Compounds and pharmaceutical compositions thereof for the treatment of diseases such as cancer

Abstract

The current invention relates to a compound of Formula (I), wherein X1, X2, X3, X4, Y, Y1, Y2, Y3, Y4, R1, R2, R16, B and A are as defined herein. The present invention relates to compounds that are inhibitors of FGFR enzymes, in particular FGFR3, methods for their production, pharmaceutical compositions comprising the same, and methods of treatment using the same, for the prophylaxis and / or treatment of a disease, disorder or condition that is associated with abnormal activity or expression of an FGFR enzyme, particularly FGFR3, by administering a compound of the invention.

Description

[0001] COMPOUNDS AND PHARMACEUTICAL COMPOSITIONS THEREOF FOR THE TREATMENT OF DISEASES SUCH ASCANCER

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to novel compounds that are inhibitors of FGFR enzymes and are useful in the treatment of FGFR-associated diseases such as cancer, and prodrugs thereof. The present invention also provides methods for the manufacture of the compounds of the invention, pharmaceutical compositions comprising the compounds of the invention, methods for the prophylaxis and / or treatment of FGFR-associated diseases, such as cancer, by administering the compounds of the invention.

[0004] BACKGROUND

[0005] The Fibroblast Growth Factor Receptors (FGFR) are receptor tyrosine kinases that bind to fibroblast growth factor (FGF) ligands. There are four FGFR proteins (FGFR1-4) that are capable of binding ligands and are involved in the regulation of many physiological processes including tissue development, angiogenesis, wound healing, and metabolic regulation. Upon ligand binding, the receptors undergo dimerization and phosphorylation leading to stimulation of the protein kinase activity and recruitment of many intracellular docking proteins. These interactions facilitate the activation of an array of intracellular signaling pathways including Ras-MAPK, AKT-PI3K, and phospholipase C that are important for cellular growth, proliferation and survival. Aberrant activation of this pathway either through overexpression of FGF ligands or FGFR or activating mutations in the FGFRs can lead to tumor development, progression, and resistance to conventional cancer therapies.

[0006] In human cancer, genetic alterations, including gene amplification, chromosomal translocations and somatic mutations that lead to ligand-independent receptor activation have been described (Knights and Cook, Pharmacology & Therapeutics, 2010, 125, 105-117; Turner and Grose, Nature Reviews Cancer, 2010, 10, 116-129). Large scale DNA sequencing of thousands of tumor samples has revealed that FGFR genes are altered in many cancers (Helsten et al., Clin Cancer Res., 2016, 22, 259-267). Some of these activating mutations are identical to germline mutations that lead to skeletal dysplasia syndromes (Gallo et al., Cytokine & Growth Factor Reviews, 2015, 26, 425-449). Mechanisms that lead to aberrant ligand-dependent signaling in human disease include overexpression of FGFs and changes in FGFR splicing that lead to receptors with more promiscuous ligand binding abilities. Therefore, development of inhibitors targeting FGFR may be useful in the clinical treatment of diseases that have elevated FGF or FGFR activity. The cancer types in which FGF / FGFRs are implicated include, inter alia, carcinomas (e.g., bladder, breast, colorectal, endometrial, gastric, head and neck, kidney, lung, ovarian, prostate), hematopoietic malignancies (e.g., multiple myeloma, acute myelogenous leukemia, and myeloproliferative neoplasms), and other neoplasms (e.g., glioblastoma and sarcomas). In addition to a role in oncogenic neoplasms, FGFR activation has also been implicated in skeletal and chondrocyte disorders including, inter alia, achondroplasia and craniosynostosis syndromes. There is a continuing need for the development of new drugs for the treatment of cancer, and the FGFR inhibitors described herein help address this need.

[0007] There is also an ongoing need for new FGFR inhibitors with improved properties for the treatment of FGFR-associated diseases, such as cancer. In the current state of the art, a variety of FGFR3 inhibitors have been explored, yet many of these compounds continue to exhibit important limitations, particularly with respect to their selectivity profile and occurrence of treatment resistance. Many current compounds show poor selectivity toward FGFR3. The selectivity index, defined as the ratio of ICso values for off-target kinases versus FGFR3, is often relatively low. As a consequence, these compounds are likely to inhibit other members of the FGFR family (e.g., FGFR1, FGFR2, or FGFR4) or unrelated kinases at therapeutically relevant concentrations. This lack of target discrimination can give rise to undesirable pharmacological effects, including toxicity and interference with unrelated signaling pathways, ultimately reducing their therapeutic window. In addition, cancer cells can develop resistance to current compounds, a.o. through mutations at the gatekeeper residue within the kinase domain. These mutations hinder the inhibitor's access to the ATP- binding pocket, reducing the drug's efficacy. It is known that acquired resistance mutations in FGFR3 can emerge in a substantial proportion of patients following relapses after treatment with FGFR inhibitors, thereby limiting the long-term efficacy of such therapies.

[0008] The present invention aims to resolve at least some of the problems and disadvantages mentioned above.

[0009] SUMMARY OF THE INVENTION

[0010] The present invention provides compounds that are FGFR inhibitors. The invention also provides compositions comprising the compounds, in particular pharmaceutical compositions, and methods of using / administering and making the compounds and compositions. The compounds provided herein are useful in treating diseases, disorders, or conditions that are mediated by FGFRs. The invention also provides compounds for use in therapy. The invention further provides compounds for use in a method of treating a disease, disorder, or condition that is mediated by a FGFR. Moreover, the invention provides uses of the compounds in the manufacture of a medicament for the treatment of a disease, disorder or condition that is mediated by a FGFR. Accordingly, in a first aspect, the present invention provides compounds according to claim 1.

[0011] More in particular, the present invention discloses compounds according to Formula (I) : wherein:

[0012] Xi is N or CH;

[0013] X2 is N and X3 is C, or, alternatively, X3 is N and X2 is C;

[0014] X4is N;

[0015] Y is a bond, O or NH;

[0016] Yi is CHR6, CH2-CHR6, CHR6-CH2, CF2, CH2-CF2 or CF2-CH2;

[0017] Y2is CR4R5or CF2;

[0018] Y3is CR3R4, CR3R4CR3R4, CH2CF2 or CF2; wherein Yi, Y2, Y3 and X4 together with the pyrazolyl ring form a 7-membered nitrogen containing ring;

[0019] B is -C(O)Y4-, -Y4, -SO2-Y4-, - CR4R9Y4-, or -CH2CH2Y4-, and A is CN, R8R9C=CR10C(=O)-, R11R12NCH2CH=CHC(=O)-, H2C=CHSC>2- or R13C=CC(=O)-; wherein R4and R9are as defined herewith, or, alternatively, R4and R9together with the carbon atoms to which they are attached form a 3-5-membered carbocyclic ring;

[0020] Y4 is a 4-10 membered monocyclic heterocycloalkyl or a 5-12 membered fused, spiro or bridged polycyclic heterocycloalkyl, said monocyclic or polycyclic heterocycloaklyl comprising a nitrogen atom, wherein the heterocycloalkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, methyl, halomethyl, hydroxyl or methoxy and wherein the nitrogen atom of the heterocycloalkyl is connected to A; R1is hydrogen, halogen, C1-5 alkyl, C1-5 alkoxy, CN, C3-10 cycloalkyl, C2-4 alkynyl, 4-10 membered heterocycloalkyl comprising 1, 2 or 3 heteroatoms independently selected from N, O and S, phenyl or a 5-6 membered heteroaryl, wherein the 3-10 membered cycloalkyl, 4-10 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl are unsubstituted or substituted with one or more substituents independently selected from halogen, methyl, halomethyl, hydroxyl or methoxy and wherein the C1-5 alkyl and C1-5 alkoxy, are unsubstituted or independently substituted with one or more substituents independently selected from halogen, CN, hydroxyl and methoxy;

[0021] R2is hydrogen, hydroxyl, Ci-4 alkyl, CN, -CONHC1-4 alkyl, or Ci-4 haloalkyl; each R3is independently selected from hydrogen, fluoro, hydroxyl, methoxy, C1-3 alkyl, cyclopropyl, or a R3is fused with R5or R6to form CH2, CH2-CH2 or CH2OCH2; each R4is independently selected from hydrogen, or C1-3 alkyl;

[0022] R5is hydrogen, fluoro, hydroxyl, methoxy, C1-3 alkyl, cyclopropyl or is fused with one R3to form CH2, CH2-CH2 or CH2OCH2, wherein the C1-3 alkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, heteroaryl (which heteroaryl is preferably pyridyl), phenyl, hydroxyl, -CO2R4or C1-3 alkoxy;

[0023] R6is hydrogen, fluoro, hydroxyl, methoxy, C1-3 alkyl or is fused with one R3to form CH2, CH2- CH2or CH2OCH2; each R8is independently selected from hydrogen and C1-6 alkyl; each R9is independently selected from hydrogen, CF3 and C1-6 alkyl, which alkyl is unsubstituted or substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxyl, and C1-6 alkoxy; and R10is hydrogen, C1-3 alkyl or halogen; or R9and R10together with the carbon atoms to which they are attached form a 4-8- membered carbocyclic ring; each of R11and R12is independently hydrogen, C1-6 alkyl, or R11and R12together with the nitrogen atom to which they are attached form a 4-6 membered heterocycloalkyl optionally having an additional ring heteroatom which is O, wherein said ring is optionally substituted with halogen;

[0024] R13is hydrogen, R14R15NCH2-, or Ci-3 alkyl which is unsubstituted or substituted with hydroxyl; R14and R15are each independently hydrogen or C1-6 alkyl;

[0025] R16is C1-3 alkyl-R17or C3-6 cycloalkyl-R17, wherein the C1-3 alkyl or C3-6 cycloalkyl are unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, CN, Ci-4 alkyl optionally substituted with one or more OH, C3-6 cycloalkyl, OH, OMe, -NR4R8wherein R4and R8are as defined above;

[0026] R17is aryl or heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O and S, wherein the aryl or heteroaryl are unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, CN, NH2, Ci-4 alkyl, and Ci-4 haloalkyl, Ci-4 alkoxy, and Ci-4 haloalkoxy; or a pharmaceutically acceptable salt and / or solvate thereof. In a second aspect, the invention provides compositions comprising the compounds, in particular pharmaceutical compositions comprising a compound of the invention and a pharmaceutical carrier, excipient or diluent. In a particular aspect, the pharmaceutical composition may additionally comprise further therapeutically active ingredients suitable for use in combination with the compounds of the invention. In a more particular aspect, the further therapeutically active ingredient is an agent for the treatment of FGFR-associated diseases, in particular cancer.

[0027] In a further aspect, the invention provides use of the compounds, or a pharmaceutical composition comprising them, in medicine, in particular in the prophylaxis and I or treatment of a disease, disorder or condition that is associated with abnormal activity or expression of an FGFR enzyme, in particular cancer.

[0028] In another aspect, the invention provides the compounds, or a pharmaceutical composition comprising them, for use in a method of treating a disease, disorder, or condition that is associated with abnormal activity or expression of an FGFR enzyme, in particular cancer.

[0029] In a yet further aspect, the invention provides uses of the compounds in the manufacture of a medicament for the treatment of a disease, disorder or condition that is associated with abnormal activity or expression of an FGFR enzyme, in particular cancer.

[0030] In another aspect, the invention provides methods of using / administering the compounds or a pharmaceutical composition comprising them.

[0031] In an additional aspect, the invention provides methods of making the compounds and compositions comprising them.

[0032] Moreover, the compounds of the invention, useful in the pharmaceutical compositions and treatment methods disclosed herein, are pharmaceutically acceptable as prepared and used.

[0033] In additional aspects, this invention provides methods for synthesizing the compounds of the invention, with representative synthetic protocols and pathways disclosed later on herein.

[0034] Other objects and advantages will become apparent to those skilled in the art from a consideration of the ensuing detailed description.

[0035] Furthermore, it has been unexpectedly demonstrated that the compounds of the invention exhibit advantageous properties for their use treating diseases, disorders, or conditions that are associated with abnormal activity or expression of an FGFR enzyme, in particular cancer. Certain compounds provided herein have superior selectivity for FGFR3 over FGFR1 compared to certain previously known FGFR inhibitors, reducing potential dose limiting toxicity caused by inhibition of FGFR1 (e.g. hyperphosphatemia). Certain compounds provided herein have superior selectivity for FGFR3 over FGFR2 compared to certain previously known FGFR inhibitors, reducing potential dose limiting toxicity caused by inhibition of FGFR2. Certain compounds provided herein have superior selectivity for FGFR3 over FGFR4 compared to certain previously known FGFR inhibitors, reducing potential dose limiting toxicity caused by inhibition of FGFR4 (e.g. diarrhea).

[0036] DEFINITIONS

[0037] The following terms are intended to have the meanings presented therewith below and are useful in understanding the description and intended scope of the present invention.

[0038] When describing the invention, which may include compounds, pharmaceutical compositions containing such compounds and methods of using such compounds and compositions, the following terms, if present, have the following meanings unless otherwise indicated. It should also be understood that when described herein any of the moieties defined forth below may be substituted with a variety of substituents, and that the respective definitions are intended to include such substituted moieties within their scope as set out below. Unless otherwise stated, the term "substituted" is to be defined as set out below. It should be further understood that the terms "groups" and "radicals" can be considered interchangeable when used herein.

[0039] The articles 'a' and 'an' may be used herein to refer to one or to more than one ( / .e. at least one) of the grammatical objects of the article. By way of example 'an analogue' means one analogue or more than one analogue.

[0040] A dash ("-") that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -CONH2 is attached through the carbon atom. A dash at the front or end of a chemical group is a matter of convenience; chemical groups may be depicted with or without one or more dashes without losing their ordinary meaning.

[0041] A wavy line drawn through a line in a structure indicates a point of attachment of a group.

[0042] Similarly, a squiggly line on a chemical group as shown, for example , indicates a point of attachment, i.e., it shows the broken bond by which the group is connected to another described group. The prefix "Cu v" indicates that the following group has from u to v carbon atoms. For example, "Ci-8 alkyl" indicates that the alkyl group has from 1 to 8 carbon atoms.

[0043] Reference to "about" a value or parameter herein includes (and describes) embodiments that are directed to that value or parameter per se. In certain embodiments, the term "about" includes the indicated amount ± 10%. In other embodiments, the term "about" includes the indicated amount ± 5%. In certain other embodiments, the term "about" includes the indicated amount ± 1%. Also, to the term "about X" includes description of "X". Also, the singular forms "a" and "the" include plural references unless the context clearly dictates otherwise. Thus, e.g., reference to "the compound" includes a plurality of such compounds and reference to "the assay" includes reference to one or more assays and equivalents thereof known to those skilled in the art.

[0044] Unless indicated explicitly otherwise, where combinations of groups are referred to herein as one moiety, e.g. arylalkyl, the last mentioned group contains the atom by which the moiety is attached to the rest of the molecule.

[0045] 'Alkyl' means straight or branched aliphatic hydrocarbon having the specified number of carbon atoms. Particular alkyl groups have 1 to 6 carbon atoms or 1 to 4 carbon atoms.

[0046] Branched means that one or more alkyl groups such as methyl, ethyl or propyl is attached to a linear alkyl chain. Particular alkyl groups are methyl (-CH3), ethyl (-CH2-CH3), n-propyl (- CH2-CH2-CH3), isopropyl (-CH(CH3)2), n-butyl (-CH2-CH2-CH2-CH3), tert-butyl (-C(CH3)3), secbutyl (-CH(CH3)-CH2CH3), isobutyl (-CH2-CH(CH3)2), n-pentyl (-CH2-CH2-CH2-CH2-CH3), n- hexyl (-CH2-CH2-CH2-CH2-CH2-CH3), and 1,2-di methyl butyl (-CH(CH3)-CH(CH3)-CH2-CH3). Particular alkyl groups have between 1 and 4 carbon atoms.

[0047] 'Alkenyl' refers to monovalent unsaturated hydrocarbon groups with the number of carbon atoms and the number of double bonds specified. Particular alkenyl groups have 2 to 8 carbon atoms, and more particularly, from 2 to 6 carbon atoms, which can be straight-chained or branched and having at least 1 and particularly from 1 to 2 double bonds. Particular alkenyl groups include ethenyl (-CH=CH2), n-propenyl (-CH2CH=CH2), isopropenyl (-C(CH3)=CH2) and the like.

[0048] 'Alkynyl' refers to monovalent unsaturated hydrocarbon groups with the number of carbon atoms and the number of triple bonds specified. Particular alkynyl groups have 2 to 8 carbon atoms, and more particularly, from 2 to 6 carbon atoms, which can be straight-chained or branched and having at least 1 and particularly from 1 to 2 triple bonds. Particular alkynyl groups include -C=CH, -CH2-OCH, -CH(CH3)-C=CH, and the like. 'Alkylene' refers to divalent alkyl radical groups having the number of carbon atoms specified, in particular having 1 to 6 carbon atoms and more particularly 1 to 4 carbon atoms which can be straight chained or branched. This term is exemplified by groups such as methylene (-CH2- ), ethylene (CH2-CH2-), or -CH(CH3)- and the like.

[0049] 'Alkenylene' refers to divalent alkenyl radical groups having the number of carbon atoms and the number of double bonds specified, in particular 2 to 6 carbon atoms and more particularly 2 to 4 carbon atoms which can be straight-chained or branched. This term is exemplified by groups such as -CH=CH-, -CH2-CH=CH-, -C(CH3)=CH-, -C(CH3)=CH-CH2-, -C(CH3)=C(CH3)- , and -CH2-C(CH3)=CH-.

[0050] 'Alkynylene' refers to divalent alkynyl radical groups having the number of carbon atoms and the number of triple bonds specified, in particular 2 to 6 carbon atoms and more particularly 2 to 4 carbon atoms which can be straight-chained or branched. This term is exemplified by groups such as -C=C-, -CH2-OC-, and -C(CH3)H-C=CH-.

[0051] 'Alkoxy' refers to the group O-alkyl, where the alkyl group has the number of carbon atoms specified. In particular the term refers to the group -O-C1-6 alkyl. Particular alkoxy groups are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n- hexoxy, and 1,2-dimethylbutoxy. Particular alkoxy groups are lower alkoxy, i.e. with between 1 and 6 carbon atoms. Further particular alkoxy groups have between 1 and 4 carbon atoms.

[0052] 'Amino' refers to the radical -NH2.

[0053] 'Aryl' refers to a monovalent aromatic hydrocarbon group derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system. In particular aryl refers to an aromatic ring structure, monocyclic or fused polycyclic, with the number of ring atoms specified. Specifically, the term includes groups that include from 6 to 10 ring members. Particular aryl groups include phenyl, and naphthyl.

[0054] 'Cycloalkyl 'refers to a non-aromatic hydrocarbyl ring structure, monocyclic, fused polycyclic, bridged polycyclic, or spirocyclic, with the number of ring atoms specified. A cycloalkyl may have from 3 to 12 carbon atoms, in particular from 3 to 10, and more particularly from 3 to 7 carbon atoms. Such cycloalkyl groups include, by way of example, single ring structures such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0055] 'Cyano' refers to the radical -CN.

[0056] 'Halo' or 'halogen' refers to fluoro (F), chloro (Cl), bromo (Br) and iodo (I). Particular halo groups are either fluoro or chloro. As used herein, term 'polycyclic' refers to chemical groups featuring several closed rings of atoms. In particular it refers to groups featuring two (bicyclic), three (tricyclic) or four (tetracyclic) rings of atoms, more particularly two (bicyclic) or three (tricyclic) rings of atoms, most particularly two (bicyclic) rings of atoms.

[0057] 'Hetero' when used to describe a compound or a group present on a compound means that one or more carbon atoms in the compound or group have been replaced by a O, N, P and S heteroatom. Hetero may be applied to any of the hydrocarbyl groups described previously such as alkyl, e.g. heteroalkyl, cycloalkyl, e.g. heterocycloalkyl, aryl, e.g. heteroaryl, and the like having from 1 to 4, and particularly from 1 to 3 heteroatoms, more typically 1 or 2 heteroatoms, for example a single heteroatom.

