WO2015094119A1 - Wnt pathway modulators - Google Patents

Wnt pathway modulators Download PDF

Info

Publication number
WO2015094119A1
WO2015094119A1 PCT/SG2014/000602 SG2014000602W WO2015094119A1 WO 2015094119 A1 WO2015094119 A1 WO 2015094119A1 SG 2014000602 W SG2014000602 W SG 2014000602W WO 2015094119 A1 WO2015094119 A1 WO 2015094119A1
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
methyl
compound
substituents
pyrimidin
Prior art date
Application number
PCT/SG2014/000602
Other languages
French (fr)
Inventor
Jenefer ALAM
Soo Yei HO
Wei Ling WANG
Athisayamani Jeyaraj DURAISWAMY
Original Assignee
Agency For Science, Technology And Research
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to US15/105,224 priority Critical patent/US10189842B2/en
Priority to CA2933683A priority patent/CA2933683A1/en
Priority to JP2016540531A priority patent/JP6452703B2/en
Priority to CN201480075676.5A priority patent/CN105992767B/en
Priority to ES14870867.0T priority patent/ES2687493T3/en
Priority to DK14870867.0T priority patent/DK3083631T3/en
Priority to KR1020167019109A priority patent/KR102383739B1/en
Priority to EP14870867.0A priority patent/EP3083631B1/en
Application filed by Agency For Science, Technology And Research filed Critical Agency For Science, Technology And Research
Priority to SG11201604839XA priority patent/SG11201604839XA/en
Priority to AU2014367284A priority patent/AU2014367284B2/en
Priority to BR112016013640A priority patent/BR112016013640A8/en
Publication of WO2015094119A1 publication Critical patent/WO2015094119A1/en
Priority to US16/226,398 priority patent/US10870653B2/en
Priority to US17/084,507 priority patent/US11434245B2/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/18—Drugs for disorders of the alimentary tract or the digestive system for pancreatic disorders, e.g. pancreatic enzymes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/06—Antipsoriatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/04—Drugs for skeletal disorders for non-specific disorders of the connective tissue
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/02—Ophthalmic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure

Definitions

  • the invention relates to WNT pathway modulators, processes for making them and methods for using them.
  • Wnt proteins are secreted glycoproteins acting as growth factors that regulate various cellular functions include proliferation, differentiation, death, migration, and polarity, by activating multiple intracellular signaling cascades, including the ⁇ -catenin-dependent and - independent pathways.
  • Wnt members There are 19 Wnt members have been found in humans and mice, and they exhibit unique expression patterns and distinct functions during development.
  • the 10 members of the Frizzled (Fz) family comprise a series of seven-pass transmembrane receptors that have been identified as Wnt receptors.
  • single-pass transmembrane proteins such as low-density lipoprotein receptor-related protein 5 (LRP5), LRP6, receptor tyrosine kinase (RTK)-like orphan receptor l(Rorl), Ror2, and receptor-like tyrosine kinase (Ryk), have been shown to function as co-receptors for Wnt signaling. Therefore, it has been assumed traditionally that the binding of different Wnts to their specific receptors selectively triggers different outcomes via distinct intracellular pathways.
  • LRP5 low-density lipoprotein receptor-related protein 5
  • RTK receptor tyrosine kinase
  • Rorl receptor tyrosine kinase
  • Ryk receptor-like tyrosine kinase
  • ⁇ -catenin adenomatous polyposis coli (APC) and Axin proteins, as well as glycogen synthase kinase 3 (GSK3) and casein kinase I (CKI).
  • APC adenomatous polyposis coli
  • GSK3 glycogen synthase kinase 3
  • CKI casein kinase I
  • Phosphorylated ⁇ -catenin is bound by the F box protein Slimb/p-TrCP and polyubiquitinated, leading to proteosomal degradation.
  • the complex acts to prevent nuclear localization of b-catenin.
  • Frizzled Frizzled
  • LRP5/6 low-density lipoprotein-related proteins 5 and 6
  • GSK3, Axin, and other destruction complex components are recruited to the receptor complex. The function of the destruction complex is inhibited, and unphosphorylated ⁇ -catenin
  • Deregulation of components of Wnt/p-catenin signaling is implied in a wide spectrum of diseases including degenerative diseases, metabolic diseases, and a number of cancers such as cervical, colon, breast, bladder, head and neck, gastric, lung, ovarian, prostate, thyroid, non- small-cell lung, as well as chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal cell carcinoma, osteosarcoma, hepatocellular carcinoma, Wilm's tumor and medulloblastoma.
  • degenerative diseases such as cervical, colon, breast, bladder, head and neck, gastric, lung, ovarian, prostate, thyroid, non- small-cell lung, as well as chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal cell carcinoma, osteosarcoma, hepatocellular carcinoma, Wilm's tumor and medullob
  • Wnt signaling plays a role both during development, and within stem cell niches in adults. This is best established in skin, hematopoietic stem cells, mammary gland and in intestinal proliferation. For example, high level expression of DK 1, an inhibitor of Wnt signaling, blocks normal stem cell proliferation in mouse intestine, suggesting there is an essential role for Wnt signaling in maintenance of stem cells in the digestive tract. Wnt roles in self renewal and expansion of stem cells have also been demonstrated for embryonic and neural stem cells, suggesting that Wnt signaling may be a general requirement of stem cell
  • Wnt signaling e.g., by overexpression of axin or an extracellular Wnt-binding protein, sFRP, reduces hematopoietic stem cell (HSC) growth in vitro and the ability to reconstitute HSCs in vivo.
  • HSC hematopoietic stem cell
  • Wnts 1, 5a, and 10b are able to stimulate expansion of HSC populations and Wnt5a acts synergistically with stem cell factor (SCF) to expand and promote self renewal of HSCs.
  • SCF stem cell factor
  • the demonstration of a role for Wnt5a in HSC self renewal and its ability to synergize with stem cell factor is particularly interesting because Wnt5a often acts in a ⁇ -catenin independent manner. While Wnt signaling is critical for stem cell maintenance, it may therefore be via signaling pathways distinct from or in parallel to the ⁇ -catenin pathway.
  • Wnt/ -catenin signaling pathway is essential to embryonic development ingeneral and organ morphogenesis, so it is not surprising that dysregulation of this pathway in adult has been linked to fibroblast biology and fibrosis. It has been demonstrated that Wnt/p-catenin signaling play a role in severe fibrotic diseases, such as pulmonary fibrosis, liver fibrosis, skin fibrosis and renal fibrosis.
  • Dysregulation of Wnt/p-catenin signaling contributes to the development of diabetic retinopathy by inducing retinal inflammation, vascular leakage, and neovascularization.
  • Wnt proteins are also key signaling proteins in joint remodeling processes. Active Wnt signaling contributes to osteophyte formation and might have an essential role in the anabolic pattern of joint remodeling that is observed in ankylosing spondylitis and osteoarthritis. By contrast, blockade of Wnt signaling facilitates bone erosion and contributes to catabolic joint remodeling, a process that is observed in rheumatoid arthritis.
  • IC 50 A measure of the binding of an inhibitor to and the subsequent release from an enzyme is the "IC 50 " value, which reflects the inhibitor concentration, which at a given substrate concentration results in 50 % enzynie activity.
  • subject refers to an animal, preferably a mammal, most preferably a human, who has been the object of treatment, observation or experiment. In one embodiment the subject is not a human. The subject may for example be a non-human mammal.
  • terapéuticaally effective amount means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal or human being sought by a researcher, veterinarian, medical doctor or other clinician, which includes alleviation of the symptoms of the disease or disorder being treated.
  • the term “pharmaceutically acceptable” embraces both human and veterinary use:
  • pharmaceutically acceptable embraces a veterinarily acceptable compound or a compound acceptable in human medicine and health care. In one embodiment, human medicine and health care is excluded.
  • alkyl denotes a C 1-12 alkyl group, suitably a C 1-6 alkyl group, e.g. C 1-4 alkyl group.
  • Alkyl groups may be straight chain or branched. Suitable alkyl groups include, for example, methyl, ethyl, propyl (e.g. n-propyl and isopropyl), butyl (e.g n-butyl, iso-butyl, sec-butyl and tert- butyl), pentyl (e.g. n-pentyl), hexyl (e.g.
  • alk for example in the expressions "alkoxy”, "haloalkyl” and “thioalkyl” should be interpreted in accordance with the definition of "alkyl”.
  • alkoxy groups include methoxy, ethoxy, propoxy (e.g. n-propoxy), butoxy (e.g. n-butoxy), pentoxy (e.g. n- pentoxy), hexoxy (e.g. n-hexoxy), heptoxy (e.g. n-heptoxy) and octoxy (e.g. n-octoxy).
  • Exemplary thioalkyl groups include methylthio-.
  • Exemplary haloalkyl groups include fluoroalkyl e.g. CF 3 .
  • alkenyl denotes a C 2-12 alkenyl group, suitably a C 2-6 alkenyl group, e.g. a C 2-4 alkenyl group, which contains at least one double bond at any desired location and which does not contain any triple bonds.
  • Alkenyl groups may be straight chain or branched.
  • Exemplary alkenyl groups including one double bond include propenyl and butenyl.
  • Exemplary alkenyl groups including two double bonds include pentadienyl, e.g. (IE, 3E)-pentadienyl.
  • alkynyl denotes a C 2-12 alkynyl group, suitably a C2-6 alkynyl group, e.g. a C 2-4 alkynyl group, which contains at least one triple bond at any desired location and may or may not also contain one or more double bonds.
  • Alkynyl groups may be straight chain or branched.
  • Exemplary alkynyl groups include propynyl and butynyl.
  • alkylene denotes a chain of formula -(CH 2 ) n - wherein n is an integer e.g. 1-12, 1-6, 2-6 or 2-5, unless specifically limited.
  • cycloalkyl denotes a C 3-10 cycloalkyl group (i.e. 3 to 10 ring carbon atoms), more suitably a C 3-8 cycloalkyl group, e.g. a C 3-6 cycloalkyl group.
  • exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.
  • a most suitable number of ring carbon atoms is three to six.
  • cycloalkenyl denotes a C 3 - 10 cycloalkenyl group (i.e. 3 to 10 ring carbon atoms), suitably a C 5 - 10 cycloalkenyl group (i.e. 5 to 10 ring carbon atoms), more suitably a C 5-8 cycloalkenyl group e.g. a C 5-6 cycloalkenyl group.
  • Exemplary cycloalkenyl groups include cyclopropenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl.
  • a most suitable number of ring carbon atoms is five to six.
  • Carbocyclyl denotes any ring system in which all the ring atoms are carbon and which contains between three and twelve ring carbon atoms, suitably between three and ten carbon atoms and more suitably between three and eight carbon atoms.
  • Carbocyclyl groups may be saturated or partially unsaturated, but do not include aromatic rings. Examples of carbocyclyl groups include monocyclic, bicyclic, and tricyclic ring systems, in particular monocyclic and bicyclic ring systems. Other carbocyclyl groups include bridged ring systems (e.g. bicyclo[2.2.1]heptenyl).
  • a specific example of a carbocyclyl group is a cycloalkyl group. A further example of a carbocyclyl group is a cycloalkenyl group.
  • heterocyclyl refers to a carbocyclyl group wherein one or more (e.g. 1, 2 or 3) ring atoms are replaced by heteroatoms.
  • the heteroatom(s) are commonly selected from N, S and O.
  • a specific example of a heterocyclyl group is a cycloalkyl group (e.g. cyclopentyl or more particularly cyclohexyl) wherein one or more (e.g. 1, 2 or 3, particularly 1 or 2, especially 1) ring atoms are replaced by heteroatoms selected from N, S or O.
  • heterocyclyl groups containing one hetero atom include pyrrolidine, tetrahydrofuran and piperidine, and exemplary heterocyclyl groups containing two hetero atoms include morpholine and piperazine.
  • a further specific example of a heterocyclyl group is a cycloalkenyl group (e.g. a cyclohexenyl group) wherein one or more (e.g. 1, 2 or 3, particularly 1 or 2, especially 1) ring atoms are replaced by heteroatoms selected from N, S and O.
  • An example of such a group is dihydropyranyl (e.g. 3,4-dihydro-2H-pyran-2-yl-).
  • the heterocyclyl group may be linked to other part or parts of the molecule by a carbon ring atom or nitrogen ring atom.
  • aryl denotes a C 6-12 aryl group, suitably a C -1 o aryl group, more suitably a C 6-8 aryl group.
  • Aryl groups will contain at least one aromatic ring (e.g. one, two or three rings).
  • Aryl groups with multiple aromatic rings include fused aromatic rings and aromatic rings connected to each other by one single bond.
  • An example of a typical aryl group with one aromatic ring is phenyl.
  • An example of a typical aryl group with two fused aromatic rings is naphthyl.
  • An example of an aromatic group with two directly connected aromatic rings is biphenyl.
  • heteroaryl denotes an aryl residue, wherein one or more (e.g. 1, 2, 3, or 4, suitably 1, 2 or 3) ring atoms are replaced by heteroatoms, said heteroatoms commonly being selected from N, S and O, or else a 5-membered aromatic ring containing one or more (e.g. 1, 2, 3, or 4, suitably 1, 2 or 3) ring atoms, said heteroatoms commonly selected from N, S and O.
  • a heteroaryl group will have no ring heteroatoms other than nitrogen.
  • Exemplary monocyclic heteroaryl groups having one heteroatom include: five membered rings (e.g.
  • pyrrole furan, thiophene
  • six membered rings e.g. pyridine, such as pyridin-2-yl, pyridin-3-yl andpyridin-4-yl
  • exemplary monocyclic heteroaryl groups having two heteroatoms include: five membered rings (e.g. pyrazole, oxazole, isoxazole, thiazole, isothiazole, imidazole, such as imidazol-l-yl, imidazol-2-yl imidazol-4-yl); and six membered rings (e.g. pyridazine, pyrimidine, pyrazine).
  • Exemplary monocyclic heteroaryl groups having three heteroatoms include: 1,2,3-triazole, 1,2,4-triazole, 1,2,3- oxadiazole and 1,2,4-oxadiazole.
  • Exemplary monocyclic heteroaryl groups having four heteroatoms include tetrazole.
  • Exemplary bi cyclic heteroaryl groups include: indole (e.g. indol- 6-yl), benzofuran, benzthiophene, quinoline, isoquinoline, indazole, benzimidazole,
  • the heteraryl group may be linked to other part or parts of the molecule by a carbon ring atom or nitrogen ring atom.
  • aryl and heteroaryl groups may, where appropriate, optionally be substituted by one or more (e.g. 1, 2 or 3, suitably 1 or 2) groups each independently selected from monovalent or multivalent (i.e. having valency greater than 1) functional groups.
  • Suitable substituent groups include alkyl, alkenyl, alkynyl, haloalkyl, -alkoxy (e.g. OMe), cycloalkyl, alkenyloxy-, alkynyloxy-, alkoxyalkyl-, nitro, halogen (e.g. fluoro, chloro and bromo), cyano, hydroxyl, oxo, -C(0)-alkyl (e.g.
  • substituents will be selected from alkyl (e.g. Me), fluoroalkyl (e.g. CF 3 and CHF 2 ), alkoxy (e.g. OMe), halogen and hydroxyl.
  • alkylaryl denotes an aryl residue which is connected via an alkylene moiety e.g. a C 1- alkylene moiety.
  • alkylene moiety e.g. a C 1- alkylene moiety.
  • An example of such a group is benzyl: PhCH 2 -.
  • halogen or “halo” comprises fluorine (F), chlorine (CI) and bromine (Br).
  • amino refers to the group -NH 2 .
  • -arylheterocyclyl refers to a heterocyclyl residue which is connected via an aryl moiety.
  • the compounds according to this invention may accordingly exist as enantiomers. Where the compounds possess two or more chiral centers, they may additionally exist as diastereomers. It is to be understood that all such isomers and mixtures thereof are encompassed within the scope of the present invention.
  • stereoisomers [0039] Where the processes for the preparation of the compounds according to the invention give rise to a mixture of stereoisomers, these isomers may be separated by conventional techniques such as preparative chromatography. The compounds may be prepared in racemic form, or individual enantiomers maybe prepared either by enantiospecific synthesis or by resolution.
  • the compounds may, for example, be resolved into their components enantiomers by standard techniques, such as the formation of diastereomeric pairs by salt formation with an optically active acid, such as (-)-di-p-toluoyl-d-tartaric acid and/or (+)-di-p-toluoyl-l-tartaric acid followed by fractional crystallization and regeneration of the free base.
  • an optically active acid such as (-)-di-p-toluoyl-d-tartaric acid and/or (+)-di-p-toluoyl-l-tartaric acid followed by fractional crystallization and regeneration of the free base.
  • optically active acid such as (-)-di-p-toluoyl-d-tartaric acid and/or (+)-di-p-toluoyl-l-tartaric acid followed by fractional crystallization and regeneration of the free base.
  • the compounds may also be resolved by formation of diaste
  • the compounds may be resolved using a chiral HPLC column.
  • Salts and solvates of the compounds of formula (I) and physiologically functional derivatives thereof which are suitable for use in medicine are those wherein the counter-ion or associated solvent is pharmaceutically acceptable.
  • salts and solvates having non- pharmaceutically acceptable counter-ions or associated solvents are within the scope of the present invention, for example, for use as intermediates in the preparation of other compounds and their pharmaceutically acceptable salts and solvates.
  • Suitable salts according to the invention include those formed with both organic and inorganic acids or bases.
  • Pharmaceutically acceptable acid addition salts include those formed from hydrochloric, hydrobromic, sulfuric, nitric, citric, tartaric, phosphoric, lactic, pyruvic, acetic, trifluoroacetic, triphenylacetic, sulfamic, sulfanilic, succinic, oxalic, fumaric, maleic, malic, mandelic, glutamic, aspartic, oxaloacetic, methanesulfonic, ethanesulfonic, arylsulfonic (for example p-toluenesulfonic, benzenesulfonic, naphthalenesulfonic or naphthalenedisulfonic), salicylic, glutaric, gluconic, tricarballylic, cinnamic, substituted cinnamic (for example, phenyl
  • Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts such as those of sodium and potassium, alkaline earth metal salts such as those of calcium and magnesium and salts with organic bases such as dicyclohexylamine and N-methyl-D-glucamine.
  • crystalline forms of the compounds may exist as polymorphs and as such are intended to be included in the present invention.
  • some of the compounds may form solvates with water (i.e. hydrates) or common organic solvents, and such solvates are also intended to be encompassed within the scope of this invention.
  • the present invention further includes within its scope prodrugs of the compounds of this invention.
  • prodrugs will be functional derivatives of the compounds which are readily convertible in vivo into the desired therapeutically active compound.
  • the term “administering” shall encompass the treatment of the various disorders described with prodrug versions of one or more of the claimed compounds, but which converts to the above specified compound in vivo after administration to the subject. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed. H.
  • Protective Groups [0050] During any of the processes for preparation of the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups, such as those described in Protective Groups in Organic Chemistry, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, fully incorporated herein by reference. The protecting groups may be removed at a convenient subsequent stage using methods known from the art.
  • composition is intended to encompass a product comprising the claimed compounds in the therapeutically effective amounts, as well as any product which results, directly or indirectly, from combinations of the claimed compounds.
  • Carriers which can be added to the mixture, include necessary and inert pharmaceutical excipients, including, but not limited to, suitable binders, suspending agents, lubricants, flavorants, sweeteners, preservatives, coatings, disintegrating agents, dyes and coloring agents.
  • suitable carriers and additives may advantageously include water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like;
  • suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like.
  • Soluble polymers as targetable drug carriers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidephenol, polyhydroxyethylaspartamide-phenol, or polyethyleneoxidepolyllysine substituted with palmitoyl residue.
  • the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polyactic acid, polyepsilon caprolactone, polyhydroxy butyeric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels.
  • Suitable binders include, without limitation, starch, gelatin, natural sugars such as glucose or betalactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
  • Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
  • R 1 represents H; optionally substituted alkyl (wherein optional substituents include one or more substituents each independently selected from G 1-6 alkoxy, NH 2 , -NHC 1-3 alkyl and - N(C 1-3 alkyl) 2 ); -C(0)Oalkyl; haloalkyl; haloalkoxy; or -alkylaryl;
  • each R 2 independently represents H; optionally substituted alkyl (wherein optional substituents include one or more substituents each independently selected from C 1-6 alkoxy, NH 2 , -NHC 1-3 alkyl and -N(C 1-3 alkyl) 2 ); -alkylaryl; optionally substituted carbocyclyl (wherein optional substituents include one or more substituents each independently selected from Q.
  • optional substituents include one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and-C(0)NHC 1-6 alkyl); -NHalkyl; - N(alkyl) 2 ; amino; hydroxyl; alkoxy or halo;
  • n 0, 1 or 2;
  • R 3 represents H or alkyl;
  • R4 represents H or alkyl
  • R 5 represents H or alkyk
  • Y represents aryl; heteroaryl; optionally substituted carbocyclyl (wherein optional substituents include one or more substituents each independently selected from C 1-6 alkyl, C ⁇ . 6 alkoxy, C 1- haloalkyl, C 1-6 haloalkoxy and halo); or optionally substituted heterocyclyl (wherein optional substituents include one or more substituents each independently selected from C ⁇ . 6 alkyl, C 1-6 alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and-C(0)NHC 1-6 alkyl); and
  • Z represents optionally substituted alkyl (wherein optional substituents include one or more substituents each independently selected from C 1-6 alkoxy, N3 ⁇ 4, -NHC 1-3 alkyl and— ⁇ ( ⁇ _ 3 alkyl) 2 ); aryl; heteroaryl; -alkylaryl;-alkylheteroaryl; optionally substituted carbocyclyl (wherein optional substituents include one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C 1-6 haloalkoxy and halo); optionally substituted heterocyclyl (wherein optional substituents include one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and-C(0)NHC 1-6 alkyl); - alkylcarbocyclyl wherein carbocyclyl is optionally substituted (wherein
  • optional substituents include one or more substituents each independently selected from Ci -6 alkyl, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and-C(0)NHC 1-6 alkyl); -arylcarbocyclyl wherein carbocyclyl is optionally substituted (wherein optional substituents include one or more substituents each independently selected from C 1- alkyl, Ci -6 alkoxy, C 1-6 haloalkoxy and halo); or -arylheterocyclyl wherein heterocyclyl is optionally substituted (wherein optional substituents include one or more substituents each independently selected from C 1-6 alkyl, -C(0)OC 1-6 alkyl, - C(0)C 1-6 alkyl and-C(0)NHCi -6 alkyl).
  • optional substituents include one or more substituents each independently selected from Ci -6 alkyl, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and-C(0)NHC
  • R ⁇ represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkoxy, NH 2 , -NHC 1-3 alkyl and -N(C 1-3 alkyl) 2 ; -C(0)Oalkyl; haloalkyl; haloalkoxy; or -alkylaryl;
  • each R 2 independently represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkoxy, NH 2 , -NHCi -3 alkyl and— N Ci.
  • n 0, 1 or 2;
  • R 3 represents H or alkyl
  • R4 represents H or alkyl
  • R 5 represents H or alkyl
  • Y represents aryl; heteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C .
  • substituents each independently selected from C 1-6 alkyl, C ⁇ alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1- 6 alkyl and-C(0)NHC 1-6 alkyl;
  • Z represents alkyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkoxy, NH 2 , -NHC 1-3 alkyl and -N(Cj -3 alkyl) 2 ; aryl; heteroaryl; -alkylaryl;-alkylheteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C ⁇ .
  • heterocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and- C(0)NHC 1-6 alkyl; -alkylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1- alkoxy, C 1-6 haloalkoxy and halo; -alkylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and- C(0)NHC 1-6 alkyl; -arylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, -C(0)OC
  • Figure 1 A bar chart illustrating inhibition of the activity of mammalian porcupine by Compound 51 and Compound 110.
  • Figure 2 A graph illustrating efficacy of Compound 51 on the MMTV-Wntl mouse model.
  • Figure 3 A graph illustrating efficacy data of Compound 110 in the MMTV-Wntl mouse model.
  • Figure 4 Images illustrating inhibition of Palmitoylation of Wnt3a by Compound 51.
  • Figures 5 to 7 Images and graphs showing results of the Soft Agar Assay.
  • Figure 5 shows the results for Cell line AsPC-1 treated with Compound 51.
  • Figure 6 shows the results for Cell line HPAF-II treated with Compound 51.
  • Figure 7 shows the results for Cell line CFPAC-1 treated with Compound 51.
  • Ri represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkoxy, NH 2 , -NHC 1-3 alkyl and -N(C 1-3 alkyl) 2 ; -C(0)Oalkyl; haloalkyl; haloalkoxy; or -alkylaryl;
  • each R 2 independently represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C 1- alkoxy, NH 2 , -NHC 1-3 alkyl and - ⁇ ( ⁇ .
  • n 0, 1 or 2;
  • R 3 represents H or alkyl;
  • R4 represents H or alkyl
  • R 5 represents H or alkyl
  • Y represents aryl; heteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C ⁇ alkoxy, C 1-6 haloalkyl, C ⁇ .
  • substituents each independently selected from C 1-6 alkyl, C ⁇ alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1- 6 alkyl and-C(0)NHC 1-6 alkyl;
  • Z represents alkyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkoxy, N3 ⁇ 4, -NHC 1-3 alkyl and -N(C 1-3 alkyl) 2 ; aryl; heteroaryl; -alkylaryl;-alkylheteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C ⁇
  • heterocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and- C(0)NHC 1-6 alkyl; -alkylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, Ci.
  • -alkylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and-C(0)NHC 1-6 alkyl
  • -arylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, C ⁇ 6 haloalkoxy and halo
  • -arylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, -C(0)OC 1-6 alkyl, -C(0)C 1- 6 alkyl and-C(0)NHCi -6 alkyl.
  • aryl is understood to mean “arylene” (e.g.
  • phenyl is understood to mean “phenylene” (i.e. C H 4 )) because Y is a linking group, not a terminal group.
  • Y rings e.g. heteroaryl
  • R] represents alkyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkoxy, N3 ⁇ 4, -NHC ⁇ alkyl and -N(C 1-3 alkyl) 2
  • substituents include methoxy, -NH 2 , -NHmethyl and -NH(methyi) 2 .
  • Examples include C 1-6 alkyl (e.g. unsubstituted C 1- alkyl), for example methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g.
  • n-butyl iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n- hexyl).
  • exemplary Q- 6 alkyl groups are methyl and ethyl, particularly methyl.
  • R 1 represents -C(0)Oalkyl
  • examples include -C(0)OC 1-6 alkyl such as - C(0)OMe, -C(0)OEt, -C(0)OPr and -C(0)OiPr.
  • a specific example is -C(0)OMe.
  • R represents haloalkyl
  • examples include C 1-6 haloalkyl such as CH 2 F, CHF 2 , CF 3 , CH 2 CH 2 F, CH 2 CHF 2 , CH 2 CF 3 , CHFCH 3 and CF 2 CH 3 .
  • a specific example is CF 3 .
  • examples include C 1-6 haloalkoxy such as OCF 3 .
  • R 2 represents alkyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkoxy, NH 2 , -NHC 1-3 alkyl and -N(C 1-3 alkyl) 2
  • exemplary substituents include methoxy, -NH 2 , -NHmethyl and -NH(methyl) 2 .
  • Examples include C 1-6 alkyl (e.g. unsubstituted C 1-6 alkyl), for example methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g.
  • n-butyl iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n- hexyl).
  • An exemplary C 1-6 alkyl group is methyl.
  • R 2 represents -alkylaryl
  • examples include benzyl.
  • R 2 represents carbocyclyl which is optionally substituted by one or more substituents each independently selected from C 1- alkyl (e.g. methyl), C 1-6 alkoxy (e.g. methoxy), C 1-6 haloalkyl (e.g. fluoromethyl such as trifluoromethyl), Ci- haloalkoxy (e.g. fluoromethoxy such as trifluoromethoxy) and halo (e.g. fluoro, e.g. chloro), examples include C 3 -C 6 cycloalkyl - such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Substituted examples include cyclohexyl substituted by methyl.
  • substituents each independently selected from C 1- alkyl (e.g. methyl), C 1-6 alkoxy (e.g. methoxy), C 1-6 haloalkyl (e.g. fluoromethyl such as trifluor
  • R 2 represents heterocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1- 6 alkyl and-C(0)NHC 1-6 alkyl
  • Y may represent heterocyclyl which is optionally substituted by Ci- alkyl (such as methyl).
  • Examples include monocyclic heterocyclyl.
  • the heterocyclyl group may be unsubstituted or may have for example one or two (e.g. one) substituent (e.g. one methyl group).
  • Examples include piperidinyl, morpholinyl, pyrrolidinyl, 4,5-dihydropyrazolyl and 4,5-dihydroisoxazolyl.
  • a specific example is pyrrolidinyl.
  • R 2 represents -NHalkyl
  • examples include -NHMe, NHEt, NHPr and
  • R 2 represents -N(alkyl) 2
  • examples include -N(Me) 2 .
  • R 2 represents alkoxy
  • examples include methoxy and ethoxy, especially methoxy.
  • examples of -(CHR ) n - include -CH 2 - and -CH(CH 3 )-
  • examples of -(CHR 2 ) n - include -CH 2 -CH 2 -, - CH 2 CH(CH 3 , -CH(CH 3 )C3 ⁇ 4- and - CH(CH 3 )-CH(CH 3 )-.
  • An example of a -(CHR 2 ) 2 - group is -CH 2 -CH 2 -.
  • R 3 represents alkyl
  • examples include C 1-6 alkyl such as methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n-hexyl).
  • An exemplary C - 6 alkyl group is methyl.
  • R 4 represents alkyl
  • examples include C 1-6 alkyl, for example methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n- pentyl) and hexyl (e.g. n-hexyl).
  • An exemplary Ci- 6 alkyl group is methyl.
  • R 5 represents alkyl
  • examples include C 1-6 alkyl such as methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n-hexyl).
  • An exemplary C 1-6 alkyl group is methyl.
  • Y represents aryl
  • examples include optionally substituted phenyl.
  • exemplary substituents include one or more (e.g. one or two, especially one) substituents each
  • C 1-6 alkyl such as methyl
  • C 1-6 alkoxy such as methoxy
  • halo and C 1-6 haloalkyl such as fluoroemethyl, e.g. trifluoromethyl
  • Examples include unsubstituted phenyl, methylphenyl, methoxyphenyl, fluorophenyl, chlorophenyl and trifluoromethylphenyl.
  • Specific examples include unsubstituted phenyl.
  • Specific examples also include methylphenyl, methoxyphenyl, fluorophenyl.
  • Z and NR 5 may be positioned on the phenyl ring at the 1 - and 4- positions relative to each other (i.e. Z and R 5 have a para relationship).
  • examples include monocyclic (e.g. 5 and 6 membered) and bicyclic (e.g. 9 and 10 membered) heteroaryl rings, especially monocyclic heteroaryl rings, particularly 6-membered monocyclic heteroaryl rings.
  • monocyclic heteroaryl may comprise one, two or three ring heteroatoms (e.g. one or two, e.g. one) including one or two nitrogen atoms (e.g. one or e.g. two) and optionally an oxygen or sulphur atom.
  • Exemplary 5- membered monocyclic heteroaryl groups include pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl and thiadiazolyl.
  • Z and NR 5 may be positioned on the ring at non-adjacent ring atoms.
  • Exemplary 6-membered monocyclic heteroaryl groups include pyridinyl.
  • Exemplary 6- membered monocyclic heteroaryl groups also include pyridazinyl, pyrimidinyl and pyrazinyl.
  • Z and NR 5 may be positioned on the ring at 1- and 4- positions relative to each other (i.e. Z and NR 5 have a para relationship).
  • aforementioned heteroaryl group may either by unsubstituted or may be substituted by one or more (e.g. one or two, particularly one) substituents.
  • substituents are independently selected from C 1-6 alkyl (such as methyl), C 1- alkoxy (such as'methoxy), halo (such as fluoro) and C 1-6 haloalkyl (such as fluoromethyl, e.g. trifluoromethyl).
  • examples include isoxazolyl (e.g.
  • isoxazolyl-5-yl (wherein NR 5 is at the 5-position and Z is at the 3-position) and isoxazolyl-3-yl (wherein NR 5 is at the 3-position and Z is at the 5-position), especially isoxazol-5-yl), oxadiazolyl, pyridinyl (e.g. pyridin-2-yl or pyridin-3-yl), pyridazinyl (e.g. pyridazin-3-yl), pyrimidinyl (e.g. pyrimidin-3-yl), pyrazinyl (e.g. pyrazin-2-yl).
  • Y When Y is substituted heteroaryl, Y may for example be substituted by methyl or fluoro.
  • examples include methylpyridinyl, fluoropyridinyl,
  • Y represents carbocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl (e.g. methyl), Ci -6 alkoxy (e.g. methoxy), C 1-6 haloalkyl (e.g. fluoromethyl such as trifluoromethyl), C ]-6 haloalkoxy (e.g. fluoromethoxy) and halo (e.g. fluoro, e.g. chloro), Y may represent carbocyclyl which is optionally substituted by C 1-6 alkyl. Examples include monocyclic carbocyclyl which is optionally substituted by C ⁇ . alkyl (such as methyl).
  • carbocyclyl examples include C 3 . 8 cycloalkyl (e.g. cyclohexyl) and C 5-8 cycloalkenyl (e.g. cyclohexenyl).
  • the carbocycyl ring is optionally substituted by one, two or three independently selected substituents (e.g. one or two, especially one, e.g. one methyl group).
  • Y represents carbocyclyl
  • Y may represent C 3-8 cycloalkyl, such as C 5-6 cycloalkyl.
  • a specific example is cyclohexyl.
  • Z and NR5 are may be positioned on the carbocyclyl ring at the 1- and 4- positions relative to each other.
  • Y represents heterocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1- 6 alkyl and-C(0)NHC 1-6 alkyl
  • Y may represent heterocyclyl which is optionally substituted by C ⁇ -6 alkyl (such as methyl). Examples include monocyclic heterocyclyl.
  • the heterocyclyl group may be unsubstituted or may have for example one or two (e.g. one) substituent (e.g. one methyl group).
  • Examples include piperidinyl, morpholinyl, pyrrolidinyl, 4,5-dihydropyrazolyl and 4,5-dihydroisoxazolyl. Specific examples include 4,5-dihydroisoxazolyl (e.g. 4,5- dihydroisoxazol-5-yl) and piperidinyl (e.g. piperidin-4-yl).
  • Y is 6-membered heterocyclyl
  • Z and NR 5 may be positioned on the heterocyclyl ring at the l- and 4- positions relative to each other.
  • Z represents heterocyclyl, in one embodiement Z does not represent -alkylaryl.
  • exemplary substituents include methoxy, -NH 2 , -NHmethyl and -NH(methyl) 2 .
  • Examples include C 1- alkyl (e.g. unsubstituted C 1- alkyl), for example methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g.
  • n-butyl iso-butyl, sec-butyl, tert-butyl
  • pentyl e.g. n-pentyl
  • hexyl e.g. n-hexyl
  • Examples also include C 3- alkyl (e.g. unsubstituted C 3-6 alkyl), for example propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n-hexyl).
  • Exemplary C 1-6 alkyl groups are methyl and tert-butyl.
  • Z represents aryl
  • examples include optionally substituted phenyl.
  • exemplary substituents include one or more (e.g. one or two, especially one) substituents each
  • fluorophenyl e.g. 2- fluorophenyl, 3 -fluorophenyl and 4-fluorophenyl
  • chlorophenyl e.g. 2-chlorophenyl, 3- chlorophenyl, 4-chlorophenyl and 3,4-dichlorophenyl
  • bromophenyl e.g.
  • aminophenyl e.g. 2-aminophenyl, 3-aminophenyl, 4- aminophenyl
  • methoxyphenyl e.g. 2-
  • Z represents heteroaryl
  • examples include monocyclic (e.g. 5 and 6 membered) and bicyclic (e.g. 9 and 10 membered) heteroaryl rings, especially monocyclic rings.
  • monocyclic heteroaryl comprise one, two or three ring heteroatoms (e.g. one or two, e.g. one) including one or two nitrogen atoms (e.g. one or e.g. two) and optionally an oxygen or sulphur atom.
  • Exemplary 5-membered monocyclic heteroaryl groups include pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl and thiadiazolyl.
  • Exemplary 6-membered monocyclic heteroaryl groups include pyridinyl. Exemplary 6- membered monocyclic heteroaryl groups also include pyridazinyl, pyrimidinyl and pyrazinyl.
  • the aforementioned heteroaryl group may either by unsubstituted or may be substituted by one or more (e.g. one or two, particularly one) substituents. Exemplary substituents are
  • pyridinyl e.g. pyridin-2-yl, pyridin-3-yl and pyridin-4-yl
  • pyrazinyl e.g. pyrazin-2-yl
  • pyridazinyl e.g. pyridazin-3-yl
  • pyrimidinyl e.g. pyrimidin-2-yl, pyrimidin-4-yl and pyrimidin-5-yl
  • oxazolyl e.g.
  • oxazol-2-yl and oxazol- 5-yl oxazol-2-yl and oxazol- 5-yl
  • thiazolyl e.g. thiazol-2-yl and thiazole-5-yl
  • pyrazolyl e.g. pyrazol-l-yl
  • Substituted examples include chloropyridinyl (e.g. 4-chloropyridin-2-yl, 5-chloropyridin-2-yl and 5-chloropyridin-3-yl), fluoropyridinyl (e.g. 4-fluoropyridin-2-yl, 5-fluoropyridin-2-yl and 5- fluoropyridin-3-yl), methylpyridinyl (e.g. 2-methylpyridin-5-yl, 6-methylpyridin-2-yl and 5- methylpyridin-3-yl), fluoromethylpyridinyl (e.g. 5-trifluoromethylpyiidin-3-yl), aminopyridinyl (e.g.
  • methylpyrazinyl e.g. 5-methylpyrazin-2-yl
  • methylthiazolyl e.g. 2- methylthiazol'4-yl
  • methylpyrazolyl e.g. l-methylpyrazol-5-yl
  • Z represents -alkylaryl
  • examples include benzyl.
  • Z represents -alkylheteroaryl (e.g. -CH 2 -heteroaryl)
  • examples include - alkylpyrrolyl, -alkylpyrazolyl, -alkylimidazolyl, -alkyloxazolyl, -alkylisoxazolyl, -alkylthiazolyl, -alkylisothiazolyl, -alkyloxadiazolyl, -alkylthiadiazolyl, -alkylpyridinyl, -alkylpyridazinyl, - alkylpyrimidinyl and -alkylpyrazinyl.
  • Z represents carbocyclyl which is optionally substituted by one or more substituents each independently selected from Ci -6 alkyl (e.g. methyl), C 1-6 alkoxy (e.g. methoxy), C 1-6 haloalkyl (e.g. fluoromethyl), C 1-6 haloalkoxy (e.g. fluoromethoxy) and halo (e.g. fluoro, e.g. chloro)
  • Z may represent carbocyclyl which is optionally substituted by Ci- 6 alkyl. Examples include monocyclic carbocyclyl which is optionally substituted by Q-6 alkyl (such as methyl). Examples of carbocyclyl include C 3-8 cycloalkyl (e.g.
  • cyclohexyl and C 5-8 cycloalkenyl (e.g. cyclohexenyl).
  • the carbocycyl ring is optionally substituted by one, two or three substituents (e.g. one or two, especially one, e.g. one methyl group).
  • Z may represent C 3-8 cycloalkyl, such as C5 -6 cycloalkyl.
  • a specific example is cyclohexyl.
  • Z represents heterocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, -C(0)OC 1-6 alkyl, -C(0)C 1- 6 alkyl,-C(0)NHC 1-6 alkyl C 1-6 alkyl (e.g. methyl), -C(0)OC 1-6 alkyl (e.g. -C(O)Omethyl), - C(0)Ci -6 alkyl (e.g. -C(0)methyl),-C(0)NHCi -6 alkyl (e.g. -C(O)NHmethyl), examples include unsubstituted heterocyclyl and heterocyclyl with one or two (e.g. one) substituent.
  • Examples include heterocyclyl groups containing one nitrogen atom (such as pyrrolidinyl and piperidinyl) or two nitrogen atoms (such as piperazinyl). Examples also include heterocyclyl groups containing one nitrogen atom and one oxygen atom(e.g. morpholinyl) or one sulfur atom (e.g. thiomorpholinyl). In one embodiment when Z is substituted, a ring nitrogen atom is substituted. Substituted examples include substituted piperazinyl, such as 4-substitued piperazinyl, e.g. 4- Boc-piperazinyl, 4-(C(0)Me)-piperazinyl, 4-(C(0)NHEt)piperazinyl and 4-methylpiperazinyl.
  • Z represents -alkylcarbocyclyl (e.g. -CH 2 -carbocyclyl) wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl (e.g. methyl), Ci -6 haloalkyl (e.g. fluoromethyl such as trifluoromethyl), C 1-6 alkoxy (e.g.
  • Ci -6 haloalkoxy e.g. fluoromethoxy
  • halo e.g. fluoro, e.g. chloro
  • examples include -CH 2 -cyclopropyl and -CH 2 -cyclobutyl.
  • Z represents -alkylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, -C(0)OC 1-6 alkyl, -C(0)Ci. 6 alkyl,-C(0)NHC ]-6 alkyl C 1-6 alkyl (e.g. methyl), -C(0)OCi -6 alkyl (e.g. -C(O)Omethyl), - C(0)C 1-6 alkyl (e.g. -C(0)methyl),-C(0)NHC 1-6 alkyl (e.g.
  • examples include -CH 2 -heterocyclyl, such as -CH 2 -morpholinyl, -CH 2 -piperazinyl, -CH 2 -piperidinyl and -CH 2 - pyrrolidinyl.
  • Z represents— aryl carbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C 1 . 6 alkyl (e.g. methyl), Ci -6 alkoxy (e.g. methoxy), C 1-6 haloalkyl (e.g. fluoromethyl such as trifluoromethyl), Ci.ghaloalkoxy (e.g.
  • the carbocyclyl ring may be unsubstituted or may be substituted by one or more C 1-6 alkyl groups.
  • the carbocyclyl ring may be monocyclic.
  • An exemplary carbocyclyl ring is cycloalkyl.
  • Examples include cyclopropylphenyl- and cyclohexylphenyl-.
  • Z represents -arylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, -C(0)OC 1- 6 alkyl, -C(0)C 1-6 alkyl,-C(0)NHC 1-6 alkyl C 1-6 alkyl (e.g. methyl), -C(0)OCi -6 alkyl (e.g. - C(O)Omethyl), -C(0)C 1-6 alkyl (e.g. -C(0)methyl),-C(0)NHC 1-6 alkyl (e.g. -C(O)NHmethyl), Z may be -phenylheterocyclyl.
  • the heterocyclyl ring may be monocyclic and may contain one or two (e.g. one) nitrogen atoms.
  • Examples include morphinolylphenyl-, piperazinylphenyl-, piperidinylphenyl-, pyrrolidinylphenyl-.
  • a specific example is 3-(morpholin-4-yl)phenyl-.
  • 3 ⁇ 4 represents H, methyl, ethyl, -C(0)OMe, CF 3 or OMe. In one embodiment represents methyl.
  • each R 2 independently represents H or alkyl. More suitably R 2 represents H or methyl. In one embodiment, R 2 represents H. In one embodiment R 2 represents methyl.
  • n 0 or 1 , for example n represents 1.
  • R 3 represents H or methyl, for example R 3 represents H.
  • R4 represents H or methyl, for example R4 represents H.
  • R5 represents H or methyl, for example R 5 represents H.
  • W and X are the same as each other.
  • Y represents aryl (e.g. phenyl) or heteroaryl (e.g. 5- or 6-membered monocyclic heteroaryl comprising one, two or three ring heteroatoms including one or two nitrogen atoms such as isoxazolyl, pyridinyl, pyridazinyl, pyrimidinyl or pyrazinyl).
  • Y represents phenyl.
  • Y represents monocyclic heteroaryl.
  • Y is substituted by one or more substituents each independently selected from Ci- 6 alkyl, C ⁇ 6 alkoxy, halo and C 1-6 haloalkyl. In one embodiment Y is unsubstituted.
  • Y is monosubstituted.
  • Z and NR 5 are positioned on the ring at non-adjacent ring atoms.
  • Z and NR 5 are positioned on the ring at 1- and 4- positions relative to each other (i.e. Z and NR 5 have a para relationship).
  • Z represents aryl (e.g.phenyl) or heteroaryl (for example 6-membered heteroaryl such as pyridinyl, pyridazinyl, pyrimidinyl or pyrazinyl especially pyridinyl; or 5- membered heteroaryl such as oxazolyl, thiazolyl, pyrazolyl).
  • Z represents heteroaryl.
  • Z represents aryl.
  • Z does not represent methyl or ethyl.
  • Z does not represent unsubstituted alkyl.
  • Ri represents alkyl
  • each R 2 independently represents H, alkyl, carbocyclyl which may be optionally substituted by one or more substituents each independently selected from C 1- alkyl, C 1- alkoxy, Ci -6 haloalkoxy and halo, -NHalkyl, -N(alkyl) , amino, hydroxyl, alkoxy or halo;
  • n 0, 1 or 2;
  • R4 represents H or alkyl
  • R 5 represents H or alkyl
  • Y represents aryl; heteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkyl, Q.
  • substituents each independently selected from C 1-6 alkyl, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and - C(0)NHC 1-6 alkyl;
  • Z represents alkyl; aryl; heteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkoxy (e.g.
  • heterocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, -C(0)OC 1-6 alkyl, -C(0)C 1-6 alkyl and -C(0)NHC 1-6 alkyl; -arylcarbocyclyl wherein carbocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, C 1-6 alkoxy, C 1-6 haloalkoxy and halo ;or -arylheterocyclyl wherein heterocyclyl which is optionally substituted by one or more substituents each independently selected from C 1-6 alkyl, -C(0)OC 1-6 alkyl, -C(0)Ci -6 alkyl and -C(0)NHC 1-6 alkyl.
  • the following compounds of fonnula (I) are excluded:
  • Z is not phenyl substituted by trifluoroalkyl.
  • the compounds of formula (I) have an IC50 against HEK293-STF3A cells of less than about 10 micromolar.
  • Rj, R 2 and n are as defined above, with a compound of formula (V) wherein R 3 and R4 are as defined above.
  • An exemplary solvent for this reaction is chloroform.
  • R 2 and n are as defined above and wherein LG represents a leaving group such as Br, I, tosylate etc.
  • the reaction may take place in the presence of triethylamine or other amine base (DBU, DBN etc.) and a solvent such as 1,4-dioxane or other dipolar aprotic solvent.
  • DBU triethylamine or other amine base
  • solvent such as 1,4-dioxane or other dipolar aprotic solvent.
  • Compounds of formula (III) maybe synthesized by a coupling reaction, such as a Suzuki coupling reaction.
  • a coupling reaction such as a Suzuki coupling reaction.
  • compounds of formula (III) may alternatively be synthesized from the corresponding oxopropanenitrile by reaction with hydroxylamine hydrochloride in the presence of sodium hydroxide.
  • the oxopropanenitrile can be obtained from the corresponding ester.
  • This conversion can be carried out, for example, by reduction using borane dimethylsulfide.
  • This conversion can be carried out, for example, by using Lawesson's reagent.
  • the present invention provides a compound of formula (I) for use as a medicament.
  • the compounds of the present invention may have an IC 50 against STF3A of less than 20 micromolar, e.g. less than 10 micromolar.
  • the IC 50 maybe less than 5, 2, 1, 0.5, 0.2 or 0.1 micromolar. It may be between about 0.01 and about 10 micromolar, or between about 0.01 and 5, 0.01 and 1, 0.01 and 0.5, 0.01 and 0.1, 0.01 and 0.05, 0.1 and 5, 0.1 and 1, 0.1 and 0.5, 0.1 and 10, 0.5 and 10, 1 and 10, 5 and 10, 1 and 5 or 0.1 and 0.5, e.g. about 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 micromolar.
  • the present invention provides compounds of formula (I) for use in modulation of the WNT pathway.
  • the present invention also provides a method of modulating WNT activity comprising exposing a WNT protein or a WNT receptor to a compound of formula (I).
  • the present invention provides use of a compound of formula (I) for modulating WNT activity.
  • the present invention additionally provides compounds of formula (I) for use in the treatment of a disease or condition associated with WNT pathway activity.
  • a method of treating a disease or condition associated with W T pathway activity comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) is also provided by the present invention.
  • the present invention further provides use of a compound of formula (I) for treatment of a disease or condition associated with WNT pathway activity.
  • Also provided by the present invention is use of a compound of formula (I) in the manufacture of a medicament for the treatment of a disease or condition associated with WNT pathway activity.
  • the aforementioned disease or condition is suitably selected from the group consisting of cancer, fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis and myocardial infarction.
  • the cancer may be a cancer characterized by high WNT activity.
  • the disease or condition may be a cancer, such as cervical, colon, breast, bladder, head and neck, gastric, lung, ovarian, prostate, thyroid, non-small-cell lung, as well as chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal cell carcinoma, osteosarcoma, hepatocellular carcinoma, Wilm's tumor or medulloblastoma, or a fibrotic disease, such as pulmonary fibrosis, liver fibrosis, skin fibrosis or renal fibrosis, or a degenerative disease, or a metabolic disease such as diabetic retinopathy.
  • a cancer such as cervical, colon, breast, bladder, head and neck, gastric, lung, ovarian, prostate, thyroid, non-small-cell lung, as well as chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal
  • the present invention also provides use of a compound of formula (I) in diagnosis.
  • compositions of this invention at least one compound of formula (I) optionally in combination with at least one of the other aforementioned agents can be used as the active ingredient(s).
  • the active ingredient(s) is intimately admixed with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques, which carrier may take a wide variety of forms depending of the form of preparation desired for administration, e.g., oral or parenteral such as intramuscular.
  • a pharmaceutical carrier may take a wide variety of forms depending of the form of preparation desired for administration, e.g., oral or parenteral such as intramuscular.
  • any of the usual pharmaceutical media may be employed.
  • suitable carriers and additives include water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like;
  • suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously employed. If desired, tablets maybe sugar coated or enteric coated by standard techniques.
  • the carrier will usually comprise sterile water, though other ingredients, for example, for purposes such as aiding solubility or for preservation, may be included.
  • Injectable suspensions may also prepared, in which case appropriate liquid carriers, suspending agents and the like may be employed.
  • the pharmaceutical compositions herein will contain, per dosage unit, e.g., tablet, capsule, powder, injection, teaspoonful and the like, an amount of the active ingredient(s) necessary to deliver an effective dose as described above.
  • the pharmaceutical compositions herein will contain, per dosage unit, e.g., tablet, capsule, powder, injection, suppository, teaspoonful and the like, from about 0.03 mg to 100 mg/kg (preferred 0.1 - 30 mg/kg) and may be given at a dosage of from about 0.1 - 300 mg/kg per day (preferred 1 - 50 mg/kg per day) of each active ingredient or combination thereof.
  • the dosages may be varied depending upon the requirement of the patients, the severity of the condition being treated and the compound being employed. The use of either daily administration or post- periodic dosing may be employed.
  • compositions are in unit dosage forms from such as tablets, pills, capsules, powders, granules, sterile parenteral solutions or suspensions, metered aerosol or liquid sprays, drops, ampoules, autoinjector devices or suppositories; for oral parenteral, intranasal, sublingual or rectal administration, or for administration by inhalation or insufflation.
  • the composition may be presented in a form suitable for once-weekly or once- monthly administration; for example, an insoluble salt of the active compound, such as the decanoate salt, may be adapted to provide a depot preparation for intramuscular injection.
  • the principal active ingredient is mixed with a pharmaceutical carrier, e.g. conventional tableting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums, and other pharmaceutical diluents, e.g. water, to form a solid preformulation composition containing a homogeneous mixture of a compound of the present invention, or a pharmaceutically acceptable salt thereof.
  • a pharmaceutical carrier e.g. conventional tableting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums, and other pharmaceutical diluents, e.g. water
  • a pharmaceutical carrier e.g. conventional tableting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate
  • the tablets or pills of the compositions of the present invention can be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action.
  • the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former.
  • the two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release.
  • enteric layers or coatings such materials including a number of polymeric acids with such materials as shellac, cetyl alcohol and cellulose acetate.
  • aqueous solutions suitably flavoured syrups, aqueous or oil suspensions, and flavoured emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar
  • Suitable dispersing or suspending agents for aqueous suspensions include synthetic and natural gums such as tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone or gelatin.
  • the pharmaceutical composition may contain between about 0.01 mg and 100 mg, preferably about 5 to 50 mg, of each compound, and maybe constituted into any form suitable for the mode of administration selected.
  • Carriers include necessary and inert pharmaceutical excipients, including, but not limited to, binders, suspending agents, lubricants, flavorants, sweeteners, preservatives, dyes, and coatings.
  • Compositions suitable for oral administration include solid forms, such as pills, tablets, caplets, capsules (each including immediate release, timed release and sustained release formulations), granules, and powders, and liquid forms, such as solutions, syrups, elixirs, emulsions, and suspensions.
  • compositions useful for parenteral administration include sterile solutions, emulsions and suspensions.
  • compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily.
  • compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal skin patches well known to those of ordinary skill in that art.
  • the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.
  • the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like.
  • suitable binders include, without limitation, starch, gelatin, natural sugars such as glucose or betalactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
  • Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
  • liquid forms in suitable flavored suspending or dispersing agents such as the synthetic and natural gums, for example, tragacanth, acacia, methyl-cellulose and the like.
  • suitable suspending or dispersing agents such as the synthetic and natural gums, for example, tragacanth, acacia, methyl-cellulose and the like.
  • tragacanth for example, tragacanth, acacia, methyl-cellulose and the like.
  • sterile suspensions and solutions are desired.
  • Isotonic preparations which generally contain suitable preservatives are employed when intravenous administration is desired.
  • the compounds or combinations of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles.
  • Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
  • Compounds or combinations of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled.
  • the compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers.
  • Such polymers can include polyvinylpyrrolidone, pyran copolymer,
  • the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polyactic acid, polyepsilon caprolactone, polyhydroxy butyeric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross- linked or amphipathic block copolymers of hydrogels.
  • biodegradable polymers useful in achieving controlled release of a drug, for example, polyactic acid, polyepsilon caprolactone, polyhydroxy butyeric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross- linked or amphipathic block copolymers of hydrogels.
  • Compounds or combinations of this invention may be administered in any of the foregoing compositions and according to dosage regimens established in the art whenever treatment of the addressed disorders is required.
  • the daily dosage of the products may be varied over a wide range from 0.01 to 1.000 mg per mammal per day.
  • the compositions are preferably provided in the form of tablets containing, 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 150, 200, 250 and 500 milligrams of each active ingredient or combinations thereof for the symptomatic adjustment of the dosage to the patient to be treated.
  • An effective amount of the drug is ordinarily supplied at a dosage level of from about 0.1 mg/kg to about 300 mg/kg of body weight per day.
  • the range is from about 1 to about 50 mg/kg of body weight per day.
  • the compounds or combinations may be administered on a regimen of 1 to 4 times per day.
  • Optimal dosages to be administered may be readily determined by those skilled in the art, and will vary with the particular compound used, the mode of administration, the strength of the preparation, the mode of administration, and the advancement of disease condition. In addition, factors associated with the particular patient being treated, including patient age, weight, diet and time of administration, will result in the need to adjust dosages.
  • the invention also provides a process for preparing a pharmaceutical composition
  • a pharmaceutical composition comprising at least one compound of formula (I), optionally in combination with at least one of the other aforementioned agents and a pharmaceutically acceptable carrier.
  • compositions are preferably in a unit dosage form in an amount appropriate for the relevant daily dosage.
  • Suitable dosages, including especially unit dosages, of the compounds of the present invention include the known dosages including unit doses for these compounds as described or referred to in reference text such as the British and US Pharmacopoeias, Remington's
  • Step 1 Preparation of methyl 2-chlorobenzoate.
  • Step 2 Preparation of 3-(2-chlorophenyl)-3-oxopropanenitrile.
  • Step 3 Preparation of 3-(2-chlorophenyl)isoxazol-5-amine.
  • Step 2 Preparation of 3-(3-fluorophenyl)isoxazol-5-amine.
  • Step 1 Preparation of 3 -oxo-3-(o-tolyl)propanenitrile.
  • Step 2 Preparation 3-(o-tolyl)isoxazol-5-amine.
  • Step 2 Preparation of 3-(5-chloropyridin-2-yl)isoxazol-5-amine.
  • hydroxylamine hydrochloride (1.2 equiv.) in a mixture of ethanol (0.02 M) and water(0.02 M) was adjusted to pH 8 using sodium hydroxide solution and heated at 60 °C for 18 h. The reaction mixture was then acidified to pH 2 using hydrochloride solution and further heated at 80 °C for 1 h. It was then allowed to cool to room temperature, basified to pH 10, diluted with water and extracted with ethyl acetate. The combined organic layers were dried over magnesium sulphate and concentrated under vacuum. The crude residue was purified by column chromatography to afford the purified product.
  • Step 1 Preparation of l-(4-nitrophenyl)pyrrolidine.
  • Triethylamine was added to a stirred solution of l-fluoro-4-nitrobenzene(1.0 equiv.) and pyrrolidine (1.3 equiv.) in 2-propanol (0.1 M). The reaction was allowed to stir at reflux for 6 h. Upon consumption of starting material, solvent was evaporated to dryness; the crude diluted with water; extracted with dichloromethane; organic layers combined; washed with brine; dried over magnesium sulphate and concentrated in vacuo. The crude was dissolved in
  • Step 2 Preparation of 4-(pyrrolidin-l-yl)aniline.
  • Step 1 Preparation of 2-(3-chlorophenyl)-5-nitropyridine.
  • Step 2 Preparation of 6-(3-chlorophenyl)pyridin-3 -amine.
  • Step-1 Preparation of 3-chloro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyridine.
  • Step-2 Preparation of 6-(5-chloropyridin-3-yl)pyridazin-3-amine.
  • Step 1 Preparation of 3-fluoro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyridine.
  • Step 2 Preparation of 5-(5-fluoropyridin-3-yl)pyrazin-2-amine.
  • Step 1 Preparation of 2-methyl-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine.
  • Step 1 Preparation of 2-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine
  • Step 2 Preparation of 4-oxo-4-(2-(trifluoromethyl)-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)butanoic acid
  • Step 1 Preparation of 2-methylpropane- 1,3 -diyl dimethanesulfonate
  • Step 2 Preparation of 2,6-dimethyl-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine
  • Step 3 Preparation of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)- 4-oxobutanoic acid
  • Step 1 Preparation of 2-methyl-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine
  • Step 2 Preparation of 2-bromo-l-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)ethanone
  • Step 3 Preparation of ethyl 2-(diethoxyphosphoryl)-4-(2-methyl-6,7- dihydropyrazolof 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanoate
  • Step 4 Preparation of ethyl 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)- 2-methylene-4-oxobutanoate
  • Step 5 Preparation of ethyl 2-methyl-4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)-4-oxobutanoate
  • Step 1 Preparation of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrmiidin-4(5H)-yl)-N-(5- (4-nitrophenyl)isoxazol-3-yl)-4-oxobutanamide.
  • Step 2 Preparation of N-[3-(4-aminophenyl)-l,2-oxazol-5-yl]-4- ⁇ 2-methyl-5H,6H,7H- pyrazolo[l,5-a]pyrimidin-4-yl ⁇ -4-oxobutanamide.
  • Step 1 Preparation of N-(4-bromophenyl)-4-(2-methyl-6,7-dihydropyrazolo[l, 5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide.
  • Step 2 Preparation of N-(2'-amino-[l,l , -biphenyl]-4-yl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide.
  • Step 1 Preparation of 4-(2-methyl-6,7-dihydropyrazolo[ l,5-a]pyrimidin-4(5H)-yl)-N-(2- methyl-[ 1 , 1 '-biphenyl] -4-yl)-4-oxobutanamide.
  • Step 2 Preparation of N-(3*-amino-[l,r-biplienyl]-4-yl)-4-(2-niethyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide.
  • oxobutanamide (1 equiv.), ammonium chloride(4 equiv.), and iron(2 equiv.) were added to a solution of ethanol (0.02 M) and water(0.08 M), and heated at 40 °C for 4 h. The reaction mixture was then filtered, diluted with water and extracted with ethyl acetate. The combined organic layers were dried over magnesium sulphate and concentrated under vacuum. The crude product was purified by reversed phase preparative HPLC to afford the purified product.
  • Step 1 Preparation of 4-(2-methyl-6,7-dihydropyrazolo[ l,5-a]pyrimidin-4(5H)-yl)-N- (4'-nitro-[ 1 , 1 '-biphenyl]-4-yl)-4-oxobutanamide.
  • Step 2 Preparation of N-(4'-amino-[l,r-bip enyl]-4-yl)-4-(2-methyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide.
  • Step 1 Preparation of tert-butyl 4-(6-(4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)-4-oxobutanamido)pyridin-3 -yl)piperazine- 1 -carboxylate
  • Step 2 Preparation of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrirnidin-4(5H)-yl)-4-oxo-N-(5- (piperazin- 1 -yl)pyridin-2-yl)butanamide
  • Step 3 Preparation of N-ethyl-4-(6-(4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)- yl)-4-oxobutanamido)pyridin-3 -yl)piperazine- 1 -carboxamide

