 2; including pure geometric and enantiomeric forms and mixtures thereof, physiologically acceptable salts thereof, and prodrugs thereof; with the proviso that the following compounds are excluded:
Λ^(1-(6-Bromoimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide;
Figure imgf000006_0001
hydrazide;
/V-(1-(6-Bromo-2-methylimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide; /V-(1 -(6-Bromo-2-methylimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-Λ/,2-dimethyl-5-nitrobenzene- sulfonohydrazide;
/V-BePZyI-Ay-(I -(6-bromo-2-methylimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-methyl-5-nitro- benzenesulfonohydrazide;
Preferably X may represent one or two of halogen, R1, OR1, OCOR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, NO2, NR1 2 or NR1COR1, placed at any of the available positions 4-, 5-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the available positions 5-, 6-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine, where R1 is defined as above.
Preferably X may represent one or two of halogen, R1, OR1, OCOR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, NO2, NR1 2 or NR1COR1, where one of X is placed at position 5- where Het represents pyrazolo[1 ,5-a]pyridine or at position 6- where Het represents imidazo[1 , 2- ajpyridine, and the other X, if present, is placed at any of the remaining available positions 4-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the remaining available positions 5-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine, where R1 is defined as above.
Preferably D is phenyl and is optionally substituted at any available position with up to two of halogen, R1, OR1, C0NR1 2, CO2R1, SO2R1, SO2NR1 2, CN, CF3, OCF3, NO2, NR1 2, or NR1COR1, where R1 is defined as above.
Preferably Z is N(R1)CO, N(R1)CR1 2, or N(R1)S(O)X where x = O, 1 , 2, where R1 is defined as above.
Preferably Z is N(R1)S(O)X where x = O, 1 , 2, where R1 is defined as above.
Preferably Het represents pyrazolo[1 ,5-a]pyridine (below);
Figure imgf000006_0002
pyrazolo[1 ,5-a]pyridine Preferably R4 is H1 C1-6 saturated or unsaturated alkyl.
Preferably, the compound of Formula I as defined above is selected from: 5 Λ^(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide
/V^I-CS-Cyanopyrazoloti .δ-aJpyridin-S-yOethylidene^Λ/^-dimethyl-S-nitrobenzenesulfono- hydrazide
^-(^(δ-Cyanopyrazoloti .S^pyridin-S-yOpropylideneJ^-methyl-δ-nitrobenzenesulfono- 0 hydrazide
^-(^(δ-CyanopyrazoloII .S-alpyridin-S-yOpropylideneJ-Λ/^-dimethyl-δ-nitrobenzenesulfono- hydrazide
Λ^1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)butylidene)-2-methyl-5- nitrobenzenesulfonohydrazide δ Λ/"-(1 -(δ-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)butylidene)-Λ/,2-dimethyl-5-nitrobenzenesulfono- hydrazide
/V-(1-(6-Cyanopyrazolo[1 ,6-a]pyridin-3-yl)ethylidene)-2-fluoro-6-nitrobenzenesulfonohydrazide
2-((2-Aminoethyl)(methyl)amino)-Λ/'-(1-(δ-cyanopyrazolo[1 ,δ-a]pyridin-3-yl)ethylidene)-δ-nitro- benzenesulfonohydrazide 0 /V-(1-(δ-Cyanopyrazolo[1 ,δ-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(methylamino)ethyl)amino)- δ-nitrobenzenesulfonohydrazide
Λ/"-(1 -(δ-Cyanopyrazolo[1 ,δ-a]pyridin-3-yl)ethylidene)-2-((2-
(dimethylaminoethyl)(methyl)amino)-δ-nitrobenzenesulfonohydrazide
Λ/'-(1-(δ-Cyanopyrazolo[1 ,δ-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)ethyl)(methyl)-δ amino)-Λ/-methyl-δ-nitrobenzenesulfonohydrazide
Λ/'-(1-(δ-Cyanopyrazolo[1 ,δ-a]pyridin-3-yl)ethylidene)-2-((3-(dimethylamino)propyl)(methyl)- amino)-δ-nitrobenzenesulfonohydrazide
/V-(1-(6-Cyanopyrazolo[1,6-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)- δ-nitrobenzenesulfonohydrazide 0 /V-(1-(6-Cyanopyrazolo[1 ,6-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-
Λ/-methyl-δ-nitrobenzenesuIfonohydrazide
/V-(1-(6-Cyanopyrazolo[1 ,6-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(piperidin-1-yl)ethyl)amino)- δ-nitrobenzenesulfonohydrazide
/V-(1-(6-Cyanopyrazolo[1 ,6-a]pyridin-3-yl)ethylidene)-A/-methyl-2-(methyl(2-(piperidin-1-5 yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
^-(i^δ-Cyanopyrazoloti .S-alpyridin-S-yOethylideneJ^methyKS-morpholinopropyOaminoJ-δ- nitrobenzenesulfonohydrazide /V-(1-(5-Cyanopyrazolo[1 ,5-a]pyndin-3-yl)ethylidene)-2-(methyl(2-(4-(methylsulfonyl)piperazin-
1-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-2-ylmethyl)amino)-5- nitrobenzenesulfonohydrazide Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(pyridin-2-ylmethyl)- amino)-5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-4-ylmethyl)amino)-5- nitrobenzenesulfonohydrazide
/V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-2-yl)ethyl)amino)-5- nitrobenzenesulfonohydrazide
/V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-A/-methyl-2-(methyl(2-(pyridin-2-yl)ethyl)- amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-4-yl)ethyl)amino)-5- nitrobenzenesulfonohydrazide 2-((2-(1H-lmidazol-1-yl)ethyl)(methyl)amino)-Λ/'-(1-(5-cyanopyrazolo[1,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide
2-((2-(1H-lmidazol-4-yl)ethyl)(methyl)amino)-Λ/'-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide
^-(^(δ-BromopyrazoloIi .δ-alpyridin-S-yOethylideneJ^-methyl-S- nitrobenzenesulfonohydrazide
^-(^(δ-BromopyrazoloII .S-alpyridin-S-yOethylideneJ-Λ/^-dimethyl-S-nitrobenzenesulfono- hydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-fIuoro-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(methylamino)ethyl)amino)- 5-nitrobenzenesulfonohydrazide
/\y-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)ethyl)(methyl)- amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-
5-nitrobenzenesulfonohydrazide /V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-
Λ/-methyl-5-nitrobenzenesulfonohydrazide
Λ/'-CI-CS-BromopyrazoloII .S-alpyridin-S-yOethylideneJ^methyKS-morpholinopropyOaminoJ-S- nitrobenzenesulfonohydrazide
Λ/l-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4-(methylsulfonyl)piperazin- 1 -yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
^-(^(δ-BromopyrazoloII .S-aJpyridin-S-yOethylideneJ^^methyKpyridin^-ylmethyOaminoJ-S- nitrobenzenesulfonohydrazide Λ/l-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(pyridin-2-ylrnethyl)- amino)-5-nitrobenzenesulfonohydrazide
^-(^(S-BromopyrazoloII .S-alpyridin-S-yOethylideneJ^-CmethyKpyridin-S-ylmethyOaminoJ-S- nitrobenzenesulfonohydrazide ^-(i-Cδ-BromopyrazoloII .S-alpyridin-S-yOethylidene^-CmethyKpyridin^-ylmethyOaminoJ-S- nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-2-yl)ethyl)amino)-5- nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(pyridin-2-yl)ethyl)- amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-4-yl)ethyl)amino)-5- nitrobenzenesulfonohydrazide
2-((2-(1H-lmidazol-1-yl)ethyl)(methyl)amino)-Λ/'-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide 2-((2-(1H-lmidazol-4-yl)ethyl)(methyl)amino)-Λ/'-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide
/^-(^(θ-BromoimidazoII ^-alpyridin-S-yOethylideneJ^-fluoro-δ-nitrobenzenesulfonohydrazide
Λ/l-(1-(6-Bromoimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-((2-
(dimethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide including pure geometric and enantiomeric forms and mixtures thereof, physiologically acceptable salts thereof, and prodrugs thereof.
Preferably the prodrugs are selected from phosphate or carboxylic acid or aminoacid ester prodrugs.
In a second aspect the compounds of Formula I may be defined wherein: Het represents pyrazolo[1 ,5-a]pyridine (below);
Figure imgf000009_0001
pyrazolo[1 ,5-a]pyrιdιne
X may represent one or two of halogen, R1, OR1, OCOR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2,
CN, NO2, NR1 2 or NR1COR1, where one of X is placed at position 5-, and the other X if present is placed at any of the remaining available positions 4-, 6-, 7-;
Y is H or Me; Z is N(R1)S(O)X where x = O, 1 , 2; D is phenyl and is optionally substituted at any available position with up to two of halogen, R1, OR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, CF3, OCF3, NO2, NR1 2, NR1COR1; R1 is H or C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R3, OR3, NR3 2, OCOR3, CONR3 2, CO2R3, CN, SO2R3, SO2NR3 2, NR3COR3 or optionally substituted aryl or heteroaryl, or in the case where R1 forms part of NR1 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR4;
R2 is C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R3, OR3, NR3 2, OCOR3, CONR3 2, CO2R3, CN, SO2R3, SO2NR3 2, NR3COR3 or optionally substituted aryl or heteroaryl;
R3 is H, C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R4, OR4, NR4 2, OCOR4, CONR4 2, CO2R4, CN, SO2R4, SO2NR4 2, NR4COR4; or optionally substituted aryl or heteroaryl, or in the case where R3 forms part of NR3 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR4;
R4 is H, C1-6 saturated or unsaturated alkyl, C(O)-alkyl, CO2-alkyl, S(O)y-alkyl where y = O, 1 ,
2; including pure geometric and enantiomeric forms and mixtures thereof, physiologically acceptable salts thereof, and prodrugs thereof.
Preferably the prodrugs are selected from phosphate or carboxylic acid or aminoacid ester prodrugs.
In a third aspect there is provided a method of cancer prevention or therapy for treating cancers including the step of administering a compound of Formula I as defined in the first or second aspects.
Preferably the method further includes administering one or more chemotherapeutic agents and/or therapies.
Preferably the agents and/or therapies are selected from:
Alkylation agents
Antimetabolites
Antitumour antibiotics Antitumour vegetable alkaloids
Antitumour hormones
Antitumour immunological agents Radiation therapy Surgery
Preferably the method further includes the step of administering one or more chemotherapeutic agents to the subject before, during or after the administration of the compound of Formula I to the subject.
Preferably the method of cancer prevention or therapy for treating cancers a compound of Formula I is administered to a human or other primate.
Preferably the method of cancer prevention or therapy for treating cancers a compound of
Formula I is administered to a farm, sports, or pet animal.
In a fourth aspect there is provided a pharmaceutical composition including a compound of Formula I as defined in the first or second aspects, and a pharmaceutically acceptable excipient, adjuvant, carrier, buffer or stabiliser.
Preferably the pharmaceutical composition is adapted for oral or parenteral administration.
Preferably the pharmaceutical composition is adapted for cutaneous, subcutaneous, or intravenous injection.
Preferably the pharmaceutical composition is in a tablet, capsule, powder, or liquid form.
Preferably the pharmaceutical composition further includes one or more chemotherapeutic agents.
Preferably the chemotherapeutic agents are selected from any one or more of:
Alkylation agents Antimetabolites
Antitumour antibiotics
Antitumour vegetable alkaloids
Antitumour hormones
Antitumour immunological agents. In a fifth aspect there is provided the use of a compound of Formula I as defined in the first or second aspects, in the manufacture of a^medicament for cancer prevention or therapy for the treatment of cancer.
Preferably the medicament is in tablet, capsule, powder or liquid form.
Preferably the medicament is suitable for oral or parenteral administration.
In a sixth aspect of the invention there is provided compounds of Formula I as defined in the first or second aspects, for use in cancer prevention or therapy for the treatment of cancer.
In a seventh aspect there is provided a method of making a compound of Formula I as defined in the first or second aspect, the method including the step of modifying a pyrazolo[1 ,5- a]pyridine-3-carbonyl compound of Formula Il or an imidazo[1 ,2-a]pyridine-3-carbonyl compound of Formula III
Figure imgf000012_0001
wherein variables X, Y and R2 are defined above as for Formula I.
In a eighth aspect there is provided a method of making a compound of Formula I as defined in the first or second aspects, according to any one or more of the steps as substantially described in Schemes 1 to 7.
In a ninth aspect the method of making a compound of Formula I as defined in the first or second aspect from a compound of Formula Il or Formula III
Figure imgf000012_0002
wherein variables X, Y and R2 are defined according to the first or second aspect, the method including any one of the following steps: (i) condensation with a sulfonohydrazide; or
(ii) condensation with hydrazine followed by reaction with a sulfonyl chloride; or (iii) condensation with hydrazine followed by reaction with a sulfonyl chloride and then nucleophilic substitution; or (iv) any of (i) to (Ni) followed by reaction with CH2N2.
In a tenth aspect there is provided a compound of Formula I according to the first or second aspects, when produced by a method according to the seventh, eighth and/or ninth aspects.
It is to be recognised that certain compounds may exist in one or more different enantiomeric or diastereomeric forms. It is to be understood that the enantiomeric or diastereomeric forms are included in the above aspects of the invention.
Further aspects of the present invention will become apparent from the following description given by way of example only and with reference to the accompanying synthetic schemes.
DETAILED DESCRIPTION OF THE INVENTION
The present invention broadly relates to a new class of compounds for use as agents or drugs for cancer prevention, therapy and related methods. In particular, it relates to a class of compounds that can be used as PI3K inhibitors. PI3K inhibitors are thought to be valuable for the treatment of cell proliferation disorders and particularly as anti tumour agents.
In one embodiment of the invention there is provided compounds of Formula I,
Figure imgf000013_0001
I wherein Het represents pyrazolo[1 ,5-a]pyridine or imidazo[1 ,2-a]pyridine (below);
Figure imgf000013_0002
pyrazolo[1,5-a]pyridine imidazo[1 ,2-a]pyridine
X may represent up to two of halogen, R1, OR1, OCOR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, NO2, NR1 2 or NR1COR1, placed at any of the available positions A-, 5-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the available positions 5-, 6-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine; Y is H or Me; Z is CO, CR1 Z, S(O)x, N(R1)CO, N(R1)CR1 2, N(R1JS(O)x where x = 0, 1 , 2; D is phenyl, naphthyl, or 5- or 6-membered heterocycle or benzoheterocycle, where the heterocyclic ring contains up to two of the atoms S, O, N and is optionally substituted at any available position with up to two of halogen, R1, OR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, CF3; OCF3, NO2, NR1 2, NR1COR1;
R1 is H or C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R3, OR3, NR3 2, OCOR3, CONR3 2, CO2R3, CN, SO2R3, SO2NR3 2, NR3COR3 or optionally substituted aryl or heteroaryl, or in the case where R1 forms part of NR1 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR4;
R2 is C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R3, OR3, NR3 2, OCOR3, CONR3 2, CO2R3, CN, SO2R3, SO2NR3 2, NR3COR3 or optionally substituted aryl or heteroaryl; R3 is H, C1-6 saturated or unsaturated alkyl and is optionally substituted with up to three of halogen, R4, OR4, NR4 2, OCOR4, CONR4 2, CO2R4, CN, SO2R4, SO2NR4 2, NR4COR4; or optionally substituted aryl or heteroaryl, or in the case where R3 forms part of NR3 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR4; R4 is H, C1-6 saturated or unsaturated alkyl, C(O)-alkyl, CO2-alkyl, S(O)y-alkyl where y = O, 1 , 2; including pure geometric and enantiomeric forms and mixtures thereof, physiologically acceptable salts thereof, and prodrugs thereof.