[0058] 'Heteroaryl' means an aromatic ring structure, monocyclic or fused polycyclic, that includes one or more heteroatoms independently selected from O, N and S and the number of ring atoms specified. In particular, the aromatic ring structure may have from 5 to 9 ring members. The heteroaryl group can be, for example, a five membered or six membered monocyclic ring or a fused bicyclic structure formed from fused five and six membered rings or two fused six membered rings or, by way of a further example, two fused five membered rings. Each ring may contain up to four heteroatoms typically selected from nitrogen, sulfur and oxygen. Typically, the heteroaryl ring will contain up to 4 heteroatoms, more typically up to 3 heteroatoms, more usually up to 2, for example a single heteroatom. In one embodiment, the heteroaryl ring contains at least one ring nitrogen atom. The nitrogen atoms in the heteroaryl rings can be basic, as in the case of an imidazole or pyridine, or essentially non- basic as in the case of an indole or pyrrole nitrogen. In general, the number of basic nitrogen atoms present in the heteroaryl group, including any amino group substituents of the ring, will be less than five.

[0059] 'Heteroaryl' also covers partially aromatic bi- or polycyclic ring systems wherein at least one ring is an aromatic ring and one or more of the other ring(s) is a non-aromatic, saturated or partially saturated ring, provided at least one ring contains one or more heteroatoms selected from nitrogen, oxygen or sulfur. Partially aromatic heteroaryl bicyclic ring systems can be vicinally fused, i.e., where the rings are linked to each other through two adjacent carbon and / or nitrogen atoms. Examples of partially aromatic heteroaryl groups include for example, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 2-oxo-l,2,3,4-tetrahydroquinolinyl, dihydrobenzothienyl, dihydrobenzofuranyl, 1,3-dihydroisobenzofuran, 2,3- dihydrobenzo[l,4]dioxinyl, benzo[l,3]dioxolyl, 2,2-dioxo-l,3-dihydro-2-benzothienyl, 4,5,6,7-tetrahydrobenzofuranyl, indolinyl, l,2,3,4-tetrahydro-l,8-naphthyridinyl, 1,2, 3, 4- tetrahydropyrido[2,3-b]pyrazinyl and 3,4-dihydro-2H-pyrido[3,2-b][l,4]oxazinyl. Examples of five membered monocyclic heteroaryl groups include but are not limited to pyrrolyl, furanyl, thiophenyl, imidazolyl, furazanyl, oxazolyl, oxadiazolyl, oxatriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl and tetrazolyl groups.

[0060] Examples of six membered monocyclic heteroaryl groups include but are not limited to pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl and triazinyl.

[0061] Particular examples of bicyclic heteroaryl groups containing a five membered ring fused to another five-membered ring include but are not limited to imidazothiazolyl and imidazoimidazolyl

[0062] Particular examples of bicyclic heteroaryl groups containing a five membered ring fused to another five-membered ring include but are not limited to imidazothiazolyl and imidazoimidazolyl.

[0063] Particular examples of bicyclic heteroaryl groups containing a six membered ring fused to a five membered ring include but are not limited to benzofuranyl, benzothiophenyl, benzoimidazolyl, benzoxazolyl, isobenzoxazolyl, benzisoxazolyl, benzothiazolyl, benzoisothiazolyl, isobenzofuranyl, indolyl, isoindolyl, indolizinyl, purinyl (e.g. adenine, guanine), indazolyl, pyrazolopyrimidinyl, triazolopyrimidinyl, and pyrazolopyridinyl groups.

[0064] Particular examples of bicyclic heteroaryl groups containing two fused six membered rings include but are not limited to quinolinyl, isoquinolinyl, pyridopyridinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl, and pteridinyl groups. Particular heteroaryl groups are those derived from thiophenyl, pyrrolyl, benzothiophenyl, benzofuranyl, indolyl, pyridinyl, quinolinyl, imidazolyl, oxazolyl and pyrazinyl.

[0065] Examples of representative heteroaryls include the following: wherein each Y is selected from >C=O, NH, O and S.

[0066] 'Heterocycloalkyl' means a non-aromatic fully or partially saturated ring structure, monocyclic, fused polycyclic, spirocyclic, or bridged polycyclic, that includes one or more heteroatoms independently selected from O, N, P and S and the number of ring atoms specified. The heterocycloalkyl ring structure may have from 4 to 12 ring members, in particular from 4 to 10 ring members and more particularly from 4 to 7 ring members. Each ring may contain up to four heteroatoms typically selected from nitrogen, phosphorus, sulfur and oxygen. Typically the heterocycloalkyl ring will contain up to 4 heteroatoms, more typically up to 3 heteroatoms, more usually up to 2, for example a single heteroatom. When partially saturated, the heterocycloalkyl may contain one or two double bonds, and more particularly one double bond. Examples of heterocyclic rings include, but are not limited to azetidinyl, oxetanyl, thietanyl, pyrrolidinyl (e.g. 1-pyrrolidinyl, 2-pyrrolidinyl and 3- pyrrolidinyl), tetra hydrofuranyl (e.g. 1-tetrahydrofuranyl, 2-tetra hydrofuranyl and 3- tetrahydrofuranyl), tetrahydrothiophenyl (e.g. 1-tetrahydrothiophenyl, 2- tetrahydrothiophenyl and 3-tetrahydrothiophenyl), piperidinyl (e.g. 1-piperidinyl, 2- piperidinyl, 3-piperidinyl and 4-piperidinyl), tetra hydropyranyl (e.g. 4-tetrahydropyranyl), tetra hydrothio pyranyl (e.g. 4-tetrahydrothiopyranyl), morpholinyl, thiomorpholinyl, dioxanyl, or piperazinyl.

[0067] Particular examples of monocyclic rings are shown in the following illustrative examples: wherein each W and Y is independently selected from -CH2-, -NH-, -O-, and -S-.

[0068] Particular examples of fused bicyclic rings are shown in the following illustrative examples: wherein each W and Y is independently selected from -CH2-, -NH-, -O- and -S-.

[0069] Particular examples of bridged bicyclic rings are shown in the following illustrative examples: wherein each W and Y is independently selected from -CH2-, -NH-, -O- and -S- and each Z is selected from N or CH.

[0070] Particular examples of spirocyclic rings are shown in the following illustrative examples: wherein each Y is selected from -CH2-, -NH-, -O- and -S-. Particular examples of monocyclic partially saturated heterocycloalkyl rings are shown in the following illustrative examples: wherein each W and Y is independently selected from -CH2-, -NH-, -O-, and -S-, and Z is P, N, or CH.

[0071] 'Hydrocarbon chain' refers to an organic molecule consisting of nothing else but carbon and hydrogen atoms arranged in a chain, which carbon and hydrogen atoms are interconnected to each other by covalent bonding. Each carbon atom in the chain is bonded to one or up to three hydrogen atoms, hydrocarbon chains may be classified as branched, linear, or cyclical. They may also be divided into alkanes, alkenes, alkynes, cycloalkanes, and aryls. A hydrocarbon chain may also be classified as either saturated or unsaturated, or aliphatic or aromatic. A saturated hydrocarbon chain is saturated with hydrogen whereas an unsaturated hydrocarbon chain is one in which hydrogen atom can still be added in the chain by breaking the double-bond (alkene) or triple bond (alkyne) between carbon atoms.

[0072] 'Hydroxyl' refers to the radical -OH.

[0073] 'Oxo' refers to the radical =0.

[0074] 'Substituted' refers to a group in which one or more hydrogen atoms are each independently replaced with the same or different substituent(s).

[0075] 'Sulfo' or 'sulfonic acid' refers to a radical such as -SO3H.

[0076] 'Thiol' refers to the group -SH.

[0077] As used herein, term 'substituted with one or more' refers to one to four substituents. In one embodiment it refers to one to three substituents. In further embodiments it refers to one or two substituents. In a yet further embodiment it refers to one substituent.

[0078] 'Thioalkoxy' refers to the group -S-alkyl where the alkyl group has the number of carbon atoms specified. In particular the term refers to the group -S-C1-6 alkyl. Particular thioalkoxy groups are thiomethoxy, thioethoxy, n-thiopropoxy, isothiopropoxy, n-thiobutoxy, tertthiobutoxy, sec-thiobutoxy, n-thiopentoxy, n-thiohexoxy, and 1,2-dimethylthiobutoxy. Particular thioalkoxy groups are lower thioalkoxy, i.e. with between 1 and 6 carbon atoms. Further particular alkoxy groups have between 1 and 4 carbon atoms. One having ordinary skill in the art of organic synthesis will recognize that the maximum number of heteroatoms in a stable, chemically feasible heterocyclic ring, whether it is aromatic or non-aromatic, is determined by the size of the ring, the degree of unsaturation and the valence of the heteroatoms. In general, a heterocyclic ring may have one to four heteroatoms so long as the heteroaromatic ring is chemically feasible and stable.

[0079] 'Pharmaceutically acceptable' means approved or approvable by a regulatory agency of the Federal or a state government or the corresponding agency in countries other than the United States, or that is listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly, in humans.

[0080] 'Pharmaceutically acceptable salt' refers to a salt of a compound of the invention that is pharmaceutically acceptable and that possesses the desired pharmacological activity of the parent compound. In particular, such salts are non-toxic may be inorganic or organic acid addition salts and base addition salts. Specifically, such salts include: (1) acid addition salts, formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or formed with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4- toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2.2]-oct-2-ene-l-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) salts formed when an acidic proton present in the parent compound either is replaced by a metal ion, e.g. an alkali metal ion, an alkaline earth ion, or an aluminium ion; or coordinates with an organic base such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine and the like. Salts further include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the compound contains a basic functionality, salts of non-toxic organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate and the like. The term 'pharmaceutically acceptable cation' refers to an acceptable cationic counter-ion of an acidic functional group. Such cations are exemplified by sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium cations, and the like.

[0081] 'Pharmaceutically acceptable vehicle' refers to a diluent, adjuvant, excipient or carrier with which a compound of the invention is administered. 'Prodrugs' refers to compounds, including derivatives of the compounds of the invention, which have cleavable groups and become by solvolysis or under physiological conditions the compounds of the invention which are pharmaceutically active in vivo. Such examples include, but are not limited to, choline ester derivatives and the like, N-alkylmorpholine esters and the like.

[0082] 'Solvate' refers to forms of the compound that are associated with a solvent, usually by a solvolysis reaction. This physical association includes hydrogen bonding. Conventional solvents include water, EtOH, acetic acid and the like. The compounds of the invention may be prepared e.g. in crystalline form and may be solvated or hydrated. Suitable solvates include pharmaceutically acceptable solvates, such as hydrates, and further include both stoichiometric solvates and non-stoichiometric solvates. In certain instances the solvate will be capable of isolation, for example when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. 'Solvate' encompasses both solution-phase and isolable solvates. Representative solvates include hydrates, ethanolates and methanolates.

[0083] 'Subject' includes humans. The terms 'human', 'patient' and 'subject' are used interchangeably herein.

[0084] 'Effective amount' means the amount of a compound of the invention that, when administered to a subject for treating a disease, is sufficient to effect such treatment for the disease. The "effective amount" can vary depending on the compound, the disease and its severity, and the age, weight, etc., of the subject to be treated.

[0085] 'Preventing' or 'prevention' refers to a reduction in risk of acquiring or developing a disease or disorder ( / .e. causing at least one of the clinical symptoms of the disease not to develop in a subject that may be exposed to a disease-causing agent, or predisposed to the disease in advance of disease onset.

[0086] The term 'prophylaxis' is related to 'prevention', and refers to a measure or procedure the purpose of which is to prevent, rather than to treat or cure a disease. Non-limiting examples of prophylactic measures may include the administration of vaccines; the administration of low molecular weight heparin to hospital patients at risk for thrombosis due, for example, to immobilization; and the administration of an anti-malarial agent such as chloroquine, in advance of a visit to a geographical region where malaria is endemic or the risk of contracting malaria is high.

[0087] 'Treating' or 'treatment' of any disease or disorder refers, in one embodiment, to ameliorating the disease or disorder ( / .e. arresting the disease or reducing the manifestation, extent or severity of at least one of the clinical symptoms thereof). In another embodiment 'treating' or 'treatment' refers to ameliorating at least one physical parameter, which may not be discernible by the subject. In yet another embodiment, 'treating' or 'treatment' refers to modulating the disease or disorder, either physically, (e.g. stabilization of a discernible symptom), physiologically, (e.g. stabilization of a physical parameter), or both. In a further embodiment, "treating" or "treatment" relates to slowing the progression of the disease.

[0088] Certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3 over another FGFR. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3 over FGFR1. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, are at least about 3 fold (e.g. at least about 4-, 5- , 6-, 7-, 8-, 9-, 10-, 15-, 20-, 30-, 40-, or more) more potent for FGFR3 than for FGFR1.

[0089] Certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3 over another FGFR. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3 over FGFR2. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, are at least about 3 fold (e.g. at least about 4-, 5- , 6-, 7-, 8-, 9-, 10-, 15-, or more) more potent for FGFR3 than for FGFR2.

[0090] Certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3 over another FGFR. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3 over FGFR4. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, are at least about 3 fold (e.g. at least about 4-, 5- , 6-, 7-, 8-, 9-, 10-, 15-, or more) more potent for FGFR3 than for FGFR4.

[0091] As used herein, the term "selectivity" of a compound refers to the compound having more potent activity at the first target than the second target. A fold selectivity can be calculated by any method known in the art. For example, a fold selectivity can be calculated by dividing the IC50 value of a compound for the second target (e.g., FGFR1) by the IC50 value of the same compound for the first target (e.g., FGFR3). An IC50 value can be determined by any method known in the art. For example, an IC50 value can be determined as described in the assays below.

[0092] As used herein the term "cancer" refers to diseases caused by an uncontrolled division of abnormal cells in a part of the body. In particular, the term refers to metastatic tumour cell types (such as but not limited to, melanoma, lymphoma, leukaemia, fibrosarcoma, rhabdomyosarcoma, and mastocytoma) and types of tissue carcinoma (such as but not limited to, colorectal cancer, prostate cancer, small cell lung cancer and non-small cell lung cancer, breast cancer, pancreatic cancer, bladder cancer, renal cancer, gastric cancer, glioblastoma, primary liver cancer, ovarian cancer, prostate cancer and uterine leiomyosarcoma). More particularly, the term refers to acute lymphoblastic leukemia, acute myeloidleukemia, adrenocortical carcinoma, anal cancer, appendix cancer, astrocytomas, atypical teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer (osteosarcoma and malignant fibrous histiocytoma), brain stem glioma, brain tumors, brain and spinal cord tumors, breast cancer, bronchial tumors, Burkitt lymphoma, cervical cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T -Cell lymphoma, embryonal tumors, endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, ewing sarcoma family of tumors, eye cancer, retinoblastoma, gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), gastrointestinal stromal cell tumor, germ cell tumor, glioma, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumors (endocrine pancreas), Kaposi sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, leukemia, Acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, liver cancer, non-small cell lung cancer, small cell lung cancer, Burkitt lymphoma, cutaneous T-celllymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, lymphoma, Waldenstrom macroglobulinemia, medulloblastoma, medulloepithelioma, melanoma, mesothelioma, mouth cancer, chronic myelogenous leukemia, myeloid leukemia, multiple myeloma, asopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma, malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, papillomatosis, parathyroid cancer, penile cancer, pharyngeal cancer, pineal parenchymal tumors of intermediate differentiation, pineoblastoma and supratentorial primitive neuroectodermal tumors, pituitary tumor, plasma cell neoplasm / multiple myeloma, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell (kidney) cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, Ewing sarcoma family of tumors, sarcoma, kaposi, Sezary syndrome, skin cancer, small cell Lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, stomach (gastric) cancer, supratentorial primitive neuroectodermal tumors, T -cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, and Wilms tumor. As used herein the term "leukemia" refers to neoplastic diseases of the blood and blood forming organs. In particular, the term refers to neoplastic diseases of the blood and blood forming organs. More particularly, the term refers to acute myeloid leukaemia (AML), and acute lymphoblastic leukemia (ALL) and chronic lymphoblastic leukaemia (CLL).

[0093] As used herein, the term "FGFR3-associated cancer" refers to cancers associated with or having a dysregulation of the FGFR3 gene, the FGFR3 kinase protein, or expression or activity, or level of any of the same. Non-limiting examples of FGFR3- associated cancer are described herein. As used herein an "FGFR3 -associated cancer" includes but is not limited to breast cancer (e.g. invasive ductal cancer, invasive lobular cancer), lung cancer (e.g. non-small-cell lung cancer, lung adenocarcinoma, squamous cell lung cancer and small-cell lung cancer), urothelial cancer, bladder cancer (e.g. urothelial bladder cancer, non-muscle invasive bladder cancer, muscle invasive bladder cancer), upper tract cancer (e.g. urothelial upper tract cancer), urethral cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, multiple myeloma, liver cancer, melanoma (e.g. cutaneous melanoma), head and neck cancer (e.g. oral cancer), thyroid cancer, renal cancer (e.g. renal pelvis cancer), glioblastoma, endometrial cancer, cervical cancer, ovarian cancer, and testicular cancer.

[0094] 'Compound(s) of the invention', and equivalent expressions, are meant to embrace compounds of the Formula(e) as herein described, which expression includes the pharmaceutically acceptable salts, and the solvates, e.g. hydrates, and the solvates of the pharmaceutically acceptable salts where the context so permits. Accordingly, 'compound(s) of the invention' may refer to a compound according to any of Formula (I) or a pharmaceutically acceptable salt and / or solvate thereof. Similarly, reference to intermediates, whether or not they themselves are claimed, is meant to embrace their salts, and solvates, where the context so permits.

[0095] When ranges are referred to herein, for example but without limitation, Ci-8 alkyl, the citation of a range should be considered a representation of each member of said range.

[0096] Other derivatives of the compounds of this invention have activity in both their acid and acid derivative forms, but in the acid sensitive form often offers advantages of solubility, tissue compatibility, or delayed release in the mammalian organism (Bundgard, H, 1985). Prodrugs include acid derivatives well known to practitioners of the art, such as, for example, esters prepared by reaction of the parent acid with a suitable alcohol, or amides prepared by reaction of the parent acid compound with a substituted or unsubstituted amine, or acid anhydrides, or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from acidic groups pendant on the compounds of this invention are particularly useful prodrugs. In some cases it is desirable to prepare double ester type prodrugs such as (acyloxy)alkyl esters or ((alkoxycarbonyl)oxy)alkylesters. Particular such prodrugs are the Ci-8 alkyl, C2-8 alkenyl, Ce-io optionally substituted aryl, and (Ce-io aryl)-(Ci-4 alkyl) esters of the compounds of the invention.

[0097] The present disclosure includes all isotopic forms of the compounds of the invention provided herein, whether in a form (i) wherein all atoms of a given atomic number have a mass number (or mixture of mass numbers) which predominates in nature (referred to herein as the "natural isotopic form") or (ii) wherein one or more atoms are replaced by atoms having the same atomic number, but a mass number different from the mass number of atoms which predominates in nature ( referred to herein as an "unnatural variant isotopic form"). It is understood that an atom may naturally exists as a mixture of mass numbers. The term "unnatural variant isotopic form" also includes embodiments in which the proportion of an atom of given atomic number having a mass number found less commonly in nature (referred to herein as an "uncommon isotope") has been increased relative to that which is naturally occurring e.g. to the level of >20%, >50%, >75%, >90%, >95% or> 99% by number of the atoms of that atomic number (the latter embodiment referred to as an "isotopically enriched variant form"). The term "unnatural variant isotopic form" also includes embodiments in which the proportion of an uncommon isotope has been reduced relative to that which is naturally occurring. Isotopic forms may include radioactive forms (i.e. they incorporate radioisotopes) and non-radioactive forms. Radioactive forms will typically be isotopically enriched variant forms.