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Diabetes (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Hematology (AREA)
  • Cardiology (AREA)
  • Oncology (AREA)
  • Biomedical Technology (AREA)
  • Rheumatology (AREA)
  • Obesity (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Epidemiology (AREA)
  • Ophthalmology & Optometry (AREA)
  • Pain & Pain Management (AREA)
  • Immunology (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Endocrinology (AREA)
  • Pulmonology (AREA)
  • Dermatology (AREA)
  • Emergency Medicine (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Hospice & Palliative Care (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Nitrogen Condensed Heterocyclic Rings (AREA)

Abstract

The present invention relates to dihydropyrazolo[l,5-a]pyrimidine compounds of formula I, defined herein, as WNT pathways modulators, processes for making them, and pharmaceutical compositions comprising them. Methods of treatment of conditions mediated by WNT pathway signalling including cancer, fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis and myocardial infarction, comprising the compounds of formula I are also provided.

Description

WNT PATHWAY MODULATORS
Field
[0001] The invention relates to WNT pathway modulators, processes for making them and methods for using them.
Priority
[0002] This application claims priority from UK application GB 1322333.4, filed 17 December 2013, the entire contents of which are incorportated herein by cross-reference.
Background
[0003] Wnt proteins are secreted glycoproteins acting as growth factors that regulate various cellular functions include proliferation, differentiation, death, migration, and polarity, by activating multiple intracellular signaling cascades, including the β-catenin-dependent and - independent pathways. There are 19 Wnt members have been found in humans and mice, and they exhibit unique expression patterns and distinct functions during development. In humans and mice, the 10 members of the Frizzled (Fz) family comprise a series of seven-pass transmembrane receptors that have been identified as Wnt receptors. In addition to Fz proteins, single-pass transmembrane proteins, such as low-density lipoprotein receptor-related protein 5 (LRP5), LRP6, receptor tyrosine kinase (RTK)-like orphan receptor l(Rorl), Ror2, and receptor-like tyrosine kinase (Ryk), have been shown to function as co-receptors for Wnt signaling. Therefore, it has been assumed traditionally that the binding of different Wnts to their specific receptors selectively triggers different outcomes via distinct intracellular pathways.
[0004] Diverse Wnts, Wnt receptors, and downstream pathway all contribute to the role of Wnt. These pathways all play a role in development, stem cell maintenance, cancer and metastasis.
[0005] In the absence of Wnt signaling, β-catenin is bound and phosphorylated by a
"destruction complex" containing the adenomatous polyposis coli (APC) and Axin proteins, as well as glycogen synthase kinase 3 (GSK3) and casein kinase I (CKI). Phosphorylated β-catenin is bound by the F box protein Slimb/p-TrCP and polyubiquitinated, leading to proteosomal degradation. In addition, the complex acts to prevent nuclear localization of b-catenin. Upon Wnt binding to Frizzled (Fz) and low-density lipoprotein-related proteins 5 and 6 (LRP5/6), GSK3, Axin, and other destruction complex components are recruited to the receptor complex. The function of the destruction complex is inhibited, and unphosphorylated β-catenin
accumulates in the cytoplasm and eventually translocates to the nucleus. There, it associates with TCF proteins, converting TCF from a repressor into an activator of Wnt-responsive gene transcription.
[0006] Wnt in Cancer & Stem Cell:
[0007] Deregulation of components of Wnt/p-catenin signaling is implied in a wide spectrum of diseases including degenerative diseases, metabolic diseases, and a number of cancers such as cervical, colon, breast, bladder, head and neck, gastric, lung, ovarian, prostate, thyroid, non- small-cell lung, as well as chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal cell carcinoma, osteosarcoma, hepatocellular carcinoma, Wilm's tumor and medulloblastoma.
[0008] Wnt signaling plays a role both during development, and within stem cell niches in adults. This is best established in skin, hematopoietic stem cells, mammary gland and in intestinal proliferation. For example, high level expression of DK 1, an inhibitor of Wnt signaling, blocks normal stem cell proliferation in mouse intestine, suggesting there is an essential role for Wnt signaling in maintenance of stem cells in the digestive tract. Wnt roles in self renewal and expansion of stem cells have also been demonstrated for embryonic and neural stem cells, suggesting that Wnt signaling may be a general requirement of stem cell
maintenance. Inhibition of Wnt signaling, e.g., by overexpression of axin or an extracellular Wnt-binding protein, sFRP, reduces hematopoietic stem cell (HSC) growth in vitro and the ability to reconstitute HSCs in vivo. Notably, while overexpression of activated β-catenin can expand HSC populations in culture for extended periods, two groups have reported that β-catenin is not required for HSC survival and serial transplantation, supporting the proposal that there is more to Wnt signaling than stabilization of β-catenin in stem cell survival. Diverse Wnts can regulate stem cell proliferation: Wnts 1, 5a, and 10b are able to stimulate expansion of HSC populations and Wnt5a acts synergistically with stem cell factor (SCF) to expand and promote self renewal of HSCs. The demonstration of a role for Wnt5a in HSC self renewal and its ability to synergize with stem cell factor is particularly interesting because Wnt5a often acts in a β-catenin independent manner. While Wnt signaling is critical for stem cell maintenance, it may therefore be via signaling pathways distinct from or in parallel to the β-catenin pathway.
[0009] Fibrosis:
[0010] Wnt/ -catenin signaling pathway is essential to embryonic development ingeneral and organ morphogenesis, so it is not surprising that dysregulation of this pathway in adult has been linked to fibroblast biology and fibrosis. It has been demonstrated that Wnt/p-catenin signaling play a role in severe fibrotic diseases, such as pulmonary fibrosis, liver fibrosis, skin fibrosis and renal fibrosis.
[0011] Others:
[0012] Dysregulation of Wnt/p-catenin signaling contributes to the development of diabetic retinopathy by inducing retinal inflammation, vascular leakage, and neovascularization.
[0013] The binding of Wnt proteins to plasma membrane receptors on mesenchymal cells induces the differentiation of these cells into the osteoblast lineage and thereby supports bone formation. Wnts are also key signaling proteins in joint remodeling processes. Active Wnt signaling contributes to osteophyte formation and might have an essential role in the anabolic pattern of joint remodeling that is observed in ankylosing spondylitis and osteoarthritis. By contrast, blockade of Wnt signaling facilitates bone erosion and contributes to catabolic joint remodeling, a process that is observed in rheumatoid arthritis.
[0014] There is therefore a need for compounds that modulate and/or inhibit the WNT pathway so as to treat diseases associated with WNT activity.
Definitions
[0015] A measure of the binding of an inhibitor to and the subsequent release from an enzyme is the "IC50" value, which reflects the inhibitor concentration, which at a given substrate concentration results in 50 % enzynie activity.
[0016] The term "subject" as used herein, refers to an animal, preferably a mammal, most preferably a human, who has been the object of treatment, observation or experiment. In one embodiment the subject is not a human. The subject may for example be a non-human mammal.
[0017] The term "therapeutically effective amount" as used herein, means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal or human being sought by a researcher, veterinarian, medical doctor or other clinician, which includes alleviation of the symptoms of the disease or disorder being treated.
[0018] As used herein, the term "pharmaceutically acceptable" embraces both human and veterinary use: For example the term "pharmaceutically acceptable" embraces a veterinarily acceptable compound or a compound acceptable in human medicine and health care. In one embodiment, human medicine and health care is excluded.
[0019] Throughout the description and the claims the expression "alkyl", unless specifically limited, denotes a C1-12 alkyl group, suitably a C1-6 alkyl group, e.g. C1-4 alkyl group. Alkyl groups may be straight chain or branched. Suitable alkyl groups include, for example, methyl, ethyl, propyl (e.g. n-propyl and isopropyl), butyl (e.g n-butyl, iso-butyl, sec-butyl and tert- butyl), pentyl (e.g. n-pentyl), hexyl (e.g. n-hexyl), heptyl (e.g. n-heptyl) and octyl (e.g. n-octyl). The expression "alk", for example in the expressions "alkoxy", "haloalkyl" and "thioalkyl" should be interpreted in accordance with the definition of "alkyl". Exemplary alkoxy groups include methoxy, ethoxy, propoxy (e.g. n-propoxy), butoxy (e.g. n-butoxy), pentoxy (e.g. n- pentoxy), hexoxy (e.g. n-hexoxy), heptoxy (e.g. n-heptoxy) and octoxy (e.g. n-octoxy).
Exemplary thioalkyl groups include methylthio-. Exemplary haloalkyl groups include fluoroalkyl e.g. CF3.
[0020] The expression "alkenyl", unless specifically limited, denotes a C2-12 alkenyl group, suitably a C2-6 alkenyl group, e.g. a C2-4 alkenyl group, which contains at least one double bond at any desired location and which does not contain any triple bonds. Alkenyl groups may be straight chain or branched. Exemplary alkenyl groups including one double bond include propenyl and butenyl. Exemplary alkenyl groups including two double bonds include pentadienyl, e.g. (IE, 3E)-pentadienyl.
[0021] The expression "alkynyl", unless specifically limited, denotes a C2-12 alkynyl group, suitably a C2-6 alkynyl group, e.g. a C2-4 alkynyl group, which contains at least one triple bond at any desired location and may or may not also contain one or more double bonds. Alkynyl groups may be straight chain or branched. Exemplary alkynyl groups include propynyl and butynyl.
[0022] The expression "alkylene" denotes a chain of formula -(CH2)n- wherein n is an integer e.g. 1-12, 1-6, 2-6 or 2-5, unless specifically limited.
[0023] The expression "cycloalkyl", unless specifically limited, denotes a C3-10 cycloalkyl group (i.e. 3 to 10 ring carbon atoms), more suitably a C3-8 cycloalkyl group, e.g. a C3-6 cycloalkyl group. Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. A most suitable number of ring carbon atoms is three to six.
[0024] The expression "cycloalkenyl", unless specifically limited, denotes a C3-10 cycloalkenyl group (i.e. 3 to 10 ring carbon atoms), suitably a C5-10 cycloalkenyl group (i.e. 5 to 10 ring carbon atoms), more suitably a C5-8 cycloalkenyl group e.g. a C5-6 cycloalkenyl group.
Exemplary cycloalkenyl groups include cyclopropenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl. A most suitable number of ring carbon atoms is five to six.
[0025] The expression "carbocyclyl", unless specifically limited, denotes any ring system in which all the ring atoms are carbon and which contains between three and twelve ring carbon atoms, suitably between three and ten carbon atoms and more suitably between three and eight carbon atoms. Carbocyclyl groups may be saturated or partially unsaturated, but do not include aromatic rings. Examples of carbocyclyl groups include monocyclic, bicyclic, and tricyclic ring systems, in particular monocyclic and bicyclic ring systems. Other carbocyclyl groups include bridged ring systems (e.g. bicyclo[2.2.1]heptenyl). A specific example of a carbocyclyl group is a cycloalkyl group. A further example of a carbocyclyl group is a cycloalkenyl group.
[0026] The expression "heterocyclyl", unless specifically limited, refers to a carbocyclyl group wherein one or more (e.g. 1, 2 or 3) ring atoms are replaced by heteroatoms. The heteroatom(s) are commonly selected from N, S and O. A specific example of a heterocyclyl group is a cycloalkyl group (e.g. cyclopentyl or more particularly cyclohexyl) wherein one or more (e.g. 1, 2 or 3, particularly 1 or 2, especially 1) ring atoms are replaced by heteroatoms selected from N, S or O. Exemplary heterocyclyl groups containing one hetero atom include pyrrolidine, tetrahydrofuran and piperidine, and exemplary heterocyclyl groups containing two hetero atoms include morpholine and piperazine. A further specific example of a heterocyclyl group is a cycloalkenyl group (e.g. a cyclohexenyl group) wherein one or more (e.g. 1, 2 or 3, particularly 1 or 2, especially 1) ring atoms are replaced by heteroatoms selected from N, S and O. An example of such a group is dihydropyranyl (e.g. 3,4-dihydro-2H-pyran-2-yl-). The heterocyclyl group may be linked to other part or parts of the molecule by a carbon ring atom or nitrogen ring atom.
[0027] The expression "aryl", unless specifically limited, denotes a C6-12 aryl group, suitably a C -1o aryl group, more suitably a C6-8 aryl group. Aryl groups will contain at least one aromatic ring (e.g. one, two or three rings). Aryl groups with multiple aromatic rings include fused aromatic rings and aromatic rings connected to each other by one single bond. An example of a typical aryl group with one aromatic ring is phenyl. An example of a typical aryl group with two fused aromatic rings is naphthyl. An example of an aromatic group with two directly connected aromatic rings is biphenyl.
[0028] The expression "heteroaryl", unless specifically limited, denotes an aryl residue, wherein one or more (e.g. 1, 2, 3, or 4, suitably 1, 2 or 3) ring atoms are replaced by heteroatoms, said heteroatoms commonly being selected from N, S and O, or else a 5-membered aromatic ring containing one or more (e.g. 1, 2, 3, or 4, suitably 1, 2 or 3) ring atoms, said heteroatoms commonly selected from N, S and O. In some embodiments a heteroaryl group will have no ring heteroatoms other than nitrogen. Exemplary monocyclic heteroaryl groups having one heteroatom include: five membered rings (e.g. pyrrole, furan, thiophene); and six membered rings (e.g. pyridine, such as pyridin-2-yl, pyridin-3-yl andpyridin-4-yl). Exemplary monocyclic heteroaryl groups having two heteroatoms include: five membered rings (e.g. pyrazole, oxazole, isoxazole, thiazole, isothiazole, imidazole, such as imidazol-l-yl, imidazol-2-yl imidazol-4-yl); and six membered rings (e.g. pyridazine, pyrimidine, pyrazine). Exemplary monocyclic heteroaryl groups having three heteroatoms include: 1,2,3-triazole, 1,2,4-triazole, 1,2,3- oxadiazole and 1,2,4-oxadiazole. Exemplary monocyclic heteroaryl groups having four heteroatoms include tetrazole. Exemplary bi cyclic heteroaryl groups include: indole (e.g. indol- 6-yl), benzofuran, benzthiophene, quinoline, isoquinoline, indazole, benzimidazole,
benzthiazole, quinazoline and purine. The heteraryl group may be linked to other part or parts of the molecule by a carbon ring atom or nitrogen ring atom.
[0029] The aforementioned aryl and heteroaryl groups may, where appropriate, optionally be substituted by one or more (e.g. 1, 2 or 3, suitably 1 or 2) groups each independently selected from monovalent or multivalent (i.e. having valency greater than 1) functional groups. Suitable substituent groups include alkyl, alkenyl, alkynyl, haloalkyl, -alkoxy (e.g. OMe), cycloalkyl, alkenyloxy-, alkynyloxy-, alkoxyalkyl-, nitro, halogen (e.g. fluoro, chloro and bromo), cyano, hydroxyl, oxo, -C(0)-alkyl (e.g. COMe), C(0)OH, -C(0)Oalkyl (e.g. -C(O)OMe), -OC(0)alkyl (e.g. -OC(O)Me), -NH2, -NHalkyl (e.g. -NHMe), -N(alkyl)2 (e.g. dimethylamino-), -C(0)NH2, -C(0) H(alkyl) (e.g. -C(O)NHMe), -NHC(0)alkyl (e.g. -NHC(O)Me), -C(NH)NH2, thioalkyl (e.g. -thiomethyl), -S02alkyl (e.g. S02Me), -SOalkyl (e.g. -SOMe), -S02cycloalkyl and - SOcycloalkyl. More typically, substituents will be selected from alkyl (e.g. Me), fluoroalkyl (e.g. CF3 and CHF2), alkoxy (e.g. OMe), halogen and hydroxyl.
[0030] The expression "-alkylaryl", unless specifically limited, denotes an aryl residue which is connected via an alkylene moiety e.g. a C1- alkylene moiety. An example of such a group is benzyl: PhCH2-.
[0031] The term "halogen" or "halo" comprises fluorine (F), chlorine (CI) and bromine (Br). [0032] The term "amino" refers to the group -NH2.
[0033] The term "oxo" refers to an oxygen atom which, together with the carbon atom which it substitutes, forms a carbonyl group C=0.
[0034] The term "-arylheterocyclyl" refers to a heterocyclyl residue which is connected via an aryl moiety.
[0035] Stereoisomers:
[0036] All possible stereoisomers of the claimed compounds are included in the present invention.
[0037] Where the compounds according to this invention have at least one chiral center, they may accordingly exist as enantiomers. Where the compounds possess two or more chiral centers, they may additionally exist as diastereomers. It is to be understood that all such isomers and mixtures thereof are encompassed within the scope of the present invention.
[0038] Preparation and isolation of stereoisomers: [0039] Where the processes for the preparation of the compounds according to the invention give rise to a mixture of stereoisomers, these isomers may be separated by conventional techniques such as preparative chromatography. The compounds may be prepared in racemic form, or individual enantiomers maybe prepared either by enantiospecific synthesis or by resolution. The compounds may, for example, be resolved into their components enantiomers by standard techniques, such as the formation of diastereomeric pairs by salt formation with an optically active acid, such as (-)-di-p-toluoyl-d-tartaric acid and/or (+)-di-p-toluoyl-l-tartaric acid followed by fractional crystallization and regeneration of the free base. The compounds may also be resolved by formation of diastereomeric esters or amides, followed by
chromatographic separation and removal of the chiral auxiliary. Alternatively, the compounds may be resolved using a chiral HPLC column.
[0040] Pharmaceutically acceptable salts:
[0041] In view of the close relationship between the free compounds and the compounds in the form of their salts or solvates, whenever a compound is referred to in this context, a
corresponding salt, solvate or polymorph is also intended, provided such is possible or appropriate under the circumstances.
[0042] Salts and solvates of the compounds of formula (I) and physiologically functional derivatives thereof which are suitable for use in medicine are those wherein the counter-ion or associated solvent is pharmaceutically acceptable. However, salts and solvates having non- pharmaceutically acceptable counter-ions or associated solvents are within the scope of the present invention, for example, for use as intermediates in the preparation of other compounds and their pharmaceutically acceptable salts and solvates.
[0043] Suitable salts according to the invention include those formed with both organic and inorganic acids or bases. Pharmaceutically acceptable acid addition salts include those formed from hydrochloric, hydrobromic, sulfuric, nitric, citric, tartaric, phosphoric, lactic, pyruvic, acetic, trifluoroacetic, triphenylacetic, sulfamic, sulfanilic, succinic, oxalic, fumaric, maleic, malic, mandelic, glutamic, aspartic, oxaloacetic, methanesulfonic, ethanesulfonic, arylsulfonic (for example p-toluenesulfonic, benzenesulfonic, naphthalenesulfonic or naphthalenedisulfonic), salicylic, glutaric, gluconic, tricarballylic, cinnamic, substituted cinnamic (for example, phenyl, methyl, methoxy or halo substituted cinnamic, including 4-methyl and 4-methoxycinnamic acid), ascorbic, oleic, naphthoic, hydroxynaphthoic (for example 1- or 3-hydroxy-2-naphthoic), naphthaleneacrylic (for example naphthalene-2-acrylic), benzoic, 4-methoxybenzoic, 2- or 4- hydroxybenzoic, 4-chlorobenzoic, 4-phenylbenzoic, benzeneacrylic (for example 1,4- benzenediacrylic), isethionic acids, perchloric, propionic, glycolic, hydroxyethanesulfonic, pamoic, cyclohexanesulfamic, salicylic, saccharinic and trifluoroacetic acid. Pharmaceutically acceptable base salts include ammonium salts, alkali metal salts such as those of sodium and potassium, alkaline earth metal salts such as those of calcium and magnesium and salts with organic bases such as dicyclohexylamine and N-methyl-D-glucamine.
[0044] All pharmaceutically acceptable acid addition salt forms of the compounds of the present invention are intended to be embraced by the scope of this invention.
[0045] Polymorph crystal forms:
[0046] Furthermore, some of the crystalline forms of the compounds may exist as polymorphs and as such are intended to be included in the present invention. In addition, some of the compounds may form solvates with water (i.e. hydrates) or common organic solvents, and such solvates are also intended to be encompassed within the scope of this invention. The
compounds, including their salts, can also be obtained in the form of their hydrates, or include other solvents used for their crystallization.
[0047] Prodrugs:
[0048] The present invention further includes within its scope prodrugs of the compounds of this invention. In general, such prodrugs will be functional derivatives of the compounds which are readily convertible in vivo into the desired therapeutically active compound. Thus, in these cases, the methods of treatment of the present invention, the term "administering" shall encompass the treatment of the various disorders described with prodrug versions of one or more of the claimed compounds, but which converts to the above specified compound in vivo after administration to the subject. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", ed. H.
Bundgaard, Elsevier, 1985.
[0049] Protective Groups: [0050] During any of the processes for preparation of the compounds of the present invention, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups, such as those described in Protective Groups in Organic Chemistry, ed. J.F.W. McOmie, Plenum Press, 1973; and T.W. Greene & P.G.M. Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, fully incorporated herein by reference. The protecting groups may be removed at a convenient subsequent stage using methods known from the art.
[0051] As used herein, the term "composition" is intended to encompass a product comprising the claimed compounds in the therapeutically effective amounts, as well as any product which results, directly or indirectly, from combinations of the claimed compounds.
[0052] Carriers and Additives for galenic formulations:
[0053] Carriers, which can be added to the mixture, include necessary and inert pharmaceutical excipients, including, but not limited to, suitable binders, suspending agents, lubricants, flavorants, sweeteners, preservatives, coatings, disintegrating agents, dyes and coloring agents.
[0054] Thus, for liquid oral preparations, such as for example, suspensions, elixirs and solutions, suitable carriers and additives may advantageously include water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like; for solid oral preparations such as, for example, powders, capsules, gelcaps and tablets, suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like.
[0055] Soluble polymers as targetable drug carriers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamidephenol, polyhydroxyethylaspartamide-phenol, or polyethyleneoxidepolyllysine substituted with palmitoyl residue. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polyactic acid, polyepsilon caprolactone, polyhydroxy butyeric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels.
[0056] Suitable binders include, without limitation, starch, gelatin, natural sugars such as glucose or betalactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
[0057] Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
Summary of Invention
[0058] According to the invention there is provided a compound of formula (I),
Figure imgf000012_0001
or a pharmaceutically acceptable salt, solvate or polymorph thereof, including all tautomers and stereoisomers thereof wherein:
[0059] R1 represents H; optionally substituted alkyl (wherein optional substituents include one or more substituents each independently selected from G1-6alkoxy, NH2, -NHC1-3alkyl and - N(C1-3alkyl)2); -C(0)Oalkyl; haloalkyl; haloalkoxy; or -alkylaryl;
[0060] each R2 independently represents H; optionally substituted alkyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and -N(C1-3alkyl)2); -alkylaryl; optionally substituted carbocyclyl (wherein optional substituents include one or more substituents each independently selected from Q. 6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy and halo); optionally substituted heterocyclyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl); -NHalkyl; - N(alkyl)2; amino; hydroxyl; alkoxy or halo;
[0061] n represents 0, 1 or 2; [0062] R3 represents H or alkyl;
[0063] R4 represents H or alkyl; [0064] R5 represents H or alkyk
[0065] W and X each independently represent C=0; C=S; or CH2;
[0066] Y represents aryl; heteroaryl; optionally substituted carbocyclyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, C\. 6alkoxy, C1- haloalkyl, C1-6haloalkoxy and halo); or optionally substituted heterocyclyl (wherein optional substituents include one or more substituents each independently selected from C\. 6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl); and
[0067] Z represents optionally substituted alkyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkoxy, N¾, -NHC1-3alkyl and— Ν(^_ 3alkyl)2); aryl; heteroaryl; -alkylaryl;-alkylheteroaryl; optionally substituted carbocyclyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy and halo); optionally substituted heterocyclyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl); - alkylcarbocyclyl wherein carbocyclyl is optionally substituted (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, Ci. 6haloalkoxy and halo); -alkylheterocyclyl wherein heterocyclyl is optionally substituted
(wherein optional substituents include one or more substituents each independently selected from Ci-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl); -arylcarbocyclyl wherein carbocyclyl is optionally substituted (wherein optional substituents include one or more substituents each independently selected from C1- alkyl, Ci-6alkoxy, C1-6haloalkoxy and halo); or -arylheterocyclyl wherein heterocyclyl is optionally substituted (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, - C(0)C1-6alkyl and-C(0)NHCi-6alkyl). [0068] Further, according to the invention there is provided a compound of formula (I),
Figure imgf000014_0001
or a pharmaceutically acceptable salt, solvate or polymorph thereof, including all tautomers and stereoisomers thereof wherein:
[0069] R\ represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and -N(C1-3alkyl)2; -C(0)Oalkyl; haloalkyl; haloalkoxy; or -alkylaryl;
[0070] each R2 independently represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, NH2, -NHCi-3alkyl and— N Ci.
3alkyl)2; -alkylaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1- alkoxy, C1-6haloalkyl, Ci-6haloalkoxy and halo; heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, Ci-6alkoxy, -C(0)OC1-6alkyl, -C(0)Ci-6alkyl and-C(0)NHCi-6alkyl; - NHalkyl; -N(alkyl)2; amino; hydroxyl; alkoxy or halo;
[0071] n represents 0, 1 or 2;
[0072] R3 represents H or alkyl;
[0073] R4 represents H or alkyl;
[0074] R5 represents H or alkyl;
[0075] W and X each independently represent C=0; C=S; or CH2; [0076] Y represents aryl; heteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C .
6haloalkoxy and halo; or heterocyclyl which is optionally substituted by one or more
substituents each independently selected from C1-6alkyl, C^alkoxy, -C(0)OC1-6alkyl, -C(0)C1- 6alkyl and-C(0)NHC1-6alkyl; and
[0077] Z represents alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and -N(Cj-3alkyl)2; aryl; heteroaryl; -alkylaryl;-alkylheteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C\.
haloalkoxy and halo; heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and- C(0)NHC1-6alkyl; -alkylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1- alkoxy, C1-6haloalkoxy and halo; -alkylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and- C(0)NHC1-6alkyl; -arylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkoxy and halo; or -arylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from Cj-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl.
Brief Description of Drawings
[0078] Figure 1 : A bar chart illustrating inhibition of the activity of mammalian porcupine by Compound 51 and Compound 110.
[0079] Figure 2: A graph illustrating efficacy of Compound 51 on the MMTV-Wntl mouse model.
[0080] Figure 3: A graph illustrating efficacy data of Compound 110 in the MMTV-Wntl mouse model.
[0081] Figure 4: Images illustrating inhibition of Palmitoylation of Wnt3a by Compound 51. [0082] Figures 5 to 7: Images and graphs showing results of the Soft Agar Assay. Figure 5 shows the results for Cell line AsPC-1 treated with Compound 51. Figure 6 shows the results for Cell line HPAF-II treated with Compound 51. Figure 7 shows the results for Cell line CFPAC-1 treated with Compound 51.
Detailed description
[0083] According to the invention there are provided compounds of formula (I),
Figure imgf000016_0001
or a pharmaceutically acceptable salt, solvate or polymorph thereof, including all tautomers and stereoisomers thereof wherein:
[0084] Ri represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and -N(C1-3alkyl)2; -C(0)Oalkyl; haloalkyl; haloalkoxy; or -alkylaryl;
[0085] each R2 independently represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C1- alkoxy, NH2, -NHC1-3alkyl and -Ν(^.
3alkyl)2; -alkylaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy and halo; heterocyclyl which is optionally substituted by one or more substituents each independently selected from Ci-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl; - NHalkyl; -N(alkyl)2; amino; hydroxyl; alkoxy or halo;
[0086] n represents 0, 1 or 2; [0087] R3 represents H or alkyl;
[0088] R4 represents H or alkyl;
[0089] R5 represents H or alkyl;
[0090] W and X each independently represent C=0; C=S; or CH2;
[0091] Y represents aryl; heteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C^alkoxy, C1-6haloalkyl, C\.
6haloalkoxy and halo; or heterocyclyl which is optionally substituted by one or more
substituents each independently selected from C1-6alkyl, C^alkoxy, -C(0)OC1-6alkyl, -C(0)C1- 6alkyl and-C(0)NHC1-6alkyl; and
[0092] Z represents alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, N¾, -NHC1-3alkyl and -N(C1-3alkyl)2; aryl; heteroaryl; -alkylaryl;-alkylheteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C^
6haloalkoxy and halo; heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and- C(0)NHC1-6alkyl; -alkylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, Ci. 6haloalkoxy and halo; -alkylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl; -arylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C^ 6haloalkoxy and halo; or -arylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)C1- 6alkyl and-C(0)NHCi-6alkyl.
[0093] In one embodiment, any one or more of, optionally all of, the following compounds are excluded from the scope of Formula (I):
Figure imgf000018_0001
Figure imgf000019_0001
Figure imgf000020_0001