Except for the following compounds: Λ/HHe-Bromoimidazoπ ,2-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide;
/V-(1-(6-Bromoimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-/\/,2-dimethyl-5-nitrobenzenesulfono- hydrazide;
/^-(^(δ-Bromo^-methylimidazoII ^-alpyridin-S-yOethylideneJ^-methyl-δ- nitrobenzenesulfonohydrazide;
^-(^(e-Bromo^-methylimidazoII ^-alpyridin-S-yOethylideneJ-Λ/^-dimethyl-δ-nitrobenzene- sulfonohydrazide;
Λ/-Benzyl-Λ/'-(1-(6-bromo-2-methylimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-methyl-5-nitro- benzenesulfonohydrazide;
In a preferred form of the embodiment, X represents one or two of halogen, R1, OR1, OCOR1,
CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, NO2, NR1 2 or NR1COR1, placed at any of the available positions A-, 5-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the available positions 5-, 6-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine, where R1 is defined as above.
More preferably, X may represent one or two of halogen, R1, OR1, OCOR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, NO2, NR1 2 or NR1COR1, where one of X is placed at position 5- where Het represents pyrazolo[1 ,5-a]pyridine or at position 6- where Het represents imidazo[1 ,2- a]pyridine, and the other X if present is placed at any of the remaining available positions 4-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the remaining available positions 5-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine, where R1 is defined as above.
In a preferred form D is phenyl and is optionally substituted at any available position with up to two of halogen, R1, OR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, CF3, OCF3, NO2, NR1 2, or NR1COR1, where R1 is defined as above.
In a preferred form Z is N(R1)CO, N(R1)CR1 2, or N(R1)S(O)X where x = O, 1 , 2, where R1 is defined as above more preferably Z is N(R1JS(O)x where x = O, 1, 2.
While the compounds of Formula I have been found by the inventors to be inhibitors of Pl 3- kinase, the prefered options for X, D and Z above tend to result in compounds with even more potent activity.
In another form it is preferred that Het represents pyrazolo[1 ,5-a]pyridine (below);
Figure imgf000015_0001
pyrazolo[1 ,5-a]pyridine
In another preferred form R4 is H, C1-6 saturated or unsaturated alkyl.
In a further preferred form, the compound of Formula I as defined above is selected from: /V-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide
^-(i^δ-Cyanopyrazolofi .S-alpyridin-S-yOethylideneJ-Λ/^-dimethyl-S-nitrobenzenesulfono- hydrazide
^-(I^S-CyanopyrazoloII .S-alpyridin-S-yOpropylideneJ^-methyl-δ-nitrobenzenesulfono- hydrazide ^-(^(S-Cyanopyrazoloti .S-alpyridin-S-ylJpropylideneJ-Λ/^-dimethyl-δ-nitrobenzenesulfono- hydrazide yV-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)butylidene)-2-methyl-5- nitrobenzenesulfonohydrazide /V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)butylidene)-/\/,2-dinnethyl-5-nitrobenzenesulfono- hydrazide
Λ^(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5-nitrobenzenesulfonohydrazide
2-((2-Aminoethyl)(rnethyl)arnino)-Λ/I-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-5-nitro- benzenesulfonohydrazide Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(methylamino)ethyl)arnino)-
5-nitrobenzenesulfonohydrazide
/V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-
(dimethylaminoethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide
Λ/1-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)ethyl)(methyl)- amino)-Λ/-methyl-5-nitrobenzenesulfonohydrazide
Λ/l-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((3-(dirnethylamino)propyl)(methyl)- amino)-5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-
5-nitrobenzenesulfonohydrazide Λ/l-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-
Λ/-methyl-5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(piperidin-1-yl)ethyl)amino)-
5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(piperidin-1- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
/^-(I^S-Cyanopyrazoloti .δ-alpyridin-S-yOethylidene^^methyKS-morpholinopropyOaminoJ-S- nitrobenzenesulfonohydrazide
/V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4-(methylsulfonyl)piperazin-
1-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide ^-(i-CS-Cyanopyrazoloti .S-aJpyridin-S-yOethylidene^-CmethyKpyridin^-ylmethyOaminoJ-S- nitrobenzenesulfonohydrazide
Λ/l-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(pyridin-2-ylmethyl)- amino)-5-nitrobenzenesulfonohydrazide
^-(i^δ-CyanopyrazoloII .S-alpyridin-S-yOethylideneJ^methyKpyridin^-ylmethyOaminoJ-S- nitrobenzenesulfonohydrazide
Λ/1-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-2-yl)ethyl)amino)-5- nitrobenzenesulfonohydrazide /V-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(pyridin-2-yl)ethyl)- amino)-5-nitrobenzenesulfonohydrazide
Λ/l-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-4-yl)ethyl)amino)-5- nitrobenzenesulfonohydrazide 2-((2-(1/-/-lmidazol-1-yl)ethyl)(methyl)amino)-Λ/'-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide
2-((2-(1/-/-lmidazol-4-yl)ethyl)(methyl)amino)-Λ/'-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide
^-(^(δ-BromopyrazoloII .S-alpyridin-S-yOethylideneJ-Λ/^-dimethyl-S-nitrobenzenesulfono- hydrazide
^-(^(S-BromopyrazoloII .S-alpyridin-S-yOethylideneJ^-fluoro-S-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(methylamino)ethyl)arnino)- 5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)ethyl)(methyl)- amino)-5-nitrobenzenesulfonohydrazide
Λ/l-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylarnino)ethyl)(methyl)amino)-
5-nitrobenzenesulfonohydrazide Λ/l-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-
Λ/-methyl-5-nitrobenzenesulfonohydrazide
^-(^(δ-BromopyrazoloIi .δ-aJpyridin-S-yOethylidene^^methyKS-morpholinopropylJaminoJ-S- nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4-(methylsulfonyl)piperazin- 1 -yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
^-(^(δ-BromopyrazoloII .S-alpyridin-S-yOethylidene^^methyKpyridin^-ylmethyOaminoJ-δ- nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-/\/-methyl-2-(methyl(pyridin-2-ylmethyl)- amino)-5-nitrobenzenesulfonohydrazide /V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-3-ylmethyl)amino)-5- nitrobenzenesulfonohydrazide
/^-(I^S-Bromopyrazoloti .S-alpyridin-S-yOethylideneJ^methyKpyridin^-ylmethyOaminoJ-S- nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-2-yl)ethyl)amino)-5- nitrobenzenesulfonohydrazide
Λ/1-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(pyridin-2-yl)ethyl)- amino)-5-nitrobenzenesulfonohydrazide Λ/'-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-4-yl)ethyl)amino)-5- nitrobenzenesulfonohydrazide
2-((2-(1 /-/-lmidazol-1 -yl)ethyl)(methyl)amino)-Λ/'-(1 -(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide 2-((2-(1/-/-lmidazol-4-yl)ethyl)(methyl)amino)-Λ/'-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide yV-CI-Cδ-BromoimidazoII ^-alpyridin-S-yOethylideneJ^-fluoro-S-nitrobenzenesulfonohydrazide /V-(1 -(6-Bromoimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-((2- (dimethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide including pure geometric and entaniomeric forms and mixtures thereof, physiologically acceptable salts thereof, and prodrugs thereof.
It will be appreciated that compounds of Formula I may occur in different geometric and enantiomeric forms, and that both pure forms and mixtures of these separate isomers are included, and any physiologically acceptable salts thereof.
The term physiologically acceptable salt used throughout the specification is to be taken as meaning any suitable acid or base derived salt and, in particular, those formed from hydrochloric, sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic, maleic, methanesulfonic, isoethonic acids and the like and potassium carbonate, sodium or potassium hydroxide, ammonia, triethylamine, triethanolamine and the like.
The term prodrug as used herein means any compound which releases an active parent drug according to formula (I) in vivo when such prodrug is administered to a subject. Prodrugs of a compound of formula (I) are prepared by modifying functional groups present in the compound of formula (I) in such a way that the modifications may be cleaved in vivo to release the parent compound. In particular embodiments of the invention, prodrugs include phosphate prodrugs of phenols or alcohols, or carboxylic acid ester or amino acid ester prodrugs. Such prodrugs may be made by any number of standard methods recognised in the art. However, by way of example, phosphate prodrugs may be prepared by methods similar to those described by G. S. Gill et a/., Org. Prep. Proc. Int. 2006, 38(6), 604, and amino acid ester prodrugs may be prepared by methods similar to those described by L. Ribeiro et a/., Arch. Pharm. 2007, 340, 32.
Examples of suitable prodrugs include phosphate or carboxylic acid or aminoacid ester derivatives of Formula I. The term halo or halogen group used throughout the specification is to be taken as meaning a fluoro, chloro, bromo or iodo group.
Suitable substitution for the optionally substituted aryl, heteroaryl and 4-7 membered saturated rings include alkyl, unsaturated alkyl, hydroxyalkyl, alkoxyalkyl, O-alkyl, C(O)-alkyl, C(O)NH2, C(O)NH(alkyl), C(O)N(alkyl)2, CO2H, CO2-alkyl, S-alkyl, S(O)-alkyl, SO2-alkyl, SO2NH2, SO2NH(alkyl), SO2N(alkyl)2, OH, CN, CF3, OCF3, NO2, NH2, NH(alkyl), N(alkyl)2, N(alkyl)C(O)(alkyl), halo.
The compounds of Formula I, in addition to the biological activity as PI3K inhibitors, have been found to have a level of metabolic stability that is unexpected.
Metabolic stability of a drug molecule is a desirable property, since extensive metabolism reduces the systemic exposure and decreases the half-life of the compound. The rate of metabolism can be assessed using a liver microsomal assay. Greater stability of a drug molecule in liver microsomes often means that the drug will have improved metabolic stability in vivo [Li, Curr. Top. Med. Chem. 2004, 4, 701].
In another embodiment of the invention there is provided a method of cancer prevention or therapy for treating cancers including the step of administering a compound of Formula I to a subject in need thereof.
In this form of the invention, the method includes administration of a compound of Formula I together with administering one or more suitable chemotherapeutic agents and/or therapies to a patient in need thereof. These agents and therapies can be of any suitable type as would be well known to a skilled person. However, a non-limiting list would include agents and therapies selected from:
Alkylation agents (for example cisplatin, carboplatin) Antimetabolites (for example methotrexate, 5-FU)
Antitumour antibiotics (for example adriamymycin, bleomycin) Antitumour vegetable alkaloids (for example taxol, etoposide) Antitumour hormones (for example dexamethasone, tamoxifen) Antitumour immunological agents (for example interferon α, β, y) Radiation therapy
Surgery. The method can further include the step of administering one or more chemotherapeutic agents to the subject before, during or after the administration of the compound of Formula I to the subject.
While these compounds of Formula I will typically be used in cancer prevention or cancer therapy of human subjects, they can be used to target cancer cells in other warm blooded animal subjects such as other primates, farm animals such as cattle, and sports animals and pets such as horses, dogs, and cats.
It will be appreciated that reference to cancer prevention or cancer therapy is not intended to be a reference to a cure for cancer or to absolute prevention. The reference is intended to include reference to a reduction in the likelihood of contraction of cancer or a mitigation of development, or like outcome.
In another embodiment there is provided a pharmaceutical composition including a compound of Formula I as defined above, and a pharmaceutically acceptable excipient, adjuvant, carrier, buffer or stabiliser. The pharmaceutical composition will preferably take the form of a tablet, capsule, powder, or liquid form. It is further preferred that the composition will be suitable for oral or parenteral administration.
The pharmaceutically acceptable excipient, adjuvant, carrier, buffer or stabiliser can be of any known type and should be non-toxic and should not interfere with the efficacy of the active ingredient. The precise nature of the carrier or other material will depend on the route of administration, which may be oral, or by injection, such as cutaneous, subcutaneous, or intravenous injection.
The pharmaceutical compositions of the invention formulated for oral administration may be in tablet, capsule, powder or liquid form. A tablet may comprise a solid carrier or an adjuvant. Liquid pharmaceutical compositions generally comprise a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil. Physiological saline solution, dextrose or other saccharide solution or glycols such as ethylene glycol, propylene glycol or polyethylene glycol may be included. A capsule may comprise a solid carrier such as gelatin. Such formulation issues will be well known to a person skilled in the formulation art.
Where pharmaceutical compositions may be formulated for intravenous, cutaneous or subcutaneous injection, the active ingredient will be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has a suitable pH, isotonicity and stability. Those of relevant skill in the art are well able to prepare suitable solutions using, for example, isotonic vehicles such as Sodium Chloride injection, Ringer's injection, Lactated Ringer's injection. Preservatives, stabilisers, buffers antioxidants and/or other additives as are known to be suitable for such use may be included as required.
The pharmaceutical compositions preferably also include one or more chemotherapeutic agents as defined above.
In another embodiment of the invention there is provided the use of a compound of Formula I in the manufacture of a medicament for cancer prevention or therapy for treatment of cancer.
The medicament is preferably in the form of a tablet, capsule, powder or liquid form. The medicament will preferably be suitable for oral or parenteral administration. Typically, the medicament will be formulated as described above.
In a further embodiment of the invention there is provided a method of making a compound of Formula I as defined above, the method including the step of modifying a pyrazolo[1 ,5-a]- pyridine-3-carbonyl compound of Formula Il or an imidazo[1 ,2-a]pyridine-3-carbonyl compound of Formula III
Figure imgf000021_0001
wherein variables X, Y and R2 are defined above as for Formula I.
The compounds of Formula I can preferably be made according to any one or more of the steps as described in Schemes 1 to 7 in the section titled "Methods for preparing compounds".
In a preferred form, the method of making a compound of Formula I from compound of Formula Il or Formula III involves one of the following steps:
(i) condensation with a sulfonohydrazide (as illustrated for example in Scheme 4 below); or (ii) condensation with hydrazine followed by reaction with a sulfonyl chloride (as illustrated for example in Scheme 4 below); or
(iii) condensation with hydrazine followed by reaction with a sulfonyl chloride and then nucleophilic substitution (as illustrated for example in Schemes 4 and 5 below); or (iv) any of the above methods followed by reaction with CH2N2 (as illustrated for example in Scheme 7 below). The reactions shown in schemes 1 to 7 are examples of the steps above. However, it will be recognised by a person skilled in the art that alternative but substantially equivalent reagents, solvents and reaction conditions to those shown in the Schemes 1 to 7 may be used. Such equivalent reagents, solvents and reaction conditions may be used without departing from the spirit and scope of the present invention and without diminishing its attendant advantages. It is therefore intended that such changes and modifications be included within the present invention.
It is to be recognised that certain compounds may exist in one or more different enantiomeric or diastereomeric forms. It is to be understood that the enantiomeric or diastereomeric forms are included in the above aspects of the invention.
Further aspects of the present invention will become apparent from the following description given by way of example only and with reference to the accompanying synthetic schemes
Methods for preparing compounds
The following methods for preparing compounds are illustrative only and a person skilled in the art will be aware of alternative reagents and reaction conditions other than those specifically described. It is to be understood that the invention includes all such variations and modifications.
The compounds described above can be prepared by routes outlined below. In Scheme 1 , ketone 3 can be made by Λ/-amination of pyridine 1 using a suitable O-substituted hydroxyl- amine such as O-(mesitylsulfonyl)hydroxylamine or O-(2,4-dinitrophenyl)hydroxylamine to form Λ/-aminopyridinium 2. Cyclisation under basic conditions with a suitable alkyne forms substituted pyrazolo[1 ,5-a]pyridine 3.
Figure imgf000022_0001
1 2 3 Scheme 1
In Scheme 2, carboxylic acid derivative 4 (where LG is a suitable leaving group) can be reacted with bis(trimethylsilyl)acetylene to form ketone 5. Then reaction with N- aminopyridinium 2 under basic conditions with the addition of a fluoride source gives pyrazolo[1 ,5-a]pyridine 6.