[0098] An unnatural variant isotopic form of a compound may thus contain one or more artificial or uncommon isotopes such as deuterium (2H or D), carbon-11 (X1C), carbon-13 (13C), carbon- 14 (14C), nitrogen-13 (13N), nitrogen-15 (15N), oxygen-15 (15O), oxygen-17 (17O), oxygen-18 (18O), phosphorus-32 (32P), sulphur-35 (35S), chlorine-36 (36CI), chlorine-37 (37CI), fluorine- 18 (18F) iodine-123 (123I), iodine-125 (125I) in one or more atoms or may contain an increased proportion of said isotopes as compared with the proportion that predominates in nature in one or more atoms.

[0099] Unnatural variant isotopic forms comprising radioisotopes may, for example, be used for drug and / or substrate tissue distribution studies. The radioactive isotopes tritium, i.e.3H, and carbon-14, i.e.14C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Unnatural variant isotopic forms which incorporate deuterium i.e2H or D may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Further, unnatural variant isotopic forms may be prepared which incorporate positron emitting isotopes, such asnC,18F,150 and13N, and would be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. It is also to be understood that compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or the arrangement of their atoms in space are termed 'isomers'. Isomers that differ in the arrangement of their atoms in space are termed 'stereoisomers'.

[0100] Stereoisomers that are not mirror images of one another are termed 'diastereomers' and those that are non-superimposable mirror images of each other are termed 'enantiomers'. When a compound has an asymmetric center, for example, it is bonded to four different groups, a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of its asymmetric center and is described by the R- and S-sequencing rules of Cahn and Prelog, or by the manner in which the molecule rotates the plane of polarized light and designated as dextrorotatory or levorotatory (i.e. as (+) or (-)-isomers respectively). A chiral compound can exist as either individual enantiomer or as a mixture thereof. A mixture containing equal proportions of the enantiomers is called a 'racemic mixture'.

[0101] 'Tautomers' refer to compounds that are interchangeable forms of a particular compound structure, and that vary in the displacement of hydrogen atoms and electrons. Thus, two structures may be in equilibrium through the movement of n electrons and an atom (usually H). For example, enols and ketones are tautomers because they are rapidly interconverted by treatment with either acid or base. Another example of tautomerism is the aci- and nitroforms of phenylnitromethane, that are likewise formed by treatment with acid or base.

[0102] Tautomeric forms may be relevant to the attainment of the optimal chemical reactivity and biological activity of a compound of interest.

[0103] The compounds of the invention may possess one or more asymmetric centers; such compounds can therefore be produced as individual (R)- or (S)- stereoisomers or as mixtures thereof.

[0104] An optical isomer with unknown absolute configuration may be depicted with an asterix (*) at the chiral center, e.g., . Likewise, when a chemical structure possessing one or more asymmetric centers is described using its chemical name, stereocenters with unknown absolute configuration may be depicted with an asterix (*), e.g. R* and / or S*.

[0105] Unless indicated otherwise, the description or naming of a particular compound in the specification and claims is intended to include both individual enantiomers and mixtures, racemic or otherwise, thereof. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art. It will be appreciated that compounds of the invention may be metabolized to yield biologically active metabolites.

[0106] DETAILED DESCRIPTION OF THE INVENTION

[0107] Compounds

[0108] The present invention relates to compounds that may be useful in the prophylaxis and / or treatment of diseases, disorders, or conditions that are associated with abnormal activity or expression of an FGFR enzyme, in particular cancer. The present invention also provides methods for the production of the compound of the invention, pharmaceutical compositions comprising the compound of the invention, methods for the prophylaxis and / or treatment of diseases, disorders, or conditions that are associated with abnormal activity or expression of an FGFR enzyme, in particular cancer, by administering a compound of the invention.

[0109] Accordingly, in a first aspect, the invention provides compounds of Formula (I): wherein:

[0110] Xi is N or CH;

[0111] X2 is N and X3 is C, or, alternatively, X3 is N and X2 is C;

[0112] X4is N;

[0113] Y is a bond, O or NH;

[0114] Yi is CHR6, CH2-CHR6, CHR6-CH2, CF2, CH2-CF2 or CF2-CH2;

[0115] Y2is CR4R5or CF2;

[0116] Y3is CR3R4, CR3R4CR3R4, CH2CF2 or CF2; wherein Yi, Y2, Y3 and X4 together with the pyrazolyl ring form a 7-membered nitrogen containing ring; B is -C(O)Y4-, -Y4, -SO2-Y4-, - CR4R9Y4-, or -CH2CH2Y4-, and A is CN, R8R9C=CR10C(=O)-, R11R12NCH2CH=CHC(=O)-, H2C=CHSO2- or R13C=CC(=O)-, wherein R4and R9are as defined herewith, or, alternatively, R4and R9together with the carbon atoms to which they are attached form a 3-5-membered carbocyclic ring;

[0117] Y4is a 4-10 membered monocyclic heterocycloalkyl or a 5-12 membered fused, spiro or bridged polycyclic heterocycloalkyl, said monocyclic or polycyclic heterocycloaklyl comprising a nitrogen atom, wherein the heterocycloalkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, methyl, halomethyl, hydroxyl or methoxy and wherein the nitrogen atom of the heterocycloalkyl is connected to A;

[0118] R1is hydrogen, halogen, C1-5 alkyl, C1-5 alkoxy, CN, C3-10 cycloalkyl, C2-4alkynyl, 4-10 membered heterocycloalkyl comprising 1, 2 or 3 heteroatoms independently selected from N, O and S, phenyl or a 5-6 membered heteroaryl, wherein the 3-10 membered cycloalkyl, 4-10 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl are unsubstituted or substituted with one or more substituents independently selected from halogen, methyl, halomethyl, hydroxyl or methoxy and wherein the C1-5 alkyl and C1-5 alkoxy, are unsubstituted or independently substituted with one or more substituents independently selected from halogen, CN, hydroxyl and methoxy;

[0119] R2is hydrogen, hydroxyl, Ci-4alkyl, CN, -CONHCI-4alkyl, or Ci-4haloalkyl; each R3is independently selected from hydrogen, fluoro, hydroxyl, methoxy, C1-3 alkyl, cyclopropyl, or a R3is fused with R5or R6to form CH2, CH2-CH2or CH2OCH2; each R4is independently selected from hydrogen, or C1-3 alkyl;

[0120] R5is hydrogen, fluoro, hydroxyl, methoxy, C1-3 alkyl, cyclopropyl or is fused with one R3to form CH2, CH2-CH2or CH2OCH2, wherein the C1-3 alkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, heteroaryl (which heteroaryl is preferably pyridyl), phenyl, hydroxyl, -CO2R4or C1-3 alkoxy;

[0121] R6is hydrogen, fluoro, hydroxyl, methoxy, C1-3 alkyl or is fused with one R3to form CH2, CH2- CH2or CH2OCH2; each R8is independently selected from hydrogen and C1-6 alkyl; each R9is independently selected from hydrogen, CF3 and C1-6 alkyl, which alkyl is unsubstituted or substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxyl, and C1-6 alkoxy; and R10is hydrogen, C1-3 alkyl or halogen; or R9and R10together with the carbon atoms to which they are attached form a 4-8- membered carbocyclic ring; each of R11and R12is independently hydrogen, C1-6 alkyl, or R11and R12together with the nitrogen atom to which they are attached form a 4-6 membered heterocycloalkyl optionally having an additional ring heteroatom which is O, wherein said ring is optionally substituted with halogen;

[0122] R13is hydrogen, R14R15NCH2-, or Ci-3 alkyl which is unsubstituted or substituted with hydroxyl;

[0123] R14and R15are each independently hydrogen or C1-6 alkyl; R16is C1-3 alkyl-R17or C3-6 cycloalkyl-R17, wherein the C1-3 alkyl or C3-6 cycloalkyl are unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, CN, Ci-4 alkyl optionally substituted with one or more OH, C3-6 cycloalkyl, OH, OMe, -NR4R8wherein R4and R8are as defined above;

[0124] R17is aryl or heteroaryl, wherein the aryl or heteroaryl are unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, CN, NH2, Ci-4 alkyl, and Ci-4 haloalkyl, Ci-4 alkoxy, and Ci-4 haloalkoxy; or a pharmaceutically acceptable salt and / or solvate thereof.

[0125] In certain embodiments, the compound of the Formula (I) is a compound of the Formula (la) : or a pharmaceutically acceptable salt and / or solvate thereof; wherein:

[0126] Z is a absent, or is CO or CR4R9; each n is independently 1 or 2;

[0127] R21is absent or is halogen, methyl, halomethyl, hydroxyl or methoxy; and

[0128] Xi, X2, X3, Y, Yi, Y2, Y3, R1, R2, R4, R9, R16and A are as defined for Formula (I). Preferred definitions of Xi, X2, X3, Y, Yi, Y2, Y3, R1, R2, R4, R9, R16and A are as defined for compounds of formula (I).

[0129] In certain embodiments, the compound of the Formula (I) is a compound of the Formula (lb) : In certain embodiments, the compound of the Formula (I) is a compound of the Formula (Ic): or a pharmaceutically acceptable salt and / or solvate thereof; wherein: a is independently 0, 1, 2, 3, or 4;

[0130] R18is halogen;

[0131] R19is methyl, hydroxymethyl, or halomethyl (preferably fluoromethyl);

[0132] R20is H or methyl; and

[0133] Xi, X2, X3, X4, Yi, Y2, Y3, R1, R2, B and A are as defined for Formula (I). Preferred definitions of Xi, X2, X3, X4, Yi, Y2, Y3, R1, R2, B and A are as defined for compounds of formula (I).

[0134] In certain embodiments, the compound of the Formula (I) is a compound of the Formula (Id): (Id) or a pharmaceutically acceptable salt and / or solvate thereof; wherein: a is independently 0, 1, 2, 3, or 4;

[0135] R18is halogen;

[0136] R19is methyl, hydroxymethyl, or OH;

[0137] R20is H or methyl; and

[0138] Xi, X2, X3, X4, Yi, Y2, Y3, R1, R2, B and A, are as defined for Formula (I). Preferred definitions of Xi, X2, X3, X4, Yi, Y2, Y3, R1, R2, B and A are as defined for compounds of formula (I).

[0139] In certain embodiments, the compound of the Formula (I) is a compound of formula (le): or a pharmaceutically acceptable salt and / or solvate thereof; and

[0140] Xi, X2, X3, Y, R1, R2, R16and B are as defined for Formula (I). Preferred definitions of Xi, X2, X3, R1, R2and B are as defined for compounds of Formula (I).

[0141] In certain embodiments, the compound of the Formula (I) is a compound of formula (If): or a pharmaceutically acceptable salt and / or solvate thereof; and

[0142] Xi, X2, X3, Y, R1, R2, R16and B are as defined for Formula (I). Preferred definitions of Xi, X2, X3, R1, R2and B are as defined for compounds of Formula (I).

[0143] In an embodiment, Xi is CH, X3 is N and X2 is C and Y is O. In an embodiment, Xi is N, X3 is N and X2 is C and Y is O.

[0144] In an embodiment, Yi is CHR6, Y2 is CR4R5or CF2 and Y3 is CH2CR3R4.

[0145] According to some embodiments, B is preferably selected from: wherein the nitrogen atom of the heterocycloalkyl is connected to A; more preferably A is CN absent or each is independently halogen (preferably fluoro), methyl or methoxy and each R22is independently hydrogen, halogen (preferably fluoro) or methyl.

[0146] In an embodiment, B is -C(O)Y4- or -Y4-.

[0147] In an embodiment, B is -CR4R9Y4 , such as -CH2Y4-, wherein R4and R9are as defined herewith, or, alternatively, R4and R9together with the carbon atoms to which they are attached form a 3-5-membered carbocyclic ring. In an embodiment, Y4is a 4-10 membered heterocycloalkyl comprising a nitrogen atom, wherein the heterocycloalkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, methyl, halomethyl, hydroxyl or methoxy and wherein the nitrogen atom of the heterocycloalkyl is connected to A

[0148] In an embodiment R1is halogen such as fluoro, chloro, bromo or iodo.

[0149] In an embodiment R1is C1-5 alkoxy, wherein the C1-5 alkoxy is unsubstituted or independently substituted with one or more substituents independently selected from halogen, CN, hydroxyl and methoxy such as halogen e.g. F, Cl, Br or I.

[0150] In an embodiment R1is -OMe or -OCHF2.

[0151] In an embodiment R2is Ci-4 alkyl such as methyl, ethyl, propyl and butyl and R3, R4, R5and R6are hydrogen.

[0152] Most preferably R2is hydrogen, methyl or ethyl.

[0153] Even more preferably R2is methyl.

[0154] Preferably each R3is independently selected from hydrogen, fluoro and C1-2 alkyl.

[0155] More preferably each R3is independently selected from hydrogen, fluoro and methyl.

[0156] Most preferably each R3is independently hydrogen.

[0157] Preferably each R4is independently selected from hydrogen and methyl.

[0158] Most preferably each R4is independently hydrogen.

[0159] Preferably R5is hydrogen, fluoro, hydroxyl, methoxy or C1-3 alkyl, wherein the C1-3 alkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, pyridyl, phenyl, hydroxyl, -CO2R4or C1-3 alkoxy.

[0160] Most preferably R5is hydrogen or C1-3 alkyl, wherein the C1-3 alkyl is unsubstituted or substituted with a substituent selected from pyridyl, phenyl, and -CO2H.

[0161] Even more preferably R5is hydrogen.

[0162] Preferably R6is hydrogen, fluoro or C1-2 alkyl. More preferably R6is hydrogen, fluoro or methyl.

[0163] Most preferably R6is hydrogen.

[0164] Preferably each R8is independently selected from hydrogen and C1-2 alkyl.

[0165] More preferably each R8is independently selected from hydrogen and methyl.

[0166] Most preferably each R8is independently hydrogen.

[0167] Preferably each R9is independently selected from hydrogen, CF3 and C1-2 alkyl, which alkyl is unsubstituted or substituted with one or more substituents independently selected from fluoro, chloro and bromo.

[0168] More preferably each R9is independently selected from hydrogen, CF3 and methyl.

[0169] Most preferably each R9is independently hydrogen.

[0170] Preferably each R10is independently selected from hydrogen, C1-2 alkyl and halogen.

[0171] More preferably each R10is independently selected from hydrogen, fluoro, chloro, and methyl.

[0172] Most preferably each R10is independently selected from hydrogen, fluoro and methyl.

[0173] In an embodiment R16is C1-3 alkyl-R17wherein the C1-3 alkyl group is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, CN, Ci-4 alkyl, C3-6 cycloalkyl, OH, OMe, -NR4R8wherein R4and R8are as defined above.

[0174] Preferably R16is C1-3 alkyl-R17, wherein the C1-3 alkyl is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, C1-2 alkyl, C3-6 cycloalkyl, OH and OMe.

[0175] In one embodiment, R16is Ci alkyl-R17, wherein the Ci alkyl is unsubstituted or substituted with one or more substituents as defined herein. In a second embodiment, R16is C2 alkyl-R17, wherein the C2 alkyl is unsubstituted or substituted with one or more substituents as defined herein. In a third embodiment, R16is C3 alkyl-R17, wherein the C3 alkyl is unsubstituted or substituted with one or more substituents as defined herein. When R16is C1-3 alkyl-R17substituted with one or more (e.g. one or two e.g. one) substituents independently selected from the group consisting of halogen, C1-2 alkyl, C3-6 cycloalkyl, OH and OMe (e.g. OH), R16is suitably -CH(R17)-CH2(substituent) e.g. -CH(R17)-CH2(OH).

[0176] In one embodiment, the C1-3 alkyl is substituted by one substituent.

[0177] Suitably, the one substituent is OH. Alternatively the one substituent is methyl.

[0178] In one embodiment, R16is selected from C2 alkyl-R17or C3 alkyl-R17, wherein the C2 alkyl or C3 alkyl is substituted with at least one OH.

[0179] In one embodiment, R16is C2 alkyl-R17, wherein the C2 alkyl is substituted with OH.

[0180] In one embodiment, R16is Ci alkyl-R17, wherein the Ci alkyl is substituted with OH or Ci alkyl- OH.

[0181] More preferably R16is selected from:

[0182] Even more preferably R16is selected from:

[0183] Most preferably R16is selected from:

[0184] In an embodiment R17is aryl or heteroaryl, such as pyrazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl and pyrazinyl, such as pyridinyl.

[0185] In one embodiment, R17is 2-pyridinyl, optionally substituted with one or more F. Preferably said 2-pyridinyl is substituted with one F in para position.

[0186] In one embodiment, A is CN.

[0187] In one embodiment, A is CH2=CR10C(=O)-.

[0188] In one embodiment, said aryl and heteroaryl are unsubstituted. In another embodiment said aryl and heteroaryl are substituted with one or more R23independently selected from the group consisting of halogen (such as F), CN, NH2, Ci-4 alkyl (such as methyl), Ci-4 haloalkyl (such as Ci haloalkyl, such as CHF2), Ci-4 alkoxy (such as OMe), and Ci-4 haloalkoxy, optionally substituted with one or more substituents independently selected from the group consisting of halogen, CN, Ci-4 alkyl, Ci-4 haloalkyl, Ci-4 alkoxy, and Ci-4 haloalkoxy.

[0189] In one embodiment, each R23is independently selected from the group consisting of halogen (such as F), CN, C1-2 alkyl (such as Me), and C1-2 haloalkyl (such as Ci haloalkyl, such as CHF2), C1-2 alkoxy (such as OMe), and C1-2 haloalkoxy. Most suitably, R23is F.

[0190] More preferably still R17is phenyl or pyridyl, wherein the phenyl or pyridyl are unsubstituted or substituted with one or more substituents independently selected from halogen. Most preferably R17is 2-pyridyl which is unsubstituted or substituted with one or two halogens (preferably fluoro). Certain compounds according to the invention may exhibit one or more benefits including, inter alia, advantageous levels of biological activity which may be useful in the prophylaxis and / or treatment of one or more disease, improved safety characteristics (e.g. relating to hERG inhibition, drug-drug interaction (DDI) or CYP-interaction characteristics, etc), improved selectivity for one or more disease-associated biological target (e.g. reduced off-target effects, etc), improved pharmacokinetic properties (e.g. relating to dosing, solubility, absorption, etc), improved pharmacodynamic properties (e.g. relating to permeability, efflux, etc) or superior properties for use as pharmaceutical active ingredients alone or in pharmaceutical compositions (e.g. stability), or advantageous physico-chemical properties useful in the manufacturability of such aforementioned pharmaceutical compositions.

[0191] Preferred values of Xi, X2, X3, X4, Y, Yi, Y2, Y3, Y4, R1, R2, R3, R4, R5, R6, R8, R9, R10, R11, R12, R13, R14, R15, R16, R17, B and A are, in any combination thereof, as set out below.

[0192] Preferably Y is O.

[0193] Preferably Yi is CHR6, most preferably Yi is CH2.

[0194] Preferably Y2is CR4R5, more preferably Y2is CHR5, most preferably Y2is CH2.

[0195] Preferably Y3is CH2CR3R4, most preferably Y3is CH2CH2.

[0196] Preferably R1is hydrogen, halogen, Ci-3alkyl, Ci-3alkoxy, CN, wherein the Ci-3alkyl and Ci-3alkoxy are unsubstituted or independently substituted with one or more substituents independently selected from halogen.

[0197] More preferably R1is hydrogen, fluoro, chloro, bromo, methyl, ethyl, methoxy, ethoxy, halomethyl, haloethyl, halomethoxy, haloethoxy, cyano, CF3, CHF2, OCH3or OCHF2.

[0198] Most preferably R1is hydrogen, fluoro, chloro, bromo, methyl, ethyl, methoxy, ethoxy, cyano, CF3, CHF2, OCH3or OCHF2(especially OCH3or OCHF2).

[0199] Preferably R2is hydrogen, hydroxyl or Ci-2alkyl.

[0200] Most preferably R2is hydrogen, methyl or ethyl.