Figure imgf000020_0002
Figure imgf000021_0001
[0094] In the context of variable Y, the term "aryl" is understood to mean "arylene" (e.g.
"phenyl" is understood to mean "phenylene" (i.e. C H4)) because Y is a linking group, not a terminal group. The terms for other Y rings (e.g. heteroaryl) are to be construed likewise.
When Y is referred to as being unsubstituted, this is understood to mean no other substituents other than Z and NR5. When Y is referred to as being monosubstitued, this is understood to mean one substituent other than Z and NR5.
[0095] When R] represents alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, N¾, -NHC^alkyl and -N(C1-3alkyl)2 exemplary substituents include methoxy, -NH2, -NHmethyl and -NH(methyi)2. Examples include C1-6 alkyl (e.g. unsubstituted C1- alkyl), for example methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n- hexyl). Exemplary Q-6 alkyl groups are methyl and ethyl, particularly methyl.
[0096] When R1 represents -C(0)Oalkyl, examples include -C(0)OC1-6alkyl such as - C(0)OMe, -C(0)OEt, -C(0)OPr and -C(0)OiPr. A specific example is -C(0)OMe.
[0097] When R represents haloalkyl, examples include C1-6haloalkyl such as CH2F, CHF2, CF3, CH2CH2F, CH2CHF2, CH2CF3, CHFCH3 and CF2CH3. A specific example is CF3. [0098] When represents haloalkoxy, examples include C1-6haloalkoxy such as OCF3.
[0099] When R2 represents alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and -N(C1-3alkyl)2, exemplary substituents include methoxy, -NH2, -NHmethyl and -NH(methyl)2. Examples include C1-6 alkyl (e.g. unsubstituted C1-6alkyl), for example methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n- hexyl). An exemplary C1-6 alkyl group is methyl.
[00100] When R2 represents -alkylaryl, examples include benzyl.
[00101 ] When R2 represents carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1- alkyl (e.g. methyl), C1-6alkoxy (e.g. methoxy), C1-6haloalkyl (e.g. fluoromethyl such as trifluoromethyl), Ci- haloalkoxy (e.g. fluoromethoxy such as trifluoromethoxy) and halo (e.g. fluoro, e.g. chloro), examples include C3-C6 cycloalkyl - such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl. Substituted examples include cyclohexyl substituted by methyl.
[00102] When R2 represents heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1- 6alkyl and-C(0)NHC1-6alkyl, Y may represent heterocyclyl which is optionally substituted by Ci- alkyl (such as methyl). Examples include monocyclic heterocyclyl. The heterocyclyl group may be unsubstituted or may have for example one or two (e.g. one) substituent (e.g. one methyl group). Examples include piperidinyl, morpholinyl, pyrrolidinyl, 4,5-dihydropyrazolyl and 4,5-dihydroisoxazolyl. A specific example is pyrrolidinyl.
[00103 ] When R2 represents -NHalkyl, examples include -NHMe, NHEt, NHPr and
NHiPr, in particular NHMe.
[00104] When R2 represents -N(alkyl)2, examples include -N(Me)2.
[00105] When R2 represents alkoxy, examples include methoxy and ethoxy, especially methoxy.
[00106] When n represents 1, examples of -(CHR )n- include -CH2- and -CH(CH3)- [00107] When n represents 2, examples of -(CHR2)n- include -CH2-CH2-, - CH2CH(CH3 , -CH(CH3)C¾- and - CH(CH3)-CH(CH3)-. An example of a -(CHR2)2- group is -CH2-CH2-.
[0100] When R3 represents alkyl, examples include C1-6 alkyl such as methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n-hexyl). An exemplary C -6 alkyl group is methyl.
[0101] When R4 represents alkyl, examples include C1-6 alkyl, for example methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n- pentyl) and hexyl (e.g. n-hexyl). An exemplary Ci-6 alkyl group is methyl.
[0102] When R5 represents alkyl, examples include C1-6 alkyl such as methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n-hexyl). An exemplary C1-6 alkyl group is methyl.
[0103] When Y represents aryl, examples include optionally substituted phenyl. Exemplary substituents include one or more (e.g. one or two, especially one) substituents each
independently selected from C1-6 alkyl (such as methyl), C1-6alkoxy (such as methoxy), halo and C1-6haloalkyl (such as fluoroemethyl, e.g. trifluoromethyl). Examples include unsubstituted phenyl, methylphenyl, methoxyphenyl, fluorophenyl, chlorophenyl and trifluoromethylphenyl. Specific examples include unsubstituted phenyl. Specific examples also include methylphenyl, methoxyphenyl, fluorophenyl. Z and NR5 may be positioned on the phenyl ring at the 1 - and 4- positions relative to each other (i.e. Z and R5 have a para relationship).
[0104] When Y represents heteroaryl, examples include monocyclic (e.g. 5 and 6 membered) and bicyclic (e.g. 9 and 10 membered) heteroaryl rings, especially monocyclic heteroaryl rings, particularly 6-membered monocyclic heteroaryl rings. Examples of monocyclic heteroaryl may comprise one, two or three ring heteroatoms (e.g. one or two, e.g. one) including one or two nitrogen atoms (e.g. one or e.g. two) and optionally an oxygen or sulphur atom. Exemplary 5- membered monocyclic heteroaryl groups include pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl and thiadiazolyl. When Y is 5-membered monocyclic heteroaryl, Z and NR5 may be positioned on the ring at non-adjacent ring atoms. Exemplary 6-membered monocyclic heteroaryl groups include pyridinyl. Exemplary 6- membered monocyclic heteroaryl groups also include pyridazinyl, pyrimidinyl and pyrazinyl. When Y is 6-membered monocyclic heteroaryl, Z and NR5 may be positioned on the ring at 1- and 4- positions relative to each other (i.e. Z and NR5 have a para relationship). The
aforementioned heteroaryl group may either by unsubstituted or may be substituted by one or more (e.g. one or two, particularly one) substituents. Exemplary substituents are independently selected from C1-6alkyl (such as methyl), C1- alkoxy (such as'methoxy), halo (such as fluoro) and C1-6haloalkyl (such as fluoromethyl, e.g. trifluoromethyl). When Y is unsubstituted heteroaryl, examples include isoxazolyl (e.g. isoxazolyl-5-yl (wherein NR5 is at the 5-position and Z is at the 3-position) and isoxazolyl-3-yl (wherein NR5 is at the 3-position and Z is at the 5-position), especially isoxazol-5-yl), oxadiazolyl, pyridinyl (e.g. pyridin-2-yl or pyridin-3-yl), pyridazinyl (e.g. pyridazin-3-yl), pyrimidinyl (e.g. pyrimidin-3-yl), pyrazinyl (e.g. pyrazin-2-yl). When Y is substituted heteroaryl, Y may for example be substituted by methyl or fluoro. When Y is substituted heteroaryl, examples include methylpyridinyl, fluoropyridinyl,
methylpyridazinyl, methylpyrazinyl methylpyrimidinyl and 1-methylpyrazolyl.
[0105] When Y represents carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl (e.g. methyl), Ci-6alkoxy (e.g. methoxy), C1-6haloalkyl (e.g. fluoromethyl such as trifluoromethyl), C]-6haloalkoxy (e.g. fluoromethoxy) and halo (e.g. fluoro, e.g. chloro), Y may represent carbocyclyl which is optionally substituted by C1-6alkyl. Examples include monocyclic carbocyclyl which is optionally substituted by C\. alkyl (such as methyl). Examples of carbocyclyl include C3.8cycloalkyl (e.g. cyclohexyl) and C5-8cycloalkenyl (e.g. cyclohexenyl). The carbocycyl ring is optionally substituted by one, two or three independently selected substituents (e.g. one or two, especially one, e.g. one methyl group). When Y represents carbocyclyl, Y may represent C3-8cycloalkyl, such as C5-6cycloalkyl. A specific example is cyclohexyl. When Y is 6-membered carbocyclyl, Z and NR5 are may be positioned on the carbocyclyl ring at the 1- and 4- positions relative to each other.
[0106] When Y represents heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1- 6alkyl and-C(0)NHC1-6alkyl, Y may represent heterocyclyl which is optionally substituted by C\-6 alkyl (such as methyl). Examples include monocyclic heterocyclyl. The heterocyclyl group may be unsubstituted or may have for example one or two (e.g. one) substituent (e.g. one methyl group). Examples include piperidinyl, morpholinyl, pyrrolidinyl, 4,5-dihydropyrazolyl and 4,5-dihydroisoxazolyl. Specific examples include 4,5-dihydroisoxazolyl (e.g. 4,5- dihydroisoxazol-5-yl) and piperidinyl (e.g. piperidin-4-yl). When Y is 6-membered heterocyclyl, Z and NR5 may be positioned on the heterocyclyl ring at the l- and 4- positions relative to each other. When Y represents heterocyclyl, in one embodiement Z does not represent -alkylaryl.
[0107] When Z represents alkyl which is optionally substituted by one or more substituents each independently selected f om C1-6alkoxy, NH2, -NHC1-3alkyl and -N(C1-3alkyl)2, exemplary substituents include methoxy, -NH2, -NHmethyl and -NH(methyl)2. Examples include C1- alkyl (e.g. unsubstituted C1- alkyl), for example methyl, ethyl, propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n-hexyl). Examples also include C3- alkyl (e.g. unsubstituted C3-6alkyl), for example propyl (e.g. n-propyl, isopropyl), butyl (e.g. n-butyl, iso-butyl, sec-butyl, tert-butyl), pentyl (e.g. n-pentyl) and hexyl (e.g. n-hexyl). Exemplary C1-6 alkyl groups are methyl and tert-butyl.
[0108] When Z represents aryl, examples include optionally substituted phenyl. Exemplary substituents include one or more (e.g. one or two, especially one) substituents each
independently selected from fluoro, chloro, bromo, amino, methoxy, methyl, haloalkyl (e.g. fluoromethyl such as trifluoromethyl), -COOH, -C(0)NMe2, dimethylamino and -NHC(0)Me. Examples include unsubstituted phenyl. Substituted examples include fluorophenyl (e.g. 2- fluorophenyl, 3 -fluorophenyl and 4-fluorophenyl), chlorophenyl (e.g. 2-chlorophenyl, 3- chlorophenyl, 4-chlorophenyl and 3,4-dichlorophenyl), bromophenyl (e.g. 2-bromophenyl, 3- bromophenyl and 4-bromophenyl), aminophenyl (e.g. 2-aminophenyl, 3-aminophenyl, 4- aminophenyl), methoxyphenyl (e.g. 2-methoxyphenyl, 3-methoxyphenyl and 4- methoxyphenyl), methylphenyl (e.g. 2-methylphenyl, 3-methylphenyl and 4-methylphenyl), fluoromethylphenyl (e.g. 3-trifluoromethylphenyl and 4-trifluoromethylphenyl), carboxyphenyl (e.g. 3-(COOH)-phenyl), 3-(C(0)NMe2)-phenyl), 3-dimethylaminophenyl and 3-(NHC(0)Me)- phenyl.
[0109] When Z represents heteroaryl, examples include monocyclic (e.g. 5 and 6 membered) and bicyclic (e.g. 9 and 10 membered) heteroaryl rings, especially monocyclic rings. Examples of monocyclic heteroaryl comprise one, two or three ring heteroatoms (e.g. one or two, e.g. one) including one or two nitrogen atoms (e.g. one or e.g. two) and optionally an oxygen or sulphur atom. Exemplary 5-membered monocyclic heteroaryl groups include pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, oxadiazolyl and thiadiazolyl.
Exemplary 6-membered monocyclic heteroaryl groups include pyridinyl. Exemplary 6- membered monocyclic heteroaryl groups also include pyridazinyl, pyrimidinyl and pyrazinyl. The aforementioned heteroaryl group may either by unsubstituted or may be substituted by one or more (e.g. one or two, particularly one) substituents. Exemplary substituents are
independently selected from methyl, fluoro, chloro, amino, halomethyl (e.g. fluoromethyl such as trifluorom ethyl). Unsubstituted examples include pyridinyl (e.g. pyridin-2-yl, pyridin-3-yl and pyridin-4-yl), pyrazinyl (e.g. pyrazin-2-yl), pyridazinyl (e.g. pyridazin-3-yl), pyrimidinyl (e.g. pyrimidin-2-yl, pyrimidin-4-yl and pyrimidin-5-yl), oxazolyl (e.g. oxazol-2-yl and oxazol- 5-yl), thiazolyl (e.g. thiazol-2-yl and thiazole-5-yl) and pyrazolyl (e.g. pyrazol-l-yl).
Substituted examples include chloropyridinyl (e.g. 4-chloropyridin-2-yl, 5-chloropyridin-2-yl and 5-chloropyridin-3-yl), fluoropyridinyl (e.g. 4-fluoropyridin-2-yl, 5-fluoropyridin-2-yl and 5- fluoropyridin-3-yl), methylpyridinyl (e.g. 2-methylpyridin-5-yl, 6-methylpyridin-2-yl and 5- methylpyridin-3-yl), fluoromethylpyridinyl (e.g. 5-trifluoromethylpyiidin-3-yl), aminopyridinyl (e.g. 5-aminopyridin-3-yl), methylpyrazinyl (e.g. 5-methylpyrazin-2-yl), methylthiazolyl (e.g. 2- methylthiazol'4-yl) and methylpyrazolyl (e.g. l-methylpyrazol-5-yl).
[0110] When Z represents -alkylaryl, examples include benzyl.
[0111] When Z represents -alkylheteroaryl (e.g. -CH2-heteroaryl), examples include - alkylpyrrolyl, -alkylpyrazolyl, -alkylimidazolyl, -alkyloxazolyl, -alkylisoxazolyl, -alkylthiazolyl, -alkylisothiazolyl, -alkyloxadiazolyl, -alkylthiadiazolyl, -alkylpyridinyl, -alkylpyridazinyl, - alkylpyrimidinyl and -alkylpyrazinyl.
[0112] When Z represents carbocyclyl which is optionally substituted by one or more substituents each independently selected from Ci-6alkyl (e.g. methyl), C1-6alkoxy (e.g. methoxy), C1-6haloalkyl (e.g. fluoromethyl), C1-6haloalkoxy (e.g. fluoromethoxy) and halo (e.g. fluoro, e.g. chloro), Z may represent carbocyclyl which is optionally substituted by Ci-6 alkyl. Examples include monocyclic carbocyclyl which is optionally substituted by Q-6 alkyl (such as methyl). Examples of carbocyclyl include C3-8cycloalkyl (e.g. cyclohexyl) and C5-8cycloalkenyl (e.g. cyclohexenyl). The carbocycyl ring is optionally substituted by one, two or three substituents (e.g. one or two, especially one, e.g. one methyl group). When Z represents carbocyclyl, Z may represent C3-8cycloalkyl, such as C5-6cycloalkyl. A specific example is cyclohexyl.
[0113] When Z represents heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1- 6alkyl,-C(0)NHC1-6alkyl C1-6alkyl (e.g. methyl), -C(0)OC1-6alkyl (e.g. -C(O)Omethyl), - C(0)Ci-6alkyl (e.g. -C(0)methyl),-C(0)NHCi-6alkyl (e.g. -C(O)NHmethyl), examples include unsubstituted heterocyclyl and heterocyclyl with one or two (e.g. one) substituent. Examples include heterocyclyl groups containing one nitrogen atom (such as pyrrolidinyl and piperidinyl) or two nitrogen atoms (such as piperazinyl). Examples also include heterocyclyl groups containing one nitrogen atom and one oxygen atom(e.g. morpholinyl) or one sulfur atom (e.g. thiomorpholinyl). In one embodiment when Z is substituted, a ring nitrogen atom is substituted. Substituted examples include substituted piperazinyl, such as 4-substitued piperazinyl, e.g. 4- Boc-piperazinyl, 4-(C(0)Me)-piperazinyl, 4-(C(0)NHEt)piperazinyl and 4-methylpiperazinyl.
[0114] When Z represents -alkylcarbocyclyl (e.g. -CH2-carbocyclyl) wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl (e.g. methyl), Ci-6haloalkyl (e.g. fluoromethyl such as trifluoromethyl), C1-6alkoxy (e.g.
methoxy), Ci-6haloalkoxy (e.g. fluoromethoxy) and halo (e.g. fluoro, e.g. chloro), examples include -CH2-cyclopropyl and -CH2-cyclobutyl.
[0115] When Z represents -alkylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)Ci. 6alkyl,-C(0)NHC]-6alkyl C1-6alkyl (e.g. methyl), -C(0)OCi-6alkyl (e.g. -C(O)Omethyl), - C(0)C1-6alkyl (e.g. -C(0)methyl),-C(0)NHC1-6alkyl (e.g. -C(O)NHmethyl), examples include -CH2-heterocyclyl, such as -CH2-morpholinyl, -CH2-piperazinyl, -CH2-piperidinyl and -CH2- pyrrolidinyl.
[0116] When Z represents— aryl carbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C1.6alkyl (e.g. methyl), Ci-6alkoxy (e.g. methoxy), C1-6haloalkyl (e.g. fluoromethyl such as trifluoromethyl), Ci.ghaloalkoxy (e.g.
fluoromethoxy) and halo (e.g. fluoro, e.g. chloro), Z maybe phenylcarbocyclyl. The carbocyclyl ring may be unsubstituted or may be substituted by one or more C1-6alkyl groups. The carbocyclyl ring may be monocyclic. An exemplary carbocyclyl ring is cycloalkyl.
Examples include cyclopropylphenyl- and cyclohexylphenyl-.
[0117] When Z represents -arylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1- 6alkyl, -C(0)C1-6alkyl,-C(0)NHC1-6alkyl C1-6alkyl (e.g. methyl), -C(0)OCi-6alkyl (e.g. - C(O)Omethyl), -C(0)C1-6alkyl (e.g. -C(0)methyl),-C(0)NHC1-6alkyl (e.g. -C(O)NHmethyl), Z may be -phenylheterocyclyl. The heterocyclyl ring may be monocyclic and may contain one or two (e.g. one) nitrogen atoms. Examples include morphinolylphenyl-, piperazinylphenyl-, piperidinylphenyl-, pyrrolidinylphenyl-. A specific example is 3-(morpholin-4-yl)phenyl-.
[0118] Suitably ¾ represents H, methyl, ethyl, -C(0)OMe, CF3 or OMe. In one embodiment represents methyl.
[0119] Suitably each R2 independently represents H or alkyl. More suitably R2 represents H or methyl. In one embodiment, R2 represents H. In one embodiment R2 represents methyl.
[0120] Suitably n represents 0 or 1 , for example n represents 1.
[0121] Suitably R3 represents H or methyl, for example R3 represents H.
[0122] Suitably R4 represents H or methyl, for example R4 represents H.
[0123] Suitably R5 represents H or methyl, for example R5 represents H.
[0124] Suitably W and X are the same as each other.
[0125] Suitably W and X each represent C=0.
[0126] Suitably Y represents aryl (e.g. phenyl) or heteroaryl (e.g. 5- or 6-membered monocyclic heteroaryl comprising one, two or three ring heteroatoms including one or two nitrogen atoms such as isoxazolyl, pyridinyl, pyridazinyl, pyrimidinyl or pyrazinyl). In one embodiment Y represents phenyl. In one embodiment Y represents monocyclic heteroaryl. In one embodiment Y is substituted by one or more substituents each independently selected from Ci-6 alkyl, C^ 6alkoxy, halo and C1-6haloalkyl. In one embodiment Y is unsubstituted. In one embodiment Y is monosubstituted. When Y is 5-membered, suitably Z and NR5 are positioned on the ring at non-adjacent ring atoms. When Y is 6-membered, suitably Z and NR5 are positioned on the ring at 1- and 4- positions relative to each other (i.e. Z and NR5 have a para relationship).
[0127] Suitably Z represents aryl (e.g.phenyl) or heteroaryl (for example 6-membered heteroaryl such as pyridinyl, pyridazinyl, pyrimidinyl or pyrazinyl especially pyridinyl; or 5- membered heteroaryl such as oxazolyl, thiazolyl, pyrazolyl). In one embodiment Z represents heteroaryl. In one embodiment Z represents aryl. In one embodiment, Z does not represent methyl or ethyl. In one embodiment Z does not represent unsubstituted alkyl.
[0128] In one embodiment there are provided compounds of formula (I) which are compounds of formula (IA)
Figure imgf000029_0001
or a pharmaceutically acceptable salt, solvate or polymorph thereof, including all tautomers and stereoisomers thereof wherein:
[0129] Ri represents alkyl;
[0130] each R2 independently represents H, alkyl, carbocyclyl which may be optionally substituted by one or more substituents each independently selected from C1- alkyl, C1- alkoxy, Ci-6haloalkoxy and halo, -NHalkyl, -N(alkyl) , amino, hydroxyl, alkoxy or halo;
[0131] n represents 0, 1 or 2;
[0132] R4 represents H or alkyl;
[0133] R5 represents H or alkyl;
[0134] W and X each independently represent C=0 or C=S (in some embodiments, W and X are both 0=0); [0135] Y represents aryl; heteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, Q.
6haloalkoxy and halo; or heterocyclyl which is optionally substituted by one or more
substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and - C(0)NHC1-6alkyl; and
[0136] Z represents alkyl; aryl; heteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkoxy (e.g. fluoromethoxy) C1-6haloalkoxy and halo; heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and -C(0)NHC1-6alkyl; -arylcarbocyclyl wherein carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkoxy and halo ;or -arylheterocyclyl wherein heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)Ci-6alkyl and -C(0)NHC1-6alkyl.
[0137] In one embodiment, compounds of formula (I) are provided in which R represents methyl; R2 represents H; n represents 1 ; W and X each represent C=0; R3 represents H; R4 represents H; Y represents isoxazolyl; and Z represents phenyl which is optionally substituted by chloro, fluoro, bromo, methyl or methoxy.
[0138] In one embodiment, compounds of formula (I) are provided in which i represents methyl; R2 represents H; n represents 1 ; W and X each represent C=0; R3 represents H; R4 represents H; Y represents phenyl; and Z represents phenyl which is optionally substituted by fluoro, chloro, amino, methyl or methoxy.
[0139] In one embodiment, compounds of formula (I) are provided in which i represents methyl; R2 represents H; n represents 1; W and X each represent C=0; R3 represents H; R4 represents H; Y represents pyridinyl; and Z represents pyridinyl which is optionally substituted by fluoro, chloro, methyl, amino or trifluoromethyl.
[0140] In one embodiment, compounds of formula (I) are provided in which R] represents methyl; R2 represents H; n represents 1 ; W and X each represent C=0; R3 represents H; R4 represents H; ; Y represents pyridazinyl, pyrimidinyl or pyrazinyl; and Z represents pyridinyl which is optionally substituted by fluoro, chloro, methyl, amino or trifluoromethyl. [0141] In one embodiment, R2, R4 and R5 are all H, n=l and W and X are both C=0. [0142] In one embodiment the following compounds of fonnula (I) are excluded:
Figure imgf000031_0001
[0146] In one embodiment, Z is not phenyl substituted by trifluoroalkyl.
[0147] In one embodiment, when Y is isoxazolyl, Z is not positioned at the 4-position of the isoxazolyl ring.
[0148] In one embodiment, the compounds of formula (I) have an IC50 against HEK293-STF3A cells of less than about 10 micromolar.
Processes
[0149] The present invention further provides a process for preparation of compounds of formula (I), wherein W and X each represent C=0, which comprises an amide coupling reaction of a compound of formula (II): 1
Figure imgf000032_0001
or a protected derivative thereof, wherein Rl5 R2, n, R3, R4, W and X are as defined above, with a compound of formula (III)
Figure imgf000032_0002
(III)
or a protected derivative thereof, wherein R5, Y and Z are as defined above.
[0150] Compounds of formula (II) may be prepared by reaction of compounds of formula (IV)
Figure imgf000032_0003
(IV)
wherein Rj, R2 and n are as defined above, with a compound of formula (V)
Figure imgf000033_0001
wherein R3 and R4 are as defined above. An exemplary solvent for this reaction is chloroform.
[0151] Compounds of formula (IV) can be prepared by reaction of compounds of formula (VI)
Figure imgf000033_0002
(VI)
wherein ¾ is defined as above; with a compound of formula (VII)
Figure imgf000033_0003
(VII)
wherein R2 and n are as defined above and wherein LG represents a leaving group such as Br, I, tosylate etc. The reaction may take place in the presence of triethylamine or other amine base (DBU, DBN etc.) and a solvent such as 1,4-dioxane or other dipolar aprotic solvent.
[0152] Compounds of formula (III) maybe synthesized by a coupling reaction, such as a Suzuki coupling reaction. When Y is isoxazole, compounds of formula (III) may alternatively be synthesized from the corresponding oxopropanenitrile by reaction with hydroxylamine hydrochloride in the presence of sodium hydroxide. The oxopropanenitrile can be obtained from the corresponding ester.
[0153] The present invention further provides a process for preparation of compounds of formula (I), wherein W and X each represent CH2 or a protected derivative thereof, which comprises conversion of the W and X groups of compounds of formula (I) from C=0 groups into CH2 groups. This conversion can be carried out, for example, by reduction using borane dimethylsulfide.
[0154] The present invention further provides a process for preparation of compounds of formula (I), wherein W and X each represent C=S, or a protected derivative thereof, which comprises conversion of the W and X groups of compounds of formula (I) from C=0 groups into C=S groups. This conversion can be carried out, for example, by using Lawesson's reagent.
Therapeutic uses
[0155] The present invention provides a compound of formula (I) for use as a medicament.
[0156] The compounds of the present invention may have an IC50 against STF3A of less than 20 micromolar, e.g. less than 10 micromolar. The IC50 maybe less than 5, 2, 1, 0.5, 0.2 or 0.1 micromolar. It may be between about 0.01 and about 10 micromolar, or between about 0.01 and 5, 0.01 and 1, 0.01 and 0.5, 0.01 and 0.1, 0.01 and 0.05, 0.1 and 5, 0.1 and 1, 0.1 and 0.5, 0.1 and 10, 0.5 and 10, 1 and 10, 5 and 10, 1 and 5 or 0.1 and 0.5, e.g. about 0.01, 0.02, 0.03, 0.04, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 micromolar.
[0157] The present invention provides compounds of formula (I) for use in modulation of the WNT pathway.
[0158] The present invention also provides a method of modulating WNT activity comprising exposing a WNT protein or a WNT receptor to a compound of formula (I).
[0159] In addition, the present invention provides use of a compound of formula (I) for modulating WNT activity.
[0160] The present invention additionally provides compounds of formula (I) for use in the treatment of a disease or condition associated with WNT pathway activity. [0161] A method of treating a disease or condition associated with W T pathway activity comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula (I) is also provided by the present invention.
[0162] The present invention further provides use of a compound of formula (I) for treatment of a disease or condition associated with WNT pathway activity.
[0163] Also provided by the present invention is use of a compound of formula (I) in the manufacture of a medicament for the treatment of a disease or condition associated with WNT pathway activity.
[0164] The aforementioned disease or condition is suitably selected from the group consisting of cancer, fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis and myocardial infarction.
[0165] The cancer may be a cancer characterized by high WNT activity.
[0166] The disease or condition may be a cancer, such as cervical, colon, breast, bladder, head and neck, gastric, lung, ovarian, prostate, thyroid, non-small-cell lung, as well as chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal cell carcinoma, osteosarcoma, hepatocellular carcinoma, Wilm's tumor or medulloblastoma, or a fibrotic disease, such as pulmonary fibrosis, liver fibrosis, skin fibrosis or renal fibrosis, or a degenerative disease, or a metabolic disease such as diabetic retinopathy.
[0167] The present invention also provides use of a compound of formula (I) in diagnosis.
Pharmaceutical compositions
[0168] To prepare the pharmaceutical compositions of this invention, at least one compound of formula (I) optionally in combination with at least one of the other aforementioned agents can be used as the active ingredient(s). The active ingredient(s) is intimately admixed with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques, which carrier may take a wide variety of forms depending of the form of preparation desired for administration, e.g., oral or parenteral such as intramuscular. In preparing the compositions in oral dosage form, any of the usual pharmaceutical media may be employed. Thus, for liquid oral preparations, such as for example, suspensions, elixirs and solutions, suitable carriers and additives include water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like; for solid oral preparations such as, for example, powders, capsules, gelcaps and tablets, suitable carriers and additives include starches, sugars, diluents, granulating agents, lubricants, binders, disintegrating agents and the like. Because of their ease in administration, tablets and capsules represent the most advantageous oral dosage unit form, in which case solid pharmaceutical carriers are obviously employed. If desired, tablets maybe sugar coated or enteric coated by standard techniques. For parenterals, the carrier will usually comprise sterile water, though other ingredients, for example, for purposes such as aiding solubility or for preservation, may be included.
[0169] Injectable suspensions may also prepared, in which case appropriate liquid carriers, suspending agents and the like may be employed. The pharmaceutical compositions herein will contain, per dosage unit, e.g., tablet, capsule, powder, injection, teaspoonful and the like, an amount of the active ingredient(s) necessary to deliver an effective dose as described above. The pharmaceutical compositions herein will contain, per dosage unit, e.g., tablet, capsule, powder, injection, suppository, teaspoonful and the like, from about 0.03 mg to 100 mg/kg (preferred 0.1 - 30 mg/kg) and may be given at a dosage of from about 0.1 - 300 mg/kg per day (preferred 1 - 50 mg/kg per day) of each active ingredient or combination thereof. The dosages, however, may be varied depending upon the requirement of the patients, the severity of the condition being treated and the compound being employed. The use of either daily administration or post- periodic dosing may be employed.
[0170] Preferably these compositions are in unit dosage forms from such as tablets, pills, capsules, powders, granules, sterile parenteral solutions or suspensions, metered aerosol or liquid sprays, drops, ampoules, autoinjector devices or suppositories; for oral parenteral, intranasal, sublingual or rectal administration, or for administration by inhalation or insufflation. Alternatively, the composition may be presented in a form suitable for once-weekly or once- monthly administration; for example, an insoluble salt of the active compound, such as the decanoate salt, may be adapted to provide a depot preparation for intramuscular injection. For preparing solid compositions such as tablets, the principal active ingredient is mixed with a pharmaceutical carrier, e.g. conventional tableting ingredients such as corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate or gums, and other pharmaceutical diluents, e.g. water, to form a solid preformulation composition containing a homogeneous mixture of a compound of the present invention, or a pharmaceutically acceptable salt thereof. When referring to these preformulation compositions as homogeneous, it is meant that the active ingredient is dispersed evenly throughout the composition so that the composition may be readily subdivided into equally effective dosage forms such as tablets, pills and capsules. This solid preformulation composition is then subdivided into unit dosage forms of the type described above containing from 0.1 to about 500 mg of each active ingredient or combinations thereof of the present invention.
[0171] The tablets or pills of the compositions of the present invention can be coated or otherwise compounded to provide a dosage form affording the advantage of prolonged action. For example, the tablet or pill can comprise an inner dosage and an outer dosage component, the latter being in the form of an envelope over the former. The two components can be separated by an enteric layer which serves to resist disintegration in the stomach and permits the inner component to pass intact into the duodenum or to be delayed in release. A variety of material can be used for such enteric layers or coatings, such materials including a number of polymeric acids with such materials as shellac, cetyl alcohol and cellulose acetate.
[0172] This liquid forms in which the compositions of the present invention may be
incorporated for administration orally or by injection include, aqueous solutions, suitably flavoured syrups, aqueous or oil suspensions, and flavoured emulsions with edible oils such as cottonseed oil, sesame oil, coconut oil or peanut oil, as well as elixirs and similar
pharmaceutical vehicles. Suitable dispersing or suspending agents for aqueous suspensions, include synthetic and natural gums such as tragacanth, acacia, alginate, dextran, sodium carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone or gelatin.
[0173] The pharmaceutical composition may contain between about 0.01 mg and 100 mg, preferably about 5 to 50 mg, of each compound, and maybe constituted into any form suitable for the mode of administration selected. Carriers include necessary and inert pharmaceutical excipients, including, but not limited to, binders, suspending agents, lubricants, flavorants, sweeteners, preservatives, dyes, and coatings. Compositions suitable for oral administration include solid forms, such as pills, tablets, caplets, capsules (each including immediate release, timed release and sustained release formulations), granules, and powders, and liquid forms, such as solutions, syrups, elixirs, emulsions, and suspensions. Forms useful for parenteral administration include sterile solutions, emulsions and suspensions. [0174] Advantageously, compounds of the present invention may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. Furthermore, compounds for the present invention can be administered in intranasal form via topical use of suitable intranasal vehicles, or via transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of transdermal delivery system, the dosage administration will, of course, be continuous rather than intermittent throughout the dosage regimen.
[0175] For instance, for oral administration in the form of a tablet or capsule, the active drug component can be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. Moreover, when desired or necessary, suitable binders; lubricants, disintegrating agents and coloring agents can also be incorporated into the mixture. Suitable binders include, without limitation, starch, gelatin, natural sugars such as glucose or betalactose, corn sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Disintegrators include, without limitation, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
[0176] The liquid forms in suitable flavored suspending or dispersing agents such as the synthetic and natural gums, for example, tragacanth, acacia, methyl-cellulose and the like. For parenteral administration, sterile suspensions and solutions are desired. Isotonic preparations which generally contain suitable preservatives are employed when intravenous administration is desired.
[0177] The compounds or combinations of the present invention can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, such as cholesterol, stearylamine or phosphatidylcholines.
[0178] Compounds or combinations of the present invention may also be delivered by the use of monoclonal antibodies as individual carriers to which the compound molecules are coupled. The compounds of the present invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer,
polyhydroxypropylmethacrylamidephenol, polyhydroxyethylaspartamid-ephenol, or polyethyl eneoxidepolyllysine substituted with palmitoyl residue. Furthermore, the compounds of the present invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polyactic acid, polyepsilon caprolactone, polyhydroxy butyeric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross- linked or amphipathic block copolymers of hydrogels.
[0179] Compounds or combinations of this invention may be administered in any of the foregoing compositions and according to dosage regimens established in the art whenever treatment of the addressed disorders is required.
[0180] The daily dosage of the products may be varied over a wide range from 0.01 to 1.000 mg per mammal per day. For oral administration, the compositions are preferably provided in the form of tablets containing, 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 150, 200, 250 and 500 milligrams of each active ingredient or combinations thereof for the symptomatic adjustment of the dosage to the patient to be treated. An effective amount of the drug is ordinarily supplied at a dosage level of from about 0.1 mg/kg to about 300 mg/kg of body weight per day. Preferably, the range is from about 1 to about 50 mg/kg of body weight per day. The compounds or combinations may be administered on a regimen of 1 to 4 times per day.