Figure imgf000023_0001
4 5 6
Scheme 2
The synthesis of halogen-containing compounds 9 is depicted in Scheme 3. Protected amine 7 can be deprotected with acid to form amine 8, which can then undergo a Sandmeyer reaction to introduce the halogen atom. HHaI
Figure imgf000023_0003
Figure imgf000023_0002
Scheme 3
Compound 12 can be made, as shown in Scheme 4, either by condensation of ketone 10 with sulfonohydrazide 11 , or by condensation with hydrazine followed by sulfonylation with a suitably substituted sulfonyl chloride 13.
SO2NHNH2
Figure imgf000023_0004
10 12
■ Il
SO2CI 13
Scheme 4
In Scheme 5, fluorobenzene 14 can be substituted by a primary or secondary amine to form amine 15, or by an alcohol with NaH to form ether 16.
Figure imgf000023_0005
Scheme 5 Amine 18 can be made according to Scheme 6. Hydroxyalkylpiperazine 17 can be reacted with a minimum of two molar equivalents of a sulfonyl chloride, and then with a primary amine to form secondary amine 18.
Figure imgf000024_0001
Scheme 6
Λ/-Methylation of 19 can be carried out using CH2N2 to afford 20, as shown in Scheme 7.
R2 H CH2N2 R2 Me
HtAr1V O2 0 HetArN- Or2 D
19 20
Scheme 7
EXAMPLES
The present invention will be described in more detail by referring to the following examples but is not deemed to be limited thereto.
Table 1 gives examples of compounds representative of the invention, and preparable by the methods outlined in Schemes 1-7.
Table 1
Figure imgf000024_0002
Figure imgf000025_0001
Figure imgf000026_0001
General experimental information
The following examples are representative of the invention, and provide detailed methods for preparing the compounds of the invention including the preparation of intermediate compounds. In these examples, elemental analyses were carried out in The Campbell Microanalytical Laboratory, University of Otago, Dunedin, New Zealand. 1H NMR spectra were obtained on a Bruker Avance-400 spectrometer at 400 MHz, referenced to Me4Si when measured in CDCI3 and to the residual DMSO when measured in d6-DMSO. Mass spectra were determined on a Thermo Finnigan MSQ single quadrupole mass spectrometer using atmospheric pressure chemical ionisation (APCI). High resolution mass spectra were obtained on a Bruker micrOTOF-QII mass spectrometer using either electrospray ionisation (ESI) or atmospheric pressure chemical ionisation (APCI). Column chromatography was carried out on silica gel (200-320 mesh, APS Finechem Ltd), unless otherwise stated.
Example 1: /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-methyl-5-nitro- benzenesulfonohydrazide (E1).
Step 1.1 : A fresh solution of 0-(mesitylsulfonyl)hydroxylamine in CH2CI2 (30 mL, 0.38 mol L"1, 11.4 mmol) [T. Eichenberger ef a/., HeIv. Chim. Acta 1986, 69(6), 1521] was added to 4- pyridinecarbonitrile (1.19 g, 11.4 mmol) in CH2CI2 (20 mL) at 0 0C. After 2 h, the solvent was removed in vacuo to leave crude 1-amino-4-cyanopyridinium 2-mesitylenesulfonate as a yellow solid. This was taken up in dry DMF (20 mL), then 3-butyn-2-one (0.89 mL, 11.4 mmol) and K2CO3 (3.16 g, 22.9 mmol) were added, and the suspension stirred at room temperature for 18 h. The reaction mixture was diluted with water and extracted twice with EtOAc. The combined organic phases were washed three times with water then with brine, dried (Na2SO4) and the solvent removed in vacuo. Chromatography (eluting with hexanes: EtOAc 4:1 to 3:1 to 2:1 to 1 :1) gave 3-acetylpyrazolo[1 ,5-a]pyridine-5-carbonitrile as a pale yellow solid (746 mg, 35%). 1H NMR δ (400 MHz, CDCI3) 8.80 (dd, J 1.8, 1.0 Hz, 1 H), 8.61 (dd, J 7.2, 1.0 Hz, 1 H), 8.45 (s, 1 H), 7.13 (dd, J 7.2, 1.9 Hz, 1 H), 2.60 (s, 3H). LCMS (APCI+) 186 (MH+, 100%).
Step 1.2: A solution of 3-acetylpyrazolo[1 ,5-a]pyridine-5-carbonitrile (50 mg, 0.27 mmol) and 2- methyl-5-nitrobenzenesulfonohydrazide (69 mg, 0.30 mmol) [I. Kh. Fel'dman et al., Zh. Obshch. Khim., 1963, 33, 38] in MeOH (5 mL) was refluxed for 18 h. After cooling to room temperature, the precipitated solid was filtered off, washed with a little MeOH and dried to leave /V-(1 -(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide (E1) as a yellow solid (82 mg, 76%). 1H NMR δ (400 MHz, d6- DMSO) 11.15 (s, 1 H), 8.91 (dd, J 7.2, 0.6 Hz, 1 H), 8.77 (d, J 2.5 Hz, 1 H), 8.55 (s, 1 H), 8.40 (dd, J 8.4, 2.5 Hz, 1 H), 7.93 (s, 1 H), 7.76 (d, J 8.4 Hz, 1 H), 7.28 (dd, J 7.2, 1.9 Hz, 1 H), 2.78 (s, 3H), 2.35 (s, 3H). LCMS (APCI+) 399 (MH+, 100%). HRMS (ESI+) Calcd for C17H15N6O4S: 399.0870; found (MH+) 399.0879.
Example 2: AT-(I -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/,2-dimethyl-5-nitro- benzenesulfonohydrazide (E2).
CH2N2 solution in Et2O was added dropwise to a solution of E1 (50 mg, 0.13 mmol) in THF (5 mL) until the reaction was complete by t.l.c. analysis. The solvents were removed in vacuo. Chromatography (eluting with CH2CI2: MeOH 99.75:0.25 to 99.5:0.5) gave /V-(1-(5-cyano- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/,2-dimethyl-5-nitrobenzenesulfonohydrazide (E2) as a yellow solid (35 mg, 67%). 1H NMR δ (400 MHz, CDCI3) 8.76 (d, J 2.4 Hz, 1 H), 8.59 (dd, J 7.2, 1.0 Hz, 1 H), 8.42 (dd, J 8.4, 2.5 Hz, 1 H), 8.38 (s, 1 H), 8.30 (dd, J 1.9, 1.0 Hz, 1 H), 7.63 (d, J 8.4 Hz, 1 H)1 7.04 (dd, J 7.2, 1.9 Hz, 1 H), 3.02 (s, 3H), 2.72 (s, 3H), 2.67 (s, 3H). LCMS (APCI+) 413 (MH+, 100%). Anal. Calcd for C18H16N6O4S.0.33 H2O: C, 51.68; H, 4.01 ; N, 20.09. Found C, 51.69; H, 4.07; N, 19.81.
Example 3: /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)propylidene)-2-methyl-5-nitro- benzenesulfonohydrazide (E3). Step 3.1 : AICI3 (921 mg, 6.91 mmol) was added to a solution of propionic anhydride (0.74 mL, 5.76 mmol) and bis(trimethylsilyl)acetylene (1.43 mL, 6.31 mmol) in dry CH2CI2 (20 mL) at 0 0C. After 30 mins, the reaction mixture was warmed to room temperature, stirred for a further 1 h, and then poured onto ice-1 M HCI (1 :1 , 80 mL). The layers were separated, and the aqueous layer was extracted with CH2CI2. The combined organic layers were dried (Na2SO4) and the solvent removed in vacuo. Chromatography (eluting with hexanes:Et2O 98:2) gave 1- (trimethylsilyl)pent-1-yn-3-one as a brown oil (739 mg, 83%). 1H NMR δ (400 MHz, CDCI3) 2.58 (q, J 7.4 Hz, 2H), 1.14 (t, J 7.4 Hz, 3H), 0.24 (s, 9H). LCMS (APCI+) 155 (MH+, 100%). Step 3.2: A solution of 1-(trimethylsilyl)pent-1-yn-3-one (520 mg, 3.38 mmol) in DMF (5 mL) was added to a suspension of 1-amino-4-cyanopyridinium 2-mesitylenesulfonate (1.62 g, 5.08 mmol), KF (589 mg, 10.1 mmol) and K2CO3 (1.4Og, 10.1 mmol) in DMF (20 mL) at 0 0C, and then slowly warmed to room temperature overnight. The reaction mixture was diluted with water and extracted twice with EtOAc. The combined extracts were washed with water, 1 M HCI, water, brine, and then dried (Na2SO4) and the solvent removed in vacuo. Chromatography (eluting with CH2CI2) gave 3-propionylpyrazolo[1 ,5-a]pyridine-5-carbonitrile as an off-white solid (479 mg, 71%). 1H NMR δ (400 MHz, CDCI3) 8.81 (dd, J 1.8, 1.0 Hz, 1 H), 8.61 (dd, J 7.2, 1.0 Hz, 1 H), 8.46 (s, 1H), 7.11 (dd, J 7.2, 1.8 Hz, 1 H), 2.96 (q, J 7.4 Hz, 2H), 1.27 (t, J 7.4 Hz, 3H). LCMS (APCI+) 200 (MH+, 100%).
Step 3.3: Reaction of 3-propionylpyrazolo[1 ,5-a]pyridine-5-carbonitrile (71 mg, 0.36 mmol) using the conditions of Step 1.2 gave ^-(^(δ-cyanopyrazoloti .δ-aJpyridin-S-yOpropylidene)^- methyl-5-nitrobenzenesulfonohydrazide (E3) as a yellow solid (105 mg, 71%). 1H NMR δ (400 MHz, d6-DMSO) 11.37 (s, 1 H)1 8.93 (dd, J 7.2, 0.9 Hz, 1 H), 8.76 (d, J 2.4 Hz, 1 H), 8.56 (s, 1 H), 8.40 (dd, J 8.4, 2.4 Hz, 1 H), 7.92 (dd, J 1.8, 0.9 Hz, 1 H), 7.76 (d, J 8.4 Hz, 1 H), 7.29 (dd, J 7.2, 1.8 Hz1 1 H), 2.82 (q, J 7.6 Hz1 2H), 2.76 (s, 3H), 1.10 (t, J 7.6 Hz1 3H). LCMS (APCI+) 413 (MH+, 100%). HRMS (ESI+) Calcd for C18H17N6O4S: 413.1027; found (MH+) 413.1021.
Example 4: /V-(1 -(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)propylidene)-Λ/,2-dimethyl-5-nitro- benzenesulfonohydrazide (E4).
Reaction of E3 (70 mg, 0.17 mmol) using the conditions of Example 2 gave Λy-(1-(5-cyano- pyrazolo[1 ,5-a]pyridin-3-yl)propylidene)-Λ/,2-dimethyl-5-nitrobenzenesulfonohydrazide (E4) as a yellow solid (40 mg, 56%). 1H NMR δ (400 MHz, CDCI3) 8.77 (d, J 2.4 Hz, 1 H), 8.59 (dd, J 7.2, 1.0 Hz, 1 H), 8.44 (dd, J 8.4, 2.4 Hz, 1 H), 8.37 (s, 1 H)1 8.26 (dd, J 1.9, 1.0 Hz, 1 H), 7.64 (d, J 8.4 Hz, 1 H), 7.04 (dd, J 7.2, 1.9 Hz, 1 H), 3.11 (q, J 7.7 Hz, 2H)1 3.01 (s, 3H), 2.71 (s, 3H), 1.37 (t, J 7.7 Hz, 3H). LCMS (APCI+) 427 (MH+, 100%). Anal. Calcd for C19H18N6O4S: C, 53.51 ; H, 4.25; N, 19.71. Found C, 53.43; H, 4.35; N1 19.36.
Example 5: /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)butylidene)-2-methyl-5-nitro- benzenesulfonohydrazide (E5).
Step 5.1 : Reaction of butyryl chloride (2.00 mL, 19.3 mmol) using the conditions of Step 3.1 gave 1-(trimethylsilyl)hex-1-yn-3-one as a brown oil (2.98 g, 92%). 1H NMR δ (400 MHz, CDCI3) 2.54 (t, J 7.3 Hz, 2H), 1.70 (m, 2H)1 0.95 (t, J 7.4 Hz, 3H), 0.24 (s, 9H). LCMS (APCI+) 169 (MH+, 100%). Step 5.2: Reaction of 1-(trimethylsilyl)hex-1-yn-3-one (147 mg, 0.88 mmol) using the conditions of Step 3.1 gave 3-butyrylpyrazolo[1 ,5-a]pyhdine-5-carbonitrile as an off-white solid (153 mg, 82%). 1H NMR δ (400 MHz, CDCI3) 8.82 (dd, J 1.9, 1.0 Hz, 1 H), 8.61 (dd, J 7.2, 1.0 Hz, 1 H), 8.46 (s, 1 H), 7.12 (dd, J 7.2, 1.9 Hz, 1 H), 2.89 (t, J 7.3 Hz, 2H), 1.82 (m, 2H), 1.04 (t, J 7.4 Hz, 3H). LCMS (APCI+) 214 (MH+, 100%).
Step 5.3: Reaction of 3-butyrylpyrazolo[1 ,5-a]pyridine-5-carbonitrile (57 mg, 0.27 mmol) using the conditions of Step 3.1 gave /V-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)butylidene)-2-methyl- 5-nitrobenzenesulfonohydrazide (E5) as a yellow solid (68 mg, 60%). 1H NMR δ (400 MHz, d6-DMSO) 11.36 (s, 1 H), 8.91 (dd, J 7.2, 0.9 Hz, 1 H), 8.76 (d, J 2.5 Hz, 1 H), 8.55 (s, 1 H), 8.39 (dd, J 8.4, 2.5 Hz, 1 H), 7.96 (dd, J 1.9, 0.9 Hz, 1 H), 7.75 (d, J 8.4 Hz, 1 H), 7.27 (dd, J 7.2, 1.9 Hz, 1 H), 2.81 - 2.75 (m, 5H), 1.54 (m, 2H), 0.96 (t, J 7.3 Hz, 3H). LCMS (APCI+) 427 (MH+, 100%). HRMS (ESI+) Calcd for C19H19N6O4S: 427.1183; found (MH+) 427.1184.
Example 6: W-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)butylidene)-A/,2-dimethyl-5-nitro- benzenesulfonohydrazide (E6).
Reaction of E5 using the conditions of Example 2 gave /V-(1-(5-cyanopyrazolo[1,5-a]pyridin-3- yl)butylidene)-Λ/,2-dimethyl-5-nitrobenzenesulfonohydrazide (E6) as a yellow solid (42 mg, 93%). 1H NMR δ (400 MHz, CDCI3) 8.75 (d, J 2.4 Hz, 1 H), 8.58 (dd, J 7.2, 1.0 Hz, 1 H), 8.43 (dd, J 8.4, 2.4 Hz, 1 H), 8.35 (s, 1 H), 8.26 (dd, J 1.9, 1.0 Hz, 1 H), 7.63 (d, J 8.4 Hz, 1 H), 7.04 (dd, J 7.2, 1.9 Hz, 1 H), 3.06 (m, 2H), 3.00 (s, 3H), 2.70 (s, 3H), 1.78 (m, 2H), 1.11 (t, J 7.4 Hz, 3H). LCMS (APCI+) 441 (MH+, 100%). Anal. Calcd for C20H20N6O4S.0.25 EtOAc: C, 54.54; H, 4.79; N, 18.17. Found C, 54.34; H, 4.73; N, 18.19.
Example 7: /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5- nitrobenzenesulfonohydrazide (E7).
Step 7.1 : A solution of 3-acetylpyrazolo[1 ,5-a]pyridine-5-carbonitrile (295 mg, 1.59 mmol) and hydrazine monohydrate (402 mg, 8.04 mmol) in MeOH (4 mL) heated at 65 0C for 3 h. After cooling to room temperature, the residue was concentrated in vacuo to leave 3-(1-hydrazono- ethyl)pyrazolo[1 ,5-a]pyridine-5-carbonitrile as a yellow solid which was used in the next step without purification. 1H NMR δ (400 MHz, CDCI3) 8.75 (dd, J 1.9, 1.0 Hz, 1 H), 8.48 (dd, J 7.2, 1.0 Hz, 1 H), 8.12 (s, 1 H), 6.91 (dd, J 7.2, 1.8 Hz, 1 H), 5.28 (s, broad, 2H), 2.20 (s, 3H). LCMS (APCI+) 200 (MH+, 100%).