[0201] Preferably each R3is independently selected from hydrogen, fluoro and Ci-2alkyl. More preferably each R3is independently selected from hydrogen, fluoro and methyl. Most preferably each R3is independently hydrogen. Preferably each R4is independently selected from hydrogen and methyl. Most preferably each R4is independently hydrogen.

[0202] Preferably R5is hydrogen, fluoro, hydroxyl, methoxy or C1-3 alkyl, wherein the C1-3 alkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, pyridyl, phenyl, hydroxyl, -CO2R4or C1-3 alkoxy.

[0203] Most preferably R5is hydrogen or C1-3 alkyl, wherein the C1-3 alkyl is unsubstituted or substituted with a substituent selected from pyridyl, phenyl, and -CO2H.

[0204] Preferably R6is hydrogen, fluoro or C1-2 alkyl. More preferably R6is hydrogen, fluoro or methyl. Most preferably R6is hydrogen.

[0205] Preferably each R8is independently selected from hydrogen and C1-2 alkyl. More preferably each R8is independently selected from hydrogen and methyl. Most preferably each R8is independently hydrogen.

[0206] Preferably each R9is independently selected from hydrogen, CF3 and C1-2 alkyl, which alkyl is unsubstituted or substituted with one or more substituents independently selected from fluoro, chloro and bromo. More preferably each R9is independently selected from hydrogen, CF3 and methyl. Most preferably each R9 is independently hydrogen.

[0207] Preferably each R10is independently selected from hydrogen, C1-2 alkyl and halogen. More preferably each R10is independently selected from hydrogen, fluoro, chloro, and methyl. Most preferably each R10is independently selected from hydrogen, fluoro and methyl.

[0208] Preferably each of R11and R12is independently hydrogen, C1-3 alkyl, or R11and R12together with the nitrogen atom to which they are attached form a 4-6 membered heterocycloalkyl optionally having an additional ring heteroatom which is O, wherein said heterocycloalkyl is unsubstituted or substituted with one or more halogen atoms. More preferably each of R11and R12is independently hydrogen, C1-2 alkyl, or R11and R12together with the nitrogen atom to which they are attached form a 4-6 membered heterocycloalkyl, wherein said heterocycloalkyl is unsubstituted or substituted with one or more halogen atoms. Most preferably each of R11and R12is independently hydrogen or methyl, or R11and R12together with the nitrogen atom to which they are attached form a 4-6 membered heterocycloalkyl.

[0209] Preferably each R13is independently selected from hydrogen, R14R15NCH2-, and C1-3 alkyl, which alkyl is unsubstituted or substituted with hydroxyl. More preferably each R13is independently selected from hydrogen and C1-2 alkyl, which alkyl is unsubstituted or substituted with hydroxyl. Most preferably each R13is independently selected from hydrogen and methyl.

[0210] Preferably R14and R15are each independently hydrogen or C1-3 alkyl. More preferably R14and R15are each independently hydrogen or C1-2 alkyl. Most preferably R14and R15are each independently hydrogen or methyl.

[0211] In some embodiments, X4is N, B is -C(O)Y4-, -Y4-, -SO2-Y4-, -CR4R9Y4CH2CH2Y4or -CH2Y4-, and preferably A is CN or R8R9C=CR10C(=O)-, R11R12NCH2CH=CHC(=O)-, H2C=CHSO2- or R13C=CC(=O)-; wherein R4and R9are as defined herewith, or, alternatively, R4and R9together with the carbon atoms to which they are attached form a 3-5-membered carbocyclic ring, wherein preferably Y4is a 4-10 membered heterocycloalkyl comprising a nitrogen atom wherein the heterocycloalkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, methyl, halomethyl, hydroxyl or methoxy and wherein the nitrogen atom of the heterocycloalkyl is connected to A.

[0212] Embodiments

[0213] Further embodiments according to the invention are provided as set out below.

[0214] Table 1 Compounds according to an embodiment of the invention

[0215]

[0216] Pharmaceutical compositions and formulations

[0217] When employed as a pharmaceutical, a compound of the invention is typically administered in the form of a pharmaceutical composition. Such compositions can be prepared in a manner well known in the pharmaceutical art and comprise at least one active compound of the invention according to Formula I. Generally, a compound of the invention is administered in a pharmaceutically effective amount. The amount of compound of the invention actually administered will typically be determined by a physician, in the light of the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound of the invention administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

[0218] The pharmaceutical compositions of this invention can be administered by a variety of routes including oral, rectal, transdermal, subcutaneous, intra-articular, intravenous, intramuscular, and intranasal. Depending on the intended route of delivery, a compound of the invention is preferably formulated as either injectable or oral compositions or as salves, as lotions or as patches all for transdermal administration.

[0219] The compositions for oral administration can take the form of bulk liquid solutions or suspensions, or bulk powders. More commonly, however, the compositions are presented in unit dosage forms to facilitate accurate dosing. The term 'unit dosage forms' refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient, vehicle or carrier. Typical unit dosage forms include prefilled, premeasured ampules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions. In such compositions, the compound of the invention according to Formula I is usually a minor component (from about 0.1 to about 50% by weight or preferably from about 1 to about 40% by weight) with the remainder being various vehicles or carriers and processing aids helpful for forming the desired dosing form.

[0220] Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous vehicle with buffers, suspending and dispensing agents, colorants, flavors and the like. Solid forms may include, for example, any of the following ingredients, or compound of the inventions of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or corn starch; a lubricant such as magnesium stearate; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint or orange flavoring.

[0221] Injectable compositions are typically based upon injectable sterile saline or phosphate- buffered saline or other injectable carriers known in the art. As before, the active compound of the invention according to Formula I in such compositions is typically a minor component, often being from about 0.05 to 10% by weight with the remainder being the injectable carrier and the like. Transdermal compositions are typically formulated as a topical ointment or cream containing the active ingredient(s), generally in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, preferably from about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight. When formulated as an ointment, the active ingredients will typically be combined with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients may be formulated in a cream with, for example an oil-in-water cream base. Such transdermal formulations are well-known in the art and generally include additional ingredients to enhance the dermal penetration of stability of the active ingredients or the formulation. All such known transdermal formulations and ingredients are included within the scope of this invention.

[0222] A compound of the invention can also be administered by a transdermal device. Accordingly, transdermal administration can be accomplished using a patch either of the reservoir or porous membrane type, or of a solid matrix variety.

[0223] The above-described components for orally administrable, injectable or topically administrable compositions are merely representative. Other materials as well as processing techniques and the like are set forth in Part 8 of Remington's Pharmaceutical Sciences, 17thedition, 1985, Mack Publishing Company, Easton, Pennsylvania, which is incorporated herein by reference.

[0224] A compound of the invention can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.

[0225] The following formulation examples illustrate representative pharmaceutical compositions that may be prepared in accordance with this invention. The present invention, however, is not limited to the following pharmaceutical compositions.

[0226] Formulation 1 - Tablets

[0227] A compound of the invention according to Formula I may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate may be added as a lubricant. The mixture may be formed into 240-270 mg tablets (80-90 mg of active compound of the invention according to Formula I per tablet) in a tablet press.

[0228] Formulation 2 - Capsules

[0229] A compound of the invention according to Formula I may be admixed as a dry powder with a starch diluent in an approximate 1 : 1 weight ratio. The mixture may be filled into 250 mg capsules (125 mg of active compound of the invention according to Formula I per capsule).

[0230] Formulation 3 - Liquid A compound of the invention according to Formula I (125 mg), may be admixed with sucrose (1.75 g) and xanthan gum (4 mg) and the resultant mixture may be blended, passed through a No. 10 mesh U.S. sieve, and then mixed with a previously made solution of microcrystalline cellulose and sodium carboxymethyl cellulose (11 :89, 50 mg) in water. Sodium benzoate (10 mg), flavor, and color may be diluted with water and added with stirring. Sufficient water may then be added with stirring. Further sufficient water may be then added to produce a total volume of 5 mL.

[0231] Formulation 4 - Tablets

[0232] A compound of the invention according to Formula I may be admixed as a dry powder with a dry gelatin binder in an approximate 1:2 weight ratio. A minor amount of magnesium stearate may be added as a lubricant. The mixture may be formed into 450-900 mg tablets (150-300 mg of active compound of the invention according to Formula I) in a tablet press.

[0233] Formulation 5 - Injection

[0234] A compound of the invention according to Formula I may be dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of approximately 5 mg / mL.

[0235] Formulation 6 - Topical

[0236] Stearyl alcohol (250 g) and a white petrolatum (250 g) may be melted at about 75°C and then a mixture of A compound of the invention according to Formula I (50 g) methylparaben (0.25 g), propylparaben (0.15 g), sodium lauryl sulfate (10 g), and propylene glycol (120 g) dissolved in water (about 370 g) may be added and the resulting mixture may be stirred until it congeals.

[0237] Methods of treatment

[0238] In one embodiment, the present invention provides compounds of the invention, or pharmaceutical compositions comprising a compound of the invention, for use in medicine. In a particular embodiment, the present invention provides compounds of the invention or pharmaceutical compositions comprising a compound of the invention, for use in the prophylaxis and / or treatment of a disease, disorder, or condition that is associated with abnormal activity or expression of an FGFR enzyme (in particular FGFR.3), in particular cancer.

[0239] In an embodiment, the present invention provides compounds of the invention, or pharmaceutical compositions comprising a compound of the invention for use in the manufacture of a medicament for use in the prophylaxis and / or treatment of a disease, disorder, or condition that is associated with abnormal activity or expression of an FGFR enzyme (in particular FGFR3), in particular cancer.

[0240] In another embodiment, this invention provides methods of prophylaxis and / or treatment of a mammal afflicted with a disease, disorder, or condition that is associated with abnormal activity or expression of an FGFR enzyme, in particular cancer, which methods comprise the administration of an effective amount of a compound of the invention or one or more of the pharmaceutical compositions described herein for the treatment or prophylaxis of said condition.

[0241] In one embodiment, the present invention provides pharmaceutical compositions comprising a compound of the invention, and another therapeutic agent. In a particular embodiment, the other therapeutic agent is a treatment agent directed to a disease, disorder, or condition that is associated with abnormal activity or expression of an FGFR enzyme (in particular FGFR3), in particular cancer.

[0242] In a particular embodiment, the present invention provides compounds of the invention or pharmaceutical compositions comprising a compound of the invention, for use in the prophylaxis and / or treatment of a disease selected from: metastatic tumours (such as but not limited to, melanoma, lymphoma, leukaemia, fibrosarcoma, rhabdomyosarcoma, and mastocytoma) and types of tissue carcinoma (such as but not limited to, colorectal cancer, prostate cancer, small cell lung cancer and non-small cell lung cancer, breast cancer, pancreatic cancer, bladder cancer, renal cancer, gastric cancer, glioblastoma, primary liver cancer, ovarian cancer, prostate cancer and uterine leiomyosarcoma), acute lymphoblastic leukemia, acute myeloidleukemia, adrenocortical carcinoma, anal cancer, appendix cancer, astrocytomas, atypical teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer (osteosarcoma and malignant fibrous histiocytoma), brain stem glioma, brain tumors, brain and spinal cord tumors, breast cancer, bronchial tumors, Burkitt lymphoma, cervical cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T -Cell lymphoma, embryonal tumors, endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, ewing sarcoma family of tumors, eye cancer, retinoblastoma, gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), gastrointestinal stromal cell tumor, germ cell tumor, glioma, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumors (endocrine pancreas), Kaposi sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, leukemia, Acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, liver cancer, non-small cell lung cancer, small cell lung cancer, Burkitt lymphoma, cutaneous T-celllymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, lymphoma, Waldenstrom macroglobulinemia, medulloblastoma, medulloepithelioma, melanoma, mesothelioma, mouth cancer, chronic myelogenous leukemia, myeloid leukemia, multiple myeloma, asopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma, malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, papillomatosis, parathyroid cancer, penile cancer, pharyngeal cancer, pineal parenchymal tumors of intermediate differentiation, pineoblastoma and supratentorial primitive neuroectodermal tumors, pituitary tumor, plasma cell neoplasm / multiple myeloma, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer, rectal cancer, renal cell (kidney) cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, Ewing sarcoma family of tumors, sarcoma, kaposi, Sezary syndrome, skin cancer, small cell Lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, stomach (gastric) cancer, supratentorial primitive neuroectodermal tumors, T -cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, and Wilms tumor.

[0243] In an embodiment, the disease or condition is a neoplastic disease of the blood and blood forming organs, including but not limited to: acute myeloid leukaemia (AML), and acute lymphoblastic leukemia (ALL) and chronic lymphoblastic leukaemia (CLL).

[0244] In an embodiment, the disease or condition is selected from breast cancer (e.g. invasive ductal cancer, invasive lobular cancer), lung cancer (e.g. non-small-cell lung cancer, lung adenocarcinoma, squamous cell lung cancer and small-cell lung cancer), urothelial cancer, bladder cancer (e.g. urothelial bladder cancer, non-muscle invasive bladder cancer, muscle invasive bladder cancer), upper tract cancer (e.g. urothelial upper tract cancer), urethral cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, multiple myeloma, liver cancer, melanoma (e.g. cutaneous melanoma), head and neck cancer (e.g. oral cancer), thyroid cancer, renal cancer (e.g. renal pelvis cancer), glioblastoma, endometrial cancer, cervical cancer, ovarian cancer, and testicular cancer.

[0245] Injection dose levels range from about 0.1 mg / kg / h to at least 10 mg / kg / h, all for from about 1 to about 120 h and especially 24 to 96 h. A preloading bolus of from about 0.1 mg / kg to about 10 mg / kg or more may also be administered to achieve adequate steady state levels. The maximum total dose is not expected to exceed about 1 g / day for a 40 to 80 kg human patient.

[0246] For the prophylaxis and / or treatment of long-term conditions, such as degenerative conditions, the regimen for treatment usually stretches over many months or years so oral dosing is preferred for patient convenience and tolerance. With oral dosing, one to four (1-4) regular doses daily, especially one to three (1-3) regular doses daily, typically one to two (1- 2) regular doses daily, and most typically one (1) regular dose daily are representative regimens. Alternatively for long lasting effect drugs, with oral dosing, once every other week, once weekly, and once a day are representative regimens. In particular, dosage regimen can be every 1-14 days, more particularly 1-10 days, even more particularly 1-7 days, and most particularly 1-3 days.

[0247] Using these dosing patterns, each dose provides from about 1 to about 1000 mg of a compound of the invention, with particular doses each providing from about 10 to about 500 mg and especially about 30 to about 250 mg.

[0248] Transdermal doses are generally selected to provide similar or lower blood levels than are achieved using injection doses.

[0249] When used to prevent the onset of a condition, a compound of the invention will be administered to a patient at risk for developing the condition, typically on the advice and under the supervision of a physician, at the dosage levels described above. Patients at risk for developing a particular condition generally include those that have a family history of the condition, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the condition.

[0250] A compound of the invention can be administered as the sole active agent or it can be administered in combination with other therapeutic agents, including other compound of the inventions that demonstrate the same or a similar therapeutic activity and that are determined to be safe and efficacious for such combined administration. In a specific embodiment, co-administration of two (or more) agents allows for significantly lower doses of each to be used, thereby reducing the side effects seen.

[0251] In one embodiment, a compound of the invention or a pharmaceutical composition comprising a compound of the invention is administered as a medicament. In a specific embodiment, said pharmaceutical composition additionally comprises a further active ingredient.

[0252] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of a disease involving inflammation, particular agents include, but are not limited to, immunoregulatory agents e.g. azathioprine, corticosteroids (e.g. prednisolone or dexamethasone), cyclophosphamide, cyclosporin A, tacrolimus, mycophenolate, mofetil, muromonab-CD3 (OKT3, e.g. Orthocolone®), ATG, aspirin, acetaminophen, ibuprofen, naproxen, and piroxicam. In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of arthritis {e.g. rheumatoid arthritis), particular agents include but are not limited to analgesics, non-steroidal anti-inflammatory drugs (NSAIDS), steroids, synthetic DMARDS (for example but without limitation methotrexate, leflunomide, sulfasalazine, auranofin, sodium aurothiomalate, penicillamine, chloroquine, hydroxychloroquine, azathioprine, tofacitinib, baricitinib, fostamatinib, and cyclosporin), and biological DMARDS (for example but without limitation infliximab, etanercept, adalimumab, rituximab, and abatacept).

[0253] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of proliferative disorders, particular agents include but are not limited to: methotrexate, leukovorin, adriamycin, prednisone, bleomycin, cyclophosphamide, 5-fluorouracil, paclitaxel, docetaxel, vincristine, vinblastine, vinorelbine, doxorubicin, tamoxifen, toremifene, megestrol acetate, anastrozole, goserelin, anti-HER2 monoclonal antibody (e.g. Herceptin™), capecitabine, raloxifene hydrochloride, EGFR inhibitors (e.g. Iressa®, Tarceva™, Erbitux™), VEGF inhibitors (e.g. Avastin™), proteasome inhibitors (e.g. Velcade™), Glivec® and hsp90 inhibitors (e.g. 17-AAG). Additionally, the compound of the invention according to Formula I may be administered in combination with other therapies including, but not limited to, radiotherapy or surgery. In a specific embodiment the proliferative disorder is selected from cancer, myeloproliferative disease or leukaemia.

[0254] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of autoimmune diseases, particular agents include but are not limited to: glucocorticoids, cytostatic agents (e.g. purine analogs), alkylating agents, (e.g nitrogen mustards (cyclophosphamide), nitrosoureas, platinum compound of the inventions, and others), antimetabolites (e.g. methotrexate, azathioprine and mercaptopurine), cytotoxic antibiotics (e.g. dactinomycin anthracyclines, mitomycin C, bleomycin, and mithramycin), antibodies (e.g. anti-CD20, anti-CD25 or anti-CD3 (OTK3) monoclonal antibodies, Atgam® and Thymoglobuline®), cyclosporin, tacrolimus, rapamycin (sirolimus), interferons (e.g. IFN-P), TNF binding proteins (e.g. infliximab, etanercept, or adalimumab), mycophenolate, fingolimod and myriocin..

[0255] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of transplant rejection, particular agents include but are not limited to: calcineurin inhibitors (e.g. cyclosporin or tacrolimus (FK506)), mTOR inhibitors (e.g. sirolimus, everolimus), anti-proliferatives (e.g. azathioprine, mycophenolic acid), corticosteroids (e.g. prednisolone, hydrocortisone), antibodies (e.g. monoclonal anti- IL-2Ro receptor antibodies, basiliximab, daclizumab), polyclonal anti-T-cell antibodies (e.g. anti-thymocyte globulin (ATG), anti-lymphocyte globulin (ALG)). In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of asthma and / or rhinitis and / or COPD, particular agents include but are not limited to: beta2-adrenoceptor agonists (e.g. salbutamol, levalbuterol, terbutaline and bitolterol), epinephrine (inhaled or tablets), anticholinergics (e.g. ipratropium bromide), glucocorticoids (oral or inhaled). Long-acting [32-agonists (e.g. salmeterol, formoterol, bambuterol, and sustained-release oral albuterol), combinations of inhaled steroids and long-acting bronchodilators (e.g. fluticasone / salmeterol, budesonide / formoterol), leukotriene antagonists and synthesis inhibitors (e.g. montelukast, zafirlukast and zileuton), inhibitors of mediator release (e.g. cromoglycate and ketotifen), biological regulators of IgE response (e.g. omalizumab), antihistamines (e.g. ceterizine, cinnarizine, fexofenadine) and vasoconstrictors (e.g. oxymethazoline, xylomethazoline, nafazoline and tramazoline).

[0256] Additionally, a compound of the invention may be administered in combination with emergency therapies for asthma and / or COPD, such therapies include oxygen or heliox administration, nebulized salbutamol or terbutaline (optionally combined with an anticholinergic (e.g. ipratropium), systemic steroids (oral or intravenous, e.g. prednisone, prednisolone, methylprednisolone, dexamethasone, or hydrocortisone), intravenous salbutamol, non-specific beta-agonists, injected or inhaled (e.g. epinephrine, isoetharine, isoproterenol, metaproterenol), anticholinergics (IV or nebulized, e.g. glycopyrrolate, atropine, ipratropium), methylxanthines (theophylline, aminophylline, bamiphylline), inhalation anesthetics that have a bronchodilatory effect (e.g. isoflurane, halothane, enflurane), ketamine and intravenous magnesium sulfate.