[0181] Optimal dosages to be administered may be readily determined by those skilled in the art, and will vary with the particular compound used, the mode of administration, the strength of the preparation, the mode of administration, and the advancement of disease condition. In addition, factors associated with the particular patient being treated, including patient age, weight, diet and time of administration, will result in the need to adjust dosages.
[0182] In a further aspect, the invention also provides a process for preparing a pharmaceutical composition comprising at least one compound of formula (I), optionally in combination with at least one of the other aforementioned agents and a pharmaceutically acceptable carrier.
[0183] The compositions are preferably in a unit dosage form in an amount appropriate for the relevant daily dosage.
[0184] Suitable dosages, including especially unit dosages, of the compounds of the present invention include the known dosages including unit doses for these compounds as described or referred to in reference text such as the British and US Pharmacopoeias, Remington's
Pharmaceutical Sciences (Mack Publishing Co.), Martindale The Extra Pharmacopoeia (London, The Pharmaceutical Press) (for example see the 31st Edition page 341 and pages cited therein) or the above mentioned publications.
Examples
Figure imgf000040_0001
Figure imgf000041_0001
Figure imgf000042_0001
Figure imgf000043_0001
Figure imgf000044_0001
Figure imgf000045_0001
Figure imgf000046_0001
STF3A
Cpd
Structure lUPAC Name IC50 ID
μΜ
N-([2/3'-bipyridin]-5-yl)-4-(2- methyl-6,7-
48 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
4-{2-methyl-5H,6H,7H- pyrazolo[l,5-a]pyrimidin-4-
49 <0.1 yl}-4-oxo-N-[5-(pyridin-3- yl)pyridin-2-yl]butanamide
4-(2-met yl-6,7- dihydropyrazolo[l,5-
50 a]pyrimidin-4(5H)-yl)-4-oxo- <0.1
N-(5-(pyrazin-2-yl)pyridin-2- yl)butanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5-
51 a]pyrimidin-4(5H)-yl)-4-oxo- <0.1
N-(6-(pyridin-3-yl)pyridazin-3- yl)butanamide
4-(2-met yl-6,7- dihydropyrazolo[l,5-
52 a]pyrimidin-4(5H)-yl)-4-oxo- <0.1
N-(5-(pyridin-3-yl)pyrazin-2- yl)butanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5-
53 a]pyrimidin-4(5H)-yl)-N-(3- >10 methyl-[l,l'-biphenyl]-4-yl)- 4-oxobutanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5-
54 a]pyrimidin-4(5H)-yl)-N-(2- <0.1 methyl-[l,l'-biphenyl]-4-yl)- 4-oxobutanamide
Figure imgf000047_0001
Figure imgf000048_0001
Figure imgf000049_0001
STF3A
Cpd
Structure lUPAC Name IC50 ID
μΜ
N-(2'-amino-[l,l'-biphenyl]-4- yl)-4-(2-methyl-6,7-
69 dihydropyrazolo[l,5- <1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(3'-amino-[l,l'-biphenyl]-4- yl)-4-(2-methyl-6,7-
70 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(4'-amino-[l, -biphenyl]-4- yl)-4-(2-methyl-6,7-
71 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(3'-(dimethylamino)-[l,l'- biphenyl]-4-yl)-4-(2-methyl-
72 6,7-dihydropyrazolo[l,5- >10 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5-
73 a]pyrimidin-4(5H)-yl)-N-(3'- <5 morpholinobiphenyl-4-yl)-4- oxobutanamide
N-(3'-acetamido-[l,l'- biphenyl]-4-yl)-4-(2-methyl-
74 6,7-dihydropyrazolo[l,5- <5 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
Figure imgf000050_0001
N-(4-cyclohexylphenyl)-4-(2- methyl-6,7-
75 dihydropyrazolo[l,5- >10 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
Figure imgf000051_0001
STF3A
Cpd
Structure lUPAC Name IC50 ID
μΜ
N-(6-(3-chlorophenyl)pyridin- 3-yl)-4-(2-methyl-6,7-
83 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(6-(3-fluorophenyl)pyridin- 3-yl)-4-(2-methyl-6,7-
84 dihydropyrazolo[l,5- <0.1
FYj N 0 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5-
85 a]pyrimidin-4(5H)-yl)-4-oxo- <0.1
IM-(6-(m-tolyl)pyridin-3- yl)butanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5-
86 a]pyrimidin-4(5H)-yl)-N-(5- <5 methyl-6-phenylpyridin-3-yl)- 4-oxobutanamide
N-(6-(4-fluorophenyl)pyridin- 3-yl)-4-(2-methyl-6,7-
87 dihydropyrazolo[l,5- <1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(6-(3- fluorophenyl)pyridazin-3-yl)- 4-(2-methyl-6,7-
88 <0.1 dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(5-(3-fluorophenyl)pyridin- 2-yl)-4-(2-methyl-6,7-
89 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
Figure imgf000052_0001
STF3A
Cpd
Structure lUPAC Name IC50 ID
μΜ
N-(5-(4-fluorophenyl)pyridin- 2-yl)-4-(2-methyl-6,7-
90 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(3-methyl-5-phenylpyridin- 2-yl)-4-(2-methyl-6,7-
91 dihydropyrazolo[l,5- <5 a]pyrimidin-4(5H)-yl)-4- oxobutana
N-(4-methyl-5-phenylpyridin- 2-yl)-4-(2-methyl-6,7-
92 dihydropyrazolo[l,5- <1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
Figure imgf000053_0001
4-(2-methyl-6,7- dihydropyrazolo[l,5-
93 a]pyrimidin-4(5H)-yl)-4-oxo- <0.1
N-(5-(m-tolyl)pyridin-2- yl)butanamide
N-(5-fluoro-2,3'-bipyridin-6'- yl)-4-(2-methyl-6,7-
94 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(4-fluoro-2/3'-bipyridin-6'- yl)-4-(2-methyl-6,7-
95 dihydropyrazolo[l,5- <0.1
XX a]pyrimidin-4(5H)-yl)-4- oxobutanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5-
96 a]pyrimidin-4(5H)-yl)-N-(6- <0.1
(X methyl-[2,3'-bipyridin]-6'-yl)- 4-oxobutanamide
Figure imgf000054_0001
STF3A
Cpd
Structure lUPAC Name IC50 ID
μΜ
4-(2-methyl-6,7- dihydropyrazolo[l,5-
104 a]pyrimidin-4(5H)-yl)-N-(6-(5- <0.1 methylpyridin-3-yl)pyridazin- 3-yl)-4-oxobutanamide
Figure imgf000055_0001
4-(2-methyl-6,7- dihydropyrazolo[l/5- a]pyrimidin-4(5H)-yl)-4-oxo-
105 <0.1
N-(6-(5- (trifluoromethyl)pyridin-3-
N yl)pyridazin-3-yl)butanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-N-(5-
106 <5 methyl-6-(pyridin-3- yl)pyridazin-3-yl)-4-
N oxobutanamide
4-(2-methyl-6,7- di ydropyrazolo[l,5-
107 a]pyrimidin-4(5H)-yl)-N-(6-(6- <0.1 methylpyridin-3-yl)pyridazin- 3-yl)-4-oxobutanamide
N-(5'-chloro-[2,3'-bipyridin]- 5-yl)-4-(2-methyl-6,7-
108 dihydropyrazolo[l,5- <0,1
C'Y N ° a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(5'-fluoro-[2,3'-bipyridin]-5- yl)-4-(2-methyl-6,7-
109 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(5'-methyl-2,3'-bipyridin-5- yl)-4-(2-methyl-6,7-
110 dihydropyrazolol S- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N
Figure imgf000056_0001
STF3A
Cpd
Structure lUPAC Name IC50 ID
μΜ
4-(2-methyl-6,7- dihydropyrazolo[l,5-
118 a]pyrimidin-4(5H)-yl)-4-oxo- <0.1
N-(5'-(trifluoromethyl)-[3,3'- bipyridin]-6-yl)butanamide
N-(4-methyl-3,3'-bipyridin-6- yl)-4-(2-methyl-6,7-
119 dihydropyrazolo[l,5- <5 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(4,5'-dimethyl-3,3'- bipyridin-6-yl)-4-(2-methyl-
120 6,7-dihydropyrazolo[l,5- <5 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(6'-methyl-3,3'-bipyridin-6- yl)-4-(2-methyl-6,7-
121 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
Figure imgf000057_0001
N-(5-(5-fluoropyridin-3- yl)pyrazin-2-yl)-4-(2-methyl-
122 6,7-dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-(5-(3-fluorophenyl)pyrazin- 2-yl)-4-(2-methyl-6,7-
123 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5-
124 a]pyrimidin-4(5H)-yl)-N-(5-(5- <0.1 methylpyridin-3-yl)pyrazin-2- yl)-4-oxobutanamide
Figure imgf000057_0002
Figure imgf000058_0001
Figure imgf000059_0001
Figure imgf000060_0001
STF3A
Cpd
Structure lUPAC Name IC50 ID
μΜ
2-methyl-4-(2-methyl-6,7- dihydropyrazolo[l,5-
146 a]pyrimidin-4(5H)-yl)-4-oxo- <0.1
N-(6-phenylpyridazin-3- yl)butanamide
N-([3,3'-bipyridin]-6-yl)-2- methyl-4-(2-methyl-6,7-
147 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-^-methyl^'-bipyridin-e'- yl)-4-(2-methyl-6,7-
148 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
4-(2-methyl-6,7- dihydropyrazolo[l,5-
149 a]pyrimidin-4(5H)-yl)-N-(5-(5- <0.1 methylpyrazin-2-yl)pyridin-2- yl)-4-oxobutanamide
2-methyl-4-(2-methyl-6,7- dihydropyrazolo[l,5-
150 a]pyrimidin-4(5H)-yl)-N-(5'- <0.1 methyl-[2,3'-bipyridin]-5-yl)- 4-oxobutanamide
2-methyl-4-(2-methyl-6,7- dihydropyrazolo[l,5-
151 a]pyrimidin-4(5H)-yl)-N-(5-(6- <0.1 methylpyrazin-2-yl)pyridin-2- yl)-4-oxobutanamide
2-methyl-4-(2-methyl-6/7- dihydropyrazolo[l,5-
152 a]pyrimidin-4(5H)-yl)-N-(5'- <0.1 methyl-[3,3'-bipyridin]-6-yl)- 4-oxobutanamide
Figure imgf000062_0001
Figure imgf000063_0001
STF3A
Cpd
Structure lUPAC Name IC50 ID
μΜ
4-(2,6-dimethyl-6,7- dihydropyrazolo[l,5-
167 a]pyrimidin-4(5H)-yl)-N-(6-(5- <0.1 fluoropyridin-3-yl)pyridazin- 3-yl)-4-oxobutanamide
Figure imgf000064_0001
N-(3,5'-dimethyl-[2,3'- bipyridin]-5-yl)-4-(2-methyl-
168 6,7-dihydropyrazolo[l,5- <1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
(S)-N-([3,3'-bipyridin]-6-yl)-4- (2,6-dimethyl-6,7-
169 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
(R)-N-([3,3'-bipyridin]-6-yl)-4- (2,6-dimethyl-6,7-
170 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
N-([2,3'-bipyridin]-6'-yl)-4- (2,6-dimet yl-6,7-
171 dihydropyrazolo[l,5- <0.1 a]pyrimidin-4(5H)-yl)-4- oxobutanamide
Figure imgf000064_0002
[0185] In one embodiment, compounds selected from Examples 1 to 171 having an IC50 against STF3A cells of 10 micromolar or more are excluded.
[0186] Synthesis of the examples:
[0187] The compounds were synthesized according to the following general synthesis schemes:
[0188] Compounds of formula (I) in which W and X each represent C=0, and
synthesised according to the following scheme:
Figure imgf000065_0001
[0189] Compounds of formula (I) in which W and X each represent CH2, and
synthesised according to the following scheme:
Figure imgf000065_0002
[0190] Compounds of formula (I) in which W and X each represent C=S, and
synthesised according to the following scheme:
Figure imgf000065_0003
[0191] For example, compounds of formula (I) in which ¾ represents methyl, R2 represents H , n represents 1 and W and X each represent C=0, and can be synthesised according to the following scheme:
Figure imgf000066_0001
[0192] Synthesis of amines
[0193] General esterifi cation procedure
[0194] To a solution of the corresponding benzoic acid (1 equiv.) in methanol (0.65 M) was added concentrated sulphuric (32 M), and the resulting reaction mixture was heated at 70 °C for 10 h. After completion, the reaction mixture concentrated, water was added and extracted with ethyl acetate. The combined organic layers were washed with aqueous sodium bicarbonate solution and brine, dried over sodium sulphate and concentrated under vacuum to give the desired ester.
[0195] General formation of 3-phenyl-3-oxopropanenitrile procedure
[0196] The corresponding ester prepared above (1 equiv.) in tetrahydrofuran (0.9 M) and acetonitiile (1 equiv.) was added dropwise to a stirred solution of sodium hydride (1.5 equiv.) in tetrahydrofuran (0.65M). The resulting reaction mixture was stirred at 70 °C for 16 h. After completion of starting material, the reaction mixture was quenched with ice cold water and acdified to pH 5 with concentrated hydrochloric acid and extracted with ethyl acetate. The combined organic layers were washed brine, dried over sodium sulphate and concentrated under vacuum to give the desired product.
[0197] General formation of amines
[0198] To a stirred solution of the corresponding 3-phenyl-3-oxopropanenitrile (1 equiv.) in water (0.7M) was added aqueous sodium hydroxide (2 equiv.) solution and hydroxylamine hydrochloride (1.1 equiv.) at room temperature. The resulting reaction mixture was stirred at 100 °C for 5 h. After completion of starting material, the reaction mixture was cooled to room temperature and precipitated solid was collected by filtration and dried to give the desired product.
[0199] Synthesis of 3-(2-chlorophenyl)isoxazol-5-amine.
[0200] Step 1 : Preparation of methyl 2-chlorobenzoate.
Figure imgf000067_0001
[0201] 1H NMR (400 MHz, DMSO-c¾): 7.84-7.82 (dd, Ji = 7.6 Hz, J2
(m, 2H), 7.34-7.26 (m, IH), 3.94 (s, 3H). LC-MS: m/z 170.9 [M+H]+.
[0202] Step 2: Preparation of 3-(2-chlorophenyl)-3-oxopropanenitrile.
Figure imgf000067_0002
[0203] LC-MS: 178.0 [M+H]+.
[0204] Step 3: Preparation of 3-(2-chlorophenyl)isoxazol-5-amine.
Figure imgf000067_0003
[0205] 1H NMR (400 MHz, DMSO-d6): 7.70-7.67 (m, IH), 7.49-7.42 (m, IH), 7.39-7.32 (m, 2H), 5.60 (s, IH), 4.52 (brs, 2H). LC-MS: 194.9 [M+H]+.
[0206] Synthesis of 3-phenylisoxazol-5-amine.
Figure imgf000068_0001
[0207] 1H NMR (400 MHz, DMSO-i 6): 7.73-7.74 (d, J= 8.4 Hz, 2H), 7.45-7.43 (m, 3H), 6.78 (brs, 2H), 5.40 (s, IH). LC-MS: m/z 161.0 [M+H]+.
[0208] Synthesis of 3-(3-chlorophenyl)isoxazol-5-amine.
Figure imgf000068_0002
[0209] LC-MS: m/z 195 [M+H]+.
[0210] Synthesis of 3-(4-chlorophenyl)isoxazol-5-amine.
Figure imgf000068_0003
[0211] 1H NMR (400 MHz, DMSO- 6): 7.76-7.74 (d, J= 8.4 Hz, 2H), 7.52-7.50 (d, J=8.4 Hz, 2H), 6.85 (brs, 2H), 5.42 (s, IH). LC-MS: m/z 194.9 [M+H]+.
[0212] Synthesis of 3-(3,4-dichlorophenyl)isoxazol-5-amine (VCW-WNT- 168B)
Figure imgf000068_0004
[0213] 1H NMR (400 MHz, DMSO-d6): 7.97 - 7.96 (m, IH), 7.73 - 7.72 (m, 2H), 6.88 (s, 2H), 5.51 (s, IH). LC-MS: m/z 230 [M+H]+.
[0214] Synthesis of 3-(3-fluorophenyl)isoxazol-5-amine. [0215] Step 1 : Preparation of 3-(3-fluorophenyl)-3-oxopropanenitrile.
Figure imgf000069_0001
[0216] LC-MS: m/z 162.0 [M+H]+.
[0217] Step 2: Preparation of 3-(3-fluorophenyl)isoxazol-5-amine.
Figure imgf000069_0002
[0218] 1H NMR (400 MHz, DMSO-c¾: 7.60-7.47 (m, 3H), 7.30-7.26 (t, J
(brs, 2H), 5.42 (s, IH). LC-MS: m/z 177.0 [M+H]+.
[0219] Synthesis of 3-(o-tolyl)isoxazol-5-amine.
[0220] Step 1 : Preparation of 3 -oxo-3-(o-tolyl)propanenitrile.
Figure imgf000069_0003
[0221] 1H NMR (400 MHz, DMSO-i¾: 7.63-7.61 (m, IH), 7.50-7.46 (m, IH), 7.34-7.31 (m, 2H), 4.03 (s, 2H), 2.57 (s, 3H).
[0222] Step 2: Preparation 3-(o-tolyl)isoxazol-5-amine.
Figure imgf000069_0004
[0223] 1H NMR (400 MHz, DMSO-i¾: 7.44-7.42 (m, IH), 7.35-7.23 (m, 3H), 6.70 (s, 2H), 5.18 (s, IH), 2.39 (s, 3H). LC-MS: m/z 175 [M+H]+. [0224] Synthesis of 3-(4-chloropyridin-2-yl)isoxazol-5-
Figure imgf000070_0001
[0225] 1H NMR (400 MHz, Methanol-^): 8.55 (d, J=4.8 Hz, IH), 7.92 (m, IH), 7.50 (dd, J=4 Hz, 2 Hz, IH), 5.61 (s, IH). LC-MS: m/z 196 [M+H]+.
[0226] Synthesis of 3-(5-chloropyridin-2-yl)isoxazol-5-amine.
[0227] Ste 1 : Preparation of 3-(5-chloropyridin-2-yl)-3-oxopropanenitrile.
Figure imgf000070_0002
[0228] 1H NMR (400 MHz, DMSO- ): 8.81 (br s, IH), 8.17 (br s, IH), 8.04 - 8.02 (m, IH), 4.70 (br s, 2H). LC-MS: m/z 182 [M+H]+.
[0229] Step 2: Preparation of 3-(5-chloropyridin-2-yl)isoxazol-5-amine.
Figure imgf000070_0003
[0230] 1H NMR (400 MHz, DMSO-c?6): 8.70 (m, IH), 8.03 - 8.00 (m, IH), 7.89 - 7.87 (m, IH), 6.88 (s, 2H), 5.44 (s, IH). LC-MS: m/z 196 [M+H]+.
[0231] Synthesis of 3-(5-chloropyridin-3-yl)isoxazol-5-amine.
Figure imgf000070_0004
[0232] 1H NMR (400 MHz, DMSO-i¾): 8.90 (s, 1H), 8.69 (s, 1H), 8.24 (s, 1H), 6.96 (s, 2H), 5.59 (s, 1H). LC-MS: m/z 196 [M+H]+.
[0233] Synthesis of 3-(3-chlorophenyl)-l-methyl-lH-pyrazol-5-amine. [0234] Step 1 : Preparation of 3-(3-chlorophenyl)-3-oxopropanenitrile.
Figure imgf000071_0001
[0235] LC-MS: m/z 178.0 [M+H]+.
[0236] Step 2: Preparation of 3-(3-chlorophenyl)-l-methyl-lH-pyrazol-5-
Figure imgf000071_0002
[0237] 1H NMR (400 MHz, DMSO- ): 7.81-7.77 (m, 1H), 7.72-7.70 (m, 1H), 7.46-7.42 (m, 3H), 5.99 (s, 2H), 3.68 (s, 3H). LC-MS: 208 [M+H]+.
[0238] Synthesis of 5-(3-chlorophenyl)isoxazol-3 -amine.
Figure imgf000071_0003
[0239] A stirred solution of 3-(3-chlorophenyl)-3-oxopropanenitrile(l equiv.) and
hydroxylamine hydrochloride (1.2 equiv.) in a mixture of ethanol (0.02 M) and water(0.02 M) was adjusted to pH 8 using sodium hydroxide solution and heated at 60 °C for 18 h. The reaction mixture was then acidified to pH 2 using hydrochloride solution and further heated at 80 °C for 1 h. It was then allowed to cool to room temperature, basified to pH 10, diluted with water and extracted with ethyl acetate. The combined organic layers were dried over magnesium sulphate and concentrated under vacuum. The crude residue was purified by column chromatography to afford the purified product.
[0240] 1H NMR (400 MHz, DMSO- 6): 7.83 (s, 1H), 7.74-7.71 (m, 1H), 7.52-7.51 (m, 2H), 6.44 (s, 1H), 5.71 (s, 2H). LC-MS: m/z 195 [M+H]+.
[0241] Synthesis of 4-(pyrrolidin-l-yl)aniline.
[0242] Step 1 : Preparation of l-(4-nitrophenyl)pyrrolidine.
Figure imgf000072_0001
[0243] Triethylamine was added to a stirred solution of l-fluoro-4-nitrobenzene(1.0 equiv.) and pyrrolidine (1.3 equiv.) in 2-propanol (0.1 M). The reaction was allowed to stir at reflux for 6 h. Upon consumption of starting material, solvent was evaporated to dryness; the crude diluted with water; extracted with dichloromethane; organic layers combined; washed with brine; dried over magnesium sulphate and concentrated in vacuo. The crude was dissolved in
isopropanol and filtered to afford the pure product as a bright yellow solid.
[0244] 1H NMR (400 MHz, DMSO- 6): 8.05 (d, J=9.2 Hz, 2H), 6.61 (d, J=9.2 Hz, 2H), 3.39 (m, 4H), 2.00 - 1.97 (m, 4H). LC-MS: m/z 193 [M+H]+.
[0245] Step 2: Preparation of 4-(pyrrolidin-l-yl)aniline.
Figure imgf000072_0002
[0246] 1H NMPv (400 MHz, DMSO-J6): 6.49 (d, J=8.4 Hz, 2H), 6.35 (d, J=8.4 Hz, 2H), 4.24 (br s, 2H), 3.07 (br s, 4H), 1.90 - 1.87 (m, 4H). LC-MS: m/z 163 [M+H]+.
[0247] Synthesis of l-phenylpiperidin-4-amine
Figure imgf000073_0001
[0248] To a stirred solution of chlorobenzene (1.0 g, 8.92 mmol) in xylene (20 mL) were added 4-amino piperidine (0.93 mL, 8.92 mmol), KC^Bu (2.0 g, 17.84 mmol), X-Phos (636 mg, 1.33 mmol) and Pd2(dba)3 (816 mg, 0.89 mmol), degassed with argon for 10 min. The reaction mixture was heated at 130 °C for 16 h. TLC indicated some starting material and formation of a new spot. The reaction mixture was filtered and concentrated under reduced pressure, poured into ice-cold water and extracted with EtOAc. The combined organic layer was washed with water and brine solution, dried over anhydrous sodium sulphate, filtered, concentrated under reduced pressure and purified by column chromatography to afford the desired product.
[0249] LC-MS: m/z 175 [M+H]+.
[0250] Suzuki Coupling
[0251] General Suzuki Procedure A
[0252] Degassed solution of bromoaniline derivative (1.0 equiv.) in 1,4-dioxane (4 mL). Add potassium carbonate (3.0 equiv.) in water (1 mL) and boronic acid/pinacol ester to solution. Degassed for 10 minutes before adding tetrakistriphenylphosphine palladium catalyst (0.1 eq). Degas for another 10 minutes and heat at reflux for 18 h. Solution is filtered through Celite® and extracted with water and dichloromethane. The organic phase combined, dried over sodium sulphate, filtered and evaporated to dryness in vacuo. Crude was dissolved in
dimethylformamide, acetonitrile and water and purified with preparative HPLC or by flash column chromatography to yield the desired product.
[0253] Synthesis of 4-(l-methyl-lH-pyrazol-5-yl)aniline.
Figure imgf000073_0002
[0254] 1H NMR (400 MHz, DMSO- 6): 7.36 (s, IH), 7.14 (d, J=8.4 Hz, 2H), 6.64 (d, J=8.4 Hz, 2H), 6,.18 (s, IH), 5.33 (s, 2H), 3.77 (s, 3H). LC-MS: m/z 174 [M+H]+.
[0255] Synthesis of 5-phenylpyrazin-2-amine.
Figure imgf000074_0001
[0256] 1H NMR (400 MHz, DMSO-i¾): 8.49 (m, IH), 7.96 (m, IH), 7.92 - 7.90 (m, 2H), 7.43 7.40 (m, 2H), 7.33 - 7.29 (m, IH), 6.53 (s, 2H). LC-MS: m/z 172 [M+H]+.
[0257] Synthesis of 3-methoxy-[l,l'-biphenyl]-4-amine.
Figure imgf000074_0002
[0258] LC-MS: m/z 200 [M+H]+.
[0259] Synthesis of 4'-nitro-[l,r-biphenyl]-4-amine.
Figure imgf000074_0003
[0260] 1H NMR (400 MHz, Chloroform-i/): 8.24 (d, J=9.2 Hz, 2H), 7.66 (d, J=9.2 Hz, 2H), 7.47 (d, J=8.4 Hz, 2H), 6.77 (d, J=8.4 Hz, 2H), 3.87 (s, 2H). LC-MS: m/z 215 [M+H]+.
[0261] Synthesis of 6-(3-chlorophenyl)pyridin-3 -amine.
[0262] Step 1: Preparation of 2-(3-chlorophenyl)-5-nitropyridine.
Figure imgf000075_0001
[0263] 1H NMR (400 MHz, Chloroform-c/): 9.49 - 9.50 (m, 1H), 8.54 - 8.56 (m, 1H), 8.11 (s, 1H), 7.95 - 7.97 (m, 1H), 7.89 - 7.91 (m, 1H), 7.48 - 7.51 (m, 2H). LC-MS: m/z 235 [M+H]+.
[0264] Step 2: Preparation of 6-(3-chlorophenyl)pyridin-3 -amine.
Figure imgf000075_0002
[0265] A solution of 2-(3-chlorophenyl)-5-nitropyridine in 2:1 ethanol and water (0.1M) was added iron powder (5 equiv.) and acetic acid (5 equiv.). The reaction was stirred vigorously at room temperature under ambient atmosphere for 30 min. After completion of starting material, the reaction mixture was basify with IN NaOH (pH 8 - 10) and extracted with ethyl acetate. The combined organic layers were washed with brine, dried with sodium sulphate and concentrated under vacuum to afford product.
[0266] 1H NMR (400 MHz, Chloroform-i 8.17 - 8.18 (m, 1H), 7.89 - 7.90 (m, 1H), 7.74 - 7.77 (m, 1H), 7.50 - 7.52 (m, 1H), 7.32 - 7.36 (m, 1H), 7.27 - 7.30 (m, 1H), 7.03 - 7.06 (m, 1H). LC-MS: m/z 205 [M+H]+.
[0267] Synthesis of 6-(3-fluorophenyl)pyridin-3 -amine.
Figure imgf000075_0003
[0268] 1H NMR (400 MHz, DMSO-c 6): 8.14 (s, IH), 8.02 (s, IH), 7.76-7.74 (m, IH), 7.68-7.66 (m, IH), 7.44-7.38 (m, 2H), 7.10-7.07 (m, IH), 7.05-6.97 (m, IH), 5.55 (s, 2H). LC-MS: m/z 189 [M+H]+.
[0269] Synthesis of 6-(m-tolyl)pyridin-3 -amine.
Figure imgf000076_0001
[0270] 1H NMR (400 MHz, DMSO- 6): 8.01 - 8.00 (m, IH), 7.74 (bs, IH), 7.69 - 7.67 (m, IH), 7.60 (d, J=8.4 Hz, IH), 7.26 (t, J=7.6 Hz, IH), 7.09 - 7.07 (m, IH), 6.99 - 6.97 (m, IH), 5.42 (s, 2H), 2.35 (s, 3H). LC-MS: m/z 185 [M+H]+.
[0271] Synthesis of 5-(3-chlorophenyl)pyridin-2-amine.
Figure imgf000076_0002
[0272] LC-MS: m/z 205 [M+H]+.
[0273] Synthesis of 5-(3-fluorophenyl)pyridin-2-
Figure imgf000076_0003
[0274] 1H NMR (400 MHz, DMSO-i¾): 8.29 (s, IH), 7.73 (dd, J=8.6 Hz, J=2.6 Hz, IH), 7.43 - 7.39 (m, 3H), 7.09 - 7.05 (m, IH), 6.52 (d, J=8.8 Hz, IH), 6.12 (s, 2H). LC-MS: m/z 189
[0275] Synthesis of 5-(4-fluorophenyl)pyridin-2-amine.
Figure imgf000077_0001
[0276] 1H NMR (400 MHz, Methanol-***): 8.20 (s, IH), 8.09 - 8.10 (m, IH), 7.87 - 7.90 (m, IH), 7.54 - 7.57 (m, 2H), 7.14 - 7.18 (m, 2H), 6.80 - 6.82 (m, IH).
[0277] Synthesis of 3-methyl-5-phenylpyridin-2-amine.
Figure imgf000077_0002
[0278] 1H NMR (400 MHz, DMSO-J6): 8.11 (s, IH), 7.56-7.54 (m, 3H), 7.39 (t, J=7.2 Hz, 2H), 7.25 (t, J=7.6 Hz, IH), 5.80 (s, 2H), 2.11 (s, 3H). LC-MS: m/z 185 [M+H]+.
[0279] Synthesis of 5-(m-tolyl)pyridin-2-amine.
Figure imgf000077_0003
[0280] 1H NMR (400 MHz, DMSO- 6): 8.22 - 8.21 (m, IH), 7.68 - 7.66 (m, IH), 7.37 - 7.32 (m, 2H), 7.27 (m, IH), 7.08 - 7.06 (m, IH), 6.51 (d, J=8.8 Hz, IH), 6.01 (s, 2H), 2.34 (s, 3H). LC-MS: m/z 185 [M+H]+.
[0281] Synthesis of 6-(3-chlorophenyl)pyridazin-3 -amine.
Figure imgf000077_0004
[0282] 1H NMR (400 MHz, Methanol-^): 7.96 (s, IH), 7.81 - 7.88 (m, 2H), 7.41 - 7.48 (m, 2H). LC-MS: m/z 206 [M+H]+.
[0283] Synthesis of 6-(3-fluorophenyl)pyridazin-3 -amine.
Figure imgf000078_0001
[0284] 1H NMR (400 MHz, DMSO- ): 7.86-7.76 (m, 3H), 7.53-7.47 (m, IH), 7.23-7.18 (m, IH), 6.85 (d, J= 9.6Hz, IH), 6.55 (s, 2H). LC-MS: m/z 190 [M+H]+.
[0285] Synthesis of 6-(m-tolyl)pyridazin-3 -amine.
Figure imgf000078_0002
[0286] 1H NMR (400 MHz, DMSO- ): 7.79 - 7.77 (m, 2H), 7.74 - 7.72 (m, IH), 7.34 (t, J=7.6 Hz, IH), 7.21 - 7.19 (m, IH), 6.84 (d, J=9.6 Hz, IH), 6.44 (s, 2H), 2.38 (s, 3H). LC-MS: m/z 186 [M+H]+.
[0287] Synthesis of 6-(5-methylpyridin-3-yl)pyridazin-3-amine.
Figure imgf000078_0003
[0288] 1H NMR (400 MHz, DMSO- ): 8.92 (d, J=2.0 Hz, IH), 8.42 (d, J=1.6 Hz, IH), 8.15 (s, IH), 7.86 (d, J=9.2 Hz, IH), 6.87 (d, J=9.2 Hz, IH), 6.56 (s, 2H), 2.37 (s, 3H). LC-MS: m/z 187 [M+H]+.
[0289] Synthesis of 6-(6-methylpyridin-3-yl)pyridazin-3 -amine.
Figure imgf000079_0001
[0290] LC-MS: m/z 187[M+H]+.
[0291] Synthesis of 5'-fluoro-[2,3'-bipyridin]-5-amine.
Figure imgf000079_0002
[0292] LC-MS: m/z 190 [M
[0293] Synthesis of 6'-methyl-[2,3'-bipyridin]-5-amine.
Figure imgf000079_0003
[0294] LC-MS: m/z 186 [M+H]+.
[0295] Synthesis of 5'-chloro-[3,3'-bipyridin]-6-amine.
Figure imgf000079_0004
[0296] LC-MS: m/z 206 [M+H]+.
[0297] Synthesis of 5'-fluoro-[3,3'-bipyridin]-6-amine.
Figure imgf000080_0001
[0298] LC-MS: m/z 190 [M+H]+.
[0299] Synthesis of 5-(m-tolyl)pyrazin-2-amine.
Figure imgf000080_0002
[0300] 1H NMR (400 MHz, DMSO-d6): 8.47 (d, J=1.6 Hz, IH), 7.94 (d, J=1.6 Hz, IH), 7.73 (s, IH), 7.68 (d, J=8.0 Hz, IH), 7.29 (t, J=7.6 Hz, IH), 7.12 (d, J=7.2 Hz, IH), 6.50 (s, 2H), 2.35 (s, 3H). LC-MS: m/z 186 [M + H]+.
[0301] General Suzuki Procedure B
[0302] To a stirred solution of the arylhalide (1 equiv.), boronic acid (1.1 equiv.) and sodium carbonate (2 equiv.) in dioxane (0.02 M) and water (0.05 M) was degassed for 10 min with nitrogen. Bis(cyclopentyldiphenylphosphane) dichloromethane palladium chloride iron (0.1 equiv.) was added to the reaction mixture and it was heated to reflux for 16 h. The reaction mixture was allowed to cool and filtered over Celite®. It was diluted with water and extracted with ethyl acetate. The combined organic layers were dried with magnesium sulphate and concentrated under vacuum. The crude compound was purified by column chromatography to afford the desired product.
[0303] Synthesis of 4-(pyridazin-3-yl)aniline.
Figure imgf000080_0003
[0304] 1H NMR (400 MHz, DMSO-i 6): 9.01-9.00 (m, IH), 7.99 (d, J= 7.2 Hz, IH), 7.87 (d, J= 8.8 Hz, 2H), Ί.62-1.59 (m, IH), 6.68 (d, J= 8.4 Hz, 2H), 5.58 (s, 2H). LC-MS: m/z 111 [M+H]+.
[0305] Synthesis of 4-(pyrimidin-2-yl)aniline.
Figure imgf000081_0001
[0306] 1H NMR (400 MHz, DMSO-i 6): 8.72 (d, J= 4.8 Hz, 2H), 8.08 (d, J= 8.8 Hz, 2H), 7.19 (t, J= 4.8 Hz, IH), 6.62 (d, J= 8.4 Hz, 2H), 5.64 (s, 2H). LC-MS: m/z 172 [M+H]+.
[0307] Synthesis of 4-(pyrimidin-5-yl)aniline.
Figure imgf000081_0002
[0308] 1H NMR (400 MHz, DMSO-d6): 9.00 (m, 3H), 7.49 (d, J= 8.8 Hz, 2H), 6.68 (d, J= 8.4 Hz, 2H), 5.45 (s, 2H). LC-MS: m/z 172 [M+H]+.
[0309] Synthesis of 5-phenylpyrimidin-2-amine.
Figure imgf000081_0003
[0310] 1H NMR (400 MHz, DMSO- ): 8.56 (s, 2H), 7.61 (d, J= 7.2 Hz, 2H), 7.43 (t, J= 7.6 Hz, 2H), 7.31 (t, J= 1.2 Hz, IH), 6.75 (s, 2H). LC-MS: m/z 172 [M+H]+.
[0311] Synthesis of [3,3'-bipyridin]-6-amine.
Figure imgf000082_0001
[0312] 1H NMR (400 MHz, DMSO- ): 8.79 (d, J= 1.6 Hz, IH), 8.47-8.45 (m, IH), 8.29 (d, J= 2.0 Hz, IH), 7.97-7.94 (m, IH), 7.77-7.74 (m, IH), 7.42-7.38 (m, IH), 6.55-6.53 (m, IH), 6.15 (s, 2H). LC-MS: m/z 172 [M+H]+.
[0313] Synthesis of 6-(pyridin-3-yl)pyridazin-3 -amine.
Figure imgf000082_0002
[0314] 1H NMR (400 MHz, DMSO-rf6): 9.14 - 9.13 (m, IH), 8.60 - 8.59 (m, IH), 8.34 - 8.31 (m, IH), 8.13 (s, IH), 7.92 (d, j=9.6 Hz, IH), 7.51 - 7.48 (m, IH), 6.92 (d, J=9.2 Hz, IH), 6.71 (bs, IH). LC-MS: m/z 173 [M+H]+.
[0315] Synthesis of 2-methyl-[ 1 , 1 '-biphenyl]-4-amine.
Figure imgf000082_0003
[0316] LC-MS: m/z 184 [M+H]+.
[0317] Synthesis of 3-methyl-[l, -biphenyl]-4-amine.
Figure imgf000082_0004
[0318] LC-MS: m/z 184 [M+H] . [0319] Synthesis of 2'-fluoro-[l,l'-biphenyl]-4-amine.
Figure imgf000083_0001
[0320] LC-MS: m/z 188 [M+H]+.
[0321] Synthesis of 3'-fluoro-[l,r-biphenyl]-4-amine.
Figure imgf000083_0002
[0322] 1H NMR (400 MHz, DMSO-d6): 7.41 - 7.32 (m, 5H), 7.02 - 6.99 (m, IH), 6.63 (d, J=8.4 Hz, 2H), 5.29 (s, 2H). LC-MS: m/z 188 [M+H]+.
[0323] Synthesis of 3'-nitro-[l,l'-biphenyl]-4-amine.
Figure imgf000083_0003
[0324] 1H NMR (400 MHz, DMSO-i¾: 8.29 (t, J=2.0 Hz, IH), 8.05-8.00 (m, 2H), 7.64 (t, J=8.0 Hz, IH), 7.48 (d, J=8.8 Hz, 2H), 6.67 (d, J=8.4 Hz, 2H), 5.41 (s, 2H).
[0325] Synthesis of 4-(pyrazin-2-yl)aniline.
Figure imgf000083_0004
[0326] MS (ESI) m/z 172 [M+H]+.
[0327] Synthesis of 2-fluorobiphenyl-4-amine.
Figure imgf000084_0001
[0328] MS (ESI) m/z 188 [M+H]
[0329] Synthesis of 4'-fluorobiphenyl-4-amine.
Figure imgf000084_0002
[0330] MS (ESI) m/z 188 [M+H]+.
[0331] Synthesis of 5-methyl-6-phenylpyridin-3-amine.
Figure imgf000084_0003
[0332] MS (ESI) m/z 185 [M+Hf.
[0333] Synthesis of 4-methyl-5-phenylpyridin-2-amine.
Figure imgf000084_0004
[0334] MS (ESI) m/z 186 [M+H]+.
[0335] Synthesis of 5-fluoro-2,3'-bipyridin-6'-amine.
Figure imgf000085_0001
[0336] MS (ESI) m/z 190 [M+H]+.
[0337] Synthesis of 6-(4-chlorophenyl)pyridazin-3 -amine.
Figure imgf000085_0002
[0338] LC-MS: m/z 206 [M+H]+.
[0339] Synthesis of 6-(5-chloropyridin-3-yl)pyridazin-3 -amine.
[0340] Step-1 : Preparation of 3-chloro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyridine.
Figure imgf000085_0003
[0341] A stirred solution of 3-bromo-5-chloropyridine (1 equiv.), bis(pinacaloto) diborane (1.2 equiv.), potassium acetate (3 equiv.) in 1,4-dioxane (0.2 M) was degassed with argon for 20 min. To the reaction mixture was added Pd(dppf)Cl2.DCM (0.05 equiv.) under argon atmosphere. The reaction mixture was heated at 100 °C for 16 h. The worked-up material was used in the next step without purification.
[0342] Step-2: Preparation of 6-(5-chloropyridin-3-yl)pyridazin-3-amine.
Figure imgf000085_0004
[0343] LC-MS: m/z 207 [M+H]+.
[0344] Synthesis of 5-methyl-6-( yridin-3-yl)pyridazin-3-amine.
Figure imgf000086_0001
[0345] 1H NMR (400 MHz, DMSO- ): 8.71 (s, IH), 8.60-8.59 (d, J= 4.8 Hz, IH), 7.94-7.92 (d, J= 7.6 Hz, IH), 7.49-7.46 (m, IH), 6.68 (s, IH), 6.36 (brs, 2H), 2.18 (s, 3H). LC-MS: 187.0 [M+H]+
[0346] Synthesis of 5'-methyl-2,3'-bipyridin-5-amine.
Figure imgf000086_0002
[0347] LC-MS: m/z 186 [M+H]+.
[0348] Synthesis of 5'-methyl-[3,3'-bipyridin]-6-
Figure imgf000086_0003
[0349] 1H NMR (400 MHz, DMSO-<¾): 8.58 (s, IH), 8.30 - 8.29 (m, IH), 8.27 - 8.26 (m, IH), 7.78 (bs, IH), 7.74 (dd, J=9.2 Hz, J=2.6 Hz, IH), 6.54 (d, J=8.4 Hz, IH), 6.12 (s, 2H), 2.33 (s, 3H). LC-MS: m/z 186 [M+H]+.
[0350] Synthesis of 4-methyl-3,3'-bipyridin-6-amine.
Figure imgf000086_0004
[0351] LC-MS: m/z 186 [M+H]+.
[0352] Synthesis of 6'-methyl-3,3'-bipyridin-6-amine.
Figure imgf000087_0001
[0353] MS (ESI) m/z 186 [M+H]+.
[0354] Synthesis of 5-(6-methylpyridin-3-yl)pyrazin-2-amine.
Figure imgf000087_0002
[0355] MS (ESI) m/z 187 [M+H]+.
[0356] Synthesis of 4-(pyrazin-2-yl)aniline.
Figure imgf000087_0003
[0357] MS (ESI) m/z 172 [M+H]+.
[0358] Synthesis of Di tert-butyl (6'-amino-[3,3'-bipyridin]-5-yl)carbamate.
Figure imgf000087_0004
[0359] 1H NMR (400 MHz, DMSO-tf6): 8.75 (s, IH), 8.31 (d, J= 8.0 Hz, 2H), 7.94 (s, IH), 7.78 (d, J= 6.8 Hz, IH), 6.54 (d, J= 8.8 Hz, IH), 6.19 (bs, 2H), 1.39 (s, 18H); MS (ESI) m/z 387.30 [M+H]+.
[0360] Synthesis of 5-(pyridin-3-yl)pyrazin-2-amine.
Figure imgf000088_0001
[0361] 1H NMR (400 MHz, DMSO-d6): 9.11 (s, IH), 8.58 (s, IH), 8.51 (d, J= 4.0 Hz, IH), 8.25 (d, J- 11.6 Hz, IH), 7.99 (s, IH), 7.46-7.40 (m, IH), 6.67 (bs, 2H); MS (ESI) m/z 173.0 [M+H]+.
[0362] Synthesis of 4,5'-dimethyl-3,3'-bipyridin-6-amine.
Figure imgf000088_0002
[0363] 1H NMR (400 MHz, DMSO-d6): 8.35 (s, IH), 8.31 (s, IH), 7.73 (s, IH), 7.55 (s, IH), 6.37 (s, IH), 5.93 (bs, 2H), 8.31 (s, IH), 2.33 (s, IH), 2.11 (s, IH); MS (ESI) m/z 200.0
[M+H]+.
[0364] Synthesis of 5-(5-fluoropyridin-3-yl)pyrazin-2-amine.
[0365] Step 1 : Preparation of 3-fluoro-5-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyridine.
Figure imgf000088_0003
[0366] A stirred solution of 3-bromo-5-fluoropyridine (1 equiv.), Bis(pinacolato) diborane (1.1 equiv.), KOAc (2 equiv.) in 1,4-dioxane (0.4 M) was degassed with argon for 20 min. To the reaction mixture, Pd(dppf)Cl2.DCM (0.1 equiv.) was added under argon atmosphere.The reaction mixture was heated at 110 °C for 4 h. The worked-up material was used in the next step without further purification.
[0367] MS (ESI) m/z 224 [M+H]+.
[0368] Step 2: Preparation of 5-(5-fluoropyridin-3-yl)pyrazin-2-amine.
Figure imgf000089_0001
[0369] 1H NMR (400 MHz, DMSO-c¾: 9.01 (t, J= 1.7 Hz, 1H), 8.65 (d, J= 1.3 Hz, 1H), 8.49 (d, J= 3.0 Hz, lH), 8.16-8.12 (m, 1H), 7.99 (d, J- 1.3 Hz, 1H), 6.79 (s, 2H); MS (ESI) m/z 191.17 [M+H]+.
[0370] Procedure for acid-intermediate:
[0371] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanoic acid.
[0372] Step 1 : Preparation of 2-methyl-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine.
Figure imgf000089_0002
[0373] A stirred solution of 3-methyl-lH-pyrazol-5-amine (1 equiv.) in 1,4-dioxane (0.5 M) was added TEA (5 equiv.) at 0 °C . After 15 minutes 1,3-dibromopropane (1.2 equiv.) was added to the reaction mixture at 0 °C. The reaction mixture was stirred at 100 °C for 5 h. After completion of the reaction, the reaction mixture was filtered and washed with ethyl acetate. The filtrate was concentrated under reduced pressure to afford the intermediate. [0374] Step 2: Preparation of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrirnidin-4(5H)-yl)-4- oxobutanoic acid
Figure imgf000090_0001
[0375] To a stirred solution of 2-methyl-4,5,6,7-tetraliydropyrazolo[l,5-a]pyrimidine (1 equiv.) in chloroform (0.5 M) was added succinic anhydride (1.3 equiv.). The reaction mixture was stirred at room temperature for 16 h. After completion, the solid which was precipitated was filtered and washed with n-pentane and dried to afford the product.
[0376] 1H NMR (400 MHz, DMSO-i 6): 12.11 (s, IH), 6.34 (bs, IH), 4.02-3.99 (t, J= 5.2 Hz, 2H), 3.82 (m, 2H), 2.77-2.75 (m, 2H), 2.41 (m, 2H), 2.08 (bs, 5H). MS (ESI) m/z 238 [M+H]+.
[0377] Synthesis of 4-(6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanoic acid
Figure imgf000090_0002
[0378] To a solution of 4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine (1 equiv.) in chloroform (0.4 M) was added succinic anhydride (1.3 equiv.) at room temperature. The reaction mixture was stirred at room temperature for 16 h. After completion, the reaction mixture was filtered and solid was washed with chloroform and dried to afford the product.
[0379] 1H NMR (400 MHz, DMSO- ): 12.14 (s, IH), 7.30 (s, IH), 6.55 (s, IH), 4.12-4.09 (t, J = 5.6 Hz, 2H), 3.88-3.86 (m, 2H), 2.80-2.77 (t, J= 6.4 Hz, 2H), 2.52-2.50 (m, 2H), 2.10-2.08 (m, 2H). LC-MS (ESI): MS m/z 224.0 [M+H]+.
[0380] Synthesis of 4-oxo-4-(2-(trifluoromethyl)-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)- yl)butanoic acid
[0381] Step 1: Preparation of 2-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine
Figure imgf000091_0001
[0382] A solution of 3-(trifluorometliyl)-lH-pyrazol-5-amine (1 equic.) in 1,4-dioxane (0.2 M) was treated with 1,3-dibromopropane (1.2 equiv.) and triethylamine (5 equiv.) at 0 C and the reaction mixture was heated to 100 °C for 12 h. After completion, reaction mixture was cooled to room temperature, filtered under reduced pressure and concentrated to afford crude product. The crude product was purified by flash chromatography to afford the product.
[0383] 1H-NMR (400 MHz; DMSO-i¾: 12.25 (brs, IH), 6.40 (s, IH), 5.52 (s, IH), 4.03-4.00 (t, 2H), 3.58 (s, IH), 3.18-3.15 (m, 2H), 2.05-1.97 (m, 2H). LC-MS: MS m/z 192 [M+H]+.
[0384] Step 2: Preparation of 4-oxo-4-(2-(trifluoromethyl)-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)butanoic acid
Figure imgf000091_0002
[0385] To a solution of compound 2-(trifluoromethyl)-4,5,6,7-tetrahydropyrazolo[l,5- a]pyrimidine (1 equiv.) in CHC13 (0.15 M) was added succinic anhydride (2 equiv.) at room temperature. The resulted reaction mixture was stirred at room temperature for 60 h. After completion, the reaction mixture was concentrated to afford crude product. The crude product was purified by column chromatography to afford the product.
[0386] 1H NMR (400 MHz, DMSO-c¾: 12.20 (s, 2H), 6.88 (s, IH), 4.20 (t, J = 6 Hz, 2H), 3.92 (t, J = 5.2 Hz, 2H), 2.82 (t, J = 6 Hz, 2H), 2.53-2.49 (m, 3H), 2.41 (s, 3H), 2.17 (brs, 2H). LC- MS: MS m/z 290 [M+H]+.
[0387] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4- oxobutanoic acid
Figure imgf000092_0001
[0388] Step 1: Preparation of 2-methylpropane- 1,3 -diyl dimethanesulfonate