Step 7.2: To a solution of 3-(1-hydrazonoethyl)pyrazolo[1 ,5-a]pyridine-5-carbonitrile in CH2CI2 (3 mL) and pyridine (0.57 mL, 7.05 mmol), was added 2-fluoro-5-nitrobenzenesulfonyl chloride (400 mg, 1.67 mmol). Reaction mixture was stirred at room temperature for 1 h. The solvents were removed in vacuo. Chromatography (eluting with hexane: EtOAc 1 :1 to CH2CI2: MeOH 95:5) gave /V-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5- nitrobenzenesulfonohydrazide (E7) as an orange solid (161 mg, 25% over 2 steps). 1H NMR δ (400 MHz1 d6-DMSO) 11.3 (s, 1 H), 8.94 (d, J 7.2 Hz, 1 H), 8.72 (dd, J 5.9, 2.9 Hz, 1H), 8.61- 8.56 (m, 2H), 8.10 (s, 1 H), 7.77 (t, J 9.2 Hz, 1 H), 7.31 (dd, J 7.2, 1.9 Hz, 1 H), 2.34 (s, 3H). LCMS (APCI+) 403 (MH+, 20%). Anal. Calcd for C16H11FN6O4SO^S EtOAc: C, 48.11 ; H, 3.09; N, 19.80. Found C, 47.92; H, 3.08; N, 19.58.
Example 8: 2-((2-Aminoethyl)(methyl)amino)-Λ/l-(1 -(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide (E8).
A solution of
Figure imgf000030_0001
sulfonohydrazide (150 mg, 0.37 mmol) and 2,2,2-trifluoro-Λ/-(2-(methylamino)ethyl)acetamide (444 mg, 2.61 mmol) in THF (20 mL) was stirred at room temperature for 4 days. The solvent was removed in vacuo. Chromatography (eluting with CH2CI2 to CH2CI2:Me0H 99.5:0.5 to 99:1) gave impure Λ/-(2-((2-(2-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3- yl)ethylidene)hydrazinylsulfonyl)-4-nitrophenyl)(methyl)amino)ethyl)-2,2,2-trifluoroacetamide as an orange oil (165 mg). This was taken up immediately in THF (9 mL) and MeOH (3 mL), and then concentrated aqueous NH3 (25 mL) was added. After stirring for 24 h, the THF and MeOH were removed in vacuo. The residue was extracted four times with CH2CI2 then the combined organic layers were dried (Na2SO4) and the solvent removed in vacuo.
Chromatography (eluting with CH2CI2 to CH2CI2: MeOH 99.1 to 98:2 to 97:3 to 96:4 to 94:6) gave 2-((2-aminoethyl)(methyl)amino)-Λ/'-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-5- nitrobenzenesulfonohydrazide (E8) as a yellow powder (29 mg, 17%). 1H NMR δ (400 MHz, de-DMSO) 8.80 (dd, J 7.2, 0.9 Hz, 1H), 8.59 (d, J 2.8 Hz1 1H), 8.33 (s, 1H), 8.22 (dd, J 1.9, 0.9 Hz, 1 H), 8.12 (dd, J 9.3, 2.8 Hz, 1 H), 7.13 (dd, J 7.2, 1.9 Hz, 1 H), 6.91 (d, J 9.3 Hz, 1 H), 3.57 (t, J 6.6 Hz1 2H), 2.96 (t, J 6.6 Hz, 2H), 2.54 (S1 3H), 2.21 (s, 3H). LCMS (APCI+) 457 (MH+, 100%). Anal. Calcd for C22H21 N9O4S.0.05 CH2CI2.0.15 hexanes: C, 52.54; H1 4.46; N1 24.02. Found C, 52.41 ; H1 4.60; N, 23.99.
Example 9: ΛP-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(methyl- amino)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E9).
A solution of E7 (137 mg, 0.34 mmol) and Λ/./V-dimethylethylenediamine (0.55 mL, 5.2 mmol) in THF (6 mL) was stirred for 18 h. The solvent was removed in vacuo. Chromatography (eluting with CH2CI2:MeOH 99:1 to 95:5) gave AT-(I -(5-cyanopyrazolo[1,5-a]pyridin-3- yl)ethylidene)-2-(methyl(2-(methylamino)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E9) as a yellow powder (69 mg, 43%). 1H NMR δ (400 MHz1 CDCI3) 9.20 (d, J 2.7 Hz, 1 H), 8.47 (dd, J 8.8, 2.7 Hz, 1 H), 8.42 (dd, J 7.2, 1.0 Hz, 1 H), 8.13 (s, 1 H), 7.57 (m, 1 H), 7.40 (d, J 8.8 Hz, 1 H), 6.85 (dd, J 7.2, 1.9 Hz, 1 H), 3.20 (m, 2H), 2.89 (m, 2H), 2.67 (s, 3H), 2.61 (s, 3H), 2.36 (s, 3H). LCMS (APCI+) 471 (MH+, 100%). Anal. Calcd for C20H22N8O4S.1.34 H2O: C, 48.56; H, 5.03; N1 22.65. Found C, 48.54; H, 4.94; N1 22.50.
Example 10: /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)- ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E10).
Reaction of E7 (155 mg, 0.39 mmol) and Λ/,Λ/,/V'-trimethylethylenediamine (0.25 ml_, 1.92 mmol) using the conditions of Example 9 gave /V-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3- yl)ethylidene)-2-((2-(dimethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E10) as an orange solid (119 mg, 64%). 1H NMR δ (400 MHz, d6-DMSO) 8.84 (dd, J 7.2, 0.9 Hz, 1 H), 8.76 (d, J 2.8 Hz, 1H), 8.47 (s, 1 H), 8.26 (dd, J 9.1 , 2.8 Hz, 1 H), 7.71 (t, J 0.8 Hz, 1 H),
7.44 (d, J 9.1 Hz, 1 H), 7.17 (dd, J 7.2, 2.0 Hz, 1 H), 3.54 (t, J 6.1 Hz, 2H), 2.92 (s, 3H), 2.76 (t, J 6.1 Hz, 2H), 2.36 (s, 6H), 2.32 (s, 3H). LCMS (APCI+) 485 (MH+, 100%). Anal. Calcd for C2iH24N8O4S.0.3 hexane.0.1 CH2CI2: C, 53.01 ; H, 5.52; N, 21.59. Found Cr 53.02; H, 5.49; N1 21.69.
Example 11 : ΛP-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)- ethyl)(methyl)amino)-Λ/-methyl-5-nitrobenzenesulfonohydrazide (E11 ).
Reaction of E10 (119 mg, 0.25 mmol) using the conditions of Example 2 gave /V-(1-(5-cyano- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)ethyl)(methyl)amino)-Λ/-methyl-5- nitrobenzenesulfonohydrazide (E11) as a yellow solid (95 mg, 76%). 1H NMR δ (400 MHz, d6- DMSO) 9.05 (dd, J 7.2, 0.9 Hz, 1 H), 8.78-8.77 (m, 2H), 8.31 (dd, J 9.2, 2.8 Hz, 1H), 8.23 (dd, J 1.8, 0.9 Hz, 1 H), 7.46 (d, J 9.3 Hz, 1 H)1 7.42 (dd, J 7.2, 1.9 Hz1 1 H)1 3.53 (t, J 6.6 Hz1 2H)1
3.45 (t, J 6.4 Hz, 2H), 3.04 (s, 3H), 2.97 (s, 3H), 2.56 (s, 3H)1 2.08 (S1 6H). LCMS (APCI+) 499 (MH+, 100%). HRMS (FAB+) Calcd for C22H27N8O4S: 499.18760; found (MH+) 499.18824.
Example 12: ΛT-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((3-(dimethylamino)- propyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E12).
Reaction of E7 (137 mg, 0.34 mmol) and Λ/,Λ/,Λ/'-trimethyl-1 ,3-propanediamine (0.25 mL, 1.7 mmol) using the conditions of Example 9 gave /V-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-((3-(dimethylamino)propyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E12) as a yellow powder (55 mg, 32%). 1H NMR δ (400 MHz, d6-DMSO) 8.82 (dd, J 7.2, 0.9 Hz, 1 H), 8.77 (d, J 2.9 Hz, 1H), 8.41 (s, 1 H), 8.18 (dd, J 9.2, 2.9 Hz1 1 H)1 7.91 (m, 1 H)1 7.34 (d, J 9.2 Hz, 1H)1 7.15 (dd, J 7.2, 1.9 Hz, 1H), 3.45 (t, J 6.7 Hz, 2H), 2.97 (s, 3H), 2.69 (t, J 6.5 Hz1 2H), 2.44 (s, 6H), 2.27 (s, 3H), 1.84 (m, 2H). LCMS (APCI+) 499 (MH+, 100%). Anal. Calcd for C22H26N8O4S. H20.0.1 CH2CI2: C, 50.09; H1 5.37; N, 21.07. Found C, 49.97; H, 5.06; N, 20.67. Example 13: ΛT-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)- ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E13).
Reaction of E7 (240 mg, 0.60 mmol) and Λ/,Λ/-diethyl-Λ/'-methylethylenediamine (0.50 mL, 3.1 mmol) using the conditions of Example 9 gave /V-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E13) as an orange powder (147 mg, 48%). 1H NMR δ (400 MHz, CDCI3) 8.92 (d, J 2.8 Hz, 1 H), 8.53 (dd, J 8.8, 2.8 Hz, 1 H), 8.46 (dd, J 7.2, 1.0 Hz, 1 H), 8.21 (s, 1 H), 7.59 (dd, J 1.9, 1.0 Hz, 1 H), 7.52 (d, J 8.8 Hz, 1 H), 6.88 (dd, J 7.2, 1.9 Hz, 1 H), 3.26 (m, 2H), 2.85 (s, 3H), 2.74 (m, 2H), 2.66 (q, J 7.1 Hz, 4H), 2.51 (s, 3H), 1.05 (t, J 7.1 Hz, 6H). LCMS (APCI+) 513 (MH+, 100%). Anal. Calcd for C23H28N8O4S-Ce CH2CI2: C, 50.30; H, 5.22; N, 19.88. Found C, 50.55; H, 5.17; N, 19.71.
Example 14: /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)- ethyl)(methyl)amino)-Λ/-methyl-5-nitrobenzenesulfonohydrazide (E14). Reaction of E13 (82 mg, 0.16 mmol) using the conditions of Example 2 gave /V-(1-(5-cyano- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-Λ/-methyl-5- nitrobenzenesulfonohydrazide (E14) as a yellow powder (52 mg, 62%). 1H NMR δ (400 MHz, CDCI3) 8.94 (d, J 2.8 Hz, 1 H), 8.56 (dd, J 7.2, 1.0 Hz, 1 H), 8.37 (dd, J 1.9, 1.0 Hz, 1 H), 8.35 (s, 1 H), 8.33 (dd, J 9.2, 2.8 Hz, 1 H), 7.29 (d, J 9.2 Hz, 1 H), 7.04 (dd, J 7.2, 1.9 Hz, 1 H), 3.55 (m, 2H), 3.11 (s, 3H), 3.03 (s, 3H), 2.70 (m, 2H), 2.64 (s, 3H), 2.47 (q, J 7.1 Hz, 4H), 0.94 (t, J 7.1 Hz, 6H). LCMS (APCI+) 527 (MH+, 100%). Anal. Calcd for C24H3oN804S.0.33 MeOH: C, 54.40; H, 5.88; N, 20.86. Found C, 54.51; H, 5.93; N, 20.58.
Example 15: /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(piperidin- 1-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E15).
Reaction of E7 (240 mg, 0.60 mmol) and Λ/-methyl-2-(piperidin-1-yl)ethanamine (435 mg, 3.1 mmol) using the conditions of Example 9 gave /V-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-(methyl(2-(piperidin-1 -yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E15) as a yellow powder (163 mg, 52%). 1H NMR δ (400 MHz, CDCI3) 8.93 (d, J 2.8 Hz, 1 H), 8.50 (dd, J 8.8, 2.8 Hz, 1 H), 8.46 (dd, J 7.2, 0.9 Hz, 1 H), 8.21 (s, 1 H), 7.56 (dd, J 1.9, 0.9 Hz, 1 H), 7.52 (d, J 8.8 Hz, 1 H), 6.88 (dd, J 7.2, 1.9 Hz, 1 H), 3.27 (m, 2H), 2.81 (s, 3H), 2.59 (m, 2H), 2.56 - 2.47 (m, 7H), 1.66 - 1.57 (m, 4H), 1.50 - 1.41 (m, 2H). LCMS (APCI+) 525 (MH+, 100%). Anal. Calcd for C24H28N8O4S: C, 54.95; H, 5.38; N, 21.36. Found C, 54.78; H, 5.37; N, 21.09. Example 16: Λf-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2- (piperidin-1-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E16).
Reaction of E15 (72 mg, 0.14 mmol) using the conditions of Example 2 gave /V-(1-(5-cyano- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(piperidin-1-yl)ethyl)amino)-5- nitrobenzenesulfonohydrazide (E16) as a yellow powder (25 mg, 34%). 1H NMR δ (400 MHz, CDCI3) 8.92 (d, J 2.8 Hz, 1 H), 8.56 (dd, J 7.2, 0.9 Hz, 1 H), 8.36 - 8.31 (m, 3H), 7.31 (d, J 9.2 Hz, 1 H)1 7.04 (dd, J 7.2, 1.9 Hz, 1H), 3.61 (t, J 6.6 Hz, 2H), 3.09 (s, 3H), 3.03 (s, 3H), 2.64 (s, 3H), 2.59 (t, J 6.6 Hz, 2H), 2.38 - 2.29 (m, 4H), 1.48 - 1.32 (m, 6H). LCMS (APCI+) 539 (MH+, 100%). Anal. Calcd for C25H30N8O4S.1.2 MeOH: C, 55.41 ; H, 6.31 ; N, 18.87. Found C, 55.47; H, 6.18; N1 18.74.
Example 17: AT-(I -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(3- morpholinopropyl)amino)-5-nitrobenzenesulfonohydrazide (E17).
Reaction of E7 (221 mg, 0.55 mmol) and Λ/-methyl-3-morpholinopropan-1-amine (230 mg, 1.5 mmol) using the conditions of Example 9 gave ΛΛ-O^S-cyanopyrazolofi.δ-aJpyridin-S-yl)- ethylidene)-2-(methyl(3-morpholinopropyl)amino)-5-nitrobenzenesulfonohydrazide (E17) as a yellow powder (162 mg, 55%). 1H NMR δ (400 MHz, CDCI3) 8.93 (d, J 2.7 Hz, 1 H), 8.47 (dd, J 7.2, 1.0 Hz, 1 H), 8.45 (dd, J 8.9, 2.7 Hz, 1 H), 8.19 (s, 1 H), 7.70 (dd, J 1.9, 1.0 Hz, 1 H)1 7.49 (d, J 8.9 Hz, 1 H)1 6.92 (dd, J 7.2, 1.9 Hz, 1 H), 3.63 (m, 4H), 3.24 (m, 2H), 2.93 (s, 3H)1 2.43 (s, 3H), 2.41 - 2.34 (m, 6H), 1.83 (s, 2H). LCMS (APCI+) 541 (MH+, 100%). Anal. Calcd for C24H28N8O5S.1.25 MeOH: C, 52.23; H, 5.73; N1 19.30. Found C, 51.87; H1 5.34; N1 19.26.