[0257] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of inflammatory bowel disease (IBD), particular agents include but are not limited to: glucocorticoids (e.g. prednisone, budesonide) synthetic disease modifying, immunomodulatory agents (e.g. methotrexate, leflunomide, sulfasalazine, mesalazine, azathioprine, 6-mercaptopurine and cyclosporin) and biological disease modifying, immunomodulatory agents (infliximab, adalimumab, rituximab, and abatacept).

[0258] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of SLE, particular agents include but are not limited to: human monoclonal antibodies (belimumab (Benlysta)), Disease-modifying antirheumatic drugs (DMARDs) such as antimalarials (e.g. plaquenil, hydroxychloroquine), immunosuppressants (e.g. methotrexate and azathioprine), cyclophosphamide and mycophenolic acid, immunosuppressive drugs and analgesics, such as nonsteroidal antiinflammatory drugs, opiates (e.g. dextropropoxyphene and co-codamol), opioids (e.g. hydrocodone, oxycodone, MS Contin, or methadone) and the fentanyl duragesic transdermal patch. In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of psoriasis, particular agents include but are not limited to: topical treatments such as bath solutions, moisturizers, medicated creams and ointments containing coal tar, dithranol (anthralin), corticosteroids like desoximetasone (Topicort™), fluocinonide, vitamin D3 analogues (for example, calcipotriol), argan oil and retinoids (etretinate, acitretin, tazarotene), systemic treatments such as methotrexate, cyclosporine, retinoids, tioguanine, hydroxyurea, sulfasalazine, mycophenolate mofetil, azathioprine, tacrolimus, fumaric acid esters or biologies such as Amevive™, Enbrel™, Humira™, Remicade™, Raptiva™ and ustekinumab (a IL-12 and IL-23 blocker). Additionally, a compound of the invention may be administered in combination with other therapies including, but not limited to phototherapy, or photochemotherapy (e.g. psoralen and ultraviolet A phototherapy (PUVA)).

[0259] In one embodiment, a compound of the invention is co-administered with another therapeutic agent for the treatment and / or prophylaxis of allergic reaction, particular agents include but are not limited to: antihistamines (e.g. cetirizine, diphenhydramine, fexofenadine, levocetirizine), glucocorticoids (e.g. prednisone, betamethasone, beclomethasone, dexamethasone), epinephrine, theophylline or anti-leukotrienes (e.g. montelukast or zafirlukast), anti-cholinergics and decongestants.

[0260] By co-administration is included any means of delivering two or more therapeutic agents to the patient as part of the same treatment regime, as will be apparent to the skilled person. Whilst the two or more agents may be administered simultaneously in a single formulation, i.e. as a single pharmaceutical composition, this is not essential. The agents may be administered in different formulations and at different times.

[0261] Synthesis Routes

[0262] The compound of the invention can be prepared from readily available starting materials using the following general methods and procedures. It will be appreciated that where typical or preferred process conditions i.e. reaction temperatures, times, mole ratios of reactants, solvents, pressures, etc.) are given, other process conditions can also be used unless otherwise stated. Optimum reaction conditions may vary with the particular reactants or solvent used, but such conditions can be determined by one skilled in the art by routine optimization procedures.

[0263] Additionally, as will be apparent to those skilled in the art, conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The choice of a suitable protecting group for a particular functional group as well as suitable conditions for protection and deprotection are well known in the art (Wuts & Greene 2006).

[0264] The following methods are presented with details as to the preparation of a compound of the invention as defined hereinabove and the comparative examples. A compound of the invention may be prepared from known or commercially available starting materials and reagents by one skilled in the art of organic synthesis.

[0265] All reagents are of commercial grade and are used as received without further purification, unless otherwise stated. Commercially available anhydrous solvents are used for reactions conducted under inert atmosphere. Reagent grade solvents are used in all other cases, unless otherwise specified. Column chromatography is performed on silica gel 60 (35-70 pm) or with Interchim® PuriFlash® Si HC flash chromatography cartridges. Thin layer chromatography is carried out using pre-coated silica gel F-254 plates (thickness 0.25 mm). Biotage® ISOLUTE® phase separators (e.g., Cat# 120-1907-E) are used for aqueous phase separation. Biotage® ISOLUTE® SCX-3 cation exchange sorbent (e.g., Cat# 533-0100-C) is used to extract basic analytes.XH NMR spectra are recorded on a Bruker Avance 400 NMR spectrometer (400 MHz). Chemical shifts (6) forXH NMR spectra are reported in parts per million (ppm) relative to tetramethylsilane (6 0.00) or the appropriate residual solvent peak, i.e. CHCh (6 7.27), as internal reference. Multiplicities are given as singlet (s), doublet (d), triplet (t), quartet (q), multiplet (m) and broad (br). Electrospray MS spectra are obtained on a Waters Acquity H- Class UPLC system coupled to a to a Waters QDA Mass detector spectrometer. Columns used: Waters Acquity UPLC BEH C18 1.7pm, 2.1mm ID x 50mm L, or Waters Acquity UPLC CSH C18 1.7pm, 2.1mm ID x 50mm L. The methods are using 5-95 % MeCN / H?© gradients in either 2 or 8 minute runs. H2O contains either 0.1% formic acid (when used with CSH column) , or 15 mM NH3 (when used with BEH column). Final compounds were analyzed on 8 minute runs, using either acidic conditions (analytical method A) or basic conditions (analytical method B). Preparative HPLC is performed on a Waters AutoPurification system with UV and MS detection using Waters XBRIDGE BEH C18 OBD 19 mm ID x 100 mm L columns and ACN / H2O gradients with either 0.1% formic acid in H2O or 0.5% NH3 in H2O. Reversed phase column chromatography is performed using a Buchi® Pure C850 FlashPrep using the same columns as for preparative HPLC. Microwave heating is performed with a Biotage® Initiator-i- .

[0266] Scheme 1

[0267] A compound of Formula (la) (R21 not shown), Z is a linker such as a bond, CO or (CH2)n can be prepared from an amine of Formula (V) following procedure depicted in Scheme 3. When Z is CO, compound of Formula (IX) can be synthesized by reaction between compound of Formula XX and an acyl chloride in presence of a base and a suitable solvent. Alternatively, compound of formula (IX) can be obtained from a carboxylic acid in the presence of a coupling agent (such as HATU) and a base (such as DIEA) in a suitable solvent (such as DMF or THF). When Z is a bond or (CH2)x, compound of Formula (IX) can be synthesized from an aldehyde or a ketone using a reducing agent (such as STAB) in a solvent (such as DCE). Alternatively, such compound can also be obtained from an alkyl halide in presence of a base and in a suitable solvent. The Boc protecting group of compound of Formula (IX) can be cleaved using acidic condition, for example, in presence of HCI in a suitable solvent (such as 1,4-dioxane). The reaction of the resulting deprotected amine of Formula (X) with a suitable agent can afford a compound of Formula (la). As an example, compounds of Formula (la) in which A is a nitrile group, can be obtained by reacting a compound of Formula (X) with BrCN in the presence of a base in a suitable solvent. Other compounds of Formula (la) in which A is an acyl moiety, can be obtained by reacting a compound of Formula (X) with an acyl chloride (such as CH2=CH-C(=O)CI) in the presence of a base in a suitable solvent or by reacting with a carboxylic acid in presence of a coupling agent (such as HATU) in the presence of a base in a suitable solvent.

[0268] Compound of Formula (XXXV) can be obtained from intermediate of Formula (XXIV) in two steps via the addition of a protecting group such as Boc and the bromination of the pyrazol intermediate of Formula (XXXIV) using a suitable halogenating agent (such as bromine or NBS). Scheme 3

[0269] Intermediate of Formula (XXIV) where Y3 is (CH?)? can be synthesized following protocol described in Scheme 3. And aldehyde of Formula (XX) where R is an alkyl can react with an hydrazine of Formula (XXI) by condensation in a suitable solvent to afford substituted pyrazol of Formula (XXII). Reduction of the ester of Formula (XXII) using a reducing agent (such as UAIH4) in a suitable solvent lead to the isolation of a bis alcohol intermediate that was subsequently tosylated to afford compound of Formula (XXIII). Other reducing agent (such as BH3) could also be used in this sequence. Compound of Formula (XXIII) can finally undergo cyclisation using ammonia in a solvent of choice to lead to the isolation of intermediate of Formula (XXIV).

[0270] Scheme 4

[0271] An alternative synthesis to obtain intermediate of Formula (XXIV) where Y3 is (CH?)? is depicted in Scheme 4. The alcohol of Formula (XXII) where R is an alkyl can be substituted by an azide using reagent (such as DPPA) and standard Mitsunobu reagent in a suitable solvent to afford compound of Formula (XXV) that can then undergo a reduction and lead to the amine intermediate of Formula (XXVI). A variety of reagents could be used for the reduction of the azide into amine such as PPhs in water but also other more standardly use reducting agents such as UAIH4 or Pd / C. Compound of Formula (XXVI) can finally undergo cyclisation using a suitable base (such as K?COs) in a solvent of choice (such as EtOH) and lead to the cyclized compound of Formula (XXVII) that can be reduced using reducing agent (such as BH3) in a suitable solvent (such as THF) and lead to the isolation of intermediate of Formula (XIV).

[0272] Scheme 5

[0273] A compound of Formula (XXXXV) can be prepared from a compound of Formula (XXXVI) according to procedure represented in Scheme 11. Protected intermediate of Formula (XXXVIII) where PG is a suitable protecting group (such as MOM ether) can be synthesized in two steps via the demethylation of starting material of Formula (XXXVI) using a suitable reagent (such as dodecanethiol), followed by protection of compound of Formula (XXXVII) with a suitable protecting group in presence of a base. Compound of Formula (XXXVIII) can be iodinated using an halogenating agent (such as NIS) in a suitable solvent. Iodinated intermediate of Formula (XXXIX) can react with Turbo Grignard (such as iPrMgCl. LiCI) and a boronic ester in a suitable solvent (such as THF) to afford compound of Formula (XXXX). Oxidation of compound of Formula (XXXX) using reagent such as sodium perborate in presence of a solvent such as THF and water lead to compound of Formula (XXXXI). Subsequent reaction with an alkylating agent (such as methyl iodide, 2-chloro-2,2- difluoroacetate or diethyl bromodifluoromethylphosphonate ) in presence of a base (such as K2CO3) afford intermediate compound of Formula (XXXXII). Subsequent deprotection in presence of a suitable agent (such as HCI) can lead to compound of Formula (XXXXIII). An alcohol of general formula R16-OH, can be introduced on the alcohol of Formula (XXXXIII), using a Mitsunobu reaction. For example DIAD and PPhs in a suitable solvent afford compound a Formula (XXXXIV). Alternatively, substitution of alcohol of Formula (XXXXIII) by an alkyl halide (such as BrCH2R17) in presence of a base (such as K2CO3 or KOH), a catalytic amount of an iodine salt (such as Nal or KI) in a suitable solvent (such as EtOH or DMSO) also afford O-substituted intermediate of Formula (XXXXIV). A Miyaura borylation reaction of brominated intermediate of Formula (XXXXIV) using a palladium catalyst (such as Pd(dppf)Cl2), a suitable base (such as AcOK) in a suitable solvent (such as 1,4-dioxane) afford compound of Formula (XXXXV). Scheme 6

[0274] An alcohol of general formula R16-OH, can be introduced on the alcohol of a compound of Formula (XXXII), where PG is a suitable protecting group (such Boc), using a Mitsunobu reaction. For example DIAD and PPhs in a suitable solvent afford compound a Formula (XXXIII) as represented in Scheme 6. Alternatively, substitution of alcohol of Formula (XXXII) by an alkyl halide (such as BrCI- R17) in presence of a base (such as K2CO3 or KOH), a catalytic amount of an iodine salt (such as Nal or KI) in a suitable solvent (such as EtOH or DMSO) also afford O-substituted intermediate of Formula (XXXIII).

[0275] Syntheses of intermediates a. Synthesis of tert-butyl 2-bromo-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l / 4]diazepine-6-carboxylate

[0276] A I L round bottom flask was charged with ethyl 3-oxopentanoate (20.0 g, 139 mmol, 1.00 equiv), [2-(dimethylamino)ethyl]dimethylamine (19.3 g, 166 mmol, 1.20 equiv) and tetra hydrofuran (300 mL). To the above mixture was added sodium hydride (6.66 g, 166 mmol, 1.20 equiv, 60% in mineral oil) in three portions at -20°C under nitrogen atmosphere. The mixture was stirred for 15 min at -20°C and n-butyllithium (83.2 mL, 208 mmol, 1.50 equiv, 2.5 M in hexane) was added dropwise at -20°C under nitrogen atmosphere. The mixture was stirred for 15 min at -20 °C. To the above solution was added rapidly methyl formate (9.16 g, 153 mmol, 1.10 equiv) in THF (50 mL) at -20°C and stirred for 15 min at - 20°C. The reaction was quenched with 10 % HCI aqueous solution and extracted with ethyl acetate (3 x 200 mL). The combined organic layers were washed with brine(3 x 100 mL), dried over anhydrous sodium sulfate, filtrated and concentrated under reduced pressure to afford ethyl ethyl 4-methyl-3,5-dioxopentanoate (23 g, crude) as a brown oil. LCMS(ESI-MS) m / z = 173.1 [M + H]+.

[0277] A 1000 mL round bottom flask was charged with ethyl 4-methyl-3,5-dioxopentanoate (20 g, 58.1 mmol, 1.00 equiv, 50% purity), 2-hydroxyethylhydrazine (4.42 g, 58.1 mmol, 1.00 equiv) and ethanol (300 mL). The resulting solution was stirred overnight at 80 °C. The reaction was concentrated under reduced pressure and the residue was diluted with water (500 mL). The mixture was extracted with ethyl acetate (3 x 200 mL). The combined organic layers were washed with brine(3 x 50 mL), dried over anhydrous sodium sulfate, filtrated and concentrated under reduced pressure. The crude was purified by silica gel column chromatography, eluted with (DCM I MeOH =10 / 1) to afford ethyl ethyl 2-(l-(2- hydroxyethyl)-4-methyl-lH-pyrazol-5-yl)acetate (8 g, crude) as a yellow oil. LCMS(ESI-MS) m / z =213.1 [M + H]+.

[0278] A 250 mL round bottom flask was charged with ethyl 2-(l-(2-hydroxyethyl)-4-methyl-lH- pyrazol-5-yl)acetate (4.0 g, 18.8 mmol, 1.00 equiv) and THF (100 mL) at 0 °C under nitrogen atmosphere. To the above mixture was added dropwise LiAIH4 (7.54 mL, 15.1 mmol, 0.80 equiv, 2 M in THF) dropwise at 0 °C. The resulting solution was stirred for 2 h at room temperature under nitrogen atmosphere. The reaction was quenched with water (10 mL) at 0 °C. The mixture was diluted with THF (300 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford 2,2'-(4-methyl-lH- pyrazole-l,5-diyl)bis(ethan-l-ol) (5.3 g, crude) as a brown oil. LCMS(ESI-MS) m / z =171.1 [M + H]+.

[0279] A 250 mL round bottom flask was charged with 2,2'-(4-methyl-lH-pyrazole-l,5- diyl)bis(ethan-l-ol) I (3 g, 17.6 mmol, 1.00 equiv), p-toluenesulfonyl chloride (3 g, 15.7 mmol, 0.89 equiv), TEA (5 g, 49.4 mmol, 2.80 equiv), DMAP (0.2 g, 1.64 mmol, 0.09 equiv) and DCM (100 mL). The resulting solution was stirred for overnight at room temperature. The reaction was quenched with water (200 mL). The resulting mixture was extracted with DCM (3 x 100 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The crude was purified by silica gel column chromatography, eluted with (PE I EA = 3 / 1) to afford (4-methyl-lH-pyrazole-l,5-diyl)bis(ethane-2,l-diyl) bis(4- methylbenzenesulfonate) (2.5 g, 30% yield) as a yellow oil. LCMS(ESI-MS) m / z = 479.1 [M + H]+. A 250 mL round bottom flask was charged with (4-methyl-lH-pyrazole-l,5-diyl)bis(ethane- 2,1-diyl) bis(4-methylbenzenesulfonate) (2.5 g, 5.22 mmol, 1.00 equiv), ammonia solution (50 mL, 80% in water) and 1,4-dioxane (20 mL). The resulting solution was stirred overnight at 80 °C. The resulting mixture was concentrated under reduced pressure to afford 3-methyl- 5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepine (2.7 g, crude) as a light yellow oil. LCMS(ESI-MS) m / z =152.1 [M + H]+.

[0280] A 100 mL round bottom flask was charged with 3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepine (2.7 g, 8.93 mmol, 1.00 equiv, 50% purity), di-tert-butyl dicarbonate (1.95 g, 8.93 mmol, 1.00 equiv), tetra hydrofuran (30 mL), TEA (1.81 g, 17.9 mmol, 2.00 equiv) and water (10 mL). The resulting solution was stirred for 3 h at room temperature. The reaction was quenched with water (200 mL). The resulting mixture was extracted with ethyl acetate (3 x 80 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EA = 1 / 2) to afford tert-butyl 3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepine-6- carboxylate (210 mg, 9% yield) as a white solid. LCMS(ESI-MS) m / z =252.2 [M+H]+.

[0281] A 100 mL round bottom flask was charged with tert-butyl 3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepine-6-carboxylate (210 mg, 0.836 mmol, 1.00 equiv), ACN (20 mL) and NBS (148.72 mg, 0.836 mmol, 1.00 equiv). The resulting solution was stirred for 2 h at 60 °C. The reaction was quenched with water (100 mL). The resulting mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (2 x 50 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EA = 1: 2) to afford tert-butyl 2-bromo-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepine- 6-carboxylate (160 mg, 58% yield) as a yellow oil. LCMS(ESI-MS) m / z =330.1 [M+H]+. b. Synthesis of tert-butyl 3-(l-(2-bromo-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)cyclopropyl)azetidine-l-carboxylate

[0282] A solution of tert-butyl 2-bromo-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepine-6-carboxylate (500 mg, 1.51 mmol, 1.00 equiv) in HCI (9.00 mL, 36.0 mmol, 23.8 equiv, 4.0 M in 1,4-dioxane), and stirred for 5 hours at room temperature. The resulting mixture was concentrated under reduced pressure to afford 2-bromo-3-methyl- 5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepine (500 mg, crude) as a yellow solid. LCMS(ESI-MS) m / z =230.0 [M + H]+. A 40 mL vial was charged with 2-bromo-3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5- d][l,4]diazepine (500 mg, 2.17 mmol, 1.00 equiv, DMF (10 mL), DIEA (1.14 mL, 6.52 mmol, 3.00 equiv) and l-(tert-butoxycarbonyl)azetidine-3-carboxylic acid (0.44 g, 2.17 mmol, 1.00 equiv), and stirred for overnight at room temperature. The reaction was quenched with water (100 mL) and extracted with EtOAc (3x50 mL). The combined organic layers were washed with water (5 x 15 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (2 / 3) to afford tert-butyl 3-(2-bromo-3-methyl- 5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepine-6-carbonyl)azetidine-l-carboxylate (800 mg, 89% yield) as a brown red solid. LCMS(ESI-MS) m / z =413.1 [M+H]+.