[0389] To a stirred solution of 2-methylpropane-l,3-diol (1 equiv.) in dichloromethane (0.45 M) was added methane sulfonyl chloride (2.8 equiv.), using triethylamine (2.8 equiv.) at 0 °C to room temperature for 16 h. Upon completion of reaction, the reaction mixture was washed with water and brine solution, dried over anhydrous sodium sulphate, filtered and concentrated under reduced pressure to afford the crude intermediate.
Figure imgf000092_0002
[0390] Step 2: Preparation of 2,6-dimethyl-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine
[0391] To a stirred solution of 2-methylpropane- 1,3 -diyl dimethanesulfonate (1.2 equiv.) in 1,4- dioxane (0.5 M) were added 3 -methyl- lH-pyrazol- 5 -amine (1 equiv.) and triethylamine (5 equiv.). The reaction mixture was heated at 100 °C for 48 h. Upon completion, the reaction mixture was filtered and concentrated under reduced pressure, purified by column
chromatography to afford the intermediate.
Figure imgf000092_0003
[0392] Step 3: Preparation of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)- 4-oxobutanoic acid
[0393] To a stirred solution of 2,6-dimethyl-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine (1 equiv.) in chloroform (0.15 M) was added succinic anhydride (1 equiv.) and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure to afford the product.
[0394] 1H MR (400 MHz, DMSO- ): 12.08 (s, IH), 6.35 (brs, IH), 4.12-4.08 (m, IH), 3.99 ( brs, IH), 3.64-3.58 (m, IH), 3.35 (brs, IH), 2.85-2.79 (m, IH), 2.74-2.70 (brs, IH), 2.49-2.48 (m, 2H), 2.25 (brs, IH), 2.08 (brs, 3H), 1.04-1.02 (d, J= 6.1 Hz, 3H). MS (ESI) m/z 252.07
[M+H]+.
[0395] Synthesis of ethyl 2-methyl-4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)- yl)-4-oxobutanoate
Figure imgf000093_0001
[0396] Step 1: Preparation of 2-methyl-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine
[0397] To a stirred solution of 3-methyl-lH-pyrazol-5-amine (1 equiv.) in 1,4-dioxane (0.1 M) was added triethylamine (5 equiv.) at 0 °C. After 15 minutes, 1,3-dibromopropane (1.2 equiv.) was added to the reaction mixture at 0 °C. The reaction mixture was stirred at 100 °C for 5 h. The reaction mixture was filtered and washed with ethyl acetate. The filtrate was concentrated under reduced pressure to afford the intermediate.
Figure imgf000093_0002
[0398] Step 2: Preparation of 2-bromo-l-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)ethanone
[0399] To a stirred solution of 2-methyl-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine (1 equiv.) in dichloromthane (0.6 M) was added 2-bromoacetyl bromide (2 equiv.) drop wise at 0 °C. The reaction mixture was stirred at room temperature for 12 h. The solid thus formed was filtered and washed with n-pentane then dried to afford the intermediate.
Figure imgf000094_0001
[0400] Step 3: Preparation of ethyl 2-(diethoxyphosphoryl)-4-(2-methyl-6,7- dihydropyrazolof 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanoate
[0401] To a stirred solution of NaH (60% suspension in mineral oil) (1.2 equiv.) in DMSO (0.5 M) was added ethyl 2-(diethoxyphosphoryl)acetate (1.1 equiv.) at room temperature. After 30 min, to the reaction mixture was added 2-bromo-l-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)ethanone (1 equiv.) portion-wise at room temperature. The reaction mixture was stirred at 70 °C for 4 h. The reaction mixture was quenched with ice-cold water and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography to afford the intermediate. LC-MS: m/z 402 [M+H]+.
Figure imgf000094_0002
[0402] Step 4: Preparation of ethyl 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)- 2-methylene-4-oxobutanoate
[0403] To a stirred solution of ethyl 2-(diethoxyphosphoryl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanoate (1 equiv.) in THF (0.4 M) was added aqueous K2C03 (1.5 equiv., 2 M) followed by aq. HCHO (20 equiv.) at room
temperature. The reaction mixture was stirred at 80 °C for 45 min. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography to afford the product. LC-MS: m/z 278 [M+H]+.
Figure imgf000095_0001
[0404] Step 5: Preparation of ethyl 2-methyl-4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)-4-oxobutanoate
[0405] To a solution of ethyl 4-(2-methyl-6, 7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-2- methylene-4-oxobutanoate (1 equiv.) in ethanol (0.1 M) was added a slurry of 10% Pd/C (10% w/w) in methanol under nitrogen and the reaction mixture was hydrogenated under H2 (50 psi) at room temperature for 8 h. The reaction mixture was filtered through celite bed and washed with methanol The filtrate was concentrated under reduced pressure to give crude product. The crude product was purified by column chromatography to afford the product. 1H NMR (400 MHz, DMSO-<¾: 6.33 (brs, 1H), 4.08-4.03 (m, 4H), 3.99-3.81 (m, 2H), 2.93-2.79 (m, 2H), 2.72-2.67 (m, 1H), 2.08 (brs, 5H), 1.19-1.13 (m, 6H). MS (ESI) m/z 280 [M+H]+.
[0406] General Procedure for amide coupling reactions:
[0407] Amide Coupling Method A:
[0408] A solution of the respective amine (1 equiv.) and acid-intermediate (1.1 equiv.) in pyridine (0.1 M) was cooled to 0 °C under nitrogen atmosphere and treated with POCl3 (3 equiv.). The reaction mixture stirred for 45 min. After consumption of starting material, the reaction mixture was quenched with saturated sodium bicarbonate and extracted with DCM. The combined organic layers were washed with brine, dried over Na2S04 and concentrated under vacuum. The crude compound was purified by reversed phase chromatography to afford desired product.
[0409] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(3- methylisoxazol-5-yl)-4-oxobutanamide, Compound 1
Figure imgf000095_0002
[0410] 1H NMR (400 MHz, Methanol-^): 6.15 (s, IH), 4.55 (s, IH), 4.10 (t, J= 6.0 Hz, 2H ), 3.91 - 3.94 (m, 2H), 2.96 - 2.98 (m, 2H), 2.78 (t, J= 6.0 Hz, 2H), 2.22 (bs, 2H), 2.17 (s, 3H). LC-MS: m/z 318 [M+H]+.
[0411] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(3- phenylisoxazol-5-yiybutanamide, Compound 2
Figure imgf000096_0001
[0412] 1H MR (400 MHz, DMSO-d6): 11.80 (s, IH), 7.84-7.83 (d, J= 2.8 Hz, 2H), 7.50-7.48 (m, 3H), 6.68 (s, IH), 6.34 (s, IH), 4.07 (m, 2H), 3.86 (m, 2H), 2.92-2.90 (m, 2H), 2.73-2.71 (t, J= 5.2 Hz, 2H), 2.07-2.05 (m, 5H). LC-MS: 380.2 [M+H]+.
[0413] Synthesis of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 3
Figure imgf000096_0002
[0414] 1H MR (400 MHz, Methanol-^): 7.66 (s, IH), 7.60-7.58 (m, IH), 7.33-7.29 (m, 2H), 6.53 (s, IH), 6.13 (brs, IH), 3.81 (brs, 2H), 3.64 (s, IH), 2.70 (brs, 2H), 2.52-2.51 (m, 2H), 2.29 (s, 2H), 1.86 (brs, 5H). LC-MS: MS m/z 414 [M+H]+.
[0415] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- phenylisoxazol-5-yl)butanamide, Compound 4
Figure imgf000096_0003
[0416] 1H MR (400 MHz, DMSO-cfc): 8.99 (s, 1H), 7.58-7.57 (d, J= 7.2 Hz, 1H), 7.38-7.34 (t, J= 7.2 Hz, 2H), 7.29-7.25 (t, J= 7.2 Hz, 1H), 6.68 (brs, 1H), 6.35 (brs, 1H), 4.00-3.97 (t, J= 6.0 Hz, 2H), 3.83-3.80 (t, J= 6.0 Hz, 2H), 2.67-2.64 (m, 2H), 2.07 (brs, 5H). LC-MS: MS m/z 380 [M+H]+.
[0417] Synthesis of N-(3-(4-chlorophenyl)isoxazol-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 5
Figure imgf000097_0001
[0418] 1H NMR (400 MHz, DMSO-d6): 11.83 (s, 1H), 7.88-7.86 (d, J= 8 Hz, 2H), 7.57-7.55 (d, J= 8.8 Hz, 2H), 6.71 (s, 1H), 6.33 (s, 1H), 4.02-4.01 (t, J= 5.2 Hz, 2H), 3.87-3.85 (m, 2H), 2.92-2.89 (m, 2H), 2.73-2.70 (t, J= 6.4 Hz, 2H), 2.05-2.03 (m, 5H). LC-MS: 414.2 [M+H]+
[0419] Synthesis of N-(3-(2-chlorophenyl)isoxazol-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 6
Figure imgf000097_0002
[0420] 1H MR (400 MHz, DMSO-<¾: H-89 (s, 1H), 7.69-7.67 (d, J= 7.2 Hz, 1H), 7.64-7.62 (d, J= 8.4 Hz, 1H), 7.55-7.53 (t, J= 7.6 Hz, 1H), 7.49-7.45 (t, J= 7.2 Hz, 1H), 6.57 (s, 1H), 6.34 (s, 1H), 4.03-3.99 (m, 2H), 3.86-3.83 (m, 2H), 2.93-2.90 (m, 2H), 2.72-2.70 (t, J= 6 Hz, 2H), 2.08-2.05 (m, 5H). LC-MS: 414.15 [M+H]+.
[0421] Synthesis of N-(3-(3,4-dichlorophenyl)isoxazol-5-yl)-4-(2-methyl-6,7- dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 7
Figure imgf000097_0003
[0422] 1H NMR (400 MHz, OMSO-d6): 11.84 (br s, IH), 8.10 (d, J=2 Hz, IH), 7.86 - 7.83 (m, IH), 7.76 (d, J=8.4 Hz, IH), 6.78 (s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (br s, 2H), 2.91 - 2.89 (m, 2H), 2.71 - 2.67 (m, 2H), 2.08 (br s, 5H). LC-MS: m/z 449 [M+H]+.
[0423] Synthesis ofN-(3-(3-fluorophenyl)isoxazol-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 8
Figure imgf000098_0001
[0424] 1H NMR (400 MHz, DMSO- ): 11.84 (s, IH), 7.71-7.67 (t, J= 7.2 Hz, 2H), 7.57-7.52 (q, J= 8.4 Hz IH), 7.37-7.32 (m, IH), 6.76(s, IH), 6.34 (s, IH), 4.03-4.01 (m, 2H), 3.88-2.85 (m, 2H), 2.92-2.90 (m, 2H), 2.73-2.70 (t, J= 5.6 Hz, 2H), 2.09-2.05 (m, 5H).LC-MS: 398.2 [M+H]+.
[0425] Synthesis of N-[3-(3-bromophenyl)-l,2-oxazol-5-yl]-4-{2-methyl-5H,6H,7H- pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxobutanamide, Compound 9
Figure imgf000098_0002
[0426] 1H NMR (400 MHz, Methanol-^) 7.97 (s, IH), 7.77 (d, J= 7.6 Hz, IH), 7.62 (d, J= 8.0 Hz, IH), 7.40 (t, J= 8.0 Hz, IH), 6.68 (s, IH), 6.44 (br s, IH), 4.10 (t, J= 6.0 Hz, 2H), 3.94 (br s, 2H), 3.00 (br s, 2H), 2.82 (t, J= 6.8 Hz, 2H), 2.22-2.17 (m, 5H). LC-MS: m/z 459 [M+H]+.
[0427] Synthesis of N-[3-(3-methoxyphenyl)-l ,2-oxazol-5-yl]-4- {2-methyl-5H,6H,7H- pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxobutanamide, Compound 10
Figure imgf000098_0003
[0428] 1H NMR (400 MHz, Methanol-^) 7.40-7.33 (m, 3H), 7.04-7.01 (m, IH), 6.65 (s, IH), 6.45 (br s, IH), 4.10 (t, J=6.0 Hz, 2H), 3.95 (t, J=5.6 Hz, 2H), 3.84 (s, 3H), 3.00 (br s, 2H), 2.82 (t, J=6.8 Hz, 2H), 2.22-2.17 (m, 5H). LC-MS: m/z 410 [M+H]+.
[0429] Synthesis of 4- {2-methyl-5H,6H,7H-pyrazolo[ 1 ,5-a]pyrimidin-4-yl} -N-[3 -(3- methylphenyl)-l,2-oxazol-5-yl]-4-oxobutanamide, Compound 11
Figure imgf000099_0001
[0430] 1H NMR (400 MHz, Methanol-^) 7.61-7.56 (m, 2H), 7.36-7.27 (m, 2H), 6.64 (s, IH), 6.46 (br s, IH) , 4.10 (t, J=Hz, 2H), 3.95-3.93 (m, 2H), 3.00 (s, 2H), 2.82 (t, J=6.4Hz, 2H), 2.39 (s, 3H), 2.22-2.17 (m, 5H). LC-MS: m/z 394 [M+H]+.
[0431] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(3-(o- tolyl)isoxazol-5-yl)butanamide, Compound 12
Figure imgf000099_0002
[0432] 1H NMR (400 MHz, DMSO-<¾ 11.79 (br s, IH), 7.50-7.49 (m, IH), 7.40-7.28 (m, 3H), 6.45 (s, IH), 6.33 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.85 (s, 2H), 2.91-2.90 (m, 2H), 2.73-2.70 (m, 2H), 2.41 (s, 3H), 2.08 (s, 5H). LC-MS: m/z 394 [M+H]+.
[0433] Synthesis of 4-{2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxo-N-{3-[3- (trifluoromethyl)phenyl]-l,2-oxazol-5-yl}butanamide, Compound 13
Figure imgf000099_0003
[0434] 1H NMR (400 MHz, Chloroform-; ) 9.34-9.25 (m, IH), 8.05 (s, IH), 7.97 (d, J=7.6Hz, IH), 7.68 (d, J=7.6Hz, IH)) , 7.56 (t, J=8.0Hz, IH), 6.71 (s, IH), 4.17 (t, J=6.4Hz, 2H), 3.99- 3.88 (m, 2H), 3.09-2.98 (m, 2H), 2.88-2.86 (m, 2H), 2.24-2.19 (m, 5H). LC-MS: m/z 448
[M+H]+.
[0435] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4'- (trifluoromethyl)-[l, -biphenyl]-4-yl)butanamide, Compound 14
Figure imgf000100_0001
[0436] 1H NMR (400 MHz, Methanol-^) 7.78 - 7.80 (m, 2H), 7.68 - 7.72 (m, 4H), 7.63 - 7.65 (m, 2H), 4.55 (s, IH), 4.10 (t, J= 6.0 Hz, 2H), 3.94 - 3.96 (m, 2H), 2.98 - 3.00 (m, 2H), 2.79 (t, J- 6.0 Hz, 2H), 2.22 (bs, 2H), 2.18 (s, 3H). LC-MS: m/z 457 [M+H]+.
[0437] Synthesis of N-[3-(4-aminophenyl)-l,2-oxazol-5-yl]-4-{2-methyl-5H,6H,7H- pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxobutanamide, Compound 15
[0438] Step 1: Preparation of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrmiidin-4(5H)-yl)-N-(5- (4-nitrophenyl)isoxazol-3-yl)-4-oxobutanamide.
Figure imgf000100_0002
[0439] 1H NMR (400 MHz, DMSO-<¾ 11.90 (s, IH), 8.33 (d, J=8.8 Hz, 2H), 8.14 (d, J=8.8 Hz, 2H),6.85 (s, IH), 6.34 (br s, IH), 4.02 (t, J=5.6 Hz, 2H), 3.86 (s, 2H), 2.92 (s, 2H), 2.73 (t, J=6.4 Hz, 2H), 2.08 (s, 5H). LC-MS: m/z 425 [M+H]+.
[0440] Step 2: Preparation of N-[3-(4-aminophenyl)-l,2-oxazol-5-yl]-4-{2-methyl-5H,6H,7H- pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxobutanamide.
Figure imgf000101_0001
[0441] 4-{2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrimidin-4-yl}-N-[3-(4-nitrophenyl)-l,2- oxazol-5-yl]-4-oxobutanamide (1 equiv.), ammonium chloride (4 equiv.), and iron (2 equiv.) were added to a solution of ethanol (0.02 M) and water (0.08 M), and heated at 40°C for 3h. The reaction mixture was then filtered, diluted with water and extracted with ethyl acetate. The combined organic layers were dried over MgS04 and concentrated under vacuum. The crude product was purified by reversed phase preparative HPLC to afford the purified product.
[0442] 1H NM (400 MHz, DMSO-i¾ 11.61 (br s, 1H), 7.46 (d, J=8.8 Hz, 2H), 6.61(d, J-8.8 Hz, 2H), 6.46 (s, 1H), 6.34 (br s, 1H), 5.50 (s, 2H), 4.02 (t, J=5.6 Hz, 2H), 3.85 (s, 2H), 2.90 (s, 2H), 2.69 (t, J=6.4 Hz, 2H), 2.08 (s, 5H). LC-MS: m/z 395 [M+H]+.
[0443] Synthesis of N-(3-(4-chloropyridin-2-yl)isoxazol-5-yl)-4-(2-methyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 16
Figure imgf000101_0002
[0444] 1H NMR (400 MHz, DMSO-d6): 11.90 (br s, 1H), 8.69 (d, J=5.6 Hz, 1H), 8.03 (d, 1=1.6 Hz, 1H), 7.67 (dd, J=4.0 Hz, 2 Hz, 1H), 6.71 (s, 1H), 6.33 (br s, 1H), 4.02 (t, J=6.0 Hz, 2H), 3.86 (br s, 2H), 2.93 - 2.89 (m, 2H), 2.72 - 2.70 (m, 2H), 2.08 (br s, 5H). LC-MS: m/z 415 [M+H]+.
[0445] Synthesis of N-(3-(5-chloropyridin-2-yl)isoxazol-5-yl)-4-(2-methyl-6,7- dihydropyrazolo [ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 17
Figure imgf000101_0003
[0446] 1H NMR (400 MHz, DMSO- ): 11.88 (br s, IH), 8.76 (d, J=2 Hz, IH), 8.09 - 8.07 (m, IH), 8.01 - 7.99 (m, IH), 6.68 (s, IH), 6.33 (br s, IH), 4.02 (t, J=5.8 Hz, 2H), 3.86 (br s, 2H), 2.91 (t, J=6 Hz, 2H), 2.70 (t, J=6.2 Hz, 2H), 2.08 (br s, 5H). LC-MS: m/z 415 [M+H]+.
[0447] Synthesis of N-[3-(5-chloropyridin-3-yl)-l,2-oxazol-5-yl]-4-{2-methyl-5H,6H,7H- pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxobutanamide, Compound 18
Figure imgf000102_0001
[0448] 1H NMR (400 MHz, DMSO-t¾) 11.83 (br s, IH), 9.01 (d, J=1.6 Hz, IH), 8.75 (d, J=2.4 Hz, IH), 4.05 (t, J=2.0 Hz, IH), 6.90 (s, IH), 6.33 (br s, IH), 4.03 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.92-2.91 (m, 2H), 2.74-2.71 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 415 [M+H]+.
Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(3-(pyridin- -yl)isoxazol-5-yl)butanamide, Compound 19
Figure imgf000102_0002
[0449] 1H NMR (400 MHz, DMSO-<¾): 11.85 (br s, IH), 9.03 (s, IH), 8.69 - 8.67 (m, IH), 8.23 (d, J=8.0 Hz, IH), 7.55 - 7.51 (m, IH), 6.78 (s, IH), 6.33 (br s, IH), 4.02 (t, J=5.8 Hz, 2H), 3.86 (br s, 2H), 2.91 - 2.90 (m, 2H), 2.71 (t, J=6.0 Hz, 2H), 2.08 (br s, 5H). LC-MS: m/z 381
[M+H]+.
Synthesis of 4-{2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxo-N-[3-(pyridin-4-yl)- l,2-oxazol-5-yl]butanamide, Compound 20
Figure imgf000102_0003
[0450] 1H MR (400 MHz, Methanol-^): 8.66 (d, J=6.4 Hz, 2H), 8.09 (s, 1H), 7.84 (d, J=6.4 Hz, 2H), 6.79 (s, 1H), 6.44 (br s, 1H), 4.10 (t, J=7.6 Hz, 2H), 3.95-3.93 (m, 2H), 3.01 (s, 2H), 2.84-2.81 (m, 2H), 2.21-2.17 (m, 5H). LC-MS: m/z 381 [M+H]+.
[0451] Synthesis of N-(3-(3-chlorophenyl)-l-methyl-lH-pyrazol-5-yl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 21
Figure imgf000103_0001
[0452] 1H NMR (400 MHz, DMSO-rf6): 10.13 (s, 1H), 7.79 (s, 1H), 7.72-7.70 (d, J= 8 Hz, 1H), 7.43-7.39 (t, J= 8 Hz, 1H), 7.34-7.32 (d, J= 8.8 Hz, 1H), 6.70 (s, 1H), 6.36 (s, 1H), 4.02- 4.00 (t, J= 5.6 Hz, 2H), 3.88-3.85 (m, 2H), 3.72 (s, 3H), 2.92-288 (m, 2H), 2.70-2.67 (t, J= 6.4 Hz, 2H), 2.09-2.07 (m, 5H). LC-MS: 427.1 [M+H]+.
[0453] Synthesis of N-(3-(3-chlorophenyl)-l,2,4-oxadiazol-5-yl)-4-(2-methyl-6,7- dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 22
Figure imgf000103_0002
[0454] 1H NMR (400 MHz, Chloroform-^ ): 8.05 (1H, t), 7.93-7.95 (1H, m), 7.46-7.52 (1H, m), 7.40 (1H, t), 4.17 (2H, t), 3.66 (2H, bs), 3.07(4H, m), 2.18(5H, m). LC-MS: m/z 415.1 [M+H]+.
[0455] Synthesis of N-[5-(3-chlorophenyl)-l ,2-oxazol-3-yl]-4- {2-methyl-5H,6H,7H- pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxobutanamide, Compound 23
Figure imgf000103_0003
[0456] 1H NMR (400 MHz, DMSO-<¾: 11.14 (s, IH), 7.97 (s, IH), 7.86-7.84 (m, IH), 7.57- 7.56 (m, 2H), 7.45 (s, IH), 6.33 (br s, IH), 4.03 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.91-2.88 (m, 2H), 2.72-2.69 (m, 2H), 2.09 (s, 5H). LC-MS: m/z 414 [M+H]+.
[0457] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(4-(oxazol- 5-yl)phenyl)-4-oxobutanamide, Compound 24
Figure imgf000104_0001
[0458] 1H NMR (400 MHz, DMSO-</6): 10.18 (s, IH), 8.38 (s, IH), 7.71 - 7.64 (m, 5H), 7.56 (s, IH), 6.34 (bs, IH), 4.04 - 4.01 (t, J=6 Hz, 2H), 3.86 (bs, 2H), 2.88 - 2.87 (m, 2H), 2.67 - 2.64 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 380 [M+H]+.
[0459] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(4-(oxazol- 5-yl)phenyl)-4-oxobutanamide, Compound 25
Figure imgf000104_0002
[0460] 1H NMR (400 MHz, DMSO-<¾: 10.25 (s, IH), 7.90 - 7.86 (m, 3H), 7.73 - 7.70 (m, 3H), 4.04 - 4.01 (m, 2H), 3.86 (br s, 2H), 2.90 - 2.87 (m, 2H), 2.69 - 2.66 (m, 2H), 2.08 (br s, 5H). LC-MS: m/z 396 [M+H]+.
[0461] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(5- (thiazol-2-yl)pyridin-2-yl)butanamide, Compound 26
Figure imgf000105_0001
[0462] 1H MR (400 MHz, DMSO- 6): 8.87 (s, IH), 8.49 (br s, IH), 8.28 - 8.21 (m, 2H), 7.88 (d, J=3.2 Hz, IH), 7.34 (d, J=3.2 Hz, IH), 6.55 (br s, 0.5H), 5.81 (br s, 0.5H), 4.16 (t, J=6.2 Hz, 2H), 3.93 (br s, 2H), 3.05 - 2.96 (m, 2H), 2.86 - 2.84 (m, 2H), 2.23 - 2.18 (m, 5H). LC-MS: m/z 397 [M+H]+.
[0463 ] Synthesis of N-(4-( 1 H-imidazol- 1 -yl)phenyl)-4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 27
Figure imgf000105_0002
[0464] 1H NMR (400 MHz, DMSO- ): 10.17 (s, IH), 8.17 (br s, IH), 7.72 - 7.68 (m, 3H), 7.57 - 7.55 (m, 2H), 7.09 (br s, IH), 6.34 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (br s, 2H), 2.88 (t, J=6.4 Hz, 2H), 2.65 (t, J=6.4 Hz, 2H), 2.08 (br s, 5H). LC-MS: m/z 379 [M+H]+.
[0465] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-N-(4-(2- methylthiazol-4-yl)phenyl)-4-oxobutanamide, Compound 28
Figure imgf000105_0003
[0466] 1H NMR (400 MHz, DMSO-< 6): 10.08 (s, IH), 7.85 (d, J=8.4 Hz, 2H), 7.78 )s, IH), 7.64 (d, J=8.8 Hz, 2H), 6.31 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (bs, 2H), 2.88 (t, J=6.2 Hz, 2H), 2.70 (s, 3H), 2.67 - 2.64 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 410 [M+H]+. [0467] Synthesis of N-(4-(l -methyl-lH-pyrazol-5-yl)phenyl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 29
Figure imgf000106_0001
[0468] 1H NMR (400 MHz, DMSO- 6): 10.17 (s, 1H), 7.71 (d, J=8.4 Hz, 2H), 7.46 - 7.43 (m, 3H), 6.34 (d, J=2 Hz, 2H), 4.02 (t, J=6 Hz, 2H), 3.87 (br s, 2H), 3.83 (s, 3H), 2.89 (t, J=6.2 Hz, 2H), 2.67 (t, J=6.4 Hz, 2H), 2.08 (br s, 5H). LC-MS: m/z 393 [M+H] +.
[0469] Synthesis of N-(4-(tert-butyl)phenyl)-4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin- 4(5H)-yl)-4-oxobutanamide, Compound 30
Figure imgf000106_0002
[0470] 1H NMR (400 MHz, D SO-ifc): 9.90 (s, 1H), 7.48 (d, J=8.4 Hz, 2H), 7.29 (d, J=8.8 Hz, 2H), 6.34 (br s, 1H), 4.01 (t, J=6.0 Hz, 2H), 3.85 (br s, 2H), 2.87 - 2.84 (m, 2H), 2.61 (t, J=6.4 Hz, 2H), 2.07 (br s, 5H), 1.25 (s, 9H). LC-MS: m/z 369 [M+H] +.
[0471] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- (pyrrolidin-l-yl)phenyl)butanamide, Compound 31
Figure imgf000106_0003
[0472] 1H NMR (400 MHz, Methanol-^): 7.30 (d, J=8.8 Hz, 2H), 6.53 (d, J=8.8 Hz, 2H), 4.10 (t, J=6.2 Hz, 2H), 3.93 (t, J=5.6 Hz, 2H), 3.26 - 3.23 (m, 4H), 2.95 (bs, 2H), 2.72 (t, J=6.6 Hz, 2H), 2.33 - 2.18 (m, 6H), 2.02 - 2.00 (m, 4H). LC-MS: m/z 382 [M+H] +. [0473] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(4-(2- methylthiazol-4-yl)phenyl)-4-oxobutanamide, Compound 32
Figure imgf000107_0001
[0474] 1H NMR (400 MHz, DMSO-i¾ 10.65 (s, IH), 8.87 (s, IH), 8.26-8.24 (m, IH), 8.12- 8.10 (m, IH), 7.96 (s, IH), 6.33 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.85 (s, 2H), 2.89-2.86 (m, 2H), 2.74-2.72 (m, 5H), 2.07 (s, 5H). LC-MS: m/z Al l [M+H] +.
[0475] Synthesis of N-([l, -biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)-4-oxobutanamide, Compound 33
Figure imgf000107_0002
[0476] 1H NMR (400 MHz, DMSO-c 6): 10.09 (s, IH), 7.70 - 7.68 (m, 2H), 7.64 - 7.60 (m, 4H), 7.43 (m, 2H), 7.33 - 7.31 (m, IH), 6.35 (bs, IH), 4.02 (t, J=6 Hz, 2H), 3.87 (bs, 2H), 2.89 (t, J=6.4 Hz, 2H), 2.68 - 2.65 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 389 [M+H]+.
[0477] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- (pyridin-2-yl)phenyl)butanamide, Compound 34
Figure imgf000107_0003
[0478] 1H NMR (400 MHz, DMSO- 6): 10.15 (s, IH), 8.62 (d, J=4.4 Hz, IH), 8.04 (d, J=8.4 Hz, 2H), 7.91 - 7.89 (m, IH), 7.86 - 7.82 (m, IH), 7.71 (d, J=8.4 Hz, 2H), 7.31 - 7.27 (m, IH), 6.34 (br s, IH), 4.03 (t, J=5.8 Hz, 2H), 3.87 (br s, 2H), 2.91 - 2.88 (m, 2H), 2.69 - 2.66 (m, 2H), 2.08 (br s, 5H). LC-MS: m/z 390 [M+H]+.
[0479] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- (pyridin-3-yl)phenyl)butanamide, Compound 35
Figure imgf000108_0001
[0480] 1H NMR (400 MHz, DMSO-J6): 10.14 (s, IH), 8.87 (s, IH), 8.52 (s, IH), 8.04 (d, J=8.0 Hz, IH), 7.70 (q, J=9.2 Hz, 4H) 7.47 - 7.44 (m, IH), 6.35 (br s, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (br s, 2H), 2.89 (t, J=6.2 Hz, 2H), 2.67 (t, J=6.2 Hz, 2H), 2.08 (br s, 5H). LC-MS: m/z 390
[M+H]+.
[0481] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- (pyridin-4-yl)phenyl)butanamide, Compound 36
Figure imgf000108_0002
[0482] 1H NMR (400 MHz, DMSO-t ): 10.20 (s, IH), 8.59 (d, J=5.4 Hz, 2H), 7.76 (q, J=8.8 Hz, 4H), 7.68 (d, J=6 Hz, 2H), 6.34 (br s, IH), 4.10 - 4.01 (t, J=6.0 Hz, 2H), 3.86 (br s, 2H), 2.91 - 2.88 (t, J=6.2 Hz, 2H), 2.67 (t, J=6.2 Hz, 2H), 2.08 (br s, 5H). LC-MS: m/z 390 [M+H]+.
[0483] Synthesis of 4-{2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxo-N-[4- (pyridazin-3-yl)phenyl]butanamide, Compound 37
Figure imgf000109_0001
[0484] 1H NMR (400 MHz, DMSO-i¾: 10.24 (s, IH), 8.18-8.11 (m, 3H), 7.79-7.73 (m, 3H), 6.35 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.87 (s, 2H), 2.90-2.88 (m, 2H), 2.69 (t, J=6.4 Hz, 2H), 2.08 (s, 5H). LC-MS: m/z 391 [M+H]+.
[0485] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- (pyrimidin-4-yl)phenyl)butanamide, Compound 38
Figure imgf000109_0002
[0486] 1H NMR (400 MHz, DMSO-i¾: 10.29 (s, IH), 9.17 (d, J=1.2 Hz, IH), 8.79 (d, J=5.2 Hz, IH), 8.19-8.17 (m, 2H), 8.03-8.01 (m, IH), 7.78-7.76 (m, 2H), 6.33 (br s, IH), 4.02 (t, J=5.6 Hz, 2H), 3.86 (s, 2H), 2.91-2.88 (m, 2H), 2.70-2.67 (m, 2H), 2.39 (s, 3H), 2.08 (s, 5H). LC-MS: m/z 391 [M+H]+.
[0487] Synthesis of 4-{2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxo-N-[4- (pyrimidin-5-yl)phenyl]butanamide, Compound 39
Figure imgf000109_0003
[0488] 1H NMR (400 MHz, DMSO-i¾: 10.20 (s, IH), 9.13-9.11 (m, 3H), 7.79-7.74 (m, 4H), 6.34 (br s, IH), 4.04-4.01 (m, 2H), 3.86 (s, 2H), 2.89 (t, J=6.4 Hz, 2H), 2.67 (t, J=6.8 Hz, 2H), 2.08 (br s, 5H). LC-MS: m/z 391 [M+H]+. [0489] Synthesis of 4-{2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrimidin-4-yl}-4- (pyrimidin-2-yl)phenyl]butanamide, Compound 40
Figure imgf000110_0001
[0490] 1H NMR (400 MHz, Methanol-^): 8.82 (d, J=4.8 Hz, 2H), 8.36 (d, J=8.8 Hz, 2H), 7.73 (d, J=8.8 Hz, 2H), 7.32 (t, J=4.8 Hz, 1H), 4.12 (t, J=6.0 Hz, 2H), 3.98-3.96 (m, 2H), 3.01 (br s, 2H), 2.82 (t, J=6.8 Hz, 2H), 2.24-2.20 (m, 5H). LC-MS: m/z 391 [M+H]+.
[0491] Synthesis of N-([2 '-bipyridin]-6'-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)-4-oxobutanamide, Compound 41
Figure imgf000110_0002
[0492] 1H NMR (400 MHz, Chloroform- d): 10.72 (1H, s), 9.02 (1H, d, J= 2.0 Hz), 8.66 (1H, d, J= 4.4 Hz), 8.43 (1H, dd, J= 8.4, 2.4 Hz), 8.17 (1H, d, J= 8.8 Hz), 8.00 (1H, d, J= 8.0 Hz), 7.86-7.91 (1H, m), 7.35-7.38 (1H, m), 6.34 (1H, bs), 4.02 (2H, t), 3.86 (2H, bs), 2.89(2H, t), 2.74 (2H, t), 2.08(5H, m). LC-MS: m/z 391.1 [M+H]+.
[0493] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- (pyrazin-2-yl)phenyl)butanamide, Compound 42
Figure imgf000110_0003
[0494] 1H NMR (400 MHz, DMSO- 6): 10.24 (s, IH), 9.21 (d, J=1.6 Hz, IH), 8.68-8.65 (m, IH), 8.55 (d, J=2.4 Hz, IH), 8.12-8.08 (m, 2H), 7.80-7.72 (m, 2H), 6.35 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.95-2.85 (m, 2H), 2.72-2.65 (m, 2H), 2.08 (bs, 5H). MS (ESI) m/z 391 [M+H] +.
[0495] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(6- phenylpyridin-3-yl)butanamide, Compound 43
Figure imgf000111_0001
[0496] 1H MR (400 MHz, Chloroform-^: 8.66 (IH, s), 8.52 (IH, bs), 8.23 (IH, bs), 8.10 (IH, s), 7.89 (2H, d, J= 7.6 Hz), 7.65 (IH, d, J= 8.8 Hz), 7.37-7.47 (3H, m), 4.17 (2H, t), 3.90 (2H, bs), 2.98 (2H, m), 2.82 (2H, t), 2.23 (5H, m). LC-MS: m/z 390.1 [M+H] +.
[0497] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(5- phenylpyridin-2-yl)butanamide, Compound 44
Figure imgf000111_0002
[0498] 1H NMR (400 MHz, Chloroform-c/): 8.66 (IH, s), 8.52 (IH, bs), 8.23 (IH, bs), 8.10 (IH, s), 7.89 (2H, d, J= 7.6 Hz), 7.65 (IH, d, J= 8.8 Hz), 7.37-7.47 (3H, m), 4.17 (2H, t), 3.90 (2H, bs), 2.98 (2H, m), 2.82 (2H, t), 2.23 (5H, m). LC-MS: m/z 390.1 [M+H]+.
[0499] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(6- phenylpyridazin-3-yl)butanamide, Compound 45
Figure imgf000112_0001
[0500] 1H NMR (400 MHz, OMSO-d6): 11.25 (IH, s), 8.36 (IH, d, J= 9.6 Hz), 8.21 (IH, d, J= 9.6 Hz), 8.09 (2H, d, J= 7.2 Hz), 7.49-7.56 (3H, m), 6.33 (IH, bs), 4.02 (2H, t), 3.86 (2H, bs), 2.91(2H, t), 2.80 (2H, t), 2.08(5H, m). LC-MS: m/z 391.1 [M+H]+.
[0501] Synthesis of 4-{2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxo-N-(5- phenylpyrimidin-2-yl)b tanamide, Compound 46
Figure imgf000112_0002
[0502] 1H NMR (400 MHz, DMSC ¾: 10.71 (s, IH), 8.98 (s, 2H), 7.77 (d, J=7.2 Hz, 2H), 7.51 (t, J=7,2 Hz, 2H), 7.43 (t, J=7.2 Hz, IH), 6.34 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.90-2.87 (s, 4H), 2.08 (s, 5H). LC-MS: m/z 391 [M+H]+.
[0503] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(5- phenylpyrazin-2-yl)butanamide, Compound 47
Figure imgf000112_0003
[0504] 1H NMR (400 MHz, DMSO-<¾): 10.93 (s, IH), 9.37 (s, IH), 9.00 (s, IH), 8.10 - 8.08 (m, 2H), 7.53 - 7.45 (m, 3H), 6.34 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.93 - 2.90 (m, 2H), 2.79 - 2.76 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 391 [M+H]+. [0505] Synthesis of N-([2,3'-bipyridin]-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyiimidin- 4(5H)-yl)-4-oxobutanamide, Compound 48
Figure imgf000113_0001
[0506] 1H NMR (400 MHz, OMSO-d6): 10.50 (s, IH), 9.21 (s, IH), 8.89 (d, J=2.0 Hz, IH), 8.58 (dd, J=4 Hz, 1.2 Hz, IH), 8.39 - 8.36 (m, IH), 8.20 - 8.17 (m, IH), 8.01 (d, J=8.8 Hz, IH), 7.50 - 7.47 (m, IH), 6.35 (bs, IH), 4.02 (t, J=5.8 Hz, 2H), 2.67 (bs, 2H), 2.93 - 2.89 (m, 2H), 2.73 - 2.69 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 391 [M+H]+.
[0507] Synthesis of 4- {2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrimidin-4-yl}-4-oxo-N-[5- (pyridin-3-yl)pyridin-2-yl]butanamide, Compound 49
Figure imgf000113_0002
[0508] 1H NMR (400 MHz, DMSC ¾ 10.68 (s, IH), 8.94 (d, J=2.0 Hz, IH), 8.71 (s, IH), 8.58-8.57 (m, IH), 8.16-8.12 (m, 3H), 7.51-7.47 (m, IH), 6.34 (br s, IH), 4.02 (t, J=5.6 Hz, 2H), 3.86 (s, 2H), 2.90-2.87 (m, 2H), 2.76-2.73 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 391 [M+H]+.
[0509] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(5- (pyrazin-2-yl)pyridin-2-yl)butanamide, Compound 50
Figure imgf000113_0003
[0510] 1H NMR (400 MHz, Chloroform-;/): 9.01 (IH, d, J= 1.2 Hz ), 8.94 (IH, s), 8.63 (lH,t), 8.53 (IH, d, J= 2.4 Hz), 8.45 (IH, s), 8.33 (2H, m), 4.16 (2H, t), 3.94 (2H, bs), 2.97 (2H, m), 2.87 (2H, t), 2.23 (4H, m), 1.25 (3H, s). LC-MS: m/z 392.1 [M+H]+.
[0511] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(5- (pyridin-3-yl)pyrazin-2-yl)butanamide, Compound 52
Figure imgf000114_0001
[0512] 1H NMR (400 MHz, DMSO-d6): 11.00 (s, IH), 9.40 (s, IH), 9.27 (m, IH), 9.08 (m, IH), 8.65 - 8.64 (m, IH), 8.45 - 8.43 (m, IH), 7.55 - 7.52 (m, IH), 6.34 (bs, IH), 4.03 (t, J=5.8 Hz, 2H), 3.87 (bs, 2H), 2.93 - 2.90 (m, 2H), 2.80 - 2.77 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 392 [M+H]+.
[0513] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-N-(3-methyl- [l,l'-biphenyl]-4-yl)-4-oxobutanamide, Compound 53
Figure imgf000114_0002
[0514] 1H NMR (400 MHz, DMSO-i¾: 9.38 (s, IH), 7.65-7.63 (m, 2H), 7.52-7.42 (m, 5H), 7.35-7.31 (m, IH), 6.36 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.89-2.88 (m, 2H), 2.71- 2.68 (m, 2H), 2.28 (s, 3H), 2.09 (s, 5H). LC-MS: m/z 403 [M+H]+.
[0515] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-N-(2-methyl- [l,l'-biphenyl]-4-yl)-4-oxobutanamide, Compound 54
Figure imgf000115_0001
[0516] 1H NMR (400 MHz, DMSO-d6): 9.99 (s, IH), 7.52-7.40 (m, 4H), 7.35-7.29 (m, 3H), 7.12-7.10 (m, IH), 6.35 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.89-2.86 (m, 2H), 2.66- 2.63 (m, 2H), 2.20 (s, 3H), 2.08 (s, 5H). LC-MS: m/z 403 [M+H]+.
[0517] Synthesis of N-(3-methoxy-[ 1 , 1 '-biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 55
Figure imgf000115_0002
[0518] 1H NMR (400 MHz, DMSO-i¾): 9.22 (s, IH), 8.07 (d, J=8.0 Hz, IH), 7.68 (d, J=7.2 Hz, 2H), 7.44 (t, J=7.2 Hz, 2H), 7.35-7.33 (m, IH), 7.28 (s, IH), 7.21-7.18 (m, IH), 6.35 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.94 (s^ 3H), 3.86 (s, 2H), 2.88-2.85 (m, 2H), 2.75-2.72 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 419 [M+H]+.
[0519] Synthesis of N-(2-methoxy-[ 1 , 1 '-biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 56
Figure imgf000115_0003
[0520] 1H NMR (400 MHz, DMSO- 6): 10.10 (s, IH), 7.51 (bs, IH), 7.46 - 7.44 (m, 2H), 7.37 (t, J=7.6 Hz, 2H), 7.29 - 7.26 (m, IH), 7.21 (s, 2H), 6.35 (bs, IH), 4.03 (t, J=5.8 Hz, 2H), 3.87 (bs, 2H), 2.90 - 2.87 (m, 2H), 2.68 - 2.64 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 419 [M+H]+. [0521] Synthesis of N-(2-fluorobiphenyl-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 57
Figure imgf000116_0001
[0522] 1H NMR (400 MHz, DMSO-i¾): 10.31 (s, IH), 7.73-7.67 (m, IH), 7.54-7.42 (m, 5H), 7.41-7.34 (m, 2H), 6.35 (bs, IH), 4.03 (t, J=6.4 Hz, 2H), 3.87 (bs, 2H), 2.92-2.83 (m, 2H), 2.70- 2.63 (m, 2H), 2.08 (bs, 5H). MS (ESI) m/z 407 [M+H]+..
[0523] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-N-(2'-rnethyl- [l,l'-biphenyl]-4-yl)-4-oxobutanamide, Compound 58
Figure imgf000116_0002
[0524] 1H NMR (400 MHz, DMSO-<¾: 10.08 (s, IH), 7.66-7.64 (m, 2H), 7.29-7.16 (m, 6H), 6.35 (br s, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (s, 2H), 2.91-2.88 (m, 2H), 2.68-2.65 (m, 2H), 2.23 (s, 3H), 2.09 (s, 5H). LC-MS: m/z 403 [M+H]+.
[0525] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(3'-methyl- [l,l'-biphenyl]-4-yl)-4-oxobutanamide, Compound 59
Figure imgf000116_0003
[0526] 1H NMR (400 MHz, DMSO-<¾: 10.09 (s, IH), 7.69-7.66 (m, 2H), 7.60-7.58 (m, 2H), 7.45-7.41 (m, 2H), 7.34-7.30 (m, IH), 7.14-7.12 (m, IH), 6.34 (br s, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (s, 2H), 2.91-2.87 (m, 2H), 2.68-2.65 (m, 2H), 2.36 (s, 3H), 2.08 (s, 5H). LC-MS: m/z 403 [M+H]+.
[0527] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-N-(4'-methyl- [l,r-biphenyl]-4-yl)-4-oxobutanamide, Compound 60
Figure imgf000117_0001
[0528] 1H NMR (400 MHz, DMSO- ): 10.07 (s, 1H), 7.67 - 7.65 (m, 2H), 7.58 - 7.51 (m, 4H), 7.24 (d, J=8.0 Hz, 2H), 6.34 (bs, 1H), 4.02 (t, J=6.0 Hz, 2H), 3.86 (bs, 2H), 2.88 (t, J=6.4 Hz, 2H), 2.67 - 2.64 (m, 2H), 2.32 (s, 3H), 2.08 (bs, 5H). LC-MS: m/z 403 [M+H]+.
[0529] Synthesis of N-(2'-methoxy-[l,l'-biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 61
Figure imgf000117_0002
[0530] 1H NMR (400 MHz, DMSO-c¾: 10.05 (s, 1H), 7.61 (d, J=8.4 Hz, 2H), 7.39 (d, J=8.8 Hz, 2H), 7.33 - 7.25 (m, 2H), 7.08 (d, J-7.6 Hz, 1H), 7.00 (t, J=4.2 Hz, 1H), 6.30 (bs, 1H), 4.02 (t, J=6.0 Hz, 2H), 3.86 (bs, 2H), 3.75 (s, 3H), 2.88 (t, J=6.4 Hz, 2H), 2.67 - 2.64 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 419 [M+H]+.
[0531] Synthesis of N-(3'-methoxy-[l,l'-biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 62
Figure imgf000118_0001
[0532] 1H NMR (400 MHz, DMSO-<¾: 10.10 (s, 1H), 7.69-7.67 (m, 2H), 7.63-7.60 (m, 6H), 7.37-7.33 (m, 1H), 7.21-7.16 (m, 2H), 6.91-6.88 (m, 1H), 6.35 (br s, 1H), 4.03 (t, J=5.6 Hz, 2H), 3.87-3.82 (m, 5H), 2.91-2.87 (m, 2H), 2.68-2.65 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 419
[M+H]+.
[0533] Synthesis of N-(4'-methoxy-[l,r-biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 63
Figure imgf000118_0002
[0534] 1H NMR (400 MHz, DMSO- ): 10.05 (s, 1H), 7.65 - 7.63 (m, 2H), 7.58 - 7.53 (m, 4H), 6.99 (d, J=8.8 Hz, 2H), 6.30 (bs, 1H), 4.02 (t, J=6 Hz, 2H), 3.87 (bs, 2H), 3.78 (s, 3H), 2.90 - 2.86 (m, 2H), 2.65 (t, J=6.2 Hz, 2H), 2.08 (bs, 5H). LC-MS: m/z 419 [M+H]+.
[0535] Synthesis of N-(3'-fluoro-[l,r-biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 65
Figure imgf000118_0003
[0536] 1H NMR (400 MHz, DMSO-J6): 10.13 (s, 1H), 7.71 - 7.65 (m, 4H), 7.51 - 7.46 (m, 3H), 7.16 - 7.12 (m, 1H), 6.35 (br, 1H), 4.03 (t, J=5.8 Hz, 2H), 3.87 (br, 2H), 2.91 - 2.88 (m, 2H), 2.68 - 2.65 (m, 2H), 2.08 (br, 5H). LC-MS: m/z 407 [M+H]+. [0537] Synthesis of N-(4'-fluorobiphenyl-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 66
Figure imgf000119_0001
[0538] 1H NMR (400 MHz, DMSO-d6): 10.10 (s, IH), 7.70-7.62 (m, 4H), 7.61-7.57 (m, 2H), 7.30-7.20 (m, 2H), 6.35 (bs, IH), 4.02 (t, J=6.4 Hz, 2H), 3.87 (bs, 2H), 2.92-2.85 (m, 2H), 2.70- 2.63 (m, 2H), 2.08 (bs, 5H); MS (ESI) m/z 407 [M+H]+.
[0539] Synthesis of 4'-(4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4- oxobutanamido)-[l, -biphenyl]-3-carboxylic acid, Compound 67
Figure imgf000119_0002
[0540] 1H NMR (400 MHz, DMSO- ): 10.12 (s, IH), 8.15 (s, IH), 7.98 - 7.94 (m, IH), 7.90 - 7.87 (m, 2H), 7.73 - 7.64 (m, 4H), 7.56 (t, J=7.6Hz, IH), 6.35 (bs, IH), 4.03 (t, j=5.8 Hz, 2H), 3.87 (bs, 2H), 2.91 - 2.88 (m, 2H), 2.67 (t, J=6.6Hz, 2H), 2.08 (s, 5H). LC-MS: m/z 433
[M+H]+.
[0541] Synthesis of N-(2'-amino-[ 1 , 1 '-biphenyl] -4-yl)-4-(2-methyl-6,7-dihydropyrazolo [1,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 69
[0542] Step 1 : Preparation of N-(4-bromophenyl)-4-(2-methyl-6,7-dihydropyrazolo[l, 5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide.
Figure imgf000119_0003
[0543] 1H NMR (400 MHz, DMSO-i¾: 10.13 (s, IH), 7.57-7.55 (m, 2H), 7.47-7.45 (m, 2H), 6.33 (br s, IH), 4.01 (t, J=6.0 Hz, 2H), 3.85 (s, 2H), 2.88-2.85 (m, 2H), 2.65-2.61 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 392 [M+H]+.
[0544] Step 2: Preparation of N-(2'-amino-[l,l,-biphenyl]-4-yl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide.
Figure imgf000120_0001
[0545] The desired product was obtained using general Suzuki procedure A.
[0546] 1H NMR (400 MHz, DMSO-<¾: 10.06 (s, IH), 7.66-7.64 (m, 2H), 7.33-7.31 (m, 2H), 7.03-6.94 (m, 2H), 6.74-6.72 (m, IH), 6.61 (t, J=7.2 Hz, IH) 6.35 (br s, IH), 4.70 (s, 2H), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.90-2.87 (m, 2H), 2.67-2.64 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 404 [M+H]+.
[0547] Synthesis of N-(3'-amino-[l,l'-biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 70
[0548] Step 1 : Preparation of 4-(2-methyl-6,7-dihydropyrazolo[ l,5-a]pyrimidin-4(5H)-yl)-N-(2- methyl-[ 1 , 1 '-biphenyl] -4-yl)-4-oxobutanamide.
Figure imgf000120_0002
[0549] 1H NMR (400 MHz, DMSO-c¾: 10.17 (s, IH), 8.41 (s, IH), 8.18-8.12 (m, 2H), 7.75- 7.71 (m, 5H), 6.35 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.87 (s, 2H), 2.91-2.88 (m, 2H), 2.69-2.66 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 434 [M+H]+. [0550] Step 2: Preparation of N-(3*-amino-[l,r-biplienyl]-4-yl)-4-(2-niethyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide.