Example 18: /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4- (methylsulfonyl)piperazin-i -yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E18). Step 18.1 : Methanesulfonyl chloride (2.98 mL, 38.5 mmol) was added dropwise to a solution of 1-(2-hydroxyethyl)piperazine (2.00 g, 15.4 mmol) and NEt3 (5.4 mL, 38.5 mmol) in CH2CI2 (20 mL) at 0 0C. After stirring for 10 mins, methylamine (10 mL, 33 wt. % solution in EtOH) was added, and the resulting solution stirred at room temperature for 24 h. The solvents were removed in vacuo. Chromatography (on alumina eluting with CH2CI2: MeOH 99:1 to 98:2 to 97:3 to 96:4 to 95:5 to 94:6) gave Λ/-methyl-2-(4-(methylsulfonyl)piperazin-1-yl)ethanamine as a yellow gum (1.20 g, 35%). 1H NMR δ (400 MHz, d6-DMSO) 3.12 (m, 4H), 2.90 (t, J 6.2 Hz, 2H), 2.88 (s, 3H), 2.56 (t, J 6.2 Hz, 2H), 2.52 - 2.47 (m, 7H). 13C NMR δ (100 MHz, d6-DMSO) 53.4 (CH2), 51.7 (CH2), 45.6 (CH2), 45.3 (CH2), 33.9 (CH3), 33.3 (CH3). LCMS (APCI+) 222 (MH+, 100%).
Step 18.2: Reaction of E7 (60 mg, 0.15 mmol) and Λ/-methyl-2-(4-(methylsulfonyl)piperazin-1- yl)ethanamine (99 mg, 0.45 mmol) using the conditions of Example 9 gave Λ/^Hδ-cyano- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4-(methylsulfonyl)piperazin-1-yl)ethyl)- amino)-5-nitrobenzenesulfonohydrazide (E18) as a yellow solid (47 mg, 52%). 1H NMR δ (400 MHz, d6-DMSO) 8.90 (d, J 7.2 Hz, 1 H), 8.72 (d, J 2.7 Hz, 1 H), 8.56 (s, 1 H), 8.24 (dd, J 9.3, 2.7 Hz, 1 H), 7.58 (m, 1 H), 7.39 (d, J 9.3 Hz, 1 H), 7.24 (dd, J 7.2, 1.8 Hz, 1 H), 3.63 (m, 2H), 3.01 (s, 3H), 2.86 (m, 4H), 2.75 (s, 3H), 2.54 (m, 2H), 2.40 - 2.33 (m, 7H). LCMS (APCI+) 604 (MH+, 100%). Anal. Calcd for C24H29N9O6S2: C, 47.75; H, 4.84; N, 20.88. Found C, 47.84; H, 5.08; N, 20.74.
Example 19: Λr-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-2-yl- methyl)amino)-5-nitrobenzenesulfonohydrazide (E19).
Reaction of E7 (300 mg, 0.75 mmol) and Λ/-methyl-2-pyridinemethanamine (453 mg, 3.7 mmol) using the conditions of Example 9 gave ΛΛ-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3- yl)ethylidene)-2-(methyl(pyridin-2-ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E19) as a yellow powder (32 mg, 9%). 1H NMR δ (400 MHz, CDCI3) 12.75 (br s, 1H), 9.25 (d, J 2.7 Hz, 1 H), 8.77 (ddd, J 5.0, 1.7, 0.9 Hz, 1 H), 8.50 (dd, J 8.9, 2.7 Hz, 1 H), 8.42 (dd, J 7.2, 1.0 Hz, 1 H), 8.12 (s, 1H), 7.79 (td, J 7.6, 1.7 Hz, 1H), 7.52 (dd, J 1.9, 1.0 Hz, 1 H), 7.43 (d, J 8.9 Hz, 1 H), 7.38 (ddd, J 7.6, 5.0, 0.9 Hz1 1 H), 7.28 (d, J 7.6 Hz, 1 H), 6.85 (dd, J 7.2, 1.9 Hz, 1 H), 4.51 (s, 2H), 2.60 (s, 3H), 2.53 (s, 3H). LCMS (APCI+) 505 (MH+, 100%). Anal. Calcd for C23H2oN804S.0.3 EtOAc.0.2 H2O: C, 54.37; H, 4.30; N, 20.96. Found C, 54.19; H, 4.39; N, 20.93.
Example 20: ΛT-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-/V-methyl-2-(methyl- (pyridin-2-ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E20).
Reaction of E19 (51 mg, 0.10 mmol) using the conditions of Example 2 gave /V-(1-(5-cyano- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(pyridin-2-ylmethyl)amino)-5-nitro- benzenesulfonohydrazide (E20) as a yellow powder (20 mg, 38%). 1H NMR δ (400 MHz, CDCI3) 9.05 (d, J 2.8 Hz, 1 H), 8.57 (dd, J 7.2, 1.0 Hz, 1 H), 8.54 (m, 1 H), 8.47 (dd, J 1.9, 1.0 Hz, 1 H), 8.37 (s, 1 H), 8.33 (dd, J 9.2, 2.8 Hz, 1 H), 7.63 (td, J 7.7, 1.8 Hz, 1 H), 7.34 (d, J 7.7 Hz, 1 H), 7.28 (d, J 9.2 Hz, 1 H)1 7.19 (ddd, J 7.7, 4.9, 1.0 Hz, 1 H), 7.05 (dd, J 7.2, 1.9 Hz, 1 H)1 4.74 (s, 2H), 3.10 (s, 3H), 3.09 (s, 3H), 2.66 (s, 3H). LCMS (APCI+) 519 (MH+, 100%). HRMS (ESI+) Calcd for C24H23N8O4S: 519.1534; found (MH+) 519.1557.
Example 21 : ΛT-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-4- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E21 ). Reaction of E7 (256 mg, 0.64 mmol) and Λ/-methyl-1-(pyridin-4-yl)methanamine (242 mg, 1.98 mmol) using the conditions of Example 9 gave /V-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-(methyl(pyridin-4-ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E21) as a yellow powder (151 mg, 45%). 1H NMR δ (400 MHz1 d6-DMSO) 11.23 (br s, 1 H), 8.91 (dd, J 7.2, 0.7 Hz, 1 H), 8.75 (d, J 2.7 Hz, 1 H), 8.59 (s, 1 H), 8.39 (m, 2H), 8.21 (dd, J 9.3, 2.7 Hz, 1 H), 7.52 (s, 1 H), 7.32 (d, J 9.3 Hz, 1 H), 7.27 (m, 2H), 7.24 (dd, J 7.2, 1.9 Hz, 1H), 4.76 (s, 2H), 3.10 (s, 3H), 2.38 (s, 3H). LCMS (APCI+) 505 (MH+, 100%). Anal. Calcd for C23H20N8O4S.0.4 H2O: C, 53.98; H, 4.10; N, 21.90. Found C, 54.00; H, 4.11 ; N1 22.02.
Example 22: ΛT-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-2- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E22).
Reaction of E7 (222 mg, 0.55 mmol) and Λ/-methyl-2-(pyridin-2-yl)ethanamine (0.27 ml_, 2.0 mmol) using the conditions of Example 9 gave /V-(1 -(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-(methyl(2-(pyridin-2-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E22) as a yellow powder (165 mg, 58%). 1H NMR δ (400 MHz, CDCI3) 10.59 (br s, 1H), 9.07 (d, J 2.7 Hz, 1 H), 8.60 (dd, J 5.3, 1.8 Hz, 1 H), 8.49 - 8.42 (m, 2H), 8.15 (s, 1 H), 7.71 (dd, J 1.9, 1.0 Hz, 1 H), 7.66 (td, J 7.7, 1.8 Hz, 1 H), 7.50 (d, J 8.9 Hz, 1 H), 7.25 - 7.20 (m, 2H), 6.88 (dd, J 7.2, 1.9 Hz, 1 H), 3.52 (t, J 6.7 Hz, 2H), 3.00 (t, J 6.7 Hz, 2H), 2.71 (s, 3H), 2.42 (s, 3H). LCMS (APCI+) 519 (MH+, 100%). Anal. Calcd for C24H22N8O4S-H2O: C, 53.72; H, 4.51 ; N, 20.88. Found C, 53.85; H, 4.54; N, 20.61.
Example 23: /V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-/V-methyl-2-(methyl(2- (pyridin-2-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E23).
Reaction of E22 (130 mg, 0.25 mmol) using the conditions of Example 2 gave /V-(1-(5-cyano- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(pyridin-2-yl)ethyl)amino)-5-nitro- benzenesulfonohydrazide (E23) as a yellow powder (98 mg, 74%). 1H NMR δ (400 MHz1 CDCI3) 8.88 (d, J 2.8 Hz1 1 H), 8.56 (dd, J 7.2, 1.0 Hz, 1 H), 8.46 (ddd, J 4.9, 1.8, 0.9 Hz, 1 H), 8.35 (S1 1 H), 8.31 (dd, J 1.9, 1.0 Hz, 1 H), 8.28 (dd, J 9.2, 2.8 Hz, 1 H), 7.54 (td, J 7.6, 1.8 Hz1 1 H), 7.22 - 7.16 (m, 2H), 7.09 (ddd, J 7.6, 4.9, 1.1 Hz, 1 H)1 7.03 (dd, J 7.2, 1.9 Hz, 1 H), 3.84 (m, 2H), 3.19 - 3.12 (m, 5H), 2.98 (s, 3H), 2.64 (s, 3H). LCMS (APCI+) 533 (MH+, 100%). Anal. Calcd for C25H24N8O4SO^ EtOAc.0.3 H2O: C1 55.77; H, 4.75; N, 20.17. Found C, 55.66; H1 4.82; N, 20.07.
Example 24: AT-(I -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-4- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E24).
Reaction of E7 (137 mg, 0.34 mmol) and Λ/-methyl-2-(pyridin-4-yl)ethanamine (250 mg, 1.84 mmol) using the conditions of Example 9 gave /V-O^δ-cyanopyrazoloπ.δ-alpyridin-S-yl)- ethylidene)-2-(methyl(2-(pyridin-4-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E24) as a yellow powder (142 mg, 80%). 1H NMR δ (400 MHz, de-DMSO) 11.00 (br s, 1 H), 8.89 (dd, J 7.2, 0.7 Hz1 1H), 8.70 (d, J 2.8 Hz, 1 H), 8.55 (s, 1 H), 8.36 (m, 2H), 8.18 (dd, J 9.3, 2.8 Hz, 1 H), 7.60 (s, 1 H), 7.34 (d, J 9.3 Hz, 1 H), 7.24 (dd, J 7.2, 1.9 Hz, 1 H), 7.20 (m, 2H), 3.77 (m, 2H), 3.11 (s, 3H), 2.90 (m, 2H), 2.36 (s, 3H). LCMS (APCI+) 519 (MH+, 100%). Anal. Calcd for C24H22N8O4S: C, 55.59; H, 4.28; N, 21.61. Found C, 55.93; H, 4.35; N, 21.58.
Example 25: 2-((2-(1W-lmidazol-1-yl)ethyl)(methyl)amino)-Λr-(1-(5-cyanopyrazolo[1,5-a]- pyridin-3-yl)ethylidene)-5-nitrobenzenesulfonohydrazide (E25).
A solution of E7 (60 mg, 0.15 mmol), 2-(1/-/-imidazol-1-yl)-Λ/-methylethanamine (42 mg, 0.34 - mmol) and NEt3 (0.06 ml_, 0.45 mmol) in DMSO (2 mL) was stirred at room temperature for 4 days. Chromatography (eluting with CH2CI2 to CH2CI2:Me0H 95:5), then recrystallisation from CH2CI2-MeOH-1Pr2O gave 2-((2-(1 H-imidazol-1 -yl)ethyl)(methyl)amino)-Λ/l-(1 -(5- cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-5-nitrobenzenesulfonohydrazide (E25) as an orange solid (43 mg, 57%). 1H NMR δ (400 MHz, d6-DMSO) 11.08 (br s, 1 H)1 8.89 (dd, J 7.2, 0.9 Hz, 1 H), 8.66 (d, J 2.8 Hz, 1 H), 8.54 (s, 1 H), 8.12 (dd, J 9.27, 2.8 Hz, 1H), 7.63 (s, 1 H), 7.58 (s, 1 H), 7.25 - 7.19 (m, 2H), 7.07 (t, J 1.1 Hz, 1 H), 6.73 (m, 1 H), 4.25 (t, J 6.3 Hz, 2H), 3.91 (t, J 6.3 Hz, 2H), 3.08 (s, 3H), 2.36 (s, 3H). LCMS (APCI+) 508 (MH+, 100%). Anal. Calcd for C22H21N9O4S: C, 52.06; H, 4.17; N, 24.84. Found C, 51.92; H, 4.23; N, 24.85.
Example 26: 2-((2-(1H-lmidazol-4-yl)ethyl)(methyl)amino)-/V-(1-(5-cyanopyrazolo[1,5-a]- pyridin-3-yl)ethylidene)-5-nitrobenzenesulfonohydrazide (E26). Reaction of E7 (70 mg, 0.17 mmol), 2-(1/-/-imidazol-4-yl)-Λ/-methylethanamine (65 mg, 0.52 mmol) using the conditions of Example 9 gave 2-((2-(1/-/-imidazol-4-yl)ethyl)(methyl)amino)-/V- (1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-5-nitrobenzenesulfonohydrazide (E26) as an orange powder (54 mg, 61%). 1H NMR δ (400 MHz, d6-DMSO) 8.88 (dd, J 7.2, 0.9 Hz, 1 H), 8.73 (d, J 2.8 Hz, 1 H), 8.53 (s, 1 H), 8.20 (dd, J 9.3, 2.8 Hz, 1 H), 7.68 (m, 1 H), 7.61 (s, 1 H), 7.33 (d, J 9.3 Hz1 1 H), 7.23 (dd, J 7.2, 1.9 Hz, 1 H), 6.78 (s, 1 H), 3.67 (m, 2H), 3.04 (s, 3H),
2.81 (m, 2H), 2.36 (s, 3H). LCMS (APCI+) 508 (MH+, 100%). Anal. Calcd for C22H2iN9O4S.0.4 CH2CI2.0.35 hexanes: C, 49.60; H, 4.98; N, 21.52. Found C, 49.49; H, 5.00; N, 21.50.
Example 27: /V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-methyl-5-nitro- benzenesulfonohydrazide (E27).
Step 27.1 : A solution of 4-(terf-butoxycarbonylamino)pyridine (4.94 g, 25.4 mmol) and O-(2,4- dinitrophenyl)hydroxylamine (5.06 g, 25.4 mmol) [C. Legault et al., J. Org. Chem. 2003, 68(18), 7119] in MeCN (100 mL) was heated at 40 0C for 18 h. The solvent was removed in vacuo, and then the residue was taken up in DMSO (150 mL) and cooled to 0 0C. K2CO3 (7.03 g, 50.9 mmol) followed by 3-butyn-2-one (1.99 mL, 25.4 mmol) were then added. After 1h, the ice bath was removed and the suspension stirred at room temperature for 18 h. The reaction was diluted with cold water and the aqueous layer was extracted three times with EtOAc. The combined organic extracts were washed with 1 N HCI, followed by water and brine. The organic layer was dried (Na2SO4) and the solvent removed in vacuo. Filtration through a plug of neutral AI2O3 (eluting with EtOAc) gave tert-butyl 3-acetylpyrazolo[1 ,5-a]pyridin-5- ylcarbamate as a pale yellow solid (4.20 g, 60%). 1H NMR δ (400 MHz, CDCI3) 8.41 (d, J 7.6 Hz, 1 H), 8.26 (s, 1H), 8.08 (d, J 2.3 Hz, 1 H), 7.58 (m, 1 H), 6.96 (s, 1 H), 2.71 (s, 3H), 1.55 (s, 9H). LCMS (APCI+) 276 (MH+, 100%).