[0283] Tetrakis(propan-2-yloxy)titanium (0.29 mL, 0.978 mmol, 2.02 equiv) in THF (2 mL) was added dropwise to a solution of ethylmagnesium bromide (1.0M in THF) (1.94 mL, 1.94 mmol, 4.01 equiv) in THF (4 mL) at -78 °C under N2, and stirred for 3 min. Tert-butyl 3-(2-bromo- 3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepine-6-carbonyl)azetidine-l- carboxylate (200 mg, 0.484 mmol, 1.00 equiv) was dissolved to THF (2 mL) and added to the above mixture at -78 °C under N2. After the reaction was stirred for 3 hours at 60 °C. Ethylmagnesium bromide (1.0M in THF) (1.94 mL, 1.941 mmol, 4.01 equiv) and tetrakis(propan-2-yloxy)titanium (0.29 mL, 0.978 mmol, 2.02 equiv) in THF (4 mL) were added at 0 °C under N2. The final reaction was stirred for overnight at room temperature under N2. The reaction was quenched with sat. NH4CI (aq.) (100 mL) at room temperature and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (3x15 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1 / 1) to afford tert-butyl 3-(l-(2-bromo-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepin-6-yl)cyclopropyl)azetidine-l-carboxylate (110 mg, 53% yield) as a light yellow solid. LCMS(ESI-MS) m / z =425.1 [M+H]+. c. Synthesis of tert-butyl 3-(l-(2-bromo-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)-2,2,2-trifluoroethyl)azetidine-l-carboxylate A 100 mL round-bottom flask was charged with 2-bromo-3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepine (1.5 g, 6.52 mmol, 1.00 equiv), ACN (40 mL) and KHF2 (1.02 g, 13.0 mmol, 2.00 equiv). TFA (1.49 g, 13.0 mmol, 2.00 equiv) and trimethyl(trifluoromethyl)silane (3.71 g, 26.1 mmol, 4.00 equiv) in ACN (8 mL) was added at room temperature under N2. The resulting mixture was stirred at room temperature for 10 min under nitrogen atmosphere. To the mixture added benzyl 3-formylazetidine-l- carboxylate (2.14 g, 9.78 mmol, 1.50 equiv) in ACN (2 mL) at room temperature under nitrogen atmosphere. The resulting mixture was stirred at room temperature for overnight under nitrogen atmosphere. The reaction was quenched by the addition of water (50 mL) at room temperature. The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with water (3 x 50 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by reversed-phase flash chromatography with the following conditions: column, C18 silica gel; mobile phase, ACN in Water (0.1% TFA), 30% to 80% gradient in 20 min; detector, UV 254 nm to afford benzyl 3-(l-(2-bromo-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)-2,2,2-trifluoroethyl)azetidine-l-carboxylate (200 mg, 6% yield) as a white solid. LCMS(ESI-MS) m / z = 501.1 [M+H]+.

[0284] A 100 mL round-bottom flask was charged with benzyl 3-(l-(2-bromo-3-methyl-4,5,7,8- tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)-2,2,2-trifluoroethyl)azetidine-l- carboxylate (150 mg, 0.299 mmol, 1 equiv), AICI3 (120 mg, 0.897 mmol, 3.00 equiv) and HFIP (20 mL). The resulting mixture was stirred at room temperature for overnight. The resulting mixture was concentrated under reduced pressure to afford 6-(l-(azetidin-3-yl)- 2,2,2-trifluoroethyl)-2-bromo-3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5- d][l,4]diazepine (150 mg, crude) as a yellow solid. LCMS(ESI-MS) m / z = 367.1 [M+H]+.

[0285] A 100 mL round-bottom flask was charged with 6-(l-(azetidin-3-yl)-2,2,2-trifluoroethyl)-2- bromo-3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepine (150 mg, 0.408 mmol, 1.00 equiv), Et3N (82.7 mg, 0.816 mmol, 2.00 equiv), (Boc)2O (178 mg, 0.816 mmol, 2.00 equiv) and DCM (20 mL). The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched by the addition of water (100 ml) at room temperature. The resulting mixture was extracted with DCM (3 x 100 mL). The combined organic layers were washed with water (3 x 10 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with (DCM / MeOH = 10 / 1) to afford tert-butyl 3-(l-(2-bromo-3- methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)-2,2,2- trifluoroethyl)azetidine-l-carboxylate (150 mg, 79% yield) as a yellow solid. LCMS(ESI-MS) m / z = 467.1 [M + H]+. d. Synthesis of tert-butyl 5-formyl-2-azabicyclo[2.2.2]octane-2-carboxylate

[0286] A 250 mL round bottom flask was charged with tert-butyl 5-oxo-2-azabicyclo[2.2.2]octane- 2-carboxylate (3.00 g, 13.3 mmol, 1.00 equiv) and THF (30 mL). To the mixture was added t-BuOK (4.48 g, 39.9 mmol, 3.00 equiv) at 0 °C. The mixture was stirred for 1 h at 0 °C. A solution of (methoxymethyl)triphenyl-l [5]-phosphane chloride (6.87 g, 20.0 mmol, 1.50 equiv) in THF (20 mL) was added to the above mixture at 0 °C. The resulting solution was stirred for 3 h at room temperature. The reaction was quenched by the addition of water (200 mL). The mixture was extracted with ethyl acetate (3 x 100 mL). The organic layers were combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EA = 5 / 1) to afford tert-butyl (E)-5- (methoxymethylene)-2-azabicyclo[2.2.2]octane-2-carboxylate (380 mg, 34% yield) as a colorless oil. LCMS(ESI-MS) m / z =254.2 [M + H] + .

[0287] A 100 mL round bottom flask was charged with tert-butyl (E)-5-(methoxymethylene)-2- azabicyclo[2.2.2]octane-2-carboxylate (380 mg, 1.50 mmol, 1.00 equiv), HCOOH (10 mL) and DCM (20 mL). The resulting solution was stirred for overnight at room temperature. The reaction was basified to pH = 8 with saturated Na?CO3 (aq.). The mixture was extracted with DCM (3 x 80 mL). The organic layers were combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by Prep-TLC (PE / EA = 3 / 1) to afford tert-butyl 5-formyl-2-azabicyclo[2.2.2]octane-2-carboxylate (260 mg, 72% yield ) as a yellow oil. LCMS(ESI-MS) m / z =240.2 [M+H]+. e. Synthesis of 2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethan-l-ol

[0288] A 500 mL round bottom flask was charged with 2-bromo-5-fluoropyridine (20 g, 114 mmol, 1.0 equiv) and toluene (150 mL). To the mixture was added isopropylmagnesium chloride (85 mL, 170 mmol, 1.5 equiv, 2.0 M in THF) at 0 °C under N2 atmosphere. The resulting solution was stirred for 1 h at 0 °C. A solution of 2-[(tert-butyldimethylsilyl)oxy]acetaldehyde (29.7 g, 170 mmol, 1.5 equiv) in toluene (50 mL) was added to the above mixture at 0 °C under N? atmosphere. The resulting solution was stirred for overnight at room temperature. The reaction was quenched with sat. NH4CI (aq., 300 mL) at 0 °C. The mixture was extracted with ethyl acetate (3 x 200 mL). The organic layers were combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (5: 1) to afford 2-[(tert-butyldimethylsilyl)oxy]- l-(5-fluoropyridin-2-yl)ethanol (20 g, 64% yield) as a yellow oil. LCMS(ESI-MS) m / z =272.1 [M + H]+. f. Synthesis of (R)-2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethan-l-ol

[0289] A 500 mL round bottom flask was charged with 2-[(tert-butyldimethylsilyl)oxy]-l-(5- fluoropyridin-2-yl)ethanol (20 g, 73.6 mmol, 1.00 equiv) and DCM (200 mL). To the mixture was added l,l-bis(acetyloxy)-3-oxo-3H-ll / K[5],2-benziodaoxol-l-yl acetate (46.88 g, 111 mmol, 1.50 equiv) at 0 °C. The resulting solution was stirred for 3 h at room temperature. The reaction was quenched with water (300 mL) at 0 °C. The mixture was extracted with DCM (3 x 150 mL). The organic layers were combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE I EA (5: 1) to afford 2-[(tert-butyldimethylsilyl)oxy]-l-(5- fluoropyridin-2-yl)ethanone (13 g, 65% yield) as a yellow solid. LCMS(ESI-MS) m / z =270.1 [M + H]+.

[0290] A 1000 mL round bottom flask was charged with (3aR)-l-methyl-3,3-diphenyl- hexahydropyrrolo[l,2-c][l,3,2]oxazaborole (40.1 g, 145 mmol, 1.00 equiv) and THF (300 mL). To the mixture was added bo rane-tetra hydrofuran complex (145 mL, 145 mmol, 1.00 equiv, 1.0 M in THF) dropwise at -78 °C and keep the temperature below -70 °C. The resulting solution was stirred for 6 h at -78 °C. Then a solution of 2-[(tert-butyldimethylsilyl)oxy]-l- (5-fluoropyridin-2-yl)ethanone (39 g, 145 mmol, 1.00 equiv) in THF (150 mL) was added to the above mixture at -78 °C. The mixture was warmed up to room temperature during two hours. The resulting solution was stirred for overnight at room temperature. The reaction was quenched by the addition of MeOH (100 mL) at 0 °C and stirred for 30 min at room temperature. The mixture was diluted with water (500 mL) and extracted with ethyl acetate (3 x 500 mL). The organic layers were combined, washed with brine (3 x 200 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (3 / 1) to afford (lR)-2-[(tert- butyldimethylsilyl)oxy]-l-(5-fluoropyridin-2-yl)ethanol (8.5 g, 20% yield, 95% purity) as a yellow oil. LCMS(ESI-MS) m / z =272.1 [M + H]+. g. Synthesis of 6-bromo-3-methoxypyrazolo[l,5-a]pyridin-4-ol

[0291] A 500 mL round-bottom flask was charged with 6-bromo-4-methoxypyrazolo[l,5-a]pyridine (10 g, 44 mmol, 1 equiv), dodecane-l-thiol (26.7 g, 132 mmol, 3 equiv), NaOH (5.28 g, 132 mmol, 3 equiv), H2O (5.28 g, 293 mmol, 6.7 equiv) and DMA (200 mL).The resulting mixture was stirred for overnight at 50 °C under nitrogen atmosphere. The reaction was quenched by the addition of water (400 mL) at room temperature. The residue was acidified to pH 4 with FA. The resulting mixture was extracted with EA (3 x 300 mL). The combined organic layers were washed with water (3 x 200 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford a crude product. The crude product was re-crystallized from (PE / EA = 9 / 1, 100 mL) to afford 6-bromopyrazolo[l,5- a]pyridin-4-ol (8 g, 85% yield) as a brown yellow solid. LCMS (ESI-MS) m / z =213.0 [M+H]+.

[0292] A 250 ml round-bottom flask was charged with 6-bromopyrazolo[l,5-a]pyridin-4-ol (6.9 g, 32 mmol, 1 equiv), bromo(methoxy)methane (6.1 g, 48.6 mmol, 1.50 equiv), K2CO3 (9.0 g, 64.8 mmol, 2 equiv) and acetone (100 mL). The resulting mixture was stirred for overnight at room temperature. The reaction was quenched by the addition of water (100 mL) at room temperature. The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with water (3 x 100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to provide 6-bromo-4- (methoxymethoxy)pyrazolo[l,5-a]pyridine (6.8 g, crude) as a white solid. LCMS(ESI-MS) m / z = 257.0 [M+H]+. A 40 mL vial were added 6-bromo-4-(methoxymethoxy)pyrazolo[l,5-a]pyridine (6.8 g, 26.5 mmol, 1 equiv), NIS (7.14 g, 31.7 mmol, 1.2 equiv) and DMF (100 mL). The resulting solution was stirred for 5 hours at room temperature. The reaction was quenched by water (100 mL). The mixture was extracted with EA (3 x 100 mL). The organic layers were combined, washed with water (3 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with (PE / EA = 2 / 1) to afford 6-bromo-3-iodo-4-(methoxymethoxy)pyrazolo[l,5-a]pyridine (6.6 g, 65% yield) as a white solid. LCMS (ESI-MS) m / z = 382.9 [M+H]+.

[0293] A 250 ml round-bottom flask was charged with 6-bromo-3-iodo-4- (methoxymethoxy)pyrazolo[l,5-a]pyridine (6.8 g, 17.8 mmol, 1 equiv), isopropylmagnesium chloride (17.8 mL, 35.5 mmol, 2 equiv, 2.0 M in diethyl ether) and THF (50 mL) . The reaction was stirred for 30 minutes at 0 °C under nitrogen atmosphere. Then 2-isopropoxy-4, 4,5,5- tetramethyl-l,3,2-dioxaborolane (4.96 g, 26.6 mmol, 1.5 equiv) was added dropwise at 0 °C. The resulting mixture was stirred for 2 hours at 0 °C under nitrogen atmosphere. The reaction was quenched by the addition of water (100 mL) at room temperature. The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with water (3 x 100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford 6-bromo-4-(methoxymethoxy)-3-(4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine (6.6 g, crude) as a colorless solid. LCMS (ESI-MS) m / z = 383.1 [M + H] + .

[0294] A 100 mL round bottom flask was charged with 6-bromo-4-(methoxymethoxy)-3-(4, 4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine (8 g, 20.9 mmol, 1 equiv), sodium perborate tetrahydrate (9.6 g, 62.7 mmol, 3 equiv), THF (200 mL) and H2O (50 mL). The resulting solution was stirred for 4 hours at room temperature. The reaction was quenched by water (100 mL). The mixture was extracted with EA (3 x 100 mL). The organic layers were combined, washed with water (3 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with (PE / EA = 1 / 10) to afford 6-bromo-4- (methoxymethoxy)pyrazolo[l,5-a]pyridin-3-ol (4 g, 70% yield) as a white solid. LCMS(ESI- MS) m / z = 273.0 [M + H] + .

[0295] A 100 mL round bottom flask was charged with 6-bromo-4-(methoxymethoxy)pyrazolo[l,5- a]pyridin-3-ol (4 g, 14.6 mmol, 1 equiv), K2CO3 (4 g, 29 mmol, 2 equiv), CH3I (1.66 g, 11.7 mmol, 0.8 equiv) and DMF (50 mL). The resulting solution was stirred for 2 hours at room temperature. The reaction was quenched by water (100 mL). The mixture was extracted with EA (3 x 100 mL). The organic layers were combined, washed with water (3 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with (PE / EA = 1 / 10) to afford 6-bromo-3-methoxy-4-(methoxymethoxy)pyrazolo[l,5-a]pyridine (3.5 g, 83% yield) as a white solid. LCMS (ESI-MS) m / z = 287.0 [M+H]+.

[0296] A 100 mL round bottom flask was charged with 6-bromo-3-methoxy-4- (methoxymethoxy)pyrazolo[l,5-a]pyridine (4 g, 13.9 mmol, 1.00 equiv), HCI in 1,4-dioxane (5 mL, 4.0 M in 1,4-dioxane) and 1,4-dioxane (2 mL). The resulting solution was stirred for 2 hours at room temperature and concentrated under reduced pressure to provide 6-bromo- 3-methoxypyrazolo[l,5-a]pyridin-4-ol (3.2 g, crude) as a white solid. LCMS (ESI-MS) m / z = 243.0 [M+H]+. h. Synthesis of 6-bromo-3-(difluoromethoxy)pyrazolo[l,5-a]pyridin-4-ol

[0297] -20°C-rt, overnight

[0298] To a stirred mixture of KOH (4.1 g, 73 mmol, 20 equiv) in ACN (50 mL) and H2O (50 mL) was added 6-bromo-4-(methoxymethoxy)pyrazolo[l,5-a]pyridin-3-ol (1.0 g, 3.66 mmol, 1 equiv) dropwise at -20°C. To the above mixture was added diethyl bromodifluoromethylphosphonate (1.96 g, 7.32 mmol, 2 equiv) dropwise over 15 mins at -20°C. The resulting mixture was stirred at -20°C room temperature for overnight. The reaction was quenched by water (100 mL). The mixture was extracted with EA (3 x 100 mL). The organic layers were combined, washed with water (3 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with (PE / EA = 2 / 1) to afford 6-bromo-3-(difluoromethoxy)-4- (methoxymethoxy)pyrazolo[l,5-a]pyridine (370 mg, 31% yield) as a white solid. LCMS (ESI- MS) m / z = 323.0 [M + H] + .

[0299] A mixture of 6-bromo-3-(difluoromethoxy)-4-(methoxymethoxy)pyrazolo[l,5-a]pyridine (720 mg, 2.23 mmol, 1 equiv) and HCI (4 M in 1,4-dioxane, 10 mL) was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure. The 6- bromo-3-(difluoromethoxy) pyrazolo[l,5-a]pyridin-4-ol (600mg) was used in the next step directly without further purification. LCMS(ESI-MS) m / z = 279.0 [M+H]+. i. Synthesis of 6-bromo-3-chloropyrazolo[l,5-a]pyridin-4-ol Cl OH N'^^^Br

[0300] 50 °C, overnight A 500 mL round bottom flask was charged with 6-bromo-4-methoxypyrazolo[l,5-a]pyridine (10 g, 44 mmol, 1.00 equiv), DMF (50 mL, 684 mmol, 15.5 equiv), PPTs (1.11 g, 4.4 mmol, 0.10 equiv) and NCS (8.82 g, 66 mmol, 1.50 equiv). The resulting solution was stirred for overnight at 50 °C. The reaction was quenched by water (100 mL). The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with H2O (3 x 100 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with EA / PE (1 I 1) to afford 6-bromo-3-chloro-4-methoxypyrazolo[l,5-a]pyridine (7.3g, crude) as a light yellow solid . LCMS(ESI-MS) m / z = 260.9 [M+H]+ .

[0301] A 500 mL round bottom flask was charged with NaOH (2.23 g, 55.8 mmol, 2.00 equiv), H2O (5 mL), DMA (200 mL), dodecane-l-thiol (11.3 g, 55.8 mmol, 2.00 equiv) and 6-bromo-3- chloro-4-methoxypyrazolo[l,5-a]pyridine (7.3 g, 27.9 mmol, 1.00 equiv). The resulting solution was stirred for overnight at 50 °C. The mixture / acidified to pH = 4 with FA at rt. The mixture was extracted with EA (3 x 200 mL). The organic layers were combined, washed with water (3 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with EA to afford 6-bromo-3-chloropyrazolo[l,5-a]pyridin-4-ol (6 g, crude) as a light yellow solid. LCMS(ESI-MS) m / z = 246.9 [M + H]+ . j. Synthesis of 4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3- (difluoromethoxy)-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazolo[l,5- a]pyridine

[0302] A mixture of 6-bromo-3-(difluoromethoxy)pyrazolo[l,5-a]pyridin-4-ol (1.4 g, 5.02 mmol, 1.00 equiv), 2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethan-l-ol (2.04 g, 7.52 mmol, 1.5 equiv) and CMBP (3.63 g, 15.1 mmol, 3.00 equiv) in toluene (50 mL) was stirred at 100 °C for overnight under nitrogen atmosphere. The reaction was quenched with water (100 mL). The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with saturated salt solution (3 x 100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with (PE / EA = 1 / 1) to afford 6-bromo- 4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3- (difluoromethoxy)pyrazolo[l,5-a]pyridine (2.1 g, 79% yield) as a light yellow solid.

[0303] LCMS(ESI-MS) m / z = 532.1 [M+H]+.