Figure imgf000121_0001
[0551] 4-{2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrim^
oxobutanamide (1 equiv.), ammonium chloride(4 equiv.), and iron(2 equiv.) were added to a solution of ethanol (0.02 M) and water(0.08 M), and heated at 40 °C for 4 h. The reaction mixture was then filtered, diluted with water and extracted with ethyl acetate. The combined organic layers were dried over magnesium sulphate and concentrated under vacuum. The crude product was purified by reversed phase preparative HPLC to afford the purified product.
[0552] 1H NMR (400 MHz, DMSO-<¾): 10.05 (s, 1H), 7.65-7.63 (m, 2H), 7.49-7.47 (m, 2H), 7.06 (t, J=7.6 Hz, 1H), 6.80 (s, 1H), 6.75-6.73 (m, 1H), 6.52-6.50 (m, 1H), 6.34 (br s, 1H), 5.09 (s, 2H), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.89-2.86 (m, 2H), 2.67-2.63 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 404 [M+H]+.
[0553] Synthesis of N-(4,-amino-[l,l'-biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 71
[0554] Step 1: Preparation of 4-(2-methyl-6,7-dihydropyrazolo[ l,5-a]pyrimidin-4(5H)-yl)-N- (4'-nitro-[ 1 , 1 '-biphenyl]-4-yl)-4-oxobutanamide.
Figure imgf000121_0002
[0555] MS: m/z 434 [M+H]+. [0556] Step 2: Preparation of N-(4'-amino-[l,r-bip enyl]-4-yl)-4-(2-methyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide.
Figure imgf000122_0001
[0557] The desired product was obtained in a similar fashion as compound 70.
[£558] MR (400 MHz, DMSO- 6): 9.97 (s, IH), 7.60-7.58 (m, 2H), 7.47-7.45 (m, 2H), 7.36-7.34 (m, 2H), 6.67 (d, J=8.0 Hz, 2H), 6.34 (br s, IH), 5.70-5.60 (br s, 2H), 4.02 (t, J=5.6 Hz, 2H), 3.86 (s, 2H), 2.89-2.86 (m, 2H), 2.65-2.62 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 404
[M+H]+.
[0559] Synthesis of N-(3 '-(dimethylamino)-[ 1 , 1 '-biphenyl]-4-yl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 72
Figure imgf000122_0002
[0560] 1H NMR (400 MHz, DMSO-<¾): 10.03(s, IH), 7.63 (d, J=8.4 Hz, 2H), 7.45 (d, J=8.4 Hz, 2H), 7.25 - 7.21 (m, IH), 7.14 - 7.12 (m, IH), 7.04 - 6.96 (m, 2H), 6.34 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.90 - 2.87 (m, 2H), 2.67 - 2.64 (m, 2H), 2.47 (s, 6H), 2.08 (bs, 5H). LC- MS: m/z 432 [M+H]+.
[0561] Synthesis of N-(3'-acetamido-[l,r-biphenyl]-4-yl)-4-(2-methyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 74
Figure imgf000123_0001
[0562] N-[4-(3-aminophenyl)phenyl]-4-{2-methyl-5H,6H,7H-pyrazolo[l,5-a]pyrimidin-4-yl}- 4-oxobutanamide(l equiv.) was dissolved in dichloromethane(0.02 M) at 0 °C. Triethylamine(l equiv.), followed by acetyl chloride(l .05 equiv.), was added to the solution, and the reaction mixture allowed to stir for 30 min. It was then diluted with water and extracted with
dichloromethane. The combined organic layers were dried over magnesium sulphate and concentrated under vacuum. The crude product was purified by reversed phase preparative HPLC to afford the purified product.
[0563] 1H NMR (400 MHz, DMSO-i¾ 10.09 (s, IH), 9.98 (s, IH), 7.83 (s, IH), 7.69-7.67 (m, 2H), 7.54-7.52 (m, 3H), 7.36-7.27 (m, 2H), 6.34 (br s, IH), 4.03 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.90-2.87 (m, 2H), 2.67-2.64 (m, 2H), 2.08-2.05 (m, 8H). LC-MS: m/z 446 [M+H]+.
[0564] Synthesis of N-(4-cyclohexylphenyl)-4-(2-methyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin- 4(5H)-yl)-4-oxobutanamide, Compound 75
Figure imgf000123_0002
[0565] 1H NMR (400 MHz, DMSO- 6): 9.88 (s, IH), 7.47 (d, J=8.4 Hz, 2H), 7.11 (d, J=8.4 Hz, 2H), 6.34 (bs, IH), 4.02 (t, J=5.6 Hz, 2H), 3.85 (bs, 2H), 2.87 - 2.84 (m, 2H), 2.61 (t, J=6.4 Hz, 2H), 2.42 (m, IH), 2.08 (bs, 5H), 1.76 - 1.67 (m, 5H), 1.41 - 1.19 (m, 5H). LC-MS: m/z 395 [M+H]+.
[0566] Synthesis of tert-butyl 4-(4-(4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)- yl)-4-oxobutanamido)phenyl)piperazine-l-carboxylate, Compound 76
Figure imgf000124_0001
[0567] 1H NMR (400 MHz, DMSO- 6): 9.77 (s, IH), 7.44 (d, J=8.8 Hz, 2H), 6.88 (d, J=9.2 Hz, 2H), 6.30 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.85 (bs, 2H), 3.45 - 3.43 (m, 4H), 3.02 - 2.99 (m, 4H), 2.87 - 2.83 (m, 2H), 2.60 - 2.57 (m, 2H), 2.08 (bs, 5H), 1.42 (s, 9H). LC-MS: m/z 497
[M+H]+.
[0568] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- (piperazin-l-yl)phenyl)butanamide, Compound 77
Figure imgf000124_0002
[0569] 4M HC1 in dioxane (15 equiv.) was added to the boc-protected amine (1 equiv.) and stirred at room temperature for 16 h. Upon completion, reaction was extracted with saturated sodium bicarbonate, organic layers combined, dried over magnesium sulphate, filtered and concentrated to dryness in vacuo. Crude was purified by recrystallisation with
dichloromethane/hexane and lyophilized to afford pure product as an off white solid.
[0570] 1H NMR (400 MHz, DMSO- 6): 9.73 (s, IH), 7.42 (d, J=8.8 Hz, 2H), 6.84 (d, J=8.8 Hz, 2H), 6.34 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.85 (bs, 2H), 2.98 - 2.95 (m, 4H), 2.84 - 2.81 (m, 6H), 2.58 (t, J=6.6 Hz, 2H), 2.08 (bs, 5H). LC-MS: m/z 397 [M+H]+.
[0571] Synthesis of N-(4-(4-acetylpiperazin-l-yl)phenyl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 78
Figure imgf000125_0001
[0572] A solution of compound 77 (1.0 equiv.) in dry tetrahydrofuran was added to a stirred mixture of acid chloride (1.0 equiv.) and triethylamine (1.2 equiv.) in tetrahydrofuran at 0 °C. The reaction mixture was allowed to warm to room temperature with continued stirring for 3 h. Upon completion of reaction, solvent was evaporated to dryness in vacuo and crude extracted with ethyl acetate and dried over magnesium sulphate. Crude was purified by crashing out solids with DCM/methano; hexane to afford a white solid.
[0573] 1H NMR (400 MHz, OMSO-d6) 9.77 (s, 1H), 7.45 (d, J=9.2 Hz, 2H), 6.89 (d, J=9.2 Hz, 2H), 6.30 (bs, 1H), 4.02 (t, J=6.0 Hz, 2H), 3.85 (bs, 2H), 3.57 - 3.54 (m, 4H), 3.08 - 3.06 (m, 2H), 3.01 - 2.99 (m, 2H), 2.87 - 2.84 (m, 2H), 2.59 (t, J=6.6 Hz, 2H), 2.08 (bs, 5H), 2.03 (s, 3H). LC-MS: m/z 439 [M+H]+.
[0574] Synthesis of N-ethyl-4-(6-(4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)- 4-oxobutanamido)pyridin-3-yl)piperazine-l-carboxamide, Compound 79
Figure imgf000125_0002
[0575] Step 1 : Preparation of tert-butyl 4-(6-(4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)-4-oxobutanamido)pyridin-3 -yl)piperazine- 1 -carboxylate
[0576] LC-MS: m/z 498 [M+H]+.
Figure imgf000126_0001
Step 2: Preparation of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrirnidin-4(5H)-yl)-4-oxo-N-(5- (piperazin- 1 -yl)pyridin-2-yl)butanamide
[0577] To a stirred solution of tert-butyl 4-(6-(4-(2-methyl-6, 7-dihydropyrazolofl ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamido)pyridin-3-yl)piperazine (1 equiv.) in dichloromethane (0.04M) was added TFA (5 equiv.) drop-wise at 0 °C and stirred at room temperature for 1 h. The reaction mixture was diluted with water and basified with saturated sodium bicarbonate solution and then extracted with dichloromethane. The organic layer was dried over anhydrous sodium sulphate and concentrated under reduced pressure to afford the intermediate.
[0578] LC-MS: m/z 398 [M+H]+.
Figure imgf000126_0002
Step 3: Preparation of N-ethyl-4-(6-(4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)- yl)-4-oxobutanamido)pyridin-3 -yl)piperazine- 1 -carboxamide
[0579] To a stirred solution of 4-(2-methyl-6, 7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4- oxo-N-(5-(piperazin-l-yl)pyridin-2-yl)butanamide (1 equiv.) in THF (0.015 M) was added ethylisocyanate (4.5 equiv.) and the reaction mixture was stirred at 80 °C in sealed tube for 16 h. The reaction mixture was concentrated under reduced pressure to give crude product. The crude product was purified by preparative TLC to afford the product. [0580] 1H NMR (400 MHz, DMSO-<¾: 10.25 (s, IH), 8.01 (d, J= 2.7 Hz, IH), 7.92 (d, J= 8.3 Hz, IH), 7.41-7.39 (dd, J= 2.7 Hz, 9.3 Hz, IH), 6.53-6.55 (m, IH), 6.32 (brs, IH), 4.01 (t, J= 5.7 Hz, 2H), 3.84 (brs, 2H), 3.39-3.41 (m, 4H), 3.05-3.07 (m, 6H), 2.82-2.84 (m, 2H), 2.64-2.66 (m, 2H), 2.07 (brs, 5H), 1.01 (t, J= 7.5 Hz, 3H). MS (ESI) m/z 469.2 [M+H]+.
[0581] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- phenylcyclohexyl)butanamide, Compound 80
Figure imgf000127_0001
[0582] 1H MR (400 MHz, DMSO-i¾: 7.78 (d, J=7.6 Hz, IH), 7.30-7.21 (m, 4H), 7.19-7.13 (m, IH), 6.35 (bs, IH), 4.01 (t, J=6.0 Hz, 2H), 3.83 (bs, 2H), 3.65-3.52 (m, IH), 2.80-2.70 (m, 2H), 2.50-2.40 (m, 2H), 2.40-2.35 (m, 2H), 2.08 (m, 5H), 1.90-1.82 (m, 2H), 1.82-1.75 (m, 2H), 1.57-1.45 (m, 2H), 1.38-1.25 (m, 2H); MS (ESI) m/z 395 [M+H]+.
[0583] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(4-(l- methylpiperidin-4-yl)phenyl)-4-oxobutanamide, Compound 82
Figure imgf000127_0002
[0584] 1H NMR (400 MHz, DMSO-c/6): 9.90 (s, IH), 7.49 (d, J=8.4 Hz, 2H), 7.14 (d, J=8.4 Hz, 2H), 6.33 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.85 (bs, 2H), 2.88 - 2.86 (m, 5H), 2.61 (t, J=6.6 Hz, 2H), 2.20 (s, 3H), 2.08 (bs, 5H), 2.01 - 1.95 (m, 2H), 1.71 - 1.60 (m, 4H). LC-MS: m/z 410 [M+H]+.
[0585] Synthesis of N-(6-(3-chlorophenyl)pyridin-3-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 83
Figure imgf000128_0001
[0586] 1H NMR (400 MHz, Methanol-^): 8.80 - 8.81 (m, IH), 8.18 - 8.20 (m, IH), 7.97 - 7.98 (m, IH), 7.82 - 7.87 (m, 2H), 7.38 - 7.47 (m, 2H), 4.55 (s, IH), 4.19 (t, J= 6.0 Hz, 2H), 3.94 - 3.96 (m, 2H), 2.99 - 3.01 (m, 2H), 2.82 (t, J= 6.0 Hz, 2H), 2.22 (bs, 2H), 2.18 (s, 3H). LC-MS: m/z 424 [M+H]+.
[0587] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(6- (m-tolyl)pyridin-3-yl)butanamide, Compound 85
Figure imgf000128_0002
[0588] 1H NMR (400 MHz, DMSO-d6): 10.31 (s, IH), 8.81 - 8.80 (m, IH), 8.14 - 8.11 (m, IH), 7.91 - 7.87 (m, 2H), 7.82 - 7.80 (m, IH), 7.34 (t, j=7.6 Hz, IH), 7.21 - 7.19 (m, IH), 6.35 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.92 - 2.89 (m, 2H), 2.71 - 2.68 (m, 2H), 2.38 (s, 3H), 2.08 (bs, 5H). LC-MS: m/z 404 [M+H]+.
[0589] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(5-]
6-phenylpyridin-3-yl)-4-oxobutanamide, Compound 86
Figure imgf000128_0003
[0590] 1H NMR (400 MHz, DMS W6): 10.24 (s, IH), 8.64 (d, J=2.0 Hz, IH), 7.97 (d, J=2.4 Hz, IH), 7.52-7.48 (m, 2H), 7.48-7.40 (m, 2H), 7.40-7.35 (m, IH), 6.35 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.95-2.85 (m, 2H), 2.75-2.65 (m, 2H), 2.31 (s, 3H), 2.08 (bs, 5H). MS (ESI) m/z 404 [M+H]+.
[0591] Synthesis of N-(6-(4-fluorophenyl)pyridin-3-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 87
Figure imgf000129_0001
[0592] 1H NMR (400 MHz, DMSO- 6): 10.33 (s, IH), 8.81 - 8.80 (m, IH), 8.13 - 8.05 (m, 3H), 7.90 (d, J=8.4 Hz, IH), 7.28 (t, 8.8 Hz, 2H), 6.34 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (bs, 2H), 2.92 - 2.88 (m, 2H), 2.70 - 2.67 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 408 [M+H]+.
[0593] Synthesis of N-(6-(3-fiuorophenyl)pyridazin-3-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 88
Figure imgf000129_0002
[0594] 1H NMR (400 MHz, DMSO-<¾: 10.67 (s, IH), 8.68 (s, 1H),8.13-8.10 (m, 2H), 7.80 (s, IH), 7.70-7.68 (m, IH), 7.52-7.48 (m, IH), 7.44-7.42 (m, IH), 6.33 (br s, IH), 4.02 (t, J=6.0Hz, 2H), 3.86 (s, 2H), 2.90-2.87 (m, 2H), 2.75-2.72 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 424 [M+H]+.
[0595] Synthesis of N-(5-(3-fluorophenyl)pyridin-2-yl)-4-(2-methyl-6,7-dihydropyrazolo[l ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 89
Figure imgf000129_0003
[0596] 1H NMR (400 MHz, DMSO-i¼): 10.67 (s, IH), 8.69 (s, IH), 8.13 (s, 2H), 7.61 - 7.48 (m, 3H), 7.23 - 7.18 (m, IH), 6.33 (bs, IH), 4.02 (t, J=5.8 Hz, 2H), 3.86 (bs, 2H), 2.91 - 2.87 (m, 2H), 2.76 - 2.72 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 408 [M+H]+.
[0597] Synthesis of N-(5-(4-fluorophenyl)pyridin-2-yl)-4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 90
Figure imgf000130_0001
[0598] 1H NMR (400 MHz, DMSO-<¾): 10.62 (s, IH), 8.62 - 8.63 (m, IH), 8.12 - 8.14 (m, IH), 8.04 - 8.07 (m, IH), 7.73 - 7.77 (m, 2H), 7.28 - 7.32 (m, 2H), 6.34 (bs, IH), 4.55 (s, IH), 4.02 (t, J= 6.0 Hz, 2H ), 3.85 - 3.87 (m, 2H), 2.87 - 2.90 (m, 2H), 2.73 (t, J= 6.0 Hz, 2H), 2.22 (bs, 2H), 2.08 (s, 3H). LC-MS: m/z 408 [M+H]+.
[0599] Synthesis of N-(3-methyl-5-phenylpyridin-2-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4~oxobutanamide, Compound 91
Figure imgf000130_0002
[0600] 1H NMR (400 MHz, DMSO-i¾: 10.07 (s, IH), 8.53 (s, IH), 7.94 (s, IH), 7.72-7.70 (m, 2H), 7.51-7.47 (m, 2H), 7.42-7.38 (m, IH), 6.35 (br s, IH), 4.01 (t, J=6.0 Hz, 2H), 3.85 (s, 2H), 2.89-2.86 (m, 2H), 2.71 -2.68 (m, 2H), 2.21 (s, 3H), 2.09 (s, 5H). LC-MS: m/z 404 [M+H]+.
[0601] Synthesis of N-(4-methyl-5-phenylpyridin-2-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 92
Figure imgf000131_0001
[0602] 1H NMR (400 MHz, DMSO-<¾): 10.55 (s, IH), 8.61 (s, 2H), 8.16 (s, IH), 8.05 (s, IH), 7.85 (d, J= 8.0 Hz, IH), 7.51-7.47 (m, IH), 6.33 (s, IH), 4.02 (t, J= 6.0 Hz, 2H), 3.86 (*, 2H), 2.88 (s, 2H), 2.73 (d, J= 6.4 Hz, 2H), 2.24 (s, 3H), 2.08 (bs, 5H). MS (ESI) m/z 404.1 [M+H]+.
[0603] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(5- (m-tolyl)pyridin-2-yl)butanamide, Compound 93
Figure imgf000131_0002
[0604] 1H NMR (400 MHz, DMSO-t¾): 10.63 (s, IH), 8.63 - 8.622 (m, IH), 8.14 - 8.12 (m, IH), 8.07 - 8.04 (m, IH), 7.53 (bs, IH), 7.49 (d, J=8.0 Hz, IH), 7.36 (t, J=7.6 Hz, IH), 7.19 (d, j=7.6 Hz, IH), 6.34 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.90 - 2.87 (m, 2H), 2.75 - 2.72 (m, 2H), 2.38 (s, 3H), 2.08 (bs, 5H). LC-MS: m/z 404 [M+H]+.
[0605] Synthesis of N-(5-fluoro-2,3'-bipyridin-6,-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 94
Figure imgf000131_0003
[0606] 1H NMR (400 MHz, DMSO--¼): 10.72 (s, IH), 8.99 (d, J=2.4 Hz, IH), 8.66 (d, J=2.8 Hz, IH), 8.43-8.38 (m, IH), 8.19-8.14 (m, IH), 8.12-8.07 (m, IH), 7.84 (td, J=8.8, 3.2 Hz, IH), 6.34 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (bs, 2H), 2.92-2.85 (m, 2H), 2.78-2.72 (m, 2H), 2.08 (bs, 5H). MS (ESI) m/z 409 [M+H]+. [0607] Synthesis of N-(4-fluoro-2,3'-bipyridin-6'-yl)-4-(2-methyl-6,7-dihydropyrazolo[l^ a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 95
Figure imgf000132_0001
[0608] 1H NMR (400 MHz, DMSO-i¾: 10.75 (s, IH), 9.07 (d, J= 1.7 Hz, IH), 8.70-8.67 (dd, J = 5.7 Hz, 9.2 Hz, IH), 8.48-8.46 (dd, J= 2.2 Hz, 8.8 Hz, IH), 8.18 (d, J= 8.8 Hz, IH), 7.99 (d, J= 8.8 Hz, IH), 7.31-7.28 (m, IH), 6.33 (brs, IH), 4.04-4.01 (t, J- 5.7 Hz, 2H), 3.86 (brs, 2H), 2.89 (d, J= 6.1 Hz, 2H), 2.76 (t, J= 6.2 Hz, 2H), 2.08 (brs, 5H). MS (ESI) m/z 409.11 [M+H]+.
[0609] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(6-rnethyl- [2,3,-bipyridin]-6,-yl)-4-oxobutanamide, Compound 96
Figure imgf000132_0002
[0610] 1H NMR (400 MHz, DMSO-c¾): 10.71 (s, IH), 9.01 - 9.00 (m, IH), 8.42 - 8.40 (m, IH), 8.17 - 8.15 (d, J=8.8 Hz, IH), 7.80 - 7.75 (m, 2H), 7.23 - 7.21 (m, IH), 6.33 (br, IH), 4.02 (t, j=6.2 Hz, 2H), 3.86 (br, 2H), 2.89 - 2.87 (m, 2H), 2.75 - 2.72 (m, 2H), 2.54 (s, 3H), 2.08 (br, 5H). LC-MS: m/z 405 [M+H]+.
[0611] Synthesis of N-(6-(3-chlorophenyl)pyridazin-3-yl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 97
Figure imgf000132_0003
[0612] 1H NMR (400 MHz, Methanol-***): 8.50 - 8.53 (m, IH), 8.08 - 8.13 (m, 2H), 7.94 - 7.95 (m, IH), 7.50 - 7.52 (m, IH), 4.55 (s, IH), 4.11 (t, ' J= 6.0 Hz, 2H), 3.95 - 3.97 (m, 2H), 3.00 - 3.02 (m, 2H), 2.91 (t, J= 6.0 Hz, 2H ), 2.22 (bs, 2H), 2.18 (s, 3H). LC-MS: m/z 425 [M+H] +.
[0613] Synthesis of N-[6-(4-chlorophenyl)pyridazin-3-yl]-4-{2-methyl-5H,6H,7H-pyrazolo[l,5- a]pyrimidin-4-yl}-4-oxobutanamide, Compound 100
Figure imgf000133_0001
[0614] 1H NMR (400 MHz, DMSO-t¾: 11.29 (s, IH), 8.37 (d, J=9.2 Hz, IH), 8.24 (d, J=9.6 Hz, IH), 8.13 (d, J=8.4 Hz, 2H), 7.60 (d, J=8.4 Hz, 2H), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.91-2.89 (m, 2H), 2.81-2.78 (m, 2H), 2.07 (s, 5H). LC-MS: m/z 426 [M+H]+.
[0615] Synthesis of N-(6-(4-fluorophenyl)pyridazin-3-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 101
Figure imgf000133_0002
[0616] 1H NMR (400 MHz, Chloroform-^): 9.17 (IH, bs), 8.50 (IH, d, J= 9.2 Hz), 8.00-8.04 (2H, m), 7.82 (IH, d, J= 9.2 Hz), 7.19 (2H, t), 4.16 (2H, t), 3.94 (2H, bs), 2.96-3.06(4H, m), 2.23 (5H, m). LC-MS: m/z 409.2 [M+H]+.
[0617] Synthesis of N-(6-(5-chloropyridin-3-yl)pyridazin-3-yl)-4-(2-methyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 102
Figure imgf000134_0001
[0618] 1H.NMR (400 MHz, DMSO-<¾: 11.36 (s, IH), 9.24 (d, J= 1.7 Hz, IH), 8.75 (d, J= 2.2 Hz, IH), 8.59 (t, J= 2.2 Hz, IH), 8.38 (q, J= 9.2 Hz, 17.1 Hz, 2H), 6.35 (br s, IH), 4.03 (t, J= 6.1 Hz, 2H), 3.86 (br s, 2H), 2.91-2.90 (m, 2H), 2.83-2.80 (m, 2H), 2.08 (br s, 5H). MS (ESI) m/z 426.24 [M+H]+.
[0619] Synthesis of N-(5'-chloro-[2,3'-bipyridin]-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 108
Figure imgf000134_0002
[0620] 1H NMR (400 MHz, Chloroform-i/): 9.02 (IH, bs), 8.68 (IH, s), 8.57 (IH, d, J= 2.4 Hz), 8.29-8.32 (3H, m), 7.73 (IH, d, J= 8.8 Hz), 4.16 (2H, t), 3.94 (2H, bs), 2.97 (2H, m), 2.89 (2H, t), 2.24 (5H, m). LC-MS: m/z 425.1 [M+H]+.
[0621] Synthesis of N-(5'-fluoro-[2,3'-bipyridin]-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 109
Figure imgf000134_0003
[0622] 1H NMR (400 MHz, DMSO-i¾: 10.43 (s, IH), 9.12 (s, IH), 8.87 (d, J=2.0 Hz, IH), 8.59 (d, j=2.8 Hz, IH), 8.29-8.25 (m, IH), 8.21-8.18 (m, IH), 8.10-8.08 (m, IH), 6.34 (br s, IH), 4.02 (t, J=6.4 Hz, 2H), 3.86 (s, 2H), 2.92-2.89 (m, 2H), 2.72-2.69 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 409 [M+H]+. [0623] Synthesis of N-(3-fluoro-2,3'-bipyTidin-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 111
Figure imgf000135_0001
[0624] 1H NMR (400 MHz, DMSO- 6): 10.68 (s, IH), 9.05 (s, IH), 8.68 (s, IH), 8.70-8.65 (m, IH), 8.30-8.15 (m, 2H), 7.55-7.50 (s, IH), 6.35 (bs, IH), 4.10-4.00 (m, 2H), 3.87 (bs, 2H), 3.00- 2.85 (m, 2H), 2.80-2.65 (m, 2H), 2.08 (bs, 5H). MS (ESI) m/z 409 [M+H]+.
[0625] Synthesis of N-(5'-amino-2,3'-bipyridin-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 112
Figure imgf000135_0002
[0626] 1H NMR (400 MHz, DMSO- 6): 10.38 (s, IH), 8.83 (d, J=2.0 Hz, IH), 8.35 (d, J=1.6 Hz, IH), 8.12-8.09 (m, IH), 7.93 (d, J=2.4 Hz, IH), 7.88-7.82 (m, IH), 7.58-7.52 (m, IH), 6.35 (bs, IH), 5.40 (s, 2H), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.95-2.85 (m, 2H), 2.75-2.65 (m, 2H), 2.08 (bs, 5H); MS (ESI) m/z 406 [M+H]+.
[0627] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(6'-methyl- [2,3'-bipyridin]-5-yl)-4-oxobutanamide, Compound 113
Figure imgf000135_0003
[0628] 1H NMR (400 MHz, DMSO-i¾): 10.36 (s, IH), 9.08 - 9.07 (m, IH), 8.83 - 8.82 (m, IH), 8.27 - 8.25 (m, IH), 8.16 - 8.13 (m, IH), 7.96 (d, J=8.8 Hz, IH), 7.34 (d, J=8.0 Hz, IH), 6.34 (bs, IH), 4.02 (t, J=5.6 Hz, 2H), 3.86 (bs, 2H), 2.92 - 2.89 (m, 2H), 2.69 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 405 [M+H]+.
[0629] Synthesis of N-(5,-chloro-[3,3'-bipyridin]-6-yl)-4-(2-methyl-6,7-dihydropyrazolo[l ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 114
Figure imgf000136_0001
[0630] 1H NMR (400 MHz, DMSO-c¾: 10.73 (s, IH), 8.92 (d, J=1.6 Hz, IH), 8.78 (d, J=1.6 Hz, IH), 8.62 (d, J=2.0 Hz, IH), 8.34-8.33 (m, IH), 8.23-8.15 (m, 2H), 6.33 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.90-2.87 (m, 2H), 2.76-2.73 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 425 [M+H]+.
[0631] Synthesis of N-(5'-fluoro-[3,3'-bipyridin]-6-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 115
Figure imgf000136_0002
[0632] 1H NMR (400 MHz, DMSO-c¾: 10.73 (s, IH), 8.86 (t, J=1.6 Hz, IH), 8.78 (d, J=2.0 Hz, IH), 8.58 (d, J=2.4 Hz, IH), 8.24 - 8.13 (m, 3H), 6.30 (bs, IH), 4.03 (t, J=5.8 Hz, 2H), 3.86 (bs, 2H), 2.91 - 2.88 (m, 2H), 2.76 - 2.73 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 409 [M+H]+.
[0633] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-N-(5'-methyl- [3,3'-bipyridin]-6-yl)-4-oxobutanamide, Compound 116
Figure imgf000137_0001
[0634] 1H NMR (400 MHz, DMSO-<¾): 10.68 (s, IH), 8.74 - 8.70 (m, 2H), 8.42 (s, IH), 8.18 - 8.12 (m, 2H), 7.96 (s, IH), 6.34 (bs, IH), 4.03 (t, J=5.8 Hz, 2H), 3.87 (bs, 2H), 2.90 - 2.87(m, 2H), 2.76 - 2.73 (m, 2H), 2.37 (s, 3H), 2.08 (bs, 5H). LC-MS: m/z 405 [M+H]+.
[0635] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- (piperazin-l-yl)phenyl)butanamide, Compound 117
[0636] Step 1: Preparation of Boc-protected 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)-4-oxo-N-(4-(piperazin- 1 -yl)phenyl)butanamide
Figure imgf000137_0002
[0637] 1H NMR (400 MHz, DMSO-i¾: 10.71 (s, IH), 8.90 (m, IH), 8.75 (bs, IH), 8.44 - 8.43 (d, J=2.4 Hz, IH), 8.19 - 8.15 (m, 3H), 6.36 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (bs, 2H), 2.91 - 2.87 (m, 2H), 2.76 - 2.73 (m, 2H), 2.08 (bs, 5H), 1.40 (s, 18H). LC-MS: m/z 606 [M+H]+.
[0638] Step 2: Preparation of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4- oxo-N-(4-(piperazin- 1 -yl)phenyl)butanamide
Figure imgf000137_0003
[0639] 1H NMR (400 MHz, DMSO-<¾: 10.64 (s, IH), 8.56 (m, IH), 8.14 (d, J=8.4 Hz, IH), 8.06 - 8.05 (m, IH), 8.01 - 7.99 (m, IH), 7.94 (d, J=2.4 Hz, IH), 7.16 (t, J=2.2 Hz, IH), 6.33 (bs, IH), 5.41 (s, 2H), 4.02 (t, J=6.0 Hz, 2H), 3.86 (bs, 2Η), 2.90 - 2.87 (m, 2H), 2.75 - 2.72 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 406 [M+H]+.
[0640] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(5'- (trifluoromethyl)-[3,3'-bipyridin]-6-yl)butananiide, Compound 118
Figure imgf000138_0001
[0641] 1H NMR (400 MHz, DMSO-d6): 10.77 (s, IH), 9.27 - 9.26 (m, IH), 8.97 (bs, IH), 8.85 - 8.84 (m, IH), 8.56 (bs, IH), 8.31 - 8.28 (m, IH), 8.20 - 8.18 (m, IH), 6.34 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.86 (bs, 2H), 2.91 - 2.88 (m, 2H), 2.77 - 2.74 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 459 [M+H]+.
[0642] Synthesis of N-(4-methyl-3,3'-bipyridin-6-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 119
Figure imgf000138_0002
[0643] 1H NMR (400 MHz, DMSO-rf6): 10.56 (s, IH), 8.61 (s, 2H), 8.16 (s, IH), 8.05 (s, IH), 7.85 (d, J= 8.0 Hz, IH), 7.51-7.47 (m, IH), 6.33 (s, IH), 4.02 (t, J= 6.0 Hz, 2H), 3.86 (s, 2H), 2.87 (d, J= 5.6 Hz, 2H), 2.73 (d, J= 6.0 Hz, 2H), 2.25 (s, 3H), 2.08 (s, 5H); MS (ESI) m/z 405.39 [M+H]+.
[0644] Synthesis of N-(4,5'-dimethyl-3,3'-bipyridin-6-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 120
Figure imgf000139_0001
[0645] 1H NMR (400 MHz, DMSO-< ): 10.57 (s, IH), 8.44 (d, J=1.2 Hz, IH), 8.40 (d, J=1.2 Hz, IH), 8.15 (s, IH), 8.05 (s, IH), 7.67 (s, IH), 6.35 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.90-2.80 (m, 2H), 2.80-2.60 (m, 2H), 2.36 (s, 3H), 2.26 (s, 3H), 2.08 (bs, 5H). MS (ESI) m/z 419 [M+H]+.
[0646] Synthesis of N-(6'-methyl-3,3'-bipyridin-6-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 121
Figure imgf000139_0002
[0647] 1H NMR (400 MHz, Methanol-^): 8.68 (d, J=2.0 Hz, IH), 8.58 (d, J=2.0 Hz, IH), 8.22- 8.18 (m, IH), 8.08-7.98 (m, 2H), 7.41 (d, J=8.0 Hz, IH), 4.56 (s, IH), 4.11 (t, J=6.8 Hz, 2H), 3.96 (bs, 2H), 3.03-2.95 (m, 2H), 2.90-2.83 (m, 2H), 2.58 (s, 3H), 2.30-2.15 (m, 5H); MS (ESI) m/z 405 [M+H]+.
[0648] Synthesis of N-(5-(5-fluoropyridin-3-yl)pyrazin-2-yl)-4-(2-methyl-6,7- dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 122
Figure imgf000139_0003
[0649] 1H NMR (400 MHz, DMSO-c¾: 11.06 (s, IH), 9.42 (d, J=1.2 Hz, IH), 9.18 (t, J=1.6 Hz, IH), 9.14 (d, J=i.2 Hz, IH), 8.66 (d, J=2.8 Hz, IH), 8.40-8.30 (m, IH), 6.33 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.95-2.85 (m, 2H), 2.85-2.75 (m, 2H), 2.08 (bs, 5H). MS (ESI) m/z 410 [M+H]+.
[0650] Synthesis of N-(5-(3-fluoropb.enyl)pyrazin-2-yl)-4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 123
Figure imgf000140_0001
[0651] 1H NMR (400 MHz, DMSO-<¾ 10.98 (s, IH), 9.37 (s, IH), 9.05 (d, j=1.2 Hz, IH), 7.97-7.89 (m, 2H), 7.58-7.52 (m, IH), 7.30-7.28 (m, IH), 6.33 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 3.27 (s, IH), 2.93-2.90 (m, 2H), 2.79-2.76 (m, 2H), 2.07 (s. 3H). LC-MS: m/z 409 [M+H]+.
[0652] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-N-(5-(5- methylpyridin-3-yl)pyrazin-2-yl)-4-oxobutanamide, Compound 124
Figure imgf000140_0002
[0653] 1H NMR (400 MHz, DMSO-i¾: 10.98 (s, IH), 9.39 (s, 2H), 9.06 (s, 2H), 8.48 (s, IH), 8.27 (s, IH), 6.33 (s, IH), 4.02 (t, J= 5.6 Hz, 2H), 3.86 (s, 2H), 2.91-2.88 (m, 2H), 2.50-2.75 (m, 2H), 2.39 (bs, 3H), 2.08 (bs, 5H); MS (ESI) m/z 404.10 [M +H]+.
[0654] Synthesis of N-(6-methyl-5-(pyridin-3-yl)pyrazin-2-yl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 125
Figure imgf000141_0001
[0655] 1H NMR (400 MHz, DMSO-<¾): 10.95 (s, IH), 9.24 (s, IH), 8.83 - 8.82 (m, IH), 8.65 - 8.63 (m, IH), 8.07 - 8.04 (m, IH), 7.54 - 7.51 (m, IH), 6.34 (bs, IH), 4.02 (t, J=6.0 z, 2H), 3.86 (bs, 2H), 2.92 - 2.89 (m, 2H), 2.78 - 2.75 (m, 2H), 2.54 (s, 3H), 2.08 (bs, 5H). LC-MS: m/z 406 [M+H]+.
[0656] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(5- (m-tolyl)pyrazin-2-yl)butanamide, Compound 126
Figure imgf000141_0002
[0657] 1H NMR (400 MHz, DMSO-J6): 10.91 (s, IH), 9.35 (s, 1H),8.97 (s, IH), 7.92-7.86 (m, 2H), 7.39 (t, J=7.6 Hz, IH), 7.26 (d, J=6.8 Hz, IH), 6.34 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.91-2.89 (m, 2H), 2.78-2.75 (m, 2H), 2.39 (s, 3H), 2.08 (s, 5H). LC-MS: m/z 405 [M+H]+.
[0658] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(5-(6- methylpyridin-3-yl)pyrazin-2-yl)-4-oxobutanamide, Compound 127
Figure imgf000141_0003
[0659] 1H NMR (400 MHz, DMSO- ): 10.97 (s, IH), 9.38 (s, IH), 9.13 (d, J=2.0 Hz, IH), 9.04 (d, J=1.2 Hz, IH), 8.33 (dd, J=8.0, 2.4 Hz, IH), 7.39 (d, J=8.0 Hz, IH), 6.35 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.95-2.85 (m, 2H), 2.80-2.70 (m, 2H), 2.53 (s, 3H), 2.08 (bs, 5H). MS (ESI) m/z 406 [M+H]+.
[0660] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(5-(6- methylpyrazin-2-yl)pyridin-2-yl)-4-oxobutanamide, Compound 128
Figure imgf000142_0001
[0661] 1H NMR (400 MHz, DMSO-d6): 10.79 (s, IH), 9.08 - 9.06 (m, 2H), 8.51 (bs, IH), 8.49 - 8.46 (m, IH), 8.20 (d, j=8.8 Hz, IH), 6.33 (bs, IH), 4.02 (t, J=5.8 Hz, 2H), 3.86 (bs, 2H), 2.91 - 2.87 (m, 2H), 2.76 - 2.73 (m, 2H), 2.57 (s, 3H), 2.08 (bs, 5H). LC-MS: m/z 406 [M+H]+.
[0662] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(4- (pyrazin-2-yl)phenyl)butanamide, Compound 129
Figure imgf000142_0002
[0663] 1H NMR (400 MHz, DMSO- ): 10.24 (s, IH), 9.21 (d, J=1.6 Hz, IH), 8.68-8.65 (m, IH), 8.55 (d, J=2.4 Hz, IH), 8.12-8.08 (m, 2H), 7.80-7.72 (m, 2H), 6.35 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.95-2.85 (m, 2H), 2.72-2.65 (m, 2H), 2.08 (bs, 5H). MS (ESI) m/z 391 [M+H]+.
[0664] Synthesis of N-(3-(3-chlorophenyl)isoxazol-5-yl)-N-methyl-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 130
Figure imgf000143_0001
[0665] Step 1 : Preparation of N-(3-(3-chlorop enyl)isoxazol-5-yl)-4-(2-metliyl-6,7- dihydropyrazolo [ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide
[0666] 1H NMR (400 MHz, Methanol-^): 7.66 (s, 1H), 7.60-7.58 (m, 1H), 7.33-7.29 (m, 2H), 6.53 (s, 1H), 6.13 (brs, 1H), 3.81 (brs, 2H), 3.64 (s, 1H), 2.70 (brs, 2H), 2.52-2.51 (m, 2H), 2.29 (s, 2H), 1.86 (brs, 5H). LC-MS: MS m/z 414 [M+H]+.
[0667] Step 2: Preparation of N-(3-(3-chlorophenyl)isoxazol-5-yl)-N-methyl-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide
Figure imgf000143_0002
[0668] To a stirred solution of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide (1 equiv.) in dry tetrahydrofuran was added K2C03 (3 equiv.) at room temperature and stirring was continued for 15 minutes.. CH3I (5 equiv.) was added to reaction mixture drop wise at 0 °C followed by stirring at room temperature for 16 h. After completion, the reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic layer was washed with water and brine solution. The organic solvent was dried over anhydrous sodium sulphate, filtered and concentrated under vacuum to give crude product. The crude product was purified by column chromatography.
[0669] 1H-NMR (400 MHz, DMSO-i¾: 7.93 (s, 1H), 7.85 (d, J= 7.1 Hz, 1H), 7.60-7.53 (m, 2H), 7.05 (s, 2H), 6.33 (bs, 1H), 4.02 (bs, 2H), 3.86 (bs, 2H), 3.40 (s, 3H), 2.87 (bs, 4H), 2.07 (bs, 5H); MS (ESI) m/z 428.20 [M+H]+. [0670] Synthesis of 3-(3-chlorophenyl)-N-(4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)butyl)isoxazol-5-amine, Compound 131
[0671] Step 1 : Preparation of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-(2-methyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide.
Figure imgf000144_0001
[0672] 1H NMR (400 MHz, DMSO-c¾): 11.88 (s, lH), 7.91 (s, 1H), 7.82 (d, 1H), 7.58-7.51 (m, 2H), 6.89 (s, 1H), 6.36 (brs, 1H), 4.02 (t, 2H), 3.86 (brs, 2H), 2.92 (t, 2H), 2.71 (q, 2H), 2.07 (brs, 5H). LC-MS: MS m/z 414 [M+H]+.
[0673] Step 2: Preparation of 3-(3-chlorophenyl)-N-(4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)butyl)isoxazol-5-amine.
Figure imgf000144_0002
[0674] A solution of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-(2-methyl-6, 7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide (1 equiv.) in THF (0.03 M) was treated with borane.DMS (5 equiv.) at 0 °C and the reaction mixture was heated to 40 °C for 10 h. After completion, the reaction mixture was quenched with methanol and concentrated under reduced pressure to obtain the crude product; water was added and extracted with ethyl acetate. The combined ethyl acetate layers were dried over sodium sulphate, filtered and concentrated under reduced pressure to obtain the crude product. The crude product was purified by preparative
HPLC.
[0675] 1H NMR (400 MHz, Chloroform- ): 7.72 (brs, 1H), 7.62 (d, J= 6.8 Hz, 1H), 7.37-7.33 (m, 2H), 5.26 (s, 1H), 5.16 (s, 1H), 4.61 (brs, 1H), 4.06 (t, J= 6.0 Hz, 2H), 3.29 (d, J= 5.6 Hz, 2H), 3.16 (brs, 4H), 2.20 (s, 3H), 2.16 (t, J= 5.6 Hz, 2H), 1.69 (brs, 4H). LC-MS: MS m/z 386 [M+H]+. [0676] Synthesis of N-(3-(3-chlorophenyl)isoxazol-5-yl)-3-methyl-4-(2-methyl-6,7- dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 132
[0677] Step 1 : Preparation of 4-methoxy-2-methyl-4-oxobutanoic acid
Figure imgf000145_0001
[0678] To a solution of 4-methoxy-2-methylene-4-oxobutanoic acid (1 equiv.) in methanol (0.3 M) was added a slurry of 10% Pd/C (0.1 equiv.) in methanol under nitrogen. The reaction mixture was hydrogenated under H2 (balloon) at room temperature for 16 h. The reaction mixture was filtered through celite bed and washed with methanol. The filtrate was concentrated under reduced pressure to give crude product. The crude product was washed with n-pentane and diethyl ether to remove solid impurities. The solvent was evaporated under reduced pressure to afford the intermediate as a pale yellow liquid.
[0679] Step 2: Preparation of methyl 3-methyl-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanoate
Figure imgf000145_0002
[0680] To a stirred solution of 4-methoxy-2-methyl-4-oxobutanoic acid (1 equiv.), 2-methyl- 4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine (1 equiv.) in DMF (0.5 M) was added HATU (1.5 equiv.), TEA (3 equiv.) and the reaction mixture was stirred at rt for 12 h. Upon completion of reaction, the reaction mixture was added to the ice-water and extracted with ethyl acetate. The combined organic layer was washed with water, brine, dried over sodium sulphate, filtered and concentrated under reduced pressure to give the crude product. The crude product was purified by column chromatography to afford the intermediate. LC-MS: MS m/z 266 [M+H]+.
[0681] Step 3: Preparation of 3-methyl-4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin- 4(5H)-yl)-4-oxobutanoic acid
Figure imgf000146_0001
[0682] To a stirred solution of methyl 3-methyl-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanoate (1 equiv.) in THF-CH3OH-H20 (1 :1 :1) (0.2 M) was added LiOH.H20 (2 equiv.) and the reaction mixture was stirred at room temperature for 2 h. After completion, the solvents were evaporated under reduced pressure and the residue was acidified with saturated KHS04 solution and extracted with 10% methanol in chloroform. The aqueous layer was evaporated under reduced pressure and stirred in 10% methanol in
chloroform, filtered and concentrated under reduced pressure to afford the intermediate. LC-MS: MS m/z 266 [M+H]+.
[0683] Step 4: Preparation of N-(3-(3-chlorophenyl)isoxazol-5-yl)-3-methyl-4-(2-methyl-6,7- dihydropyrazolo [ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide
Figure imgf000146_0002
[0684] 1H NMR (400 MHz, DMSO-i¾: 11.84 (s, 1H), 7.91 (s, 1H), 7.83 (d, J= 7 Hz, 1H), 7.57-7.49 (m, 2H), 6.77 (s, 1H), 6.35 (bs, 1H), 4.06^1.03 (t, J= 5.7 Hz, 2H), 3.96-3.92 (m, 2H), 2.89-2.86 (m, 1H), 2.60-2.55 (dd, J= 4.8 Hz, 16.7 Hz, 1H), 2.13-2.07 (bs, 5H), 1.13 (d, J = 6.6 Hz, 3H). MS (ESI) m/z 428.20 [M+H]+.
[0685] Synthesis of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-(6,7-dihydropyrazolo[ 1 ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 133
Figure imgf000146_0003
[0686] 1H NMR (400 MHz, DMSO-rf6): 11.86 (s, 1H), 7.90 (s, 1H), 7.83-7.82 (d, J
1H), 7.58-7.53 (m, 2H), 7.29 (br s, 1H), 6.78 (s, 1H), 6.54 (br s, 1H), 4.14-4.11 (t, J 2H), 3.92-3.90 (s, 2H), 2.95-2.91 (m, 2H), 2.74-2.71 (t, J= 6 Hz, 2H), 2.16-2.12 (m, 2H). LC- MS: MS m/z 400.1 [M+H]+.
[0687] Synthesis of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-oxo-4-(2-(trifluoromethyl)-6,7- dihyo^opyrazolo[l,5-a]pyrimidin-4(5H)-yl)butananiide, Compound 134
Figure imgf000147_0001
[0688] 1H NMR (400 MHz, DMSO-d6): 11.88 (s, IH), 7.90 (s, IH), 7.82 (d, J= 7.6 Hz, IH), 7.58-7.51 (m, 2H), 6.89 (brs, IH), 6.77 (s, IH), 4.22 (t, J= 5.6 Hz, 2H), 3.95 (brs, 2H), 2.96 (m, 2H), 2.74 (t, J= 6 Hz, 2H), 2.19 (brs, 2H). LC-MS: MS m/z 467 [M-H]+.
[0689] Synthesis of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-(2-methoxy-6,7- dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 135
Figure imgf000147_0002
[0690] 1H NMR (400 MHz, DMSO-<¾: 7.87 (s, IH), 7.79 (d, J=6.8 Hz, IH), 7.56 - 7.50 (m, 2H), 6.70 (s, IH), 5.96 (b s, IH), 3.93 (t, J=6.2 Hz, 2H), 3.85 (b s, 2H), 3.71 (s, 3H), 2.90 - 2.88 (m, 2H), 2.67 (b s, 2H), 2.11 (b s, 2H). LC-MS: m/z 430 [M+H]+.
[0691] Synthesis of methyl 4-(4-(3-(3-chlorophenyl)isoxazol-5-ylamino)-4-oxobutanoyl)- 4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine-2-carboxylate, Compound 136
Figure imgf000147_0003
[0692] 1H NMR (400 MHz, DMSO-c¾: 9.3 (brs, 1H), 7.78 (s, 1H), 7.65 (d, J= 7.2 Hz, 1H), 7.41-7.34 (m, 2H), 6.63 (s, 1H), 4.31 (t, J= 6.4 Hz, 2H), 3.96 (brs, 2H), 3.92 (s, 3H), 3.03 (brs, 2H), 2.90 (brs, 2H), 2.28 (brs, 2H).
[0693] Synthesis of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-(2-ethyl-6,7-dihydropyrazolo[l ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 137
Figure imgf000148_0001
[0694] 1H NMR (400 MHz, Chloroform-i/) 7.77 (1H, bs), 7.64 (1H, d, J= 8.0 Hz), 7.33-7.40 (2H, m), 7.25 (1H, s), 6.64 (1H, s), 4.18 (2H, t), 4.00 (2H, m), 2.90 (2H, m), 2.88 (2H, m, ), 2.60 (2H, t), 2.19 (2H, m), 0.88 (3H, m).
[0695] Synthesis of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-(6-methyl-2,3-dihydro-lH- imidazo[l,2-b]pyrazol-l-yl)-4-oxobutanamide, Compound 138
[0696] Step 1: Preparation of 6-methyl-2,3-dihydro-lH-imidazo[l,2-b]pyrazole
Figure imgf000148_0002
[0697] To a stirred solution of 3-methyl-lH-pyrazol-5-amine (1 equiv.) in 1,4-dioxane (0.5 M) was added triethylamine (5 equiv.) at room temperature. After 15 minutes, 1 ,2-dibromoethane (1.2 equiv.) was added to the reaction mixture at room temperature. The reaction mixture was stirred at 110 °C for 16 h. After completion, the reaction mixture was filtered and washed with ethyl acetate. The filtrate was concentrated under reduced pressure to give crude product. The crude product was purified by column chromatography to afford the product as an off-white solid. LC-MS: m/z 124 [M+H]+.