Step 27.2: A solution of tert-butyl 3-acetylpyrazolo[1 ,5-a]pyridin-5-ylcarbamate (81 mg, 0.29 mmol) and trifluoroacetic acid (0.43 ml_, 5.8 mmol) in CH2CI2 (5 mL) was stirred at room temperature for 18h. The solvents were removed in vacuo to leave the trifluoroacetate salt of 1-(5-aminopyrazolo[1 ,5-a]pyridin-3-yl)ethanone as a brown solid (86 mg, 100%). 1H NMR δ (400 MHz, d6-DMSO) 8.33 (d, J 7.4 Hz, 1 H), 8.22 (s, 1 H), 7.51 (d, J 2.6 Hz, 1 H), 6.46 (dd, J 7.4, 2.6 Hz, 1 H), 2.52 (s, 3H). LCMS (APCI+) 176 (MH+, 100%).
Step 27.3: A solution of NaNO2 (0.378 g, 5.5 mmol) in H2O (4 mL) was added dropwise to a suspension of the trifluoroacetate salt of 1-(5-aminopyrazolo[1 ,5-a]pyridin-3-yl)ethanone (1.7g, 4.2 mmol) in 48% HBr (5 mL) in an ice-salt bath at a rate so that the internal temperature of the reaction was maintained below -5 °C. The reaction was stirred for 10 mins, then a solution of CuBr (1.51 g, 10.5 mmol) in 48% HBr (5 mL) was added dropwise to maintain the internal temperature of the reaction between -5 and 0 °C. After complete addition of the CuBr, the reaction was stirred at 0 0C for a further 30 mins, and then at room temperature for 1 h. The reaction was cooled and carefully neutralised to pH 8 with 2N NaOH. A solution of EDTA (3 g) in water (100 mL), followed by CH2CI2 (100 mL) were added and the reaction was stirred vigorously for 15 mins. The solution was filtered and the layers separated. The aqueous layer was extracted twice with EtOAc, washed with water and brine. The combined organic extracts were dried (Na2SO4) and the solvent removed in vacuo. Chromatography (eluting with hexanes to hexanes: EtOAc 85:15) gave 1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethanone as a pale yellow solid (677 mg, 51%). 1H NMR δ (400 MHz, CDCI3) 8.61 (dd, J 2.2, 0.8 Hz, 1H), 8.37 (dd, J 7.3, 0.8 Hz, 1 H), 8.32 (s, 1 H), 7.10 (dd, J 7.3, 2.2 Hz1 1 H), 2.55 (s, 3H). LCMS (APCI+) 239 (MH+ with 79Br, 100%), 241 (MH+ with 81Br, 80%).
Step 27.4: Reaction of 1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethanone (60 mg, 0.25 mmol) using the conditions of Step 1.2 gave /^(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2- methyl-5-nitrobenzenesulfonohydrazide (E27) as an orange solid (88 mg, 78%). 1H NMR δ (400 MHz, d6-DMSO) 10.97 (br s, 1 H), 8.79 (d, J 2.5 Hz, 1 H), 8.67 (d, J 7.3 Hz, 1 H), 8.44 (dd, J 8.4, 2.5 Hz, 1 H), 8.39 (s, 1 H), 7.77 (d, J 8.4 Hz, 1H), 7.70 (d, J 2.2 Hz, 1 H), 7.11 (dd, J 7.3, 2.2 Hz, 1 H), 2.77 (s, 3H), 2.32 (s, 3H). LCMS (APCI+) 452 (MH+ with 79Br, 100%), 454 (MH+ with 81Br, 100%). Anal. Calcd for C16H15BrN5θ4S.0.04 hexanes: C, 43.16; H, 3.30; N, 15.74. Found C1 42.80; H, 3.22; N, 15.37.
Example 28: /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-/V,2-dimethyl-5-nitro- benzenesulfonohydrazide (E28).
Reaction of E27 (49 mg, 0.11 mmol) using the conditions of Example 2 gave /V-(1-(5-bromo- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/,2-dimethyl-5-nitrobenzenesulfonohydrazide (E28) as a cream-coloured solid (27 mg, 53%). 1H NMR δ (400 MHz, CDCI3) 8.78 (d, J 2.4 Hz, 1 H), 8.38 (dd, J 8.4, 2.4 Hz, 1H), 8.36 (dd, J 7.3, 0.7 Hz, 1 H), 8.24 (s, 1H), 8.06 (d, J 2.2 Hz, 1 H), 7.60 (d, J 8.4 Hz, 1 H), 7.02 (dd, J 7.3, 2.2 Hz, 1 H), 3.00 (s, 3H), 2.71 (s, 3H), 2.63 (s, 3H). LCMS (APCI+) 466 (MH+ with 79Br, 90%), 468 (MH+ with 81Br, 100%). Anal. Calcd for C17H16BrN5O4S.0.33 EtOAc: C, 44.42; H, 3.79; N1 14.14. Found C, 44.36; H, 3.77; N, 14.14.
Example 29: /V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5-nitro- benzenesulfonohydrazide (E29).
Step 29.1 : Reaction of 1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethanone (155 mg, 0.65 mmol) using the conditions of Step 7.1 gave 5-bromo-3-(1-hydrazonoethyl)pyrazolo[1 ,5-a]pyridine as a yellow solid. 1H NMR δ (400 MHz, CDCI3) 8.49 (dd, J 2.2, 0.7 Hz, 1 H), 8.28 (dd, J 7.3, 0.7 Hz, 1 H), 8.00 (s, 1H), 6.89 (dd, J 7.3, 2.2 Hz, 1 H), 5.22 (br s, 2H), 2.18 (s, 3H). LCMS (APCI+) 253 (MH+ with 79Br, 100%), 255 (MH+ with 81Br, 90%).
Step 29.2: Reaction of crude 5-bromo-3-(1-hydrazonoethyl)pyrazolo[1 ,5-a]pyridine using the conditions of Step 7.2 gave /V-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5- nitrobenzenesulfonohydrazide (E29) as a yellow solid (66 mg, 22% over 2 steps). 1H NMR δ (400 MHz, ds-DMSO) 11.13 (br s, 1 H), 8.72 (dd, J 5.9, 2.9 Hz, 1 H), 8.69 (dd, J 7.3, 0.6 Hz, 1 H), 8.63 (m, 1 H), 8.41 (s, 1 H), 7.83 - 7.77 (m, 2H), 7.14 (dd, J 7.3, 2.3 Hz, 1 H), 2.32 (s, 3H). LCMS (APCI+) 456 (MH+ with 79Br, 95%), 458 (MH+ with 81Br, 100%). Anal. Calcd for C15H11BrFN5O4S: C, 39.49; H, 2.43; N, 15.35. Found C, 39.83; H, 2.46; N, 15.10.
Example 30: /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(methyl- amino)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E30).
Reaction of E29 (196 mg, 0.43 mmol) and Λ/./V-dimethylethylenediamine (0.70 mL, 6.6 mmol) using the conditions of Example 9 gave /^-(^(δ-bromopyrazoloti .δ-aJpyridin-S-yOethylidene)- 2-(methyl(2-(methylamino)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E30) as a yellow powder (184 mg, 82%). 1H NMR δ (400 MHz1 CDCI3) 9.18 (d, J 2.8 Hz, 1 H), 8.39 (dd, J 8.9, 2.8 Hz, 1 H), 8.20 (dd, J 7.3, 0.7 Hz, 1 H), 8.00 (s, 1 H)1 7.35 (d, J 8.9 Hz1 1 H)1 7.30 (dd, J 2.2, 0.7 Hz, 1 H), 6.82 (dd, J 7.3, 2.2 Hz, 1 H), 3.19 (m, 2H), 2.87 (m, 2H), 2.64 (s, 3H), 2.59 (s, 3H), 2.31 (S1 3H). LCMS (APCI+) 524 (MH+ with 79Br, 95%), 526 (MH+ with 81Br, 100%). HRMS (ESI+) Calcd for C19H23 79BrN7O4S: 524.0710; found (MH+) 524.0715.
Example 31 : Λf-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2- (dimethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E31).
Reaction of E29 (183 mg, 0.40 mmol) and Λ/.Λ/./V-trimethylethylenediamine (0.27 mL, 2.1 mmol) using the conditions of Example 9 gave
Figure imgf000039_0001
.S-alpyridin-S- yl)ethylidene)-2-((2-(dimethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E31) as a yellow powder (130 mg, 61 %). 1H NMR δ (400 MHz, CDCI3) 9.05 (d, J 2.7 Hz, 1 H), 8.47 (dd, J 8.8, 2.7 Hz, 1 H), 8.23 (dd, J 7.3, 0.7 Hz1 1 H), 8.06 (s, 1 H), 7.45 (d, J 8.8 Hz, 1 H), 7.42 (dd, J 2.2, 0.7 Hz, 1H), 6.85 (dd, J 7.3, 2.2 Hz, 1 H), 3.20 (m, 2H), 2.79 (s, 3H), 2.56 (m, 2H), 2.44 (S, 3H), 2.28 (s, 6H). LCMS (APCI+) 538 (MH+ with 79Br, 100%), 540 (MH+ with 81Br, 95%). Anal. Calcd for C20H24BrN7O4SOOS hexane: C, 44.93; H, 4.59; N, 18.07. Found C, 45.07; H, 4.60; N, 17.87.
Example 32: Λf-(1 -(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)- ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E32).
Reaction of E29 (211 mg, 0.46 mmol) and A/,A/-diethyl-/V-methylethylenediamine (0.23 mL, 1.4 mmol) using the conditions of Example 9 gave /^-(^(δ-bromopyrazoloIi .δ-alpyridin-S-yl)- ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E32) as a yellow powder (174 mg, 66%). 1H NMR δ (400 MHz, CDCI3) 9.01 (d, J 2.7 Hz, 1 H), 8.46 (dd, J 8.8, 2.7 Hz, 1 H), 8.23 (dd, J 7.3, 0.7 Hz, 1 H), 8.06 (s, 1 H), 7.47 (d, J 8.8 Hz, 1H), 7.41 (d, J 2.2 Hz, 1 H), 6.86 (dd, J 7.3, 2.2 Hz, 1 H), 3.20 (m, 2H), 2.80 (s, 3H), 2.68 (m, 2H), 2.63 (q, J 7.1 Hz, 4H), 2.45 (s, 3H), 1.03 (t, J 7.1 Hz, 6H). LCMS (APCI+) 566 (MH+ with 79Br, 100%), 568 (MH+ with 81Br, 90%). Anal. Calcd for C22H28BrN7O4S-CI hexane: C, 47.20; H, 5.15; N, 17.05. Found C, 47.26; H, 5.15; N, 17.11.
Example 33: /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)- ethyl)(methyl)amino)-Λ/-methyl-5-nitrobenzenesulfonohydrazide (E33). Reaction of E32 (112 mg, 0.20 mmol) using the conditions of Example 2 gave /^-(^(δ-bromo- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-(diethylamino)ethyl)(methyl)amino)-Λ/-methyl-5- nitrobenzenesulfonohydrazide (E33) as a yellow powder (35 mg, 31%). 1H NMR δ (400 MHz, CDCI3) 8.99 (d, J 2.8 Hz, 1 H), 8.32 (d, J 7.3 Hz, 1 H), 8.29 (dd, J 9.2, 2.8 Hz, 1 H), 8.21 (s, 1 H), 8.16 (d, J 2.2 Hz, 1H), 7.26 (d, J 9.2 Hz, 1H), 7.01 (dd, J 7.3, 2.2 Hz, 1H), 3.53 (m, 2H), 3.09 (s, 3H), 3.05 (s, 3H), 2.68 (m, 2H), 2.61 (s, 3H), 2.46 (q, J 7.1 Hz, 4H), 0.93 (t, J 7.1 Hz, 6H). LCMS (APCI+) 580 (MH+ with 79Br, 100%), 582 (MH+ with 81Br, 100%). Anal. Calcd for C23H30BrN7O4S.0.5 MeOH: C, 47.76; H, 5.51 ; N, 16.25. Found C, 47.93; H, 5.59; N, 16.38. Example 34: /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(3- morpholinopropyl)amino)-5-nitrobenzenesulfonohydrazide (E34).
Reaction of E29 (202 mg, 0.44 mmol) and Λ/-methyl-3-morpholinopropan-1 -amine (119 mg, 0.75 mmol) using the conditions of Example 9 gave /V-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-(methyl(3-morpholinopropyl)amino)-5-nitrobenzenesulfonohydrazide (E34) as a yellow powder (171 mg, 65%). 1H NMR δ (400 MHz, CDCI3) 9.07 (d, J 2.7 Hz, 1 H), 8.45 (dd, J 8.8, 2.7 Hz, 1 H), 8.23 (dd, J 7.3, 0.8 Hz, 1 H), 8.04 (s, 1 H), 7.49 - 7.45 (m, 2H), 6.88 (dd, J 7.3, 2.2 Hz, 1 H), 3.61 (m, 4H), 3.14 (m, 2H), 2.84 (s, 3H), 2.37 (s, 3H), 2.36 - 2.29 (m, 6H), 1.76 (m, 2H). LCMS (APCI+) 594 (MH+ with 79Br, 100%), 596 (MH+ with 81Br, 95%). Anal. Calcd for C23H28BrN7O5S-O-S MeOH: C, 46.23; H, 4.95; N, 16.06. Found C, 46.20; H, 4.91 ; N, 15.75.
Example 35: /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4- (methylsulfonyl)piperazin-1-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E35). Reaction of E29 (80 mg, 0.18 mmol) and Λ/-methyl-2-(4-(methylsulfonyl)piperazin-1-yl)- ethanamine (116 mg, 0.53 mmol) using the conditions of Example 25 gave /V-(1-(5-bromo- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4-(methylsulfonyl)piperazin-1-yl)ethyl)- amino)-5-nitrobenzenesulfonohydrazide (E35) as a yellow powder (71 mg, 62%). 1H NMR δ (400 MHz, d6-DMSO) 8.75 (d, J 2.7 Hz, 1H), 8.66 (d, J 7.3 Hz, 1 H), 8.40 (s, 1H), 8.30 (dd, J 9.2, 2.7 Hz, 1 H), 7.43 (d, J 9.2 Hz, 1 H), 7.38 (s, 1 H), 7.08 (dd, J 7.3, 2.2 Hz, 1 H), 3.58 (m, 2H), 2.97 (s, 3H), 2.87 (m, 4H), 2.76 (s, 3H), 2.47 (m, 2H), 2.38 - 2.31 (m, 7H). LCMS (APCI+) 657 (MH+ with 79Br, 100%), 659 (MH+ with 81Br, 100%). Anal. Calcd. for C23H29BrN8O6S-CIS CH2CI2.0.15 hexanes: C, 42.28; H, 4.63; N, 16.40. Found C, 42.31 ; H, 4.64; N, 16.20.
Example 36: ΛP-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-2- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E36).
Reaction of E29 (210 mg, 0.46 mmol) and Λ/-methyl-1-(pyhdin-2-yl)methanamine (194 mg, 1.59 mmol) using the conditions of Example 9 gave /V-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-(methyl(pyridin-2-ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E36) as a yellow foam (149 mg, 58%). 1H NMR δ (400 MHz, CDCI3) 12.51 (br s, 1 H), 9.26 (d, J 2.8 Hz, 1 H), 8.77 (m, 1 H), 8.44 (dd, J 8.8, 2.8 Hz, 1 H), 8.19 (dd, J 7.3, 0.8 Hz, 1 H), 7.99 (s, 1 H), 7.77 (td, J 7.7, 1.8 Hz, 1 H), 7.40 - 7.35 (m, 2H), 7.33 (dd, J 2.2, 0.8 Hz, 1 H), 7.25 (m, 1 H), 6.83 (dd, J 7.3, 2.2 Hz, 1 H), 4.50 (s, 2H), 2.59 (s, 3H), 2.49 (s, 3H). LCMS (APCI+) 558 (MH+ with 79Br, 90%), 560 (MH+ with 81Br, 100%). Anal. Calcd for C22H20BrN7O4S: C, 47.32; H, 3.61; N, 17.56. Found C, 47.27; H, 3.57; N, 17.45. Example 37: /V-(1 -(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl- (pyridin-2-ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E37).