[0304] A 50-mL round bottom flask was charged with 6-bromo-4-(2-((tert-butyldimethylsilyl)oxy)- l-(5-fluoropyridin-2-yl)ethoxy)-3-(difluoromethoxy)pyrazolo[l,5-a]pyridine (2.1 g, 3.94 mmol, 1.0 equiv), bis(pinacolato)diboron (1.50 g, 5.92 mmol, 1.5 equiv), Pd(dppf)Cl2 (0.29 g, 0.394 mmol, 0.10 equiv), AcOK (0.78 g, 7.93 mmol, 2.0 equiv) and 1,4-dioxane (50 mL). The resulting mixture was stirred at 80 °C for 5 h under nitrogen atmosphere. The reaction was quenched with water (50 mL). The resulting mixture was extracted with EA (3 x 100 mL). The combined organic layers were washed with saturated salt solution (3 x 100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure to afford 4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3- (difluoromethoxy)-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine (2.2 g, crude) as a black solid. LCMS(ESI-MS) m / z = 580.3 [M+H]+. k. Synthesis of (S)-4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-

[0305] 3-methoxy-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine

[0306] Into a 1 L round-bottom flask was added PPhs (32.4 g, 123 mmol, 3.00 equiv) and THF (300 mL) at room temperature. DIAD (24.9 g, 123 mmol, 3.00 equiv) in THF (50 mL) was added to the above mixture at 0 °C under nitrogen atmosphere. The resulting mixture was stirred for 1 h at 0 °C. Then a solution of 6-bromo-3-methoxypyrazolo[l,5-a]pyridin-4-ol (10.0 g, 41.1 mmol, 1.00 equiv) and (lR)-2-[(tert-butyldimethylsilyl)oxy]-l-(5-fluoropyridin-2- yl)ethanol (16.7 g, 61.7 mmol, 1.5 equiv) in THF(100 mL) was added dropwise to the above mixture at 0 °C. The resulting mixture was stirred for overnight at room temperature. The reaction was quenched with water (200 mL) at room temperature. The resulting mixture was extracted with EA (3 x 200 mL). The combined organic layers were dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (3 / 1) to afford 2- [(lS)-l-({6-bromo-3-methoxypyrazolo[l,5-a]pyridin-4-yl}oxy)-2-[(tert- butyldimethylsilyl)oxy]ethyl]-5-fluoropyridine (11 g, 48% yield) as a light yellow solid. LCMS(ESI-MS) m / z =496.1 [M + H]+. A 500 mL round bottom flask was charged with 2-[(lR)-l-({6-bromo-3- methoxypyrazolo[l,5-a]pyridin-4-yl}oxy)-2-[(tert-butyldimethylsilyl)oxy]ethyl]-5- fluoropyridine (10.0 g, 20.1 mmol, 1.00 equiv), bis(pinacolato)diboron (15.4 g, 60.4 mmol, 3.00 equiv), Pd(dppf)Cl2 (0.82 g, 1.01 mmol, 0.05 equiv), AcOK (5.93 g, 60.4 mmol, 3.00 equiv), 1,4-dioxane (200 mL). The resulting solution was stirred for overnight at 80 °C under nitrogen atmosphere. The reaction was quenched by the addition of water (200 mL). The mixture was extracted with ethyl acetate(3 x 200 mL). The organic layers were combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure to afford 2- [(lR)-2-[(tert-butyldimethylsilyl)oxy]-l-{[3-methoxy-6-(4, 4,5, 5-tetra methyl- 1,3,2- dioxaborolan-2-yl)pyrazolo[l,5-a]pyridin-4-yl]oxy}ethyl]-5-fluoropyridine (11.0 g, crude Product) as a brown oil. LCMS(ESI-MS) m / z =544.3 [M+H]+-

[0307] I. Synthesis of (S)-4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-

[0308] 3-chloro-6-(4, 4, 5, 5-tetra methyl- 1, 3, 2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine

[0309] Starting from 6-bromo-3-methoxypyrazolo[l,5-a]pyridin-4-ol and (R)-2-((tert- butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethan-l-ol, intermediate (S)-4-(2-((tert- butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3-chloro-6-(4,4,5,5-tetramethyl- l,3,2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine was synthesized following the same procedure as used for intermediate (S)-4-(2-((tert-butyldimethylsilyl)oxy)-l-(5- fluoropyridin-2-yl)ethoxy)-3-methoxy-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)pyrazolo[l,5-a]pyridine.

[0310] EXAMPLES

[0311] The present invention will be now described in more details, referring to examples that are not limitative. The following abbreviations are used throughout the document: 2-Me-THF stands for 2-methyl tetra hydrofuran; ACN for acetonitrile; AcOH for acetic acid; ANOVA for analysis of variance; aq. for aqueous; ATP for adenosine 5'-triphosphate; b.i.d. means bis in die (twice a day); Boc stands for tert-butoxycarbonyl; BUN for blood urea nitrogen; Cpd for compound; d for doublet; DBU for l,8-Diazabicyclo[5.4.0]undec-7-ene; DCE for 1,2- dichloroethane; DCM for dichloromethane; dd for doublet of doublets; DIPEA for N,N- diisopropylethylamine; DMA for dimethylacetamide; DMEM for Dulbecco's Modified Eagle Medium; DMF for dimethylformamide; DMSO for dimethylsulfoxide; dppf for 1,1'- bis(diphenylphosphino)ferrocene (CAS# 12150-46-8); DSS for dextran sodium sulfate; dt for doublet of triplets; DTT for dithiothreitol; EDC for N-(3-dimethylaminopropyl)-N'- ethylcarbodiimide hydrochloride (CAS# 25952-53-8); EDTA for ethylenediaminetetraacetic acid; EGTA for ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid; ELISA for enzyme-linked immunosorbent assay; eq. for equivalent; EtOAc for ethyl acetate; EtOH for ethanol; h for hour; HATU for (l-[bis(dimethylamino)methylene]-lH-l,2,3-triazolo[4,5- b]pyridinium 3-oxide hexafluorophosphate (CAS# 148893-10-1); HEPES for 4-(2- hydroxyethyl)-l-piperazineethanesulfonic acid (CAS# 7365-45-9); His for histidine; HOBt for hydroxybenzotriazole; HPLC for high-performance liquid chromatography; i.n. for intranasal; Int for intermediate; i.p. for intraperitoneal; i-PrOH for isopropanol; i.v. for intravenous; LCMS for liquid chromatography-mass spectrometry; LPS for lipopolysaccharide; m for multiplet; MeOH for methanol; min for minute; mmol for millimole; MS for mass spectrometry; MTBE for methyl tert-butyl ether; Mtd for method; MW for molecular weight; MW (calcd) for molecular weight calculated; MW (obsd) for molecular weight observed; n.d. for not determined; N-ter for N-terminal; NA for not available; NBS for N-bromosuccinimide; NIS for N-iodosuccinimide; NMP for N-methyl-2-pyrrolidone; PBS for phosphate buffered saline; Pd2(dba)3 for tris(dibenzylideneacetone) dipalladium(O) (CAS# 51364-51-3); Pd / C for palladium on carbon; Pd(dppf)Cl2 for [1,1'- bis(diphenylphosphino)ferrocene]dichloropalladium(II) (CAS# 72287-26-4); Pd(dppf)Cl2-DCM for the same complex with dichloromethane (CAS# 95464-05-4); PIN for percentage inhibition; p.o. for per os; ppm for parts-per-million; q for quartet; q.d. for quaque die (once a day); RB for round bottom; RT for room temperature; RU for resonance units; s for singlet; SA for streptavidine; sat. for saturated; sc for subcutaneous; SEM for standard error of the mean; SM for starting material; STAB for sodium triacetoxyborohydride; t for triplet; td for triplet of doublets; TBAF for tetrabutylammonium fluoride; t-BuOH for tertbutanol; TEA for triethylamine; TFA for trifluoroacetic acid; TFE for 2,2,2-trifluoroethanol; THF for tetra hydrofuran; Xantphos for 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (CAS# 161265-03-8); and Xantphos Pd G4 for methanesulfonato9,9-dimethyl-4,5- bis(diphenylphosphino)xanthenepalladium(II) (CAS# 1621274-19-8).

[0312] Example 1 (cpd 107)

[0313] Example 1-1 (cpd 107-1) l-(3-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)azetidin-l- yl)prop-2-en-l-one, Isomer 1

[0314] Example 1-2 (cpd 107-2) l-(3-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)azetidin-l- yl)prop-2-en-l-one, Isomer 2

[0315] To a solution of tert-butyl 2-bromo-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l / 4]diazepine-6-carboxylate (128 mg, 0.388 mmol, 1.00 equiv) in dioxane (5 mL) was added 4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3-methoxy-6- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine (211 mg, 0.388 mmol, 1.00 equiv), Pd(PPh3)4 (44.79 mg, 0.039 mmol, 0.10 equiv), K2CO3 (160.71 mg, 1.164 mmol, 3.00 equiv) and H2O (1.25 mL). The reaction was stirred at 100 °C for 2 h under nitrogen atmosphere. The reaction mixture was cooled to room temperature and added water (50 mL). The resulting mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (3 x 100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (DCM I MeOH = 20 / 1) to afford tert-butyl 2-(4-(2-((tert-butyldimethylsilyl)oxy)- l-(5-fluoropyridin-2-yl)ethoxy)-3-methoxypyrazolo[ 1, 5-a]pyridin-6-yl)-3-methyl-4, 5,7,8- tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepine-6-carboxylate (210 mg, 81% yield) as a green solid. LCMS(ESI-MS) m / z = 667.3 [M+H]+.

[0316] To a solution of tert-butyl tert-butyl 2-(4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin- 2-yl)ethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepine-6-carboxylate (210 mg, 0.315 mmol, 1.00 equiv) in 1,4- dioxane (6 mL) was added HCI in 1,4-dioxane (4.0 M) (6 mL). The mixture was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure to afford 2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol (141 mg, 99% yield, crude) as a yellow solid. LCMS(ESI-MS) m / z = 453.2 [M+H]+.

[0317] To a solution of 2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol (141 mg, 0.31 mmol, 1 equiv) in DCE (6 mL) was added EtsN (158 mg, 1.56 mmol, 5 equiv) and tert-butyl 3-formylazetidine-l-carboxylate (87 mg, 0.47 mmol, 1.5 equiv). The mixture was stirred for 1 h at room temperature. Then STAB (198 mg, 0.94 mmol, 3 equiv) was added in portions. The reaction was stirred for overnight at room temperature. The reaction was quenched by water (50 mL). The mixture was extracted with DCM (3 x 50 mL). The organic layers were combined, washed with brine (3 x 50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by Prep-TLC (DCM I MeOH = 20 I 1) and concentrated to afford tert-butyl 3-((2-(4-(l-(5-fluoropyridin-2-yl)-2- hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)azetidine-l-carboxylate (180 mg, 93% yield) as a solid. LCMS (ESI-MS) m / z = 622.3 [M+H]+.

[0318] To a solution of tert-butyl tert-butyl 3-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)methyl)azetidine-l-carboxylate (180 mg, 0.29 mmol, 1 equiv) in 1,4- dioxane (10 mL) was added HCI (4 M in 1,4-dioxane, 10 mL). The mixture was stirred for 2 h at room temperature. The resulting mixture was concentrated under reduced pressure to afford 2-((6-(6-(azetidin-3-ylmethyl)-3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5- d][l,4]diazepin-2-yl)-3-methoxypyrazolo[l,5-a]pyridin-4-yl)oxy)-2-(5-fluoropyridin-2- yl)ethan-l-ol (150 mg, crude) as a yellow solid. LCMS(ESI-MS) m / z = 522.3 [M+H]+.

[0319] To a solution of 2-((6-(6-(azetidin-3-ylmethyl)-3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)-3-methoxypyrazolo[l,5-a]pyridin-4-yl)oxy)-2-(5- fluoropyridin-2-yl)ethan-l-ol (150 mg, 0.29 mmol, 1 equiv) in DCM (10 mL) was added DIEA (223 mg, 1.73 mmol, 6 equiv) and acryloyl chloride (23.4 mg, 0.26 mmol, 0.9 equiv) at 0 °C. The mixture was stirred for 1 h at 0 °C and added water (20 mL). The resulting mixture was extracted with ethyl acetate (3 x 50 mL). The combined organic layers were washed with brine (3 x 100 mL), dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (DCM I MeOH = 10 I 1) to afford crude product. The crude product was purified by Prep-Chiral-HPLC with the following conditions Column: CHIRALPAK IM, 2*25 cm, 5 pm; Mobile Phase A: Hex— HPLC, Mobile Phase B: EtOH: DCM = 1: 1-HPLC; Flow rate: 20ML / MIN mL / min; Gradient (B%): isocratic 30; Wave Length: 254 / 220nm nm to afford l-(3-((2-(4-(l-(5-fluoropyridin-2-yl)-2- hydroxy-H3-ethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro- 6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)azetid in- l-yl)prop-2-en- 1-one.

[0320] Isomer 1 ((36.9 mg, 22% yield) as a white solid and isomer 2 (36.1 mg, 22%yield) as a white solid. LCMS (ESI-MS) m / z = 576.3 [M+H]+.

[0321] Example 2 (cpd 108)

[0322] Example 2-1 (cpd 108-1) l-(3-((2-(3-(difluoromethoxy)-4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)pyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)methyl)azetidin-l-yl)prop-2-en-l-one, Isomer 1

[0323] Example 2-2 (cpd 108-2) l-(3-((2-(3-(difluoromethoxy)-4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)pyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)methyl)azetidin-l-yl)prop-2-en-l-one, Isomer 2

[0324] Starting from 4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3- (difluoromethoxy)-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine, examples 108 were synthesized following a similar procedure as used for example 107.

[0325] The crude product was purified by Prep-HPLC with the following conditions: Column: InfinityLab Poroshell 120 HPH-C18, 21.2*150 mm, 4pm; Mobile Phase A: Water(10mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 25 mL / min mL / min; Gradient (B%): 25% B to 45% B in lOmin; Wave Length: 254nm / 220nm to afford a crude product. The crude product was purified by Prep-chiral-HPLC with the following conditions: Column: Chiral ART Cellulose- SA, 2*25 cm, 5 pm; Mobile Phase A: Hex(0.5% 2M NHs-MeOH)— HPLC, Mobile Phase B: MeOH: DCM = 1 : 1— HPLC; Flow rate: 20 mL / min; Gradient (B%): isocratic 45; Wave Length: 254 / 220 nm to afford l-(3-((2-(3-(difluoromethoxy)-4-(l-(5-fluoropyridin-2-yl)-2- hydroxyethoxy)pyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)methyl)azetidin-l-yl)prop-2-en- 1-one.

[0326] Isomer 1 (12.6 mg, 16% yield) as a white solid and isomer 2 (12.8 mg, 17% yield) as a white solid.

[0327] LCMS(ESI-MS) m / z = 612.4 [M+H]+.

[0328] Example 3 (cpd 109)

[0329] Example 3-1 (cpd 109-1) l-(3-(2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepine-6-carbonyl)azetidin-l- yl)prop-2-en-l-one, Isomer 1

[0330] Example 3-2 (cpd 109-2) l-(3-(2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepine-6-carbonyl)azetidin-l- yl)prop-2-en-l-one, Isomer 2

[0331] To a solution of 2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol (135 mg, 0.298 mmol, l.OO equiv), l-(tert-butoxycarbonyl)azetidine-3-carboxylic acid (60.0 mg, 0.298 mmol, 1.00 equiv) and HATU (147 mg, 0.387 mmol, 1.30 equiv) in DMF (5 mL) was added DIEA (231 mg, 1.79 mmol, 6.00 equiv). The reaction mixture was stirred for 2 h at rt. The reaction was added water (50 mL), extracted with EA (3 x 50 mL), washed with brine (3x 100 mL), dried over anhydrous sodium sulfate and concentrated to get the residue. The residue was purified by Pre-TLC (MeOH I DCM = 1 / 15) to afford tert-butyl 3-(2-(4-(l-(5- fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl- 5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepine-6-carbonyl)azetidine-l-carboxylate (145 mg, 76% yield) as a brown solid. LCMS(ESI-MS) m / z =636.3 [M+H]+.

[0332] To a solution of tert-butyl 3-(2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5- d][l,4]diazepine-6-carbonyl)azetidine-l-carboxylate (105 mg, 0.165 mmol, 1.00 equiv) in DCM (2.5 mL) was TFA (0.25 mL). The reaction mixture was stirred for 2 h at rt and concentrated to afford azetidin-3-yl(2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)methanone (88 mg, crude) as a brown solid. LCMS(ESI-MS) m / z =536.3 [M+H]+.

[0333] To a solution of azetidin-3-yl(2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)methanone (88 mg, 0.164 mmol, 1.00 equiv) and DIEA (1.75 mL) in DCM (5 mL) was added prop-2-enoyl prop-2-enoate (20.7 mg, 0.164 mmol, 1.00 equiv). The reaction mixture was stirred for 2 h at 0 °C. The reaction was added water (50 mL), extracted with DCM (3 x 50 mL), washed with brine, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by Prep-TLC (MeOH I DCM = 1 1 15) to afford racemate product. The racemate was separated by Pre-chiral-HPLC Column: Chiral ART Cellulose-SA, 2*25 cm, 5 pm; Mobile Phase A: Hex— HPLC, Mobile Phase B: MeOH: DCM = 1 : 1— HPLC; Flow rate: 20 mL / min; Gradient (B%) : isocratic 40; Wave Length : 254 / 220 nm to afford l-(3-(2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepine-6- carbonyl)azetidin-l-yl)prop-2-en- 1-one.

[0334] Isomer 1 (10.1 mg, 10% yield) as a white solid and isomer 2 (8.1 mg, 8% yield) as a white solid.

[0335] LCMS(ESI-MS) m / z = 590.6 [M + H] + .

[0336] Example 4 (cpd 110)

[0337] Example 4-1 (cpd 110-1) l-(4-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)piperidin-l- yl)prop-2-en-l-one, Isomer 1

[0338] Example 4-2 (cpd 110-2) l-(4-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)piperidin-l- yl)prop-2-en-l-one, Isomer 2

[0339]

[0340] Starting from 2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol and tert-butyl 4-formylpiperidine-l-carboxylate, examples 110 were synthesized following a similar procedure as used for example 107.

[0341] The residue was purified by Prep-TLC (DCM I MeOH = 10 / 1) to afford crude product. The crude product was purified by Prep-Chiral-HPLC with the following conditions Column: CHIRAL ART Cellulose-SB, 2*25 cm, 5 pm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)-HPLC, Mobile Phase B: MeOH: DCM = 1: 1— HPLC; Flow rate: 20 mL / min; Gradient (B%): isocratic 20; Wave Length: 254 / 220 nm to afford l-(4-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)methyl)piperidin-l-yl)prop-2-en- 1-one.

[0342] Isomer 1 (12.7 mg, 23% yield) as a white solid and isomer 2 (12.9 mg, 23%yield) as a white solid.

[0343] LCMS(ESI-MS) m / z = 465.2 [M+H]+.

[0344] Example 5 (cpd 111)

[0345] Example 5-1 (cpd 111-1) l-((lR,5S,6r)-6-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)-

[0346] 3-azabicyclo[3.1.0]hexan-3-yl)prop-2-en-l-one, Isomer 1

[0347] Example 5-2 (cpd 111-2) l-((lR,5S,6r)-6-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)-

[0348] 3-azabicyclo[3.1.0]hexan-3-yl)prop-2-en-l-one, Isomer 2

[0349] Starting from 2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol and tert-butyl (lR,5S,6s)-6-formyl-3-azabicyclo[3.1.0]hexane-3-carboxylate, examples 111 were synthesized following a similar procedure as used for example 107.

[0350] The residue was purified by Prep-TLC (EA / MeOH = 5 / l)to afford crude product. The crude product was purified by Prep-HPLC with the following conditions: Column: Chiral ART Cellulose-SA, 2*25 cm, 5 pm; Mobile Phase A: Hex(0.5%2M NHs-MeOH)— HPLC, Mobile Phase B: MeOH: DCM = 1 : 1— HPLC; Flow rate: 20 mL / min; Gradient (B%) : isocratic 30; Wave Length : 254 / 220 nm to afford l-((lR,5S,6r)-6-((2-(4-(l-(5-fluoropyridin-2-yl)-2- hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)-3-azabicyclo[3.1.0]hexan-3-yl)prop-2-en- 1-one. Isomer 1 (42.4 mg, 24% yield) as a white solid and isomer 2 (39.2 mg, 22% yield)) as a white solid. LCMS(ESI-MS) m / z = 602.3 [M+H]+.

[0351] Example 6 (cpd 112)

[0352] Example 6-1 (cpd 112-1) l-((lR,5S,6s)-6-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)-

[0353] 3-azabicyclo[3.1.0]hexan-3-yl)prop-2-en-l-one, Isomer 1

[0354] Example 6-2 (cpd 112-2) l-((lR,5S,6s)-6-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)-

[0355] 3-azabicyclo[3.1.0]hexan-3-yl)prop-2-en-l-one, Isomer 2

[0356] Starting from 2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol and tert-butyl (lR,5S,6r)-6-formyl-3-azabicyclo[3.1.0]hexane-3-carboxylate, examples 112 were synthesized following a similar procedure as used for example 107.