[0698] Step 2: Preparation of 4-(6-methyl-2,3-dihydro-lH-imidazo[l,2-b]pyrazol-l-yl)-4- oxobutanoic acid
Figure imgf000149_0001
[0699] To a stirred solution of 6-methyl-2,3-dihydro-lH-imidazo[l,2-b]pyrazole (1 equiv.) in chloroform (0.5 M) was added dihydrofuran-2,5-dione (1.3 equiv.). The reaction mixture was stirred at room temperature for 12 h. After completion, the solid thus precipitated was filtered and washed with n-pentane and dried to afford the product as an off-white solid. LC-MS: m/z 224 [M+H]+.
[0700] Step 3: Preparation of N-(3-(3-chlorophenyl)isoxazol-5-yl)-4-(6-methyl-2,3-dihydro-lH- imidazo[ 1 ,2-b]pyrazol- 1 -yl)-4-oxobutanamide
Figure imgf000149_0002
[0701] 1H NMR (400 MHz, DMSO-i¾: 11.81 (brs, 1H), 7.89 (s, 1H), 7.82 (d, J = 7.0 Hz, 1H), 7.57-7.54 (m, 2H), 6.77 (s, 1H), 5.71 (s, 1H), 4.55 (t, J = 7.8 Hz, 1H), 4.32-4.24 (m, 2H), 4.20- 4.16 (m, 1H), 2.84 (d, J = 6.1 Hz, 1H), 2.75 (s, 3H), 2.15 (s, 2H), 2.11 (s, 1H). LC-MS: m/z 400.22 [M+H]+.
[0702] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N- (6-phenylpyridazin-3-yl)butanamide, Compound 139
[0703] Step 1 : Preparation of 2-methylpropane-l,3-diyl dimethanesulfonate
Figure imgf000149_0003
[0704] To a stirred solution of 2-methylpropane-l ,3-diol (1 equiv.) in dichloromethane (0.45 M) was added methane sulfonyl chloride (2.7 equiv.), using triethylamine (2.7 equiv.) at 0 °C to room temperature for 16 h. Upon completion of reaction, the reaction mixture was washed with water and brine solution, dried over anhydrous Na2S04, filtered and concentrated under reduced pressure to the product as a brown solid.
[0705] Step 2: Preparation of 2,6-dimethyl-4,5,6,7-tetrahydropyrazolo[l,5-a]pyrimidine
Figure imgf000150_0001
[0706] To a stirred solution of 2-methylpropane-l ,3-diyl dimethanesulfonate (1.2 equiv.) in 1,4- dioxane (0.5 M) were added 3 -methyl- lH-pyrazol-5 -amine (1 equiv.) and triethylamine (5 equiv.). The reaction mixture was heated at 100 °C for 48 h. Upon completion of reaction, the reaction mixture was filtered and concentrated under reduced pressure, purified by column chromatography to afford the product as a brown solid. LC-MS: m/z 152 [M+H]+.
[0707] Step 3: Preparation of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)- 4-oxobutanoic acid
Figure imgf000150_0002
[0708] To a stirred solution of 2, 6-dimethyl-4,5,6, 7-tetrahydropyrazolo[l,5-a]pyrimidine (1 equiv.) in chloroform (0.1 M) was added succinic anhydride (1 equiv.) and the reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated under reduced pressure to afford the product as an off-white solid. LC-MS: m/z 252 [M+H]+.
[0709] Step 4: Preparation of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)- 4-oxo-N-(6-phenylpyridazin-3-yl)butanamide
Figure imgf000150_0003
[0710] 1H NMR (400 MHz, DMSO-c¾): 8.37 (d, J= 9.2 Hz, IH), 8.23-8.17 (m, IH), 8.09 (d, J = 7.2 Hz, 2H), 7.60-7.47 (m, 5H), 4.32-3.97 (m, 3H), 3.65-3.60 (m, IH), 2.95-2.75 (bs, 4H), 2.08 (s, 3H), 1.46 (s, IH), 1.05 (s, 3H); LC-MS: m/z 405.0 [M+H]+.
[0711] Synthesis of N-(2,3'-bipyridin-5-yl)-4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 140
Figure imgf000151_0001
[0712] 1H NMR (400 MHz, DMSO-<¾: 10.40 (s, IH), 9.22 (d, J=2.0 Hz, IH), 8.86 (d, J=2.0 Hz, IH), 8.62-8.57 (m, IH), 8.40-8.35 (m, IH), 8.20-8.15 (m, IH), 8.05-7.99 (m, IH), 7.52-7.47 (m, IH), 6.35 (bs, IH), 4.15-4.00 (m, 2H), 3.67-3.60 (m, IH), 3.50-3.35 (m, IH), 3.02-2.80 (m, 2H), 2.75-2.65 (m, 2H), 2.35-2.20 (m, IH), 2.08 (bs, 3H), 1.05 (d, J=4.4 Hz, 3H). MS (ESI) m/z 405 [M+H]+.
[0713] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-4- (6-(pyridin-3-yl)pyridazin-3-yl)butanamide, Compound 141
Figure imgf000151_0002
[0714] 1H NMR (400 MHz, DMSO-d6): 11.33 (s, IH), 9.27 (d, J=l .6 Hz, IH), 8.69 (dd, J=4.8, 1.6 Hz, IH), 8.50-8.43 (m, IH), 8.43-8.37 (m, IH), 8.34-8.19 (m, IH), 7.70-7.55 (m, IH), 6.35 (bs, IH), 4.18-4.00 (m, 2H), 3.67-3.60 (m, IH), 3.50-3.35 (m, IH), 3.02-2.85 (m, 2H), 2.85-2.78 (m, 2H), 2.35-2.20 (m, IH), 2.08 (bs, 3H), 1.05 (d, J=5.6 Hz, 3H). MS (ESI) m/z 406 [M+H]+.
[0715] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(5'- methyl-[3,3'-bipyridin]-6-yl)-4-oxobutanamide, Compound 142
Figure imgf000152_0001
[0716] 1H NMR (400 MHz, DMSO- 6): 10.69 (s, IH), 8.74 - 8.70 (m, 2H), 8.42 - 8.42 (m, IH), 8.16 - 8.14 (m, 2H), 7.97 (s, IH), 6.34 (br, IH), 4.14 - 4.04 (m, 2H), 3.66 - 3.60 (q, IH), 2.97 - 2.87 (m, 2H), 2.76 - 2.73 (m, 2H), 2.37 (s, 3H), 2.08 (s, 3H), 1.06 - 1.04 (m, 3H). LC-MS: m/z 419 [M+H]+.
[0717] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-N-(6-methyl- [2,3'-bipyridin]-6'-yl)-4-oxobutanamide, Compound 143
Figure imgf000152_0002
[0718] JH NMR (400 MHz, DMSO-c/6): 10.71 (s, IH), 9.01 - 9.00 (m, IH), 8.42 - 8.40 (m, IH), 8.17 - 8.15 (d, J=8.8 Hz, IH), 7.80 - 7.75 (m, 2H), 7.23 - 7.21 (m, IH), 6.33 (br, IH), 4.02 (t, J=6.2 Hz, 2H), 3.86 (br, 2H), 2.89 - 2.87 (m, 2H), 2.75 - 2.72 (m, 2H), 2.54 (s, 3H), 2.08 (br, 5H). LC-MS: m/z 405 [M+H]+.
[0719] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(5'- methyl-[2,3'-bipyridin]-5-yl)-4-oxobutanamide, Compound 144
Figure imgf000152_0003
[0720] 1H NMR (400 MHz, DMSO-< 6): 10.39 (s, IH), 9.01 (m, IH), 8.85 (br, IH), 8.43 (m, IH), 8.21 (br, IH), 8.18 - 8.15 (m, IH), 8.00 (d, J=8.8 Hz, IH), 6.33 (br, IH), 4.14 - (m, 2H), 3.66 - 3.61 (m, IH), 2.99 - 2.89 (m, 2H), 2.72 - 2.67 (m, 2H), 2.38 (s, 3H), 2.08 (s, 3H), 1.06 - 1.05 (m, 3H). LC-MS: m/z 419 [M+H]+.
[0721] Synthesis of N-([2,3'-bipyridin]-5-yl)-2-methyl-4-(2-niethyl-6,7-diliydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 145
Figure imgf000153_0001
[0722] 1H NMR (400 MHz, DMSO-d6): 10.38 (s, IH), 9.22 (d, J=1.6 Hz, IH), 8.88 (d, J=2.4 Hz, IH), 8.59 (dd, J=4.8, 1.6 Hz, IH), 8.38 (dt, J=8.0, 2.0 Hz, IH), 8.22-8.18 (m, IH), 8.03-8.00 (m, IH), 7.53-7.48 (m, IH), 6.35 (bs, IH), 4.05-4.00 (m, 2H), 3.90-3.84 (m, 2H), 3.10-3.00 (m, 2H), 2.75-2.60 (m, IH), 2.07 (bs, 5H), 1.21 (d, J=6.4 Hz, 3H). MS (ESI) m/z 405 [M+H]+.
[0723] Synthesis of N-([3,3'-bipyridin]-6-yl)-2-methyl-4-(2-methyl-6,7-dihydropyrazolo[l ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 147
Figure imgf000153_0002
[0724] 1H NMR (400 MHz, DMSO-d6): 10.68 (s, IH), 8.94 (s, IH), 8.72 (s, IH), 8.58 - 8.57 (m, IH), 8.19 - 8.12 (m, 3H), 7.51 - 7.48 (m, IH), 6.27 (br, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (br, 2H), 3.06 - 2.99 (m, IH), 2.06 (br, 5H), 1.19 (d, j=7.2 Hz, 3H). LC-MS: m/z 405 [M+H]+.
[0725] Synthesis of N-(4-methyl-2,3'-bipyridin-6'-yl)-4-(2-methyl-6,7-dihydropyrazolo[l ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 148
Figure imgf000154_0001
[0726] 1H NMR (400 MHz, DMSO-i¾: 10.70 (s, IH), 9.01 (d, J= 2.1 Hz, IH), 8.51 (d, J= 4.8 Hz, IH), 8.43-8.40 (dd, J= 2.6 Hz, 8.7 Hz, IH), 8.15 (d, J= 8.8 Hz, IH), 7.85 (s, IH), 7.19 (d, J = 4.8 Hz, IH), 6.33 (brs, IH), 4.02 (t, J= 5.7 Hz, 2H), 3.86 (brs, 2H), 2.89 (t, J- 5.7 Hz, 2H), 2.74 (t, J= 6.1 Hz, 2H), 2.39 (s, 3H), 2.08 (brs, 5H); MS (ESI) m/z 405.18 [M+H]+.
[0727] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-N-(5-(5- methylpyrazin-2-yl)pyridin-2-yl)-4-oxobutanamidei Compound 149
Figure imgf000154_0002
[0728] 1H NMR (400 MHz, DMSO-<¾: 10.75 (s, IH), 9.14 (d, J= 0.8 Hz, IH), 9.04 (d, J= 2.2 Hz, IH), 8.60 (s, IH), 8.46-8.43 (dd, J= 2.2 Hz, 2.2 Hz, IH), 8.19 (d, J= 8.7 Hz, IH), 6.33 (brs, IH), 4.02 (t, J= 5.7 Hz, 2H), 3.86 (brs, 2H), 2.89 (t, J= 6.1 Hz, 2H), 2.75 (t, J= 6.6 Hz, 2H), 2.53 (s, 3H), 2.07 (brs, 5H). MS (ESI) m/z 406.16 [M+H]+.
[0729] Synthesis of 2-methyl-4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-N- (5'-methyl-[2,3,-bipyridin]-5-yl)-4-oxobutanamide, Compound 150
Figure imgf000154_0003
[0730] 1H NMR (400 MHz, DMSO- 6): 10.37 (s, IH), 9.01 (s, IH), 8.87 (s, IH), 8.42 (s, IH), 8.21 - 8.17 (m, 2H), 7.99 (d, J=8.8 Hz, IH), 6.32 (br, IH), 4.03 - 4.00 (m, 2H), 3.86 (br, 2H), 3.04 - 3.01 (m, 2H), 2.69 - 2.66 (m, IH), 2.38 (s, 3H), 2.07 (br, 5H), 1.21 (d, J=6.0 Hz, 3H). LC- MS: m/z 419 [M+H]+.
[0731 ] Synthesis of 2-methyl-4-(2-methyl-6,7-dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-N- (5-(6-methylpyrazin-2-yl)pyridin-2-yl)-4-oxobutanamide, Compound 151
Figure imgf000155_0001
[0732] H NMR (400 MHz, DMSO- 6): 10.78 (s, IH), 9.08 (s, 2H), 8.56 - 8.46 (m, 2H), 8.21 (d, J=8.4 Hz, IH), 6.29 (br, IH), 4.02 (m, 2H), 3.86 (br, 2H), 3.18 (br, 2H), 3.05 - 2.99 (m, IH), 2.57 (s, 3H), 2.06 (br, 5H), 1.19 (d, J=6.8 Hz, 3H). LC-MS: m/z 420 [M+H]+.
[0733] Synthesis of 2-methyl-4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N- (5'-methyl-[3,3'-bipyridin]-6-yl)-4-oxobutanamide, Compound 152
Figure imgf000155_0002
[0734] 1H NMR (400 MHz, DMSO-d6): 10.67 (s, IH), 8.75-8.69 (m, 2H), 8.42 (s, IH), 8.20- 8.10 (m, 2H), 7.96 (s, IH), 6.35 (bs, IH), 4.05-4.00 (m, 2H), 3.90-3.80 (m, 2H), 3.20-3.10 (m, 2H), 3.10-2.95 (m, IH), 2.37 (s, 3H), 2.07 (bs, 5H), 1.19 (d, J=6.8 Hz, 3H). MS (ESI) m/z 419 [M+H]+.
[0735] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(5- (pyrimidin-5-yl)pyridin-2-yl)butanamide, Compound 153
Figure imgf000156_0001
[0736] 1H NMR (400 MHz, DMSO-ife): 10.73 (s, IH), 9.19 (d, J= 2.2 Hz, 3H), 8.79 (d, J= 2.2 Hz, IH), 8.25-8.18 (m, 2H), 6.32 (brs, IH), 4.03 (t, J= 5.7 Hz, 2H), 3.86 (s, 2H), 2.89 (t, J= 6.2 Hz, 2H), 2.75 (t, J= 6.6 Hz, 2H), 2.08 (s, 5H). MS (ESI) m/z 392.15 [M+H]+.
[0737] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(6- (pyrimidin-5-yl)pyridin-3-yl)butanamide, Compound 154
Figure imgf000156_0002
[0738] 1H NMR (400 MHz, DMSO-i¾: 10.44 (s, IH), 9.39 (s, 2H), 9.20 (s, IH), 8.89 (s, IH), 8.22-8.20 (dd, J= 2.2 Hz, 8.8 Hz, IH ), 8.10 (d, J= 8.4 Hz, IH), 6.33 (brs, IH), 4.02 (t, J= 5.7 Hz, 2H), 3.86 (s, 2H), 2.90 (d, J= 6.2 Hz, 2H), 2.71 (t, J= 6.2 Hz, 2H), 2.08 (s, 5H). MS (ESI) m/z 392.15 [M+H]+.
[0739] Synthesis of N-(5-fluoro-6-phenylpyridin-3-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 156
Figure imgf000156_0003
[0740] 1H NMR (400 MHz, DMSO-i¾: 10.62 (s, IH), 8.64 (s, IH), 8.15 (dd, J=13.6, 2.0Hz, IH), 7.92-7.85 (m, 2H), 7.55-7.56 (m, 2H), 7.56-7.40 (m, IH), 6.35 (bs, IH), 4.03 (t, J=5.6 Hz, 2H), 3.87 (bs, 2H), 2.95-2.85 (m, 2H), 2.75-2.65 (m, 2H), 2.08 (bs, 5H); MS (ESI) m/z 408 [M+H]+. [0741] Synthesis of 4 2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(5-(6- methylpyrazin-2-yl)pyridin-2-yl)-4-oxobutanamide, Compound 157
Figure imgf000157_0001
[0742] 1H MR (400 MHz, DMSO- ): 10.80 (s, IH), 9.08 - 9.07 (m, 2H), 8.54 - 8.47 (m, 2H), 8.21 - 8.19 (m, IH), 6.34 (br, IH), 4.13 - 4.04 (m, 3H), 3.66 - 3.61 (m, 2H), 2.95 - 2.86 (m, 2H), 2.77 - 2.75 (m, 2H), 2.57 (s, 3H), 2.08 (s, 3H), 1.06 - 1.05 (m, 3H). LC-MS: m/z 420 [M+H]+.
[0743] Synthesis of N-(3-fluoro-5'-methyl-[2,3,-bipyridin]-5-yl)-4-(2-methyl-6,7- dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 158
Figure imgf000157_0002
[0744] 1H NMR (400 MHz, DMSO-<¾): 10.69 (s, IH), 8.85 (s, IH), 8.67 (s, IH), 8.47 (m, IH), 8.20 - 8.17 (m, IH), 8.07 (s, IH), 6.32 (br, IH), 4.03 (t, J=5.6 Hz, 2H), 3.87 (br, 2H), 2.93 - 2.90 (m, 2H), 2.73 - 2.70 (m, 2H), 2.38 (s, 3H), 2.08 (br, 5H). LC-MS: m/z 423 [M+H]+.
[0745] Synthesis of N-(3-(3-chlorophenyl)isoxazol-5-yl)-2-methyl-4-(2-methyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 159
Figure imgf000157_0003
[0746] 1H NMR (400 MHz, DMSO-i¾: 11.86 (br, IH), 7.84 - 7.77 (m, 2H), 7.53 - 7.51 (m, 2H), 6.66 (br, IH), 6.34 (br, IH), 4.03 - 4.00 (m, 2H), 3.84 (br, 2H), 3.07 - 2.95 (m, 2H), 2.07 (br, 5H), 1.74 (br, IH), 1.18 - 1.16 (d, J=4.8 Hz, 3H). LC-MS: m/z 428 [M+H] +.
[0747] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(4- methyl-2,3'-bipyridin-6'-yl)-4-oxobutanamide, Compound 160 and Compound 161
Figure imgf000158_0001
[0748] The product was subjected to chiral chromatography to afford enantiomeric pure compound 160 and compound 161.
[0749] 1H NMR (400 MHz, DMSO-<¾: 10.66 (s, IH), 8.73 (s, IH), 8.69 (s, IH), 8.42 (s, IH), 8.17-8.14 (m, 2H), 7.95 (s, IH), 6.33 (brs, IH), 4.14-4.07 (m, 2H), 3.65-3.60 (dd, J= 9.2 Hz, 11.8 Hz IH), 3.4 (s, IH) 2.97-2.86 (m, 2H), 2.74 (t, J= 6.1 Hz, 2H), 2.37 (s, 3H), 2.32-2.28 (m, IH), 2.08 (brs, 3H), 1.05 (d, J= 5.7 Hz, 3H). MS (ESI) m/z 419.15 [M+H]+.
[0750] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(5'- fluoro-[2,3'-bipyridin]-5-yl)-4-oxobutanamide, Compound 162
Figure imgf000158_0002
[0751] 1H NMR (400 MHz, DMSO- ): 10.46(s, IH), 9.13 (s, IH), 8.88 (br, IH), 8.60 - 8.59 (m, IH), 8.30 - 8.26 (m, IH), 8.21 - 8.18 (m, IH), 8.11 - 8.08 (m, IH), 6.34 (br, IH), 4.13 - 4.04 (m, 2H), 3.66 - 3.61 (m, IH), 3.36 (br, IH), 3.00 - 2.80 (m, 2H), 2.73 - 2.66 (m, 2H), 2.28 (br, IH), 2.08 (s, 3H), 1.06 - 1.05 (m, 3H). LC-MS: m/z 423 [M+H]+. [0752] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-N-(5'- fluoro-[3,3'-bipyridin]-6-yl)-4-oxobutanamide, Compound 163
Figure imgf000159_0001
[0753] 1H NMR (400 MHz, DMSO-d6): 10.74 (s, IH), 8.86 (s, IH), 8.79 - 8.78 (m, IH), 8.59 - 8.58 (m, IH), 8.24 - 8.21 (m, IH), 8.18 - 8.14 (m, 2H), 6.34 (br, IH), 4.13 - 4.05 (m, 2H), 3.66 - 3.60 (m, IH), 3.40 (br, IH), 2.95 - 2.86 (m, 2H), 2.76 - 2.73 (m, 2H), 2.28 (br, IH), 2.08 (s, 3H), 1.06 - 1.04 (m, 3H). LC-MS: m/z 423 [M+H]+.
[0754] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(5- fluoro-[2,3'-bipyridin]-6'-yl)-4-oxobutanamide, Compound 164
Figure imgf000159_0002
[0755] 1H NMR (400 MHz, DMSO- 6): 10.73 (s, IH), 8.99 (d, J=2.0 Hz, IH), 8.66 (d, J=2.8 Hz, IH), 8.42-8.38 (m, IH), 8.20-8.12 (m, IH), 8.12-8.06 (m, IH), 7.89-7.80 (m, IH), 6.33 (bs, IH), 4.15-4.00 (m, 2H), 3.67-3.60 (m, IH), 3.55-3.35 (m, IH), 3.00-2.80 (m, 2H), 2.80-2.70 (m, 2H), 2.30-2.20 (m, IH), 2.08 (bs, 3H), 1.05 (d, J=5.6 Hz, 3H). MS (ESI) m/z 423 [M+H]+.
[0756] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-N-(4- fluoro-[3,3'-bipyridin]-6-yl)-4-oxobutanamide, Compound 165
Figure imgf000159_0003
[0757] 1H NMR (400 MHz, DMSO- ): 10.99 (s, IH), 8.80 (s, IH), 8.65-8.55 (m, 2H), 8.06- 7.98 (m, 2H), 7.56-7.50 (m, IH), 6.33 (bs, IH), 4.15-4.00 (m, 2H), 3.67-3.60 (m, IH), 3.50-3.35 (m, IH), 3.00-2.80 (m, 2H), 2.80-2.70 (m, 2H), 2.35-2.20 (m, IH), 2.08 (bs, 3H), 1.05 (d, J=4.8 Hz, 3H). MS (ESI) m/z 423 [M+H]+.
[0758] Synthesis of N-(3,5,-dimethyl-[2,3,-bipyridin]-5-yl)-4-(2-methyl-6,7- dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 168
Figure imgf000160_0001
[0759] 1H NMR (400 MHz, DMSO-i 6): 10.29 (s, IH), 8.68 (s, IH), 8.53 (d, J=2.0 Hz, IH), 8.43 (d, J=1.6 Hz, IH), 8.01 (d, J=2.0 Hz, IH), 7.76 (s, IH), 6.35 (bs, IH), 4.03 (t, J=5.6 Hz, 2H), 3.87 (bs, 2H), 2.95-2.85 (m, 2H), 2.72-2.65 (m, 2H), 2.36 (s, 3H), 2.33 (s, 3H), 2.08 (bs, 5H); MS (ESI) m/z 419 [M+H]+.
[0760] Synthesis ofN-(3,3'-bipyridin-6-yl)-4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 169 and Compound 170
Figure imgf000160_0002
[0761] The product was subjected to chiral chromatography to afford enantiomeric pure compound 169 and compound 170.
[0762] 1H NMR (400 MHz, DMSO-<¾: 10.67 (s, IH), 8.94 (d, J= 2.0 Hz, IH), 8.71 (s, IH), 8.58 (d, J= 4.0 Hz, IH), 8.16-8.11 (m, 3H), 7.50-7.47 (dd, J = 4.4 Hz, 7.6 Hz, IH), 6.3 (brs, IH), 4.14-4.06 (m, 2H), 3.65-3.60 (m, IH), 3.4 (s, IH), 2.97-2.87 (m, 2H), 2.75 (t, J= 12.8 Hz, 2H), 2.30 (d, J= 14.8 Hz, IH), 2.08 (s, 3H), 1.05 (d, J= 6.4 Hz, 3H); MS (ESI) m/z 405.18 [M+H]+. [0763] Synthesis of Ν-([2,3'-Ηρ}τϊ(ϋη]-6'-^)-4-(2,6-(1ΐηΐ6 1-6,7-ά (ΐΓορ ταζο1ο[1 ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 171
Figure imgf000161_0001
[0764] 1H NMR (400 MHz, DMSO- ): 10.72 (s, IH), 9.03 (m, IH), 8.67 - 8.66 (m, IH), 8.45 - 8.42 (m, IH), 8.17 (d, J=8.4 Hz, IH), 8.01 - 7.99 (m, IH), 7.91 - 7.87 (m, IH), 7.38 - 7.35 (m, IH), 6.34 (br, IH), 4.14 - 4.06 (m, 2H), 3.66 - 3.60 (m, IH), 2.97 - 2.86 (m, 2H), 2.76 - 2.73 (m, 2H), 2.29 (br, IH), 2.08 (s, 3H), 1.05 (m, 3H). MS (ESI) m/z 405 [M+H]+.
[0765] Amide Coupling Method B:
[0766] Triethylamine (1.3 eq) and isobutyl chloroformate (1.0 equiv.) were added dropwise to a stirred solution of acid (1.1 equiv.) in dichloromethane (0.05 M) at 0 °C and stirred for 15 minutes before adding the respective amine (1.0 equiv.). The reaction was stirred from 0 °C to room temperature for 3 hours. Upon completion the reaction was diluted with dichloromethane and basified to pH 9.0 with saturated sodium carbonate solution. The aqueous layer was extracted with dichloromethane, organic phase combined, washed with brine, dried over sodium sulphate, filtered and concentrated in vacuo. Crude was purified with reversed phase HPLC to afford the pure product.
[0767] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(6- ( yridin-3-yl)pyridazin-3-yl)butanamide, Compound 51
Figure imgf000161_0002
[0768] 1H NMR (400 MHz, DMSO- 6): 11.32 (S, IH), 9.26 (d, J=2 Hz, IH), 8.70 - 8.68 (m, IH), 8.48 - 8.46 (m, IH), 8.40 (d, J=9.2 Hz, IH), 8.30 (d, J=9.6 Hz, IH), 7.59 - 7.56 (m, IH), 6.33 (bs, IH), 4.02 (t, J=5.8 Hz, 2H), 3.86 (bs, 2H), 2.91 - 2.90 (m, 2H), 2.83 - 2.80 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 392 [M+H]+.
[0769] Synthesis of N-(6-(3-fluorophenyl)pyridin-3-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 84
Figure imgf000162_0001
[0770] 1H NMR (400 MHz, DMSO-i¾: 10.73 (s, IH), 8.83 (d, J=2.4 Hz, IH), 8.16-8.14 (m, IH), 7.99-7.97 (m, IH), 7.90-7.88 (m, IH), 7.85-7.82 (m, IH), 7.53-7.47 (m, IH), 7.24-7.19 (m, IH), 6.34 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.87 (s, 2H), 2.92-2.89 (m, 2H), 2.71-2.68 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 408 [M+H]+.
[0771] Synthesis of N-(6-(3-fluorophenyl)pyridazin-3-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 98
Figure imgf000162_0002
[0772] 1H NMR (400 MHz, DMSO-c¾: 11.30 (s, IH), 8.39-8.37 (m, IH), 8.28-8.26 (m, IH), 7.97-7.91 (m, 2H), 7.61-7.56 (m, IH), 7.36-7.31 (m, IH), 6.34 (br s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.91-2.90 (m, 2H), 2.82-2.79 (m, 2H), 2.08 (s, 5H). LC-MS: m/z 409 [M+H]+.
[0773] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(6-(5- methylpyridin-3-yl)pyridazin-3-yl)-4-oxobutanamide, Compound 104
Figure imgf000163_0001
[0774] 1H NMR (400 MHz, DMSO-i¾: 11.31 (s, 1H), 9.06 (s, 1H), 8.53 (s, 1H), 8.40-8.38 (m, 1H), 8.30-8.27 (m, 2H), 6.33 (br s, 1H), 4.02 (t, J=6.0 Hz, 2H), 3.86 (s, 2H), 2.91-2.90 (m, 2H), 2.82-2.79 (m, 2H), 2.41 (s, 3H), 2.08 (s, 5H). LC-MS: m/z 406 [M+H]+.
[0775] Amide Coupling Method C:
[0776] Amine (1.1 equiv.) was added to a stirred solution of acid, Ν,Ν-diisopropyl ethyl amine (3.0 equiv.) and HATU (1.5 equiv.) in DMF (0.2 M) after 5 minutes. Solution is left to stir at room temperature for 3 h._Upon completion the reaction was diluted with water and extracted with dichloromethane. The organic phase was combined, washed with brine, dried over sodium sulphate, filtered and concentrated in vacuo. Crude was purified by Shimadzu semi-prep and lyophilized to afford a white solid.
[0777] Synthesis of N-(2'-fluoro-[ 1 , 1 '-biphenyl]-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[ 1,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 64
Figure imgf000163_0002
[0778] 1H NMR (400 MHz, DMSO-i¾): 10.14 (s, 1H), 7.70 (d, J=8.4 Hz, 2H), 7.53 - 7.48 (m, 3H), 7.39 - 7.35 (m, 1H), 7.31 - 7.26 (m, 2H), 6.35 (br, 1H), 4.02 (t, J=6.0 Hz, 2H), 3.87 (br, 2H), 2.91 - 2.88 (m, 2H), 2.68 - 2.65 (m, 2H), 2.08 (br, 5H). LC-MS: m/z 407 [M+H]+.
[0779] Synthesis of N,N-dimethyl-4'-(4-(2-methyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin-4(5H)- yl)-4-oxobutanamido)-[l,l '-biphenyl]-3-carboxamide, Compound 68
Figure imgf000164_0001
[0780] 1H NMR (400 MHz, DMSO-t¼): 10.11 (s, IH), 7.72 - 7.62 (m, 6H), 7.49 (t, J=7.6 Hz, IH), 7.34 - 7.32 (m, IH), 6.33 (bs, IH), 4.02 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.97 (d, J=23.6 Hz, 6H), 2.91 - 2.87 (m, 2H), 2.68 - 2.65 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 460 [M+H]+.
[0781] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(3'- morpholinobiphenyl-4-yl)-4-oxobutanamide, Compound 73
Figure imgf000164_0002
[0782] 1H NMR (400 MHz, DMSO- 6): 10.05 (s, IH), 7.66 (d, J= 8.8 Hz, 2H), 7.59 (d, J= 8.8 Hz, 2H), 7.27 (t, J= 8.0 Hz, IH), 7.13 (s, IH), 7.06 (d, J= 7.6 Hz, IH), 6.90 (d, J= 6.4 Hz, IH), 6.32 (bs, IH), 4.02 (t, J= 6.4 Hz, 2H), 3.86 (bs, 2H), 3.75 (t, J= 4.8 Hz, 4H), 3.17 (t, J= 4.8 Hz, 4H), 2.88 (t, J= 6.4 Hz, 2H), 2.65 (t, J= 6.4 Hz, 2H), 2.08 (bs, 5H). MS (ESI) m/z 474.33 [M+H]+.
[0783] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(l- phenylpiperidin-4-yl)butanamide, Compound 81
Figure imgf000164_0003
[0784] 1H NMR (400 MHz, DMSO- ): 7.82 (d, J= 7.6 Hz, IH), 7.18 (t, J= 6.8 Hz, 2H), 6.92 (d, J= 8.0 Hz, 2H), 6.73 (t, J= 7.6 Hz, IH), 6.20 (s, IH), 4.00 (t, J= 6.0 Hz, 2H), 3.82 (s, 2H), 3.80-3.61 (m, 3H), 2.80-2.74 (m, 4H), 2.38 (t, J= 6.8 Hz, 2H), 2.08 (bs, 5H), 1.79 (d, J= 9.6 Hz, 2H), 1.55-1.44 (m, 2H). MS (ESI) m/z 396.40 [M+H]+.
[0785] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(6- (m-tolyl)pyridazin-3-yl)butanamide, Compound 99
Figure imgf000165_0001
[0786] 1H NMR (400 MHz, DMSO- ): 11.25 (s, IH), 8.35 (d, J=9.6 Hz, IH), 8.19 (d, J=9.6 Hz, IH), 7.93 (bs, IH), 7.87 (d, J=7.6 Hz, IH), 7.42 (t, J=7.6 Hz, IH), 7.32 - 7.30 (m, IH), 6.30 (bs, IH), 4.04 - 4.01 (t, J=5.8 Hz, 2H), 3.87 (bs, 2H), 2,91 - 2.90 (m, 2H), 2.82 - 2.79 (m, 2H), 2.41 (s, 3H), 2.08 (bs, 5H). LC-MS: m/z 405 [M+H]+.
[0787] Synthesis of N-(6-(5-fluoropyridin-3-yl)pyridazin-3-yl)-4-(2-methyl-6,7- dihydropyrazolo[ 1 ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 103
Figure imgf000165_0002
[0788] 1H NMR (400 MHz, DMSO-i¾): 11.38 (s, IH), 9.18 (s, IH), 8.71 (d, J=2.8 Hz, IH), 7.45-7.33 (m, 3H), 6.34 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.95-2.85 (m, 2H), 2.85- 2.75 (m, 2H), 2.08 (bs, 5H). MS (ESI) m/z 410 [M+H]+.
[0789] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(6-(5- (trifluoromethyl)pyridin-3-yl)pyridazin-3-yl)butanamide, Compound 105
Figure imgf000166_0001
[0790] 1H MR (400 MHz, DMSO-< 6): 11.41 (s, IH), 9.58 (m, IH), 9.10 (m, IH), 8.83 (m, IH), 8.46 (m, 2H), 6.33 (bs, IH), 4.03 (t, J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.93 - 2.91 (m, 2H), 2.84 - 2.81 (m, 2H), 2.08 (bs, 5H). LC-MS: m/z 460 [M+H]+.
[0791] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(5-methyl- 6-(pyridin-3-yl)pyridazin-3-yl)-4-oxobutanamide, Compound 106
Figure imgf000166_0002
[0792] 1H NMR (400 MHz, DMSO-d6): 11.21 (s, IH), 8.81 - 8.80 (m, IH), 8.69 - 8.68 (m, IH), 8.28 (s, IH), 8.07 - 8.04 (m, IH), 7.57 - 7.54 (m, IH), 6.33 (s, IH), 4.03 (t, J=5.8 Hz, 2H), 3.87 (bs, 2H), 2.93 - 2.89 (m, 2H), 2.81 - 2.78 (m, 2H), 2.34 (s, 3H), 2.08 (bs, 5H). LC-MS: m/z 406 [M+H]+.
[0793] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(6-(6- methylpyridin-3-yl)pyridazin-3-yl)-4-oxobutanamide, Compound 107
Figure imgf000166_0003
[0794] 1H NMR (400 MHz, DMSO- 6): 11.29 (s, IH), 9.14 - 9.13(m, IH), 8.39 - 8.34 (m, 2H), 8.28 - 8.25 (m, IH), 7.42 (d, J=8.0 Hz, IH), 6.33 (bs, IH), 4.03 (t,. J=6.0 Hz, 2H), 3.87 (bs, 2H), 2.93 - 2.90 (m, 2H), 2.82 - 2.79 (m, 2H), 2.55 (s, 3H), 2.08 (bs, 5H). LC-MS: m/z 406 [M+H]+. [0795] Synthesis of N-(5'-methyl-2,3'-bipyridin-5-yl)-4-(2-methyl-6,7-dihydropyrazolo[l,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 110
Figure imgf000167_0001
[0796] 1H NMR (400 MHz, DMSO- ): 10.35 (s, IH), 9.01 (s, IH), 8.85 (d, J= 2.0 Hz, IH), 8.42 (s, IH), 8.21 (s, IH), 8.16 (d, J= 10.8 Hz, IH), 7.99 (d, J= 8.8 Hz, IH), 6.34 (s, IH), 4.02 (t, J= 5.6 Hz, 2H), 3.86 (bs, 2H), 2.93-2.88 (m, 2H), 2.73-2.67 (m, 2H), 2.37 (s, 3H), 2.08 (bs, 5H). MS (ESI) m/z 405.33 [M+H]+.
[0797] Synthesis of 2-methyl-4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4- oxo-N-(6-phenylpyridazin-3-yl)butanamide, Compound 146
Figure imgf000167_0002
[0798] 1H NMR (400 MHz, DMSO-d6): 11.25 (s, IH), 8.37 (d, J=9.6 Hz, IH), 8.21 (d, J=9.6 Hz, IH), 8.11 - 8.09 (m, 2H), 7.55 - 7.50 (m, 3H), 6.31 (s, IH), 4.02 (t, J=6.0 Hz, 2H), 3.86 (br, 2H), 3.25 - 3.21 (m, IH), 3.07 - 3.00 (m, IH), 2.06 (br, 5H), 1.22 (d, J=7.2 Hz, 3H). LC-MS: m/z 405 [M+H]+.
[0799] Synthesis of N-(2-fluorobiphenyl-4-yl)-4-(2-methyl-6,7-dihydropyrazolo[l ,5- a]pyrimidin-4(5H)-yl)-4-oxobutanamide, Compound 155
Figure imgf000167_0003
[0800] 1H NMR (400 MHz, DMSO-c 6): 11.32 (s, IH), 9.27 (d, J=2.0 Hz, IH), 8.69 (dd, J=4.8, 1.6 Hz, IH), 8.47 (dt, J=8.4, 1.6 Hz, IH), 8.45-8.38 (m, IH), 8.32-8.28 (m, IH), 7.53-7.60 (m, IH), 6.35 (bs, IH), 4.05-4.00 (m, IH), 3.95-3.80 (m, 2H), 3.25-3.15 (m, 2H), 3.10-2.95 (m, IH), 2.75-2.55 (m, IH), 2.06 (bs, 5H), 1.22 (d, J=7.2 Hz, 3H). MS (ESI) m/z 406 [M+H]+.
[0801] Synthesis of 4-(2-methyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-4-oxo-N-(6- (pyrazin-2-yl)pyridin-3-yl)butanamide, Compound 166
Figure imgf000168_0001
[0802] 1H NMR (400 MHz, DMSO-c¾): 10.46 (s, IH), 9.47 (d, J= 1.6 Hz, IH), 8.89 (s, IH), 8.70 (t, J= 2.8 Hz, IH), 8.65 (d, J=2.4 Hz, IH), 8.31-8.29 (m, IH), 8.26-8.23 (dd, J= 2.4 Hz, 8.8 Ηζ,ΙΗ), 6.3 (brs, IH), 4.06-4.01 (m, 2H), 3.87 (brs, IH), 3.17 (d, J =5.2 Hz, IH), 2.92 (t, J= 6.4 Hz, 2H), 2.73-2.70 (t, J= 6.4 Hz, 2H), 2.08 (s, 5H); MS (ESI) m/z 392.2 [M+H]+.
[0803] Synthesis of 4-(2,6-dimethyl-6,7-dihydropyrazolo[l,5-a]pyrimidin-4(5H)-yl)-N-(6-(5- fluoropyridin-3-yl)pyridazin-3-yl)-4-oxobutanamide, Compound 167
Figure imgf000168_0002
[0804] 1H NMR (400 MHz, DMSO-rf6): Π -38 (s, 1H), 9.18 (s, IH), 8.71 (d, J=2.8 Hz, IH), 8.55-8.45 (m, 3H), 6.34 (bs, IH), 4.15-4.00 (m, 2H), 3.68-3.60 (m, IH), 3.45-3.30 (m, IH), 3.00-2.85 (m, 2H), 2.85-2.80 (m, 2H), 2.35-2.20 (m, IH), 2.07 (bs, 3H), 1.05 (d, J=4.8 Hz, 3H); MS (ESI) m/z 424 [M+H]+.
[0805] Materials and Methods
[0806] Cell lines and culture conditions: [0807] HEK293-STF cell line was modified from Human embryonic kidney cell line HEK293 transfected with the STF reporter. HEK293-STF3 A cell line was further modified from
HEK293-STF cell line to express Wnt3A. This cell line was used to identify compounds that regulate either early or late signaling components of the Wnt pathway. These two cell lines were obtained from David Virshup's laboratory, Duke-NUS. L-Wnt3 A (ATCC, #CRL-2647) cell line was used for providing Wnt3 A conditioned media. The three cell lines were grown in DMEM with 10% FBS incubated in 37°C with 5% C02.
[0808] Cell viability assay:
[0809] 5000 cells in 75 μΐ culture media were seeded in each well of black 96 well plates (Greiner #655090) and incubated overnight at 37°C. 25 μΐ of serially diluted compound was added to the cells giving final concentration of 50 μΜ to 1.5 nM. After 1 day of treatment, 100 μΐ of CellTiter-Glo Luminescent Cell Viability Assay reagent (#G7571, Promega) was added to each well and incubated for 10 minutes at room temperature. Luminescence was measured using Tecan Safire2 microplate reader.
[0810] STF3A assay:
[0811] 2xl04 HEK293-STF3A cells in 75 μΐ culture media were seeded in each well of white 96 well plates (Greiner #655098) and incubated overnight at 37°C. 25 μΐ serially diluted compound was added to the cells to give final concentration of 50 μΜ to 1.5 nM. After 1 day of treatment, 100 μΐ of Steady-Glo Luciferase Assay reagent (#E2520, Promega) was added to each well and incubated for 10 minutes at room temperature. Luminescence was measured using Tecan Safire2 plate reader.
[0812] STF/WNT3A conditioned medium (STF/WNT3A CM) assay:
[0813] L-Wnt3A cells were cultured in three T-175 flasks at 3*104 cells/ml in 30 ml culture medium per flask. After 4 days of incubation, the Wnt3 A conditioned media were harvested and then centrifuged at 2000 rpm for 10 minutes to remove the debris. The Wnt3A conditioned media were stored at -20 °C if not used immediately.
[0814] 2xl04 HEK293-STF cells in 25 μΐ culture media were added in each well of white 96 well plates (Greiner #655098). 25 μΐ serially diluted compound was added to the cells. After 4 hours of incubation, 100 μΐ Wnt-3A conditioned medium was added to the cells. The final concentration of compound ranged from 33 μΜ to 1 nM. After incubation for 1 day at 37 °C, 100 μΐ of Steady-Glo® Luciferase Assay reagent (#E2520, Promega) was added to each well and incubated for 10 minutes at room temperature. Luminescence was measured using Tecan Safire2 microplate reader.
[0815] Western blot:
[0816] 8.0 x 105 cells in 2.5 ml media were seeded in T-25 flasks. Compounds were diluted to 600 nM in 1 ml medium and 0.5 ml was added to the T-25 flask to give a final concentration of 100 nM. After incubation in 37 °C for two days, the culture media were collected and centrifuged at 2000 rpm for 10 min. The supernatants were collected and 32 μΐ from each sample was used for SDS PAGE gel electrophoresis. After transferring the separated proteins to the membrane it was incubated with the primary Wnt3A antibody (1 : 1000, #09-162, Millipore) overnight. After washing with Tris-Buffered Saline containing 0.05% Tween 20 (TBST), the membrane was incubated with polyclonal Goat anti-Rabbit IgG HRP-conjugated secondary antibody (1 :3000, #P0448) for 1 hour at room temperature. After washing with TBST, the membrane was developed with Amersham™ ECL™ Select Western Blotting Detection Reagent (#RPN2235, GE Healthcare Life Sciences) and documented with Bio-Rad Molecular Imager VersaDoc MP.
[0817] MMTV-Wntl Mouse Model 1 :
[0818] 8-10 weeks old Female BALBc mice were anesthetised with 150mg kg Ketamine +75mg/kg Xylazine. Under aseptic conditions, skin near the 4 mammary fat pad was incised. Mammary fat pad was tweaked with forceps and tumor fragment ~ 2mm3 was implanted. The incision was closed using a tissue adhesive. Animals were randomised into groups of eight and treated daily with the test compounds for 14 days. Tumors were measured in two dimensions using calipers, and volume was calculated using the formula: Tumor Volume (mm3) = w2 x 1 12. Maximum tumor volume limit was 2000mm .
[0819] Wnt3A palmitoylation assay:
[0820] The assay used to determine the inhibition of the palmitoylation of Wnts by compound was described by Yap et al, (Yap MC, Kostiuk MA, Martin DD, Perinpanayagam MA, Hak PG, Siddam A, Majjigapu JR, Rajaiah G, Keller BO, Prescher JA, Wu P, Bertozzi CR, Falck JR, Berthiaume LG. 2010. Rapid and selective detection of fatty acylated proteins using omega- alkynyl-fatty acids and click chemistry. J Lipid Res. 51(6):1566-1580) with some modification. 3 x 106 HeLa cells were seeded in 10 cm culture dish and incubated at 37°C overnight. The cells were transfected with 5 μg pCDNA3.2/V5-Wnt3a vector (Najdi R, Proffitt K, Sprowl S, Kaur S, Yu J, Covey TM, Virshup DM, Waterman ML. 2012. A uniform human Wnt expression library reveals a shared secretory pathway and unique signaling activities. Differentiation, 84(2), 203- 213. doi:10.1016/j.diff.2012.06.004) to over-express V5-tagged Wnt3a. After six hours, the cells were washed with PBS and treated with 100 μΜ alkyne palmitate in medium with 5% BSA. ΙΟΟηΜ compound or DMSO was added and the cells were incubated overnight at 37°C. The cells were lyzed and 600 μg cell lysate was collected and incubated with anti-V5 antibody (Invitrogen) followed by the pull down of V5-Wnt3a with the addition of Protein A/G agarose beads (Thermo scientific). The pulled down lysates containing V5-Wnt3a was click-reacted with biotin-azide (Invitrogen). The biotin-labelled protein lysate was then separated on SDS-PAGE and transferred to a nitrocellulose membrane. The membrane was incubated with primary anti- V5 antibody, followed by secondary anti-mouse Dylight 680 (Thermo scientific) to detect V5- Wnt3a. The membrane was then incubated with streptavidin-Dylight 800 (Thermo scientific) to detect biotin labelled Wnt3a. The signals were captured on the Odyssey CLx Infrared Imaging System (LI-COR Bioscience).
[0821] Soft agar Assay:
[0822] AsPC-1 cells were maintained in RPMI164 supplemented with 10% FBS, 2 mM L- glutamine and P/S (100 units/ml penicillin and 100 μ§ ιη1 streptomycin). HPAF-II cells were maintained in MEM (Eagles') supplemented with 10% FBS, 2 mM L-glutamine and P/S (100 units/ml penicillin and 100 μg/ml streptomycin). CFPAC-1 cells were maintained in Iscove MEM supplemented with 10% FBS, 2 mM L-glutamine and P/S (100 units/ml penicillin and 100 μg/ml streptomycin).
[0823] 600 μΐ of 0.6% agar was added to 24-well plate to form the base layer. Then a middle layer of 0.36% agar (containing 5000 cells and serially diluted compound) was added on to the base layer. Finally 500 μΐ of fresh growth medium was added to the top of the middle layer. The plates were incubated at 37 °C with 5% carbon dioxide in a humidified incubator for 2 weeks. Formation of colonies was observed using a light microscope. When the colony size was larger than 500 μηι, 70 μΐ MTT (5 mg/ml) was added to each well and the plates were incubated at 37 °C for at least 2 hours. Colonies were counted with GelCount® instrument. The colony counts were plotted against compound concentrations using the GraphPad Prism software. The software was also used to perform non-linear curve fitting and calculation of compound concentration that inhibited 50% colony formation.
[0824] Results:
[0825] Compounds of the present invention specifically inhibit mammalian PORCN.
[0826] PORCN-null HT1080 cells were transfected with mammalian or Xenopus PORCN expression plasmids, along with WNT3 A, STF reporter and mCherry as transfection control. 6 hours after transfection, cells were treated with the compounds or DMSO as indicated, and the following day assayed for luciferase. Xenopus PORCN was resistant to the inhibitory effects of compounds. The two compounds, Compound 51 and Compound 110 inhibit the activity of mammalian porcupine (Figure 1).
[0827] Treatment with Compounds 51 and 110 decreased tumor growth in all the treated mice (Figure 2 and Figure 3).
[0828] Palmitoylation of Wnt3a is inhibited by Compound 51 (Figure 4). The Wnt3a-V5 was visualized using anti-V5 antibody, followed by anti-mouse Dylight 680. B. Biotin-azide clicked palmitate in V5-Wnt3a was detected with streptavidin-Dylight 800. A non-specific band was observed above the Wnt3a-V5 protein. Biotin-azide clicked palmitate was detected with streptavidin-Dylight 800 (lower band). DMSO was used as negative control. Lane 1: Without Alkyne palmitate; Lane 2: untreated control, DMSO + Alkyne palmitate; Lane 3: lOOnM Compound 51 + Alkyne palmitate.
[0829] Results of the Soft Agar Assay are shown in Table 1 and Figures 5 to 7. [0830] Table 1: Fifty percent colony growth inhibitory concentration (IC50, μΜ) of Compound 51 on pancreatic cell lines AsPC-1 and HPAF-II. Results shown below are mean ± standard deviation (SD) from two independent experiments.
Figure imgf000173_0001
Table 1
[0831] The results of the MMTV-Wntl Mouse Model are shown in Figure 2 and Table 2:
Figure imgf000173_0002
Table 2
[0832] Throughout the specification and the claims which follow, unless the context requires otherwise, the word 'comprise', and variations such as 'comprises' and 'comprising', will be understood to imply the inclusion of a stated integer, step, group of integers or group of steps but not to the exclusion of any other integer, step, group of integers or group of steps.
[0833] All patents and patent applications mentioned throughout the specification of the present invention are herein incorporated in their entirety by reference. [0834] The invention embraces all combinations of preferred and more preferred groups and embodiments of groups recited above.