Reaction of E36 (106 mg, 0.19 mmol) using the conditions of Example 2 gave /V-(1-(5-bromo- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(pyridin-2-ylmethyl)amino)-5-nitro- benzenesulfonohydrazide (E37) as a yellow solid (82 mg, 76%). 1H NMR δ (400 MHz, CDCI3) 9.05 (d, J 2.8 Hz, 1 H), 8.52 (ddd, J 4.9, 1.8, 1.0 Hz, 1 H), 8.31 (dd, J 7.2, 0.7 Hz, 1 H), 8.30 (dd, J 9.2, 2.8 Hz, 1 H), 8.21 (s, 1 H), 8.20 (dd, J 2.2, 0.7 Hz, 1 H), 7.59 (td, J 7.7, 1.8 Hz, 1 H), 7.34 (d, J 7.7 Hz, 1 H), 7.27 (d, J 9.2 Hz, 1 H), 7.16 (ddd, J 7.7, 4.9, 1.0 Hz, 1 H), 7.00 (dd, J 7.2, 2.2 Hz, 1 H), 4.70 (s, 2H)1 3.11 (s, 3H), 3.07 (s, 3H), 2.62 (s, 3H). LCMS (APCI+) 572 (MH+ with 79Br, 95%), 574 (MH+ with 81Br, 100%). Anal. Calcd for C23H22BrN7O4S-O^ EtOAc.0.5 H2O: C, 47.72; H, 4.14; N, 16.37. Found C, 47.70; H, 4.10; N, 16.05.
Example 38: /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-3- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E38). Reaction of E29 (80 mg, 0.18 mmol) and Λ/-methyl-1-(pyridin-3-yl)methanamine (107 mg, 0.88 mmol) using the conditions of Example 9 gave /V-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-(methyl(pyridin-3-ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E38) as a yellow powder (62 mg, 63%). 1H NMR δ (400 MHz, d6-DMSO) 10.89 (s, 1H), 8.78 (d, J 2.8 Hz, 1 H), 8.64 (dd, J 7.3, 0.6 Hz, 1 H), 8.47 (m, 1 H), 8.40 (s, 1 H), 8.37 (dd, J 4.8, 1.6 Hz, 1 H), 8.28 (dd, J 9.2, 2.8 Hz, 1 H), 7.63 (dt, J 7.8, 1.9 Hz, 1 H), 7.41 - 7.34 (m, 2H), 7.22 (ddd, J 7.8, 4.8, 0.6 Hz, 1 H), 7.06 (dd, J 7.3, 2.3 Hz, 1 H), 4.70 (s, 2H), 2.97 (s, 3H), 2.35 (s, 3H). LCMS (APCI+) 558 (MH+ with 79Br, 90%), 560 (MH+ with 81Br, 100%). Anal. Calcd. for C22H20BrN7O4S: C, 47.32; H, 3.61 ; N, 17.56. Found C, 47.17; H, 3.64; N, 17.44.
Example 39: /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-4- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E39).
Reaction of E29 (247 mg, 0.54 mmol) and Λ/-methyl-1-(pyridin-4-yl)methanamine (228 mg, 1.87 mmol) using the conditions of Example 9 gave /V-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-(methyl(pyridin-4-ylmethyl)amino)-5-nitrobenzenesulfonohydrazide (E39) as a yellow solid (100 mg, 33%). 1H NMR δ (400 MHz, d6-DMSO) 10.97 (s, 1 H), 8.77 (d, J 2.8 Hz1 1 H), 8.65 (dd, J 7.3, 0.6 Hz1 1 H)1 8.43 - 8.39 (m, 3H), 8.25 (dd, J 9.3, 2.8 Hz, 1 H), 7.43 (m, 1 H), 7.33 (d, J 9.3 Hz, 1H), 7.27 (m, 2H), 7.06 (dd, J 7.3, 2.2 Hz1 1 H)1 4.73 (s, 2H), 3.06 (s, 3H), 2.36 (s, 3H). LCMS (APCI ) 556 (M-H+ with 79Br, 90%), 558 (M-H+ with 81Br, 100%). Anal. Calcd for C22H20BrN7O4S.0.2 H2O: C, 47.02; H, 3.66; N, 17.45. Found C, 46.91 ; H, 3.81 ; N, 17.44. Example 40: AT-(I -(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin- 2-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E40).
Reaction of E29 (205 mg, 0.45 mmol) and Λ/-methyl-2-(pyridin-2-yl)ethanamine (197 mg, 1.45 mmol) using the conditions of Example 9 gave ^-(^(δ-bromopyrazoloII .S-alpyridin-S-yl)- ethylidene)-2-(methyl(2-(pyridin-2-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E40) as a yellow solid (210 mg, 82%). 1H NMR δ (400 MHz1 CDCI3) 10.19 (br s, 1 H), 9.13 (d, J 2.8 Hz, 1 H), 8.62 (m, 1 H), 8.43 (dd, J 8.9, 2.8 Hz, 1 H), 8.22 (dd, J 7.3, 0.7 Hz, 1 H), 8.01 (s, 1 H), 7.65 (td, J 7.7, 1.8 Hz, 1 H), 7.51 (dd, J 2.2, 0.7 Hz, 1 H), 7.47 (d, J 8.9 Hz, 1 H), 7.24 - 7.18 (m, 2H), 6.86 (dd, J 7.3, 2.2 Hz, 1 H), 3.48 (t, J 6.7 Hz, 2H), 2.99 (t, J 6.7 Hz, 2H), 2.68 (s, 3H), 2.38 (s, 3H). LCMS (APCI+) 572 (MH+ with 79Br, 100%), 574 (MH+ with 81Br, 95%). Anal. Calcd for C23H22BrN7O4SO^ EtOAc: C, 48.71 ; H, 4.25; N, 15.90. Found C, 49.07; H, 4.22; N, 15.92.
Example 41: /V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-/V-methyl-2-(methyl(2- (pyridin-2-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E41). Reaction of E40 (138 mg, 0.24 mmol) using the conditions of Example 2 gave /V-(1-(5-bromo- pyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(pyridin-2-yl)ethyl)amino)-5-nitro- benzenesulfonohydrazide (E41) as a yellow powder (107 mg, 76%). 1H NMR δ (400 MHz, CDCI3) 8.92 (d, J 2.7 Hz, 1 H), 8.46 (d, J 4.9 Hz, 1 H), 8.31 (d, J 7.3 Hz, 1 H), 8.25 (dd, J 9.2, 2.7 Hz, 1 H), 8.21 (s, 1 H), 8.10 (d, J 2.1 Hz, 1 H), 7.53 (td, J 7.6, 1.8 Hz, 1 H), 7.21 - 7.15 (m, 2H), 7.08 (dd, J 7.6, 4.9 Hz, 1 H), 6.99 (dd, J 7.3, 2.1 Hz, 1 H), 3.81 (m, 2H)1 3.15 (m, 2H), 3.11 (s, 3H), 3.00 (s, 3H), 2.60 (s, 3H). LCMS (APCI+) 586 (MH+ with 79Br, 90%), 588 (MH+ with 81Br, 100%). Anal. Calcd for C24H24BrN7O4S.0.4 EtOAcH2O: C, 48.06; H, 4.60; N, 15.33. Found C, 48.08; H, 4.41 ; N, 15.34.
Example 42: Λf-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin- 4-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E42).
Reaction of E29 (80 mg, 0.18 mmol) and Λ/-methyl-2-(pyridin-4-yl)ethanamine (71 mg, 0.53 mmol) using the conditions of Example 25 gave /V-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-2-(methyl(2-(pyridin-4-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide (E42) as a yellow powder (39 mg, 39%). 1H NMR δ (400 MHz, d6-DMSO) 10.74 (s, 1H)1 8.73 (d, J 2.7 Hz, 1H)1 8.64 (d, J 7.3 Hz, 1H), 8.42 - 8.32 (m, 3H), 8.23 (dd, J 9.13, 2.7 Hz, 1H), 7.46 - 7.31 (m, 2H), 7.18 (d, J 6.0 Hz1 2H)1 7.07 (dd, J 7.3, 2.2 Hz, 1 H), 3.74 (m, 2H), 3.06 (s, 3H), 2.86 (m, 2H), 2.33 (s, 3H). LCMS (APCI+) 572 (MH+ with 79Br, 100%), 574 (MH+ with 81Br, 90%). Anal. Calcd for C23H22BrN7O4S: C, 48.26; H, 3.87; N, 17.13. Found C, 48.07; H, 3.84; N, 16.95. Example 43: 2-((2-(1W-lmidazol-1-yl)ethyl)(methyl)amino)-W-(1-(5-bromopyrazolo[1,5-a]- pyridin-3-yl)ethylidene)-5-nitrobenzenesulfonohydrazide (E43).
Reaction of E29 (80 mg, 0.18 mmol) and 2-(1H-imidazol-1-yl)-Λ/-methylethanamine (44 mg, 0.35 mmol) using the conditions of Example 25 gave 2-((2-(1H-imidazol-1- yl)ethyl)(methyl)amino)-Λ/'-(1 -(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-5- nitrobenzenesulfonohydrazide (E43) as an orange powder (51 mg, 52%). 1H NMR δ (400 MHz, de-DMSO) 10.8 (br s, 1 H), 8.70 (d, J 2.8 Hz, 1 H), 8.64 (d, J 7.3 Hz, 1 H), 8.38 (s, 1 H), 8.17 (dd, J 9.2, 2.8 Hz, 1 H), 7.54 (s, 1 H), 7.44 (s, 1 H), 7.23 (d, J 9.2 Hz, 1 H), 7.08 - 7.03 (m, 2H), 6.73 (s, 1 H), 4.23 (t, J 6.4 Hz, 2H), 3.89 (t, J 6.4 Hz, 2H), 3.02 (s, 3H), 2.33 (s, 3H). LCMS (APCI+) 561 (MH+ with 79Br, 100%), 563 (MH+ with 81Br, 80%). Anal. Calcd for C2i H2IBrN8O4S: C, 44.93; H, 3.77; N, 19.96. Found C, 44.68; H, 3.84; N, 19.79.
Example 44: 2-((2-(1 W-lmidazol-4-yl)ethyl)(methyl)amino)-/V-(1 -(5-bromopyrazolo[1 ,5-a]- pyridin-3-yl)ethylidene)-5-nitrobenzenesulfonohydrazide (E44). Reaction of E29 (80 mg, 0.18 mmol) and 2-(1/-/-imidazol-4-yl)-Λ/-methylethanamine (44 mg, 0.35 mmol) using the conditions of Example 9 gave 2-((2-(1H-imidazol-4- yl)ethyl)(methyl)amino)-/V-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-5- nitrobenzenesulfonohydrazide (E44) as orange crystals (48 mg, 49%). 1H NMR δ (400 MHz, d6-DMSO) 8.76 (d, J 2.8 Hz, 1 H), 8.63 (dd, J 7.3, 0.6 Hz, 1 H)1 8.36 (s, 1 H), 8.25 (dd, J 9.2, 2.8 Hz, 1 H), 7.56 (s, 1 H), 7.46 (d, J 1.9 Hz, 1 H), 7.37 (d, J 9.2 Hz, 1 H), 7.06 (dd, J 7.3, 2.3 Hz, 1H), 6.75 (S, 1 H), 3.63 (m, 2H), 3.01 (s, 3H), 2.77 (m, 2H), 2.34 (s, 3H). LCMS (APCI+) 561 (MH+ with 79Br, 95%), 563 (MH+ with 81Br, 100%). Anal. Calcd for C21H21BrN8O4S: C, 44.93; H, 3.77; N, 19.96. Found C, 44.73; H, 3.91; N, 19.95.
Example 45: /V-(1-(6-Bromoimidazo[1,2-a]pyridin-3-yl)ethylidene)-2-fluoro-5-nitro- benzenesulfonohydrazide (E45).
Reaction of 1-(6-bromoimidazo[1 ,2-a]pyridin-3-yl)ethanone (300 mg, 1.25 mmol) using the conditions of Example 7 gave /V-(1-(6-bromoimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-fluoro-5- nitrobenzenesulfonohydrazide (E45) as a yellow powder (195 mg, 34%). 1H NMR δ (400 MHz, d6-DMSO) 11.50 (br s, 1 H), 9.41 (s, 1H), 8.72 (dd, J 5.9, 2.9 Hz, 1 H), 8.57 (m, 1 H), 8.16 (s, 1 H), 7.76 (t, J 9.1 Hz, 1 H), 7.67 (d, J 9.5 Hz, 1 H), 7.51 (dd, J 9.5, 1.9 Hz, 1H), 2.38 (s, 3H). LCMS (APCI+) 456 (MH+ with 79Br, 100%), 458 (MH+ with 81Br, 95%). Anal. Calcd for C15H11BrFN5O4S: C, 39.49; H1 2.43; N, 15.35. Found C, 39.35; H, 2.61 ; N1 15.22. Example 46: /V-(1-(6-Bromoimidazo[1,2-a]pyridin-3-yl)ethylidene)-2-((2-(dimethylamino)- ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E46).
Reaction of E45 (150 mg, 0.33 mmol) and Λ/,Λ/,Λ/'-trimethylethylenediamine (0.13 mL, 0.99 mmol) using the conditions of Example 9 gave Λ^1-(6-bromoimidazo[1 ,2-a]pyridin-3-yl)- ethylidene)-2-((2-(dimethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide (E46) as a yellow solid (96 mg, 54%). 1H NMR δ (400 MHz, d6-DMSO) 9.38 (s, 1 H), 8.83 (d, J 2.8 Hz, 1 H), 8.25 (dd, J 9.0, 2.8 Hz, 1 H), 7.93 (s, 1 H), 7.56 (d, J 9.5 Hz, 1 H), 7.44 (d, J 9.0 Hz, 1 H), 7.34 (dd, J 9.5, 2.0 Hz, 1H), 3.59 (m, 2H), 3.04 (m, 2H), 2.86 (s, 3H), 2.62 (s, 6H), 2.31 (s, 3H). LCMS (APCI+) 538 (MH+ with 79Br, 90%), 540 (MH+ with 81Br, 100%). Anal. Calcd for C20H24BrN7O4S: C, 44.61 ; H, 4.49; N, 18.21. Found C, 44.75; H, 4.61 ; N, 17.81.
BIOLOGICAL ACTIVITY OF COMPOUNDS OF THE INVENTION
A. Inhibition of isolated enzyme Compounds were evaluated for their ability to inhibit the Class I Pl 3-kinase enzymes p110α/p85 and p110δ/p85. Reaction mixtures comprising 0.1 μg of recombinant enzyme, 10 μg of L-α-phosphatidylinositol, the compound (DMSO only or DMSO + compound to a final concentration of 1%), 2X Lipid Kinase Buffer (40 mM Tris-HCI pH 7.4, 200 mM NaCI, 1 mM EDTA) were activated by the addition of an ATP mix (5 mM MgCI2, 100 μM ATP, 0.1 μL [Y33P]ATP). Reactions were incubated at room temperature for 1 h, then stopped by the addition of 1M HCI. The lipids were then extracted using a two step procedure. Firstly, 200 μL of chloroform/methanol (1 :1) was added, the biphasic reactions mixed and centrifuged briefly, and the inorganic phase was removed and discarded. Following this, 80 μL of methanokHCI (1 :1) was added and the same procedure followed. The organic phase (70 μL) was then transferred to a clean 1.6 mL tube and the reactions dried using a Speedvac, with no heating, for 30 mins. The reaction were spotted onto TLC plates (Merck Ltd) and developed for 1 h in propanol-1 :2M acetic acid (13:7). The TLC plates were then dried at room temperature and quantified using a phosphorimager (Stormlmager, Amersham). Nine inhibitor concentrations were used to determine the IC50.
Results for compounds of Formula I, obtained using the above methods are shown in Table 2 below.
B. Cellular growth inhibition The compounds were evaluated against two early passage cell lines NZB5 and NZOV9, whose development and culture have been described [Marshall et al, Oncol. Res. 2004, 14, 297]. The cells were grown in ITS medium (α-modified minimal essential medium supplemented with insulin, transferring, selenite and 5% fetal bovine serum) and grown on a 96-well tissue culture plates under an atmosphere of 5% O2, 5%CO2 and 90% N2. Individual wells contained 500-1000 cells (depending on the growth rate) in a volume of 150 μL Compounds were added at 10-fold concentration steps to a maximum of 20 μM and plates were incubated for five days, with 3H-thymidine being added over the last 6 h. Cells were harvested and incorporated radioactivity measured. Duplicate samples were analysed for each compound dose with multiple control samples. Data were fitted by a least-squares method to an exponential of the form y = yo + ae'bx, where y is the radioactivity (corrected for background and normalised to 100% of the control), x is the radiation dose, and y0, a and b are variables, and the IC50 value defined as the compound concentration reducing 3H- thymidine levels by 50%.
Results for compounds of Formula I, obtained using the above methods are shown in Table 2 below.
C. Mouse liver microsome metabolism.
Incubation mixtures were prepared by combining 32 μL of the test compound solution (50 μL of 0.5 mM stock solution diluted to 5 mL with MiIIi Q water), 32 μL of MgCI2 - EDTA solution (24 mM MgCI2.6H2O and 5 mM EDTA tetra sodium salt in MiIIi Q water), 64 μL microsome solution (1 part pooled male CD1 mouse liver microsomes supplied by Gentest: 1 part MiIIi Q water: 6 parts 400 mM pH 7.4 phosphate buffer), and 32 μL of 5 mM NADPH in MiIIi Q water. However, to allow for temperature equilibration of the microsomes at 37 0C, the addition of NADPH was delayed by 5 min. For the 0 min reference sample, water was added instead of NADPH to ensure no metabolism occurred. The incubation mixtures were left for 0, 5, 10, 15, and 20 min at 37 0C before 50 μL aliquots were taken and mixed with ice cold 200 μL internal standard solution. Using an orbital shaker they were vortexed and centrifuged offline. The resulting supernatants were diluted with water 1:1 v/v (100 μL aliquots of each) prior to LCMS- QQQ analysis (Agilent Zorbax SB-C18 2.1x50mm δmicron column using a gradient of 20% of solution A (5 mM NH4CO2H and 0.1% HCO2H in water) and 80% of solution B (5 mM NH4CO2H and 0.1% HCO2H in methanol) to 100% of solution B over 5 mins, at a flow rate of 0.4 mL/min).
Results for compounds of Formula I, obtained using the above methods A, B and C are shown in Table 2 below. Table 2. Biological data for selected compounds of Table 1.
Figure imgf000046_0001
Figure imgf000047_0001
IC50 values are an average of two or more determinations, except for those in italics which are a single determination.
The compounds described in Table 1 are all inhibitors of Pl 3-kinase. In particular, they inhibit at least one of the Pl 3-kinase isoforms with IC50 < 1 μM. Some of these examples have IC50 <10 nM. In addition, they show inhibition of cellular proliferation in at least one of the two cell lines described above with IC50 <20 μM.

Claims

CLAIMS:
1. A compound of Formula I,
Figure imgf000048_0001
I wherein Het represents pyrazolo[1 ,5-a]pyridine or imidazo[1 ,2-a]pyridine (below);
Figure imgf000048_0002
pyrazolo[1 ,5-a]pyridine imidazo[1 ,2-a]pyridine
wherein:
X may represent up to two of halogen, R1, OR1, OCOR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, NO2, NR1 2 or NR1COR1, placed at any of the available positions A-, 5-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the available positions 5-, 6-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine; Y is H or Me;
Z is CO, CR1 2, S(O)x, N(R1)CO, N(R1)CR1 2, N(R1)S(O)X where x = O, 1 , 2; D is phenyl, naphthyl, or 5- or 6-membered heterocycle or benzoheterocycle, where the heterocyclic ring contains up to two of the atoms S, O, N; optionally substituted at any available position which is up to two of halogen, R1, OR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, CF3, OCF3, NO2, NR1 2, NR1COR1;
R1 is H or C1-6 saturated or unsaturated alkyl, optionally substituted with up to three of halogen, R3, OR3, NR3 2, OCOR3, CONR3 2, CO2R3, CN, SO2R3, SO2NR3 2, NR3COR3 or optionally substituted aryl or heteroaryl, or in the case where R1 forms part of NR1 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR4;
R2 is C1-6 saturated or unsaturated alkyl, optionally substituted with up to three of halogen, R3, OR3, NR3 2, OCOR3, CONR3 2, CO2R3, CN, SO2R3, SO2NR3 2, NR3COR3 or optionally substituted aryl or heteroaryl; R3 is H, C1-6 saturated or unsaturated alkyl, optionally substituted with up to three of halogen, R4, OR4, NR4 2, OCOR4, CONR4 2, CO2R4, CN, SO2R4, SO2NR4 2, NR4COR4; or optionally substituted aryl or heteroaryl, or in the case where R3 forms part of NR3 2 this may form an optionally substituted 4-7 membered saturated ring optionally containing one additional heteroatom from the group O, S, NR4; R4 is H, C1-6 saturated or unsaturated alkyl, C(O)-alkyl, CO2-alkyl, S(O)y-alkyl where y
= 0, 1 , 2; including pure geometric and enantiomeric forms and mixtures thereof, physiologically acceptable salts thereof, and prodrugs thereof; with the proviso that the following compounds are excluded:
Λ/χi-(6-Bromoimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide;
Λ/χi-(6-Bromoimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-Λ/,2-dimethyl-5- nitrobenzenesulfonohydrazide; /V-(1 -(6-Bromo-2-methylimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide;
/^-(^(δ-Bromo^-methylimidazoti ^-alpyridin-S-yOethylideneJ-Λ/^-dimethyl-δ- nitrobenzenesulfonohydrazide;
Λ/-Benzyl-Λ/l-(1-(6-bromo-2-methylimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide.
2. A compound according to claim 1 where X may represent one or two of halogen, R1, OR1, OCOR1, CONR1 Z, CO2R1, SO2R1, SO2NR1 2, CN, NO2, NR1 2 Or NR1COR1, placed at any of the available positions 4-, 5-, 6-, 7- when Het represents pyrazolo[1 ,5- a]pyridine or at any of the available positions 5-, 6-, 7-, 8- when Het represents imidazo[1 ,2-a]pyridine.
3. A compound according to claim 1 or 2 where X may represent one or two of halogen, R1, OR1, OCOR1, CONR1 2, CO2R1, SO2R1, SO2NR1 2, CN, NO2, NR1 2 or NR1COR1, where one of X is placed at position 5- where Het represents pyrazolo[1 ,5-a]pyridine or at position 6- where Het represents imidazo[1 ,2-a]pyridine, and the other X if present is placed at any of the remaining available positions 4-, 6-, 7- when Het represents pyrazolo[1 ,5-a]pyridine or at any of the remaining available positions 5-, 7-, 8- when Het represents imidazo[1,2-a]pyridine.
4. A compound according to any one of the preceding claims where D is phenyl; optionally substituted at any available position with up to two of halogen, R1, OR1, CONR1 Z, CO2R1, SO2R1, SO2NR1 2, CN, CF3, OCF3, NO2, NR1 2, NR1COR1.
5. A compound according to any one of the preceding claims where Z is N(R1JCO, N(R1)CR1 2, or N(R1JS(O)x where x = O, 1 , 2.
6. A compound according to any one of the preceding claims where Z is N(R1)S(O)X where x = 0, 1 , 2.
7. A compound according to any one of the preceding claims where Het represents 5 pyrazolo[1 ,5-a]pyridine (below);
Figure imgf000050_0001
pyrazolo[1 ,5-a]pyridine
8. A compound according to any one of the preceding claims where R4 is H, C1-6 0 saturated or unsaturated alkyl.
9. A compound according to claim 1 wherein, the compound of Formula I is selected from: /V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-methyl-5- nitrobenzenesulfonohydrazide 5 /V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/,2-dimethyl-5- nitrobenzenesulfonohydrazide
^-(^(δ-CyanopyrazoloIi .δ-alpyridin-S-yOpropylideneJ^-methyl-δ-nitrobenzenesulfono- hydrazide
^-(i^δ-Cyanopyrazoloti .δ-alpyridin-S-yOpropylideneJ-Λ/^-dimethyl-δ-0 nitrobenzenesulfonohydrazide
/V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)butylidene)-2-methyl-5- nitrobenzenesulfonohydrazide
Λ^(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)butylidene)-Λ/,2-dimethyl-5- nitrobenzenesulfonohydrazide δ /V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5- nitrobenzenesulfonohydrazide
2-((2-Aminoethyl)(methyl)amino)-Λ/'-(1-(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-
5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-0 (methylamino)ethyl)amino)-5-nitrobenzenesulfonohydrazide
/V-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-
(dimethylaminoethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Cyanopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-((2-
(dimethylamino)ethyl)(methyl)amino)-Λ/-methyl-5-nitrobenzenesulfonohydrazide /V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethyliclene)-2-((3-
(dimethylamino)propyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide
Λ^1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-
(diethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide /V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-
(diethylamino)ethyl)(methyl)amino)-Λ/-methyl-5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(piperidin-1- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(piperidin- 1 -yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(3- morpholinopropyl)amino)-5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4-
(methylsulfonyl)piperazin-1-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide /V-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-2- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide
Λ/l-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-πnethyl-2-(methyl(pyridin-2- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide
^-(^(S-CyanopyrazoloII.S-alpyridin-S-yOethylidene^^methyKpyridin^- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-2- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(pyridin-2- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide /V-(1 -(5-Cyanopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-4- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
2-((2-(1 /-/-lmidazol-1 -yl)ethyl)(methyl)amino)-/V-(1 -(5-cyanopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide
2-((2-(1H-lmidazol-4-yl)ethyl)(methyl)amino)-/V-(1-(5-cyanopyrazolo[1,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide
^-(^(S-BromopyrazoloII .S-alpyridin-S-yOethylideneJ^-methyl-S- nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-/\/,2-dimethyl-5- nitrobenzenesulfonohydrazide /V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-fluoro-5- nitrobenzenesulfonohydrazide /V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-
(methylamino)ethyl)amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-
(dimethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide /V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-
(diethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-((2-
(diethylamino)ethyl)(methyl)amino)-Λ/-methyl-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(3- morpholinopropyl)amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(4-
(methylsulfonyl)piperazin-1-yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
Λ/'-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-2- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide Λ/'-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(pyridin-2- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide
Λ^(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-3- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1,5-a]pyridin-3-yl)ethylidene)-2-(methyl(pyridin-4- ylmethyl)amino)-5-nitrobenzenesulfonohydrazide
/V-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-2- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
Λ/l-(1-(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-Λ/-methyl-2-(methyl(2-(pyridin-2- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide /V-(1 -(5-Bromopyrazolo[1 ,5-a]pyridin-3-yl)ethylidene)-2-(methyl(2-(pyridin-4- yl)ethyl)amino)-5-nitrobenzenesulfonohydrazide
2-((2-(1H-lmidazol-1-yl)ethyl)(methyl)amino)-/V-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide
2-((2-(1H-lmidazol-4-yl)ethyl)(methyl)amino)-Λ/'-(1-(5-bromopyrazolo[1 ,5-a]pyridin-3-yl)- ethylidene)-5-nitrobenzenesulfonohydrazide
/V-(1-(6-Bromoimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-fluoro-5- nitrobenzenesulfonohydrazide and
/V-(1 -(6-Bromoimidazo[1 ,2-a]pyridin-3-yl)ethylidene)-2-((2-
(dimethylamino)ethyl)(methyl)amino)-5-nitrobenzenesulfonohydrazide including pure geometric and enantiomeric forms and mixtures thereof, physiologically acceptable salts thereof, and prodrugs thereof.
10. A compound according to any one of the previous claims wherein the prodrugs are selected from phosphate or carboxylic acid or aminoacid ester prodrugs.
11. A method of cancer prevention or therapy for treating cancers, the method including the step of administering a compound of Formula I as defined in any one of claims 1 to
10.
12. The method according to claim 11 further including the administration of one or more additional chemotherapeutic agents and/or therapies.
13. The method according to claim 12 wherein the additional chemotherapeutic agents and/or therapies are selected from any one or more of:
Alkylation agents
Antimetabolites Antitumour antibiotics
Antitumour vegetable alkaloids
Antitumour hormones
Antitumour immunological agents
Radiation therapy Surgery
14. The method according to claim 12 or claim 13 including the the step of administering one or more chemotherapeutic agents to the subject before, during or after the administration of the compound of Formula I as defined in any one of claims 1 to 10.
15. The method according any one claims 11 to 14 wherein a compound of Formula I is administered to a human or other primate.
16. The method according any one claims 11 to 14 wherein a compound of Formula I is administered to a farm, sports, or pet animal.
17. A pharmaceutical composition including a compound of Formula I according to any one of claims 1 to 10 together with a pharmaceutically acceptable excipient, adjuvant, carrier, buffer or stabiliser.
18. A pharmaceutical composition according to claim 17 wherein the composition is adapted for oral or parenteral administration.
19. A pharmaceutical composition according to claim 17 wherein the composition is adapted for cutaneous, subcutaneous, or intravenous injection.
20. A pharmaceutical composition according to claim 17 wherein the composition is in a tablet, capsule, powder, or liquid form.
21. A pharmaceutical composition according to any one of claims 17 to 20 wherein the composition further includes one or more chemotherapeutic agents.
22. A pharmaceutical composition according to claim 21 wherein the chemotherapeutic agents are selected from any one or more of:
Alkylation agents Antimetabolites Antitumour antibiotics
Antitumour vegetable alkaloids Antitumour hormones Antitumour immunological agents.
23. The use of a compound according to any one of claims 1 to 10 in the manufacture of a medicament for cancer prevention or therapy for the treatment of cancer.
24. The use according to claim 23 wherein the medicament is in tablet, capsule, powder or liquid form.
25. The use according to claim 23 wherein the medicament is suitable for oral or parenteral administration.
26. A compound according to any one of claims 1 to 10 for use in cancer prevention or therapy for the treatment of cancer.
27. A method of making a compound of Formula I according to any one of claims 1 to 10, the method including the step of modifying a pyrazolo[1 ,5-a]pyridine-3-carbonyl compound of Formula Il or an imidazo[1 ,2-a]pyridine of Formula III
Figure imgf000055_0001
wherein variables X, Y and R2 are defined according to any one of claims 1 to 10.
28. A method of making a compound according any one of claims 1 to 10 substantially as herein described with particular reference any one or more of the steps as described in Schemes 1 to 7 herein.
29. A method of making a compound of Formula I according to any one of claims 1 to 10 from compound of Formula Il or Formula III
Figure imgf000055_0002
wherein variables X, Y and R2 are defined according to any one of claims 1 to 10 the method including any one of the following steps: (i) condensation with a sulfonohydrazide; or
(ii) condensation with hydrazine followed by reaction with a sulfonyl chloride; or
(iii)condensation with hydrazine followed by reaction with a sulfonyl chloride and then nucleophilic substitution; or
(iv)any one of (i) to (iii) followed by reaction with CH2N2.
30. A compound according to any one of claims 1 to 10 when produced by a method according to any one of claims 27 to 29.
PCT/NZ2009/000298 2008-12-23 2009-12-21 Pyrazolo[1,5-a]pyridine and imidazo[1,2-a]pyridine derivatives and their use in cancer therapy WO2010074586A1 (en)

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US8871754B2 (en) 2012-11-19 2014-10-28 Irm Llc Compounds and compositions for the treatment of parasitic diseases
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