Claims

1. A compound of formula (I),
Figure imgf000175_0001
or a pharmaceutically acceptable salt, solvate or polymorph thereof, including all tautomers and stereoisomers thereof wherein:
Ri represents H; optionally substituted alkyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and - N(C1-3alkyl)2); -C(0)Oalkyl; haloalkyl; haloalkoxy; or -alkylaryl; each R2 independently represents H; optionally substituted alkyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and -N(C1-3alkyl)2); -alkylaryl; optionally substituted carbocyclyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy and halo); optionally substituted heterocyclyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1- alkyl and- C(0)NHC1-6alkyl); -NHalkyl; -N(alkyl)2; amino; hydroxyl; alkoxy or halo; n represents 0, 1 or 2;
R3 represents H or alkyl;
R4 represents H or alkyl;
R5 represents H or alkyl; W and X each independently represent C=0; C=S; or CH2;
Y represents aryl; heteroaryl; optionally substituted carbocyclyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, Q. alkoxy, C1-6haloalkyl, C1-6haloalkoxy and halo); or optionally substituted heterocyclyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, Ci-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHCi-6alkyl); and
Z represents optionally substituted alkyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and -N^Q. 3alkyl)2); aryl; heteroaryl; -alkylaryl;-alkylheteroaryl; optionally substituted carbocyclyl (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy and halo); optionally substituted heterocyclyl (wherein optional substituents include one or more substituents each
independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1- alkyl and- C(0)NHC1- alkyl); -alkylcarbocyclyl wherein carbocyclyl is optionally substituted (wherein optional substituents include one or more substituents each independently selected from Ci_ 6alkyl, C1-6alkoxy, C1-6haloalkoxy and halo); -alkylheterocyclyl wherein heterocyclyl is optionally substituted (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1- 6alkyl); -arylcarbocyclyl wherein carbocyclyl is optionally substituted (wherein optional substituents include one or more substituents each independently selected from C1-6alkyl, C . 6alkoxy, C1-6haloalkoxy and halo); or -arylheterocyclyl wherein heterocyclyl is optionally substituted (wherein optional substituents include one or more substituents each
independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1- 6alkyl).
2. A compound of formula (I) according to claim 1
Figure imgf000177_0001
or a pharmaceutically acceptable salt, solvate or polymorph thereof, including all tautomers and stereoisomers thereof wherein:
Ri represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and -N(C1-3alkyl)2; -C(0)Oalkyl; haloalkyl; haloalkoxy; or -alkylaryl;
each R2 independently represents H; alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, NH2, -NHC1-3alkyl and -Ν(^.
3alkyl)2; -alkylaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1- alkoxy, C1-6haloalkyl, C ^haloalkoxy and halo;
heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, d-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl; - NHalkyl; -N(alkyl)2; amino; hydroxyl; alkoxy or halo;
n represents 0, 1 or 2;
R3 represents H or alkyl;
R4 represents H or alkyl;
R5 represents H or alkyl;
W and X each independently represent C=0; C=S; or C¾;
Y represents aryl; heteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, Q.
6haloalkoxy and halo; or heterocyclyl which is optionally substituted by one or more substituents each independently selected from
Figure imgf000177_0002
C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)Ci-6alkyl and- C(0)NHCi-6alkyl; and
Z represents alkyl which is optionally substituted by one or more substituents each independently selected from C1-6alkoxy, NH2, - HC1-3alkyl and -N(C1-3alkyl)2; aryl; heteroaryl; -alkylaryl; - alkylheteroaryl; carbocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy and halo;
heterocyclyl which is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl; - alkylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy and halo; - alkylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl; - arylcarbocyclyl wherein carbocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, C1-6alkoxy, C1-6haloalkyl, C1-6haloalkoxy and halo; and - arylheterocyclyl wherein heterocyclyl is optionally substituted by one or more substituents each independently selected from C1-6alkyl, -C(0)OC1-6alkyl, -C(0)C1-6alkyl and-C(0)NHC1-6alkyl.
3. A compound according to claim 1 or claim 2 wherein ¾ represents H, methyl, ethyl, - C(0)OMe, CF3 or OMe.
4. A compound according to any one of claims 1 to 3 wherein R\ represents methyl.
5. A compound according to any one of claims 1 to 4 wherein each R2 independently represents H or alkyl.
6. A compound according to any one of claims 1 to 5 wherein each R2 represents H.
7. A compound according to any one of claims 1 to 6 wherein n represents 1.
8. A compound according to any one of claims 1 to 7 wherein R3 represents H.
9. A compound according to any one of claims 1 to 8 wherein R4 represents H.
A compound according to any one of claims 1 to 9 wherein R5 represents H.
11. A compound according to any one of claims 1 to 10 wherein W and X each represent C=0.
12. A compound according to any one of claims 1 to 11 wherein Y represents monocyclic heteroaryl or phenyl.
13. A compound according to claim 11 wherein Y represents monocyclic heteroaryl comprising one, two or three ring heteroatoms including one or two nitrogen atoms.
14. A compound according to claim 11 wherein Y represents phenyl.
15. A compound according to any one of claims 12 to 14 wherein Y is substituted by one or more substituents each independently selected from C -e alkyl, C1-6alkoxy, halo and Q.
6haloalkyl.
16. A compound according to any one of claims 1 to 15 wherein Y is monosubstituted.
17. A compound according to any one of claims 1 to 15 wherein Y is unsubstituted.
18. A compound according to any one of claims 1 to 17 wherein Z represents aryl or heteroaryl.
19. A compound according to claim 1, as defined in any one of examples 1 to 171.
20. A compound according to claim 19, wherein compounds having an IC50 against STF3A cells of 10 micromolar or more are excluded.
21. A compound according to claim 1 or claim 2 in which n=l, R1 is methyl, R2 to R5 are all H, W and X are both C=0, Y is 6-membered heterocyclyl having 1 or 2 ring nitrogens and no other heteroatoms and Z is pyridinyl, optionally substituted by methyl.
22. The compound of claim 21 which is 4-(2-methyl-6,7-dihydropyrazolo[l ,5-a]pyrimidin- 4(5H)-yl)-4-oxo-N-(6-(pyridin-3-yl)pyridazin-3-yl)butanamide:
Figure imgf000179_0001
23. The compound of claim 21 which is N-(5'-methyl-2,3'-bipyridin-5-yl)-4-(2-methyl-6,7- dihydropyrazolo[l ,5-a]pyrimidin-4(5H)-yl)-4-oxobutanamide:
Figure imgf000180_0001
24. The compound of any one of claims 1 to 23 wherein the following compounds are excluded:
Figure imgf000180_0002
Figure imgf000181_0001
Figure imgf000182_0001
Figure imgf000183_0001
25. A compound according to any one of claims 1 to 24 for use as a medicament.
26. The compound of any one of claims 1 to 25 having an IC50 against HEK293-STF3A cells of less than about 10 micromolar.
27. A pharmaceutical composition comprising a compound according to any one of claims 1 to 26 in combination with one or more therapeutically acceptable adjuvents, diluents or carriers.
28. The compound of any one of claims 1 to 26 for use in modulation of the WNT pathway.
29. A method of modulating WNT activity comprising exposing a WNT protein or a WNT receptor to a compound according to any one of claims 1 to 26.
30. A method according to claim 29 wherein the method is an in vivo method.
31. Use of a compound according to any one of claims 1 to 26 for modulating WNT activity.
32. The compound of any one of claims 1 to 26 for use in the treatment of a disease or condition associated with WNT pathway activity.
33. A method of treating a disease or condition associated with WNT pathway activity comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 26.
34. Use of a compound according to any one of claims 1 to 26 for treatment of a disease or condition associated with WNT pathway activity.
35. Use of a compound according to any one of claims 1 to 26 in the manufacture of a medicament for the treatment of a disease or condition associated with WNT pathway activity.
36. The compound of claim 32, method of claim 33, use of claim 34 or claim 35 wherein the disease or condition is selected from the group consisting of cancer, fibrosis, stem cell and diabetic retinopathy, rheumatoid arthritis, psoriasis and myocardial infarction.
37. The compound, method or use of claim 36 wherein the cancer is a cancer characterised by high WNT activity.
38. The compound, method or use of claim 36 wherein the disease or condition is a cancer, such as cervical, colon, breast, bladder, head and neck, gastric, lung, ovarian, prostate, thyroid, non-small-cell lung, chronic lymphocytic leukemia, mesothelioma, melanoma, pancreatic adenocarcinoma, basal cell carcinoma, osteosarcoma, hepatocellular carcinoma, Wilm's tumor or medulloblastoma, or a fibrotic disease, such as pulmonary fibrosis, liver fibrosis, skin fibrosis or renal fibrosis, or a degenerative disease, or a metabolic disease such as diabetic retinopathy.
39. Use of a compound according to any one of claims 1 to 26 in diagnosis.
40. A process for preparation of a compound according to any one of claims 1 to 26, wherein W and X each represent C=0, which process comprises reaction of a compound of formula (II)
Figure imgf000185_0001
wherein Rls R2, R2, n, R3 and R4 are as defined in any one of claims 1 to 26, with a compound of formula (III)
Figure imgf000185_0002
(III)
or a protected derivative thereof, wherein R5, Y and Z are as defined any one of claims 1 to 26.
41. A process for preparation of a compound according to any one of claims 1 to 10, wherein W and X each represent CH2, which process comprises conversion of the W and X groups of compounds of formula (I) from C=0 groups into C¾ groups.
42. A process for preparation of a compound according to any one of claims 1 to 10, wherein W and X each represent C=S, which process comprises conversion of the W and X groups of compounds of formula (I) from C=0 groups into C=S groups.
PCT/SG2014/000602 2013-12-17 2014-12-17 Wnt pathway modulators WO2015094119A1 (en)

Priority Applications (13)

Application Number Priority Date Filing Date Title
KR1020167019109A KR102383739B1 (en) 2013-12-17 2014-12-17 Wnt pathway modulators
JP2016540531A JP6452703B2 (en) 2013-12-17 2014-12-17 WNT pathway modulator
CN201480075676.5A CN105992767B (en) 2013-12-17 2014-12-17 WNT pathway modulators
ES14870867.0T ES2687493T3 (en) 2013-12-17 2014-12-17 WNT route modulators
DK14870867.0T DK3083631T3 (en) 2013-12-17 2014-12-17 WNT ROAD MODULARS
US15/105,224 US10189842B2 (en) 2013-12-17 2014-12-17 Wnt pathway modulators
EP14870867.0A EP3083631B1 (en) 2013-12-17 2014-12-17 Wnt pathway modulators
CA2933683A CA2933683A1 (en) 2013-12-17 2014-12-17 Wnt pathway modulators
SG11201604839XA SG11201604839XA (en) 2013-12-17 2014-12-17 Wnt pathway modulators
AU2014367284A AU2014367284B2 (en) 2013-12-17 2014-12-17 WNT pathway modulators
BR112016013640A BR112016013640A8 (en) 2013-12-17 2014-12-17 wnt reaction series modulators, their uses, pharmaceutical composition and process for their preparation
US16/226,398 US10870653B2 (en) 2013-12-17 2018-12-19 WNT pathway modulators
US17/084,507 US11434245B2 (en) 2013-12-17 2020-10-29 WNT pathway modulators

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GBGB1322333.4A GB201322333D0 (en) 2013-12-17 2013-12-17 WNT pathway modulators
GB1322333.4 2013-12-17

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US15/105,224 A-371-Of-International US10189842B2 (en) 2013-12-17 2014-12-17 Wnt pathway modulators
US16/226,398 Division US10870653B2 (en) 2013-12-17 2018-12-19 WNT pathway modulators

Publications (1)

Publication Number Publication Date
WO2015094119A1 true WO2015094119A1 (en) 2015-06-25

Family

ID=50031066

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SG2014/000602 WO2015094119A1 (en) 2013-12-17 2014-12-17 Wnt pathway modulators

Country Status (13)

Country Link
US (3) US10189842B2 (en)
EP (1) EP3083631B1 (en)
JP (1) JP6452703B2 (en)
KR (1) KR102383739B1 (en)
CN (1) CN105992767B (en)
AU (1) AU2014367284B2 (en)
BR (1) BR112016013640A8 (en)
CA (1) CA2933683A1 (en)
DK (1) DK3083631T3 (en)
ES (1) ES2687493T3 (en)
GB (1) GB201322333D0 (en)
SG (1) SG11201604839XA (en)
WO (1) WO2015094119A1 (en)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016026772A1 (en) * 2014-08-19 2016-02-25 Iomet Pharma Ltd Pharmaceutical compound
US10189842B2 (en) 2013-12-17 2019-01-29 Agency For Science, Technology And Research Wnt pathway modulators
WO2019180244A1 (en) * 2018-03-23 2019-09-26 Step Pharma S.A.S. Aminopyrimidine derivatives as ctps1 inhibitors
WO2019179652A1 (en) * 2018-03-23 2019-09-26 Step Pharma S.A.S. Aminopyrimidine derivatives as ctps1 inhibitors
TWI804545B (en) * 2018-03-23 2023-06-11 法商史坦普醫藥簡式股份公司 Compounds
US11987573B2 (en) 2018-10-23 2024-05-21 Step Pharma S.A.S. Aminopyrimidine/pyrazine derivatives as CTPS1 inhibitors

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3204381B1 (en) * 2014-10-08 2022-05-25 Redx Pharma Plc N-pyridinyl acetamide derivatives as wnt signalling pathway inhibitors
JP6824526B2 (en) * 2015-04-14 2021-02-03 国立大学法人京都大学 How to induce cerebral cortex neurons
CN111620874B (en) * 2019-02-28 2022-11-15 汇瀚医疗科技有限公司 Substituted 5, 6-bis-heterocyclic amino compounds as WNT signaling pathway inhibitors and therapeutic uses thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070219183A1 (en) * 2006-03-17 2007-09-20 Wyeth Dihydropyrazolo[1,5-A]pyrimidine and dihydroimidazo[1,5-A]pyrimidine derivatives and methods of use thereof
WO2013151708A1 (en) * 2012-04-04 2013-10-10 Samumed, Llc Indazole inhibitors of the wnt signal pathway and therapeutic uses thereof

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1918291A1 (en) 2006-10-30 2008-05-07 Novartis AG 3-Aminocarbonyl-substituted fused pyrazolo-derivatives as protein kinase modulators
CN101906105B (en) 2009-06-08 2013-01-16 河北医科大学 Pyrazolo (1,5-a) pyrimidone derivative as well as drug combination thereof and application thereof
WO2011121096A1 (en) * 2010-03-31 2011-10-06 Dkfz Deutsches Krebsforschungszentrum PYRAZOL[1,5-a]PYRIMIDINE DERIVATIVES AS WNT PATHWAY ANTAGONISTS
WO2011156632A2 (en) * 2010-06-09 2011-12-15 Georgetown University Compositions and methods of treatment for tumors in the nervous system
US9493444B2 (en) * 2012-01-28 2016-11-15 Merck Patent Gmbh Azaheterocyclic compounds
US20150157633A1 (en) * 2012-05-11 2015-06-11 The Board Of Regents Of The University Of Texas System Wnt protein signalling inhibitors
GB201322333D0 (en) 2013-12-17 2014-01-29 Agency Science Tech & Res WNT pathway modulators

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070219183A1 (en) * 2006-03-17 2007-09-20 Wyeth Dihydropyrazolo[1,5-A]pyrimidine and dihydroimidazo[1,5-A]pyrimidine derivatives and methods of use thereof
WO2013151708A1 (en) * 2012-04-04 2013-10-10 Samumed, Llc Indazole inhibitors of the wnt signal pathway and therapeutic uses thereof

Non-Patent Citations (26)

* Cited by examiner, † Cited by third party
Title
"Martindale The Extra Pharmacopoeia. 31st ed.", THE PHARMACEUTICAL PRESS, pages: 341
"Protective Groups in Organic Chemistry", 1973, PLENUM PRESS
"Remington's Pharmaceutical Sciences", MACK PUBLISHING CO.
DATABASE REGISTRY [online] 1 July 2008 (2008-07-01), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, 6,7-dihydro-2-methyl-N-(2-methylphenyl)-y-oxo", XP055351350, retrieved from STN Database accession no. 1031966-13-8 *
DATABASE REGISTRY [online] 1 July 2008 (2008-07-01), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(3,5-dimethylpheny))-6,7-dihydro-2-methyl-y-oxo", XP055351457, retrieved from STN Database accession no. 1031966-21-8 *
DATABASE REGISTRY [online] 22 June 2008 (2008-06-22), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-[3-(3-chlorophenyl)-5-isoxazolyl]-6,7-dihydro-2- methyl-y-oxo", XP055351432, retrieved from STN Database accession no. 1029768-94-2 *
DATABASE REGISTRY [online] 24 June 2008 (2008-06-24), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(3-chloro-2-methylphenyl)-6,7-dihydro-2-methyl-y- oxo", XP055351447, retrieved from STN Database accession no. 1030101-44-0 *
DATABASE REGISTRY [online] CAS; 1 July 2008 (2008-07-01), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(2-chloro-4-methylphenyt)-6,7-dihydro-2-methy1-y- oxo", XP055351440, retrieved from STN Database accession no. 1031966-17-2 *
DATABASE REGISTRY [online] CAS; 1 July 2008 (2008-07-01), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(5-chloro-2-methylphenyl)-6,7-dihydro-2-methy1-y- oxo", XP055351443, retrieved from STN Database accession no. 1031965-97-5 *
DATABASE REGISTRY [online] CAS; 11 October 2007 (2007-10-11), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, 6,7-dihydro-N-(2-methoxy-5-methylphenyl)-2-methyl- y-oxo", XP055351476, retrieved from STN Database accession no. 950287-32-8 *
DATABASE REGISTRY [online] CAS; 11 October 2007 (2007-10-11), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-[4-[(4-ethyl-1-piperazinyl)methyl]phenyl]-6,7- dihydro-2-methyl-y-oxo", XP055351356, retrieved from STN Database accession no. 950287-37-3 *
DATABASE REGISTRY [online] CAS; 22 June 2008 (2008-06-22), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, 6,7-dihydro-2-methyl-N-(4-methylcyclohexyl)-y-oxo", XP055351490, retrieved from STN Database accession no. 1029768-58-8 *
DATABASE REGISTRY [online] CAS; 22 June 2008 (2008-06-22), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, 6,7-dihydro-2-methyt-y-oxo-N-[ 1-(phenylmethyl)-4- piperidinyl", XP055351485, retrieved from STN Database accession no. 1029768-60-2 *
DATABASE REGISTRY [online] CAS; 22 June 2008 (2008-06-22), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(2,4-dimethylphenyl)-6,7-dihydro-2-methyl-y-oxo", XP055351466, retrieved from STN Database accession no. 1029768-50-0 *
DATABASE REGISTRY [online] CAS; 22 June 2008 (2008-06-22), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(2,6-dimethylphenyl)-6,7-dihydro-2-methyl-y-oxo", XP055351454, retrieved from STN Database accession no. 1029768-41-9 *
DATABASE REGISTRY [online] CAS; 22 June 2008 (2008-06-22), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(2-ethyl-6-methylphenyl)-6,7-dihydro-2-methyl-y- oxo", XP055351471, retrieved from STN Database accession no. 1029783-29-6 *
DATABASE REGISTRY [online] CAS; 22 June 2008 (2008-06-22), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(2-ethylphenyl)-6,7-dihydro-2-methyl-y-oxo", XP055351480, retrieved from STN Database accession no. 1029783-11-6 *
DATABASE REGISTRY [online] CAS; 22 June 2008 (2008-06-22), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(4-chloro-2-methoxy-5-methylphenyl)-6,7-dihydro- 2-methyl-y-oxo", XP055351364, retrieved from STN Database accession no. 1029783-45-6 *
DATABASE REGISTRY [online] CAS; 22 June 2008 (2008-06-22), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(4-chloro-2-methylphenyl)-6,7-dihydro-2-methyl-y- oxo", XP055351450, retrieved from STN Database accession no. 1029783-05-8 *
DATABASE REGISTRY [online] CAS; 24 June 2008 (2008-06-24), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, 6,7-dihydro-2-methyl-y-oxo-N-(2,4,6- trimethylphenyl", XP055351439, retrieved from STN Database accession no. 1030101-56-4 *
DATABASE REGISTRY [online] CAS; 24 June 2008 (2008-06-24), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, N-(2,3-dimethylphenyl)-6,7-dihydro-2-methyl-y-oxo", XP055351361, retrieved from STN Database accession no. 1030101-27-9 *
DATABASE REGISTRY [online] CAS; 25 April 2007 (2007-04-25), "Pyrazolo[1,5- a]pyrimidine-4(5H)-butanamide, 6,7-dihydro-2-methyl-N-(4-methylphenyl)-y-oxo", XP055351348, retrieved from STN Database accession no. 932481-41-9 *
NAJDI R; PROFFITT K; SPROWL S; KAUR S; YU J; COVEY TM; VIRSHUP DM; WATERMAN ML: "A uniform human Wnt expression library reveals a shared secretory pathway and unique signaling activities", DIFFERENTIATION, vol. 84, no. 2, 2012, pages 203 - 213, XP028450043, DOI: doi:10.1016/j.diff.2012.06.004
See also references of EP3083631A4
T.W. GREENE; P.G.M. WUTS: "Protective Groups in Organic Synthesis", 1991, JOHN WILEY & SONS
YAP MC; KOSTIUK MA; MARTIN DD; PERINPANAYAGAM MA; HAK PG; SIDDAM A; MAJJIGAPU JR; RAJAIAH G; KELLER BO; PRESCHER JA: "Rapid and selective detection of fatty acylated proteins using omega-alkynyl-fatty acids and click chemistry", J LIPID RES., vol. 51, no. 6, 2010, pages 1566 - 1580, XP055042918, DOI: doi:10.1194/jlr.D002790

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10189842B2 (en) 2013-12-17 2019-01-29 Agency For Science, Technology And Research Wnt pathway modulators
US10870653B2 (en) 2013-12-17 2020-12-22 Agency For Science, Technology And Research WNT pathway modulators
US11434245B2 (en) 2013-12-17 2022-09-06 Agency For Science, Technology And Research WNT pathway modulators
WO2016026772A1 (en) * 2014-08-19 2016-02-25 Iomet Pharma Ltd Pharmaceutical compound
US10047074B2 (en) 2014-08-19 2018-08-14 Merck Sharp & Dohme Corp. Pharmaceutical compound
WO2019180244A1 (en) * 2018-03-23 2019-09-26 Step Pharma S.A.S. Aminopyrimidine derivatives as ctps1 inhibitors
WO2019179652A1 (en) * 2018-03-23 2019-09-26 Step Pharma S.A.S. Aminopyrimidine derivatives as ctps1 inhibitors
AU2018413791B2 (en) * 2018-03-23 2023-05-18 Step Pharma S.A.S. Aminopyrimidine derivatives as CTPS1 inhibitors
TWI804545B (en) * 2018-03-23 2023-06-11 法商史坦普醫藥簡式股份公司 Compounds
IL277455B1 (en) * 2018-03-23 2024-01-01 Step Pharma S A S Aminopyrimidine derivatives as ctps1 inhibitors
IL277455B2 (en) * 2018-03-23 2024-05-01 Step Pharma S A S Aminopyrimidine derivatives as ctps1 inhibitors
US11987573B2 (en) 2018-10-23 2024-05-21 Step Pharma S.A.S. Aminopyrimidine/pyrazine derivatives as CTPS1 inhibitors

Also Published As

Publication number Publication date
BR112016013640A2 (en) 2017-08-08
AU2014367284A1 (en) 2016-06-30
CN105992767A (en) 2016-10-05
US20190185478A1 (en) 2019-06-20
EP3083631A1 (en) 2016-10-26
US20170240550A9 (en) 2017-08-24
CN105992767B (en) 2019-06-21
DK3083631T3 (en) 2018-09-24
ES2687493T3 (en) 2018-10-25
KR102383739B1 (en) 2022-04-05
AU2014367284B2 (en) 2019-01-17
EP3083631A4 (en) 2017-05-31
US11434245B2 (en) 2022-09-06
EP3083631B1 (en) 2018-06-13
JP2016540803A (en) 2016-12-28
SG11201604839XA (en) 2016-07-28
US20210047336A1 (en) 2021-02-18
GB201322333D0 (en) 2014-01-29
BR112016013640A8 (en) 2020-05-19
US10189842B2 (en) 2019-01-29
US20160311829A1 (en) 2016-10-27
KR20160124083A (en) 2016-10-26
JP6452703B2 (en) 2019-01-16
US20180093988A9 (en) 2018-04-05
CA2933683A1 (en) 2015-06-25
US10870653B2 (en) 2020-12-22

Similar Documents

Publication Publication Date Title
US11434245B2 (en) WNT pathway modulators
ES2791315T3 (en) Triazole agonists of the APJ receptor
AU2014367283B2 (en) Maleimide derivatives as modulators of WNT pathway
WO2012138648A1 (en) Compositions and methods for modulating lpa receptors
AU2018257203B2 (en) Novel tetrahydronaphthyl urea derivatives
CN111526877B (en) Compounds and compositions for IRE1 inhibition
JP6057907B2 (en) Pharmaceuticals containing thiazolidine derivatives or their salts as active ingredients
JP2022532718A (en) ACSS2 inhibitor and its usage
WO2013100672A1 (en) 3,6-disubstituted indazole derivative having protein kinase inhibiting activity
JP2021522253A (en) Compounds and their use
WO2014175832A1 (en) Wnt pathway modulators
WO2015187094A1 (en) Phthalimide derivatives as modulators of wnt pathway

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 14870867

Country of ref document: EP

Kind code of ref document: A1

DPE1 Request for preliminary examination filed after expiration of 19th month from priority date (pct application filed from 20040101)
ENP Entry into the national phase

Ref document number: 2933683

Country of ref document: CA

ENP Entry into the national phase

Ref document number: 2016540531

Country of ref document: JP

Kind code of ref document: A

WWE Wipo information: entry into national phase

Ref document number: 15105224

Country of ref document: US

NENP Non-entry into the national phase

Ref country code: DE

REEP Request for entry into the european phase

Ref document number: 2014870867

Country of ref document: EP

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112016013640

Country of ref document: BR

WWE Wipo information: entry into national phase

Ref document number: 2014870867

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2014367284

Country of ref document: AU

Date of ref document: 20141217

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 20167019109

Country of ref document: KR

Kind code of ref document: A

ENP Entry into the national phase

Ref document number: 112016013640

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20160613