[0357] The residue was purified by Prep-TLC (DCM I MeOH 6: 1) to afford crude product. Then the crude product was separated by Prep-Chiral-HPLC with the following conditions: Column: CHIRALPAK IF, 2*25 cm, 5 pm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)-HPLC, Mobile Phase B: MeOH: DCM = 1: 1— HPLC; Flow rate: 20 mL / min; Gradient (B%): isocratic 60; Wave Length: 254 / 220 nm to afford l-((lR,5S,6s)-6-((2-(4-(l-(5-fluoropyridin-2-yl)-2- hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)-3-azabicyclo[3.1.0]hexan-3-yl)prop-2-en- 1-one.

[0358] Isomer 1 (53.5 mg, 29% yield) as a white solid and isomer 2 (52.5 mg, 28% yield) as a white solid. LCMS(ESI-MS) m / z = 602.2 [M + H] + .

[0359] Example 7 (cpd 13)

[0360] (S)-l-(3-(l-(2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)cyclopropyl)azetidin-l-yl)prop-2-en- 1-one

[0361] A 40 mL glass sample vial was charged with (S)-4-(2-((tert-butyldimethylsilyl)oxy)-l-(5- fluoropyridin-2-yl)ethoxy)-3-methoxy-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)pyrazolo[l,5-a]pyridine (211 mg, 0.388 mmol, 1.50 equiv), 1,4-dioxane (4 mL), tert-butyl 3-(l-(2-bromo-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)cyclopropyl)azetidine-l-carboxylate (110 mg, 0.259 mmol, 1.00 equiv), H2O (1 mL), K2CO3 (71.48 mg, 0.517 mmol, 2.00 equiv) and Pd(PPh3)4 (14.9 mg, 0.013 mmol, 0.05 equiv), and stirred for overnight at 100 °C under N2. The reaction was quenched with water (30 mL) at room temperature and extracted with EtOAc (3x10 mL). The combined organic layers were washed with brine (3x5 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with PE / EA (1 / 9) to afford tert-butyl (S)-3-(l-(2-(4-(2-((tert- butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)cyclopropyl)azetidine-l-carboxylate (110 mg, 56% yield) as a light yellow solid. LCMS(ESI- MS) m / z =762.4 [M+H]+.A solution of tert-butyl tert-butyl (S)-3-(l-(2-(4-(2-((tert- butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)cyclopropyl)azetidine-l-carboxylate (110 mg, 0.144 mmol, 1.00 equiv) in HCI (2.5 mL, 10.0 mmol, 69.3 equiv, 4.0 M inl,4-dioxane), and stirred for 2 hours at room temperature. The resulting mixture was concentrated under reduced pressure to afford (S)-2-((6-(6-(l- (azetidin-3-yl)cyclopropyl)-3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepin-2- yl)-3-methoxypyrazolo[l,5-a]pyridin-4-yl)oxy)-2-(5-fluoropyridin-2-yl)ethan-l-ol (110 mg, crude) as a yellow solid. LCMS(ESI-MS) m / z =548.3 [M+H]+.

[0362] A 8 mL glass sample vial was charged with (S)-2-((6-(6-(l-(azetidin-3-yl)cyclopropyl)-3- methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepin-2-yl)-3-methoxypyrazolo[l,5- a]pyridin-4-yl)oxy)-2-(5-fluoropyridin-2-yl)ethan-l-ol (110 mg, 0.20 mmol, 1.00 equiv) and DIEA (104 mg, 0.80 mmol, 4.00 equiv). Acryloyl chloride (18.2 mg, 0.20 mmol, 1.00 equiv) was added and stirred for 30 min at room temperature. The reaction was poured into water (10 mL) and extracted with DCM (3x5 mL). The combined organic layers were washed with brine (3 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (EA / MeOH = 7 / 1) to afford crude product. Then the crude product was re-purified by Prep-HPLC with the following conditions: Column: XBridge Prep Phenyl OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water(10nmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient (B%): 21% B to 51 % B in 9 min; Wave Length: 254nm / 220nm to afford (S)-l-(3-(l-(2-(4- (l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3- methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)cyclopropyl)azetidin-l- yl)prop-2-en-l-one (25.8 mg, 21% yield) as a white solid. LCMS(ESI-MS) m / z =602.2 [M + H] + .

[0363] Example 8 (cpd 114)

[0364] Example 8-1 (cpd 114-1) l-(5-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)-2- azabicyclo[2.2.2]octan-2-yl)prop-2-en-l-one, Isomer 1

[0365] Example 8-2 (cpd 114-2) l-(5-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)-2- azabicyclo[2.2.2]octan-2-yl)prop-2-en-l-one, Isomer 2

[0366] Starting from 2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol and tert-butyl 5-formyl-2-azabicyclo[2.2.2]octane-2-carboxylate, examples 114 were synthesized following a similar procedure as used for example 107. The residue was purified by The residue was purified by Prep-TLC (EA I MeOH = 8 / 1) to afford crude product. The residue was purified by Prep-Chiral-HPLC with the following conditions: Column: XBridge Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water(10nmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient (B%): 26% B to56% B in8 min; Wave Length: 254nm / 220nm. Then chiral separation by Prep-Chiral-HPLC with the following conditions: Column: Chiral ART Cellulose-SA, 2*25 cm, 5 pm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)-HPLC, Mobile Phase B: MeOH: DCM=1: 1- -HPLC; Flow rate: 20 mL / min; Gradient (B%): isocratic 30; Wave Length: 254 / 220 nm; to afford l-(5-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)- 2-azabicyclo[2.2.2]octan-2-yl)prop-2-en-l-one.

[0367] Isomer 1 (64.8 mg, 20% yield) as a white solid and isomer 2 (65.5 mg, 20% yield) as a white solid. LCMS(ESI-MS) m / z =630.3 [M + H] + .

[0368] Example 9 (cpd 115)

[0369] (S)-l-(3-((2-(4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)sulfonyl)azetidin-l-yl)prop-2-en-l-one

[0370] Into a 100 mL round-bottom flask were added (S)-2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6- (3-methyl-5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin- 4-yl)oxy)ethan-l-ol (120 mg, 0.265 mmol, 1.00 equiv), DIEA (171 mg, 1.33 mmol, 5.00 equiv) and DCM (10 mL) at room temperature. To the above mixture was added tert-butyl 3- (chlorosulfonyl)azetidine-l-carboxylate (54 mg, 0.212 mmol, 0.80 equiv) in portions at 0°C. The resulting mixture was stirred at room temperature for additional 5 hours. The residue was purified by Prep-TLC (EA I MeOH = 10 / 1) to afford tert-butyl (S)-3-((2-(4-(l-(5- fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl- 4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)sulfonyl)azetidine-l-carboxylate (80 mg, 45% yield) as an off-white solid. LCMS(ESI-MS) m / z = 672.2 [M+H]+. Into a 100 mL round-bottom flask were tert-butyl (S)-3-((2-(4-(l-(5-fluoropyridin-2-yl)-2- hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepin-6-yl)sulfonyl)azetidine-l-carboxylate (80 mg, 0.119 mmol, 1.00 equiv) and 1,4-dioxane (5 mL) at room temperature. To the above mixture was added HCI (10 mL, 4.0 M in 1,4-dioxane) dropwise at room temperature. The resulting mixture was stirred at room temperature for additional 2 hours. The resulting mixture was concentrated under reduced pressure to afford (S)-2-((6-(6-(azetidin-3-ylsulfonyl)-3-methyl-5,6,7,8- tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepin-2-yl)-3-methoxypyrazolo[l,5-a]pyridin-4- yl)oxy)-2-(5-fluoropyridin-2-yl)ethan-l-ol (70 mg, crude) as a brown solid. LCMS(ESI-MS) m / z = 572.2 [M+H]+.

[0371] Into a 100 mL round-bottom flask was added (S)-2-((6-(6-(azetidin-3-ylsulfonyl)-3-methyl- 5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepin-2-yl)-3-methoxypyrazolo[l,5- a]pyridin-4-yl)oxy)-2-(5-fluoropyridin-2-yl)ethan-l-ol (70 mg, 0.122 mmol, 1.00 equiv),, DIEA (79.14 mg, 0.610 mmol, 5.00 equiv) and DCM (10 mL) at 0°C. Then acrylic anhydride (15 mg, 0.122 mmol, 1.00 equiv) in DCM was added dropwise. The resulting mixture was stirred at 0 °C for 20 minutes. The reaction was quenched with water at room temperature. The resulting mixture was extracted with CH2CI2 (3 x 20 mL). The combined organic layers were dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (EA I MeOH 10: 1) to afford crude product. The residue was purified by Prep-HPLC with following conditions: Column: XBridge Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water(10mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient (B%): 21% B to51 % B in 10 min; Wave Length: 254nm / 220nm to afford (S)-l-(3-((2-(4-(l-(5-fluoropyridin-2-yl)-2- hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepin-6-yl)sulfonyl)azetidin-l-yl)prop-2-en-l-one (25.2 mg, 33% yield over 2 steps) as a white solid. LCMS(ESI-MS) m / z = 626.3 [M + H]+.

[0372] Example 10 (cpd 116)

[0373] Example 10-1 (cpd 116-1) l-(3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)pyrrolidin-l-yl)prop-2-en-l-one, Isomer 1

[0374] Example 10-2 (cpd 116-2) l-(3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)pyrrolidin-l-yl)prop-2-en-l-one, Isomer 2

[0375] A 40 mL vial was charged with (S)-2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl- 5,6,7,8-tetrahydro-4H-pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4- yl)oxy)ethan-l-ol (300 mg, 0.663 mmol, 1.00 equiv), tert-butyl 3-oxopyrrolidine-l- carboxylate (246 mg, 1.33 mmol, 2.00 equiv), NaBHsCN (125 mg, 1.99 mmol, 3.00 equiv) and MeOH (20 mL). The resulting solution was stirred for 48 h at 60 °C. The reaction was quenched by water (100 mL). The mixture was extracted with EA (3 x 100 mL). the organic layers were combined, washed with water (3 x 100 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluted with DCM / MeOH (10 I 1) to afford tert-butyl 3-(2-(4-((S)- l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3- methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)pyrrolidine-l-ca rboxylate (150 mg, 36% yield) as a white solid. LCMS(ESI-MS) m / z =622.3 [M+H]+.

[0376] A solution of tert-butyl 3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)pyrrolidine-l-carboxylate (120 mg, 0.193 mmol, 1.00 equiv) in HCI (5 mL, 4.0 M in 1,4-dioxane), and stirred for 2 hours at room temperature. The resulting mixture was concentrated under reduced pressure to afford (2S)-2-(5-fluoropyridin-2-yl)-2-((3- methoxy-6-(3-methyl-6-(pyrrolidin-3-yl)-5,6,7,8-tetrahydro-4H-pyrazolo[l,5- d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol (120 mg, crude) as a yellow solid. LCMS(ESI-MS) m / z =522.3 [M + H]+.

[0377] Acrylic anhydride (44 mg, 0.345 mmol, 1.00 equiv) was added to a solution of (2S)-2-(5- fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-6-(pyrrolidin-3-yl)-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol (180 mg, 0.345 mmol, 1.00 equiv) and DIEA (178 mg, 1.38 mmol, 4.00 equiv) DCM (5 mL). The resulting solution was stirred for 30 min at 0 °C. The reaction was quenched with water (25 mL) and extracted with DCM (3 x 10 mL). The combined organic layers were washed with water (6 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (EA / MeOH = 2 / 1) to afford crude product. Then the crude product was re-purified by Prep-HPLC with the following conditions: Column: XBridge Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water (10 mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient (B%): 24% B to 34% B in 9 min; Wave Length: 254nm / 220nm to afford the mixture of diastereoisomers. Prep-Chiral-HPLC with the following conditions Column: CHIRAL ART Cellulose-SC, 2*25 cm, 5 pm; Mobile Phase A: MtBE(0.5% 2M NH3-MeOH)-HPLC, Mobile Phase B: EtOH: DCM = 1 : 1— HPLC; Flow rate: 20 mL / min; Gradient (B%): isocratic 60% B; Wave Length: 254 / 220 nm afford l-(3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2- hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepin-6-yl)pyrrolidin-l-yl)prop-2-en-l-one.

[0378] Isomer 1 (12.8 mg, 6% yield) as a white solid and isomer 2 (3.6 mg, 7% yield) as a white solid. LCMS(ESI-MS) m / z = 576.2 [M + H]+.

[0379] Example 11 (cpd 117) l-(3-((2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6- yl)methyl)pyrrolidin-l-yl)prop-2-en-l-one, Isomer 1

[0380] Starting from (S)-2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-5,6,7,8-tetrahydro- 4H-pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol and tertbutyl 3-formylpyrrolidine-l-carboxylate, examples 117 were synthesized following a similar procedure as used for example 107.

[0381] The crude was separated by Prep-Chiral-SFC with the following conditions: Column: CHIRALPAK IH 3*25 cm, 5um; Mobile Phase A: CO2, Mobile Phase B: MeOH: THF=l: 1(0.3%- 7M-NHs-MeOH); Flow rate: 85 mL / min; Gradient (B%): isocratic 40% B; Column Temperature (°C): 35; Back Pressure(bar): 100; Wave Length: 220 nm to afford crude product. The crude product was re-purified by Prep-HPLC with the following conditions: Column: XBridge Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water (lOmmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient (B%): 21% B to 51% B in 8min; Wave Length: 254nm / 220nm nm to afford l-(3-((2-(4-((S)-l-(5- fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl- 4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)pyrrolidin-l-yl)prop-2-en- 1-one, isomer 1 (7.0 mg, 6%yield) as an off-white solid. LCMS(ESI-MS) m / z =590.3 [M+H]+.

[0382] Example 12 (cpd 118)

[0383] (S)-l-(3-((2-(3-chloro-4-(l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)pyrazolo[l,5-a]pyridin- 6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)azetidin-l- yl)prop-2-en-l-one

[0384] Starting from (S)-4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3- chloro-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine, example 118 was synthesized following the same procedure as used for example 107.

[0385] The crude product was purified by Prep-HPLC with the following conditions: Column: XBridge Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water(10mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient (B%): 25% B to 55% B in 8min; Wave Length: 254nm / 220nm to afford (S)-l-(3-((2-(3-chloro-4-(l-(5-fluoropyridin-2- yl)-2-hydroxyethoxy)pyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepin-6-yl)methyl)azetidin-l-yl)prop-2-en-l-one (26.8 mg, 17% yield over 2 steps) as a white solid. LCMS(ESI-MS) m / z = 580.2 [M+H]+ .

[0386] Example 13 (cpd 119)

[0387] Example 13-1 (cpd 119-1) l-(3-(2,2,2-trifluoro-l-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)ethyl)azetidin-l-yl)prop-2-en- 1-one, Isomer 1

[0388] Example 13-2 (cpd 119-2) l-(3-(2,2,2-trifluoro-l-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)ethyl)azetidin-l-yl)prop-2-en- 1-one, Isomer 2

[0389] Starting from (S)-4-(2-((tert-butyldimethylsilyl)oxy)-l-(5-fluoropyridin-2-yl)ethoxy)-3- methoxy-6-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)pyrazolo[l,5-a]pyridine and tertbutyl 3-(l-(2-bromo-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)- 2,2,2-trifluoroethyl)azetidine-l-carboxylate, examples 119 were synthesized following the same procedure as used for example 113.

[0390] The crude product was purified by Prep-HPLC with the following conditions: Column: CHIRALPAK IG, 2*25 cm, 5 pm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)-HPLC, Mobile Phase B: EtOH: DCM = 1 : 1— HPLC; Flow rate: 20 mL / min; Gradient (B%): isocratic 60% B; Wave Length: 254 / 220 nm to afford l-(3-(2,2,2-trifluoro-l-(2-(4-((S)-l-(5-fluoropyridin-2- yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro- 6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)ethyl)azetidin-l-yl)prop-2-en- 1-one.

[0391] Isomer 1 (26.7 mg, 12% yield) as a white solid and isomer 2 (27.4 mg, 12% yield) as a white solid. LCMS(ESI-MS) m / z = 644.3 [M + H]+.

[0392] Example 14 (cpd 120)

[0393] Example 14-1 (cpd 120-1)

[0394] 3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)pyrrolidine-l- carbonitrile, Isomer 1

[0395] Example 14-2 (cpd 120-2)

[0396] 3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)pyrrolidine-l- carbonitrile, Isomer 2

[0397] Cyanogen bromine (49 mg, 0.460 mmol, 2.00 equiv) was added to a solution (2S)-2-(5- fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-6-(pyrrolidin-3-yl)-5,6,7,8-tetrahydro-4H- pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol (120 mg, 0.230 mmol, 1.00 equiv) and DIEA (119 mg, 0.920 mmol, 4.00 equiv) in DCM (5 mL). The final reaction was stirred for 1 hour at room temperature. The reaction was quenched with water (25 mL) and DCM (3x10 mL). The combined organic layers were washed with water (5 mL), dried over anhydrous Na2SC>4. After filtration, the filtrate was concentrated under reduced pressure. The residue was purified by Prep-TLC (EA / MeOH = 3 / 1) to afford crude product. The crude product was separated by Prep-Chiral-HPLC with the following conditions: Column: Lux 5um Cellulose-4, 2.12*25 cm, 5 pm; Mobile Phase A: Hex (0.5% 2M NH3- MeOH)— HPLC, Mobile Phase B: MeOH: EtOH=l: 1— HPLC; Flow rate: 20 mL / min; Gradient (B%): isocratic 80% B; Wave Length: 254 / 220 nm to afford two isomers. Then the first elution was re-purified by Prep-HPLC with the following conditions: Column: XBridge Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water(10mmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient (B%): 20% B to 50% B in 8min; Wave Length: 254nm / 220nm to afford 3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2- hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H- pyrazolo[l,5-d][l,4]diazepin-6-yl)pyrrolidine-l-carbonitrile.

[0398] Isomer 1 (2.1 mg, 2% yield) as a white solid and isomer 2 (1.9 mg, 1.5% yield) as a white solid. LCMS(ESI-MS) m / z =547.3 [M + H] + .

[0399] Example 15 (cpd 121)

[0400] Example 15-1 (cpd 121-1)

[0401] 3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)piperidine-l- carbonitrile, Isomer 1

[0402] Example 15-2 (cpd 121-2) 3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5-a]pyridin-6- yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)piperidine-l- carbonitrile, Isomer 2

[0403] Starting from (S)-2-(5-fluoropyridin-2-yl)-2-((3-methoxy-6-(3-methyl-5,6,7,8-tetrahydro- 4H-pyrazolo[l,5-d][l,4]diazepin-2-yl)pyrazolo[l,5-a]pyridin-4-yl)oxy)ethan-l-ol and tertbutyl 3-oxopiperidine-l-carboxylate, examples 121 were synthesized following the same procedure as used for example 116 and 120.

[0404] The crude product was purified by Prep-HPLC with the following conditions: Column: CHIRAL ART Cellulose-SB, 2*25 cm, 5 pm; Mobile Phase A: Hex(0.5% 2M NH3-MeOH)-HPLC, Mobile Phase B: IPA: DCM = 1: 1— HPLC; Flow rate: 20 mL / min; Gradient (B%): isocratic 35% B; Wave Length: 254 / 220 nm to 3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)piperidine-l-carbonitrile.

[0405] Isomer 1 (1.6 mg, 1.5%yield) as a white solid and isomer 2 (3.3 mg, 3%yield) as a white solid. LCMS(ESI-MS) m / z =561.2 [M + H] + .

[0406] Example 16 (cpd 122)

[0407] Example 16-1 (cpd 122-1) l-(3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)piperidin- l-yl)prop-2-en-l-one, isomer 1

[0408] Example 16-2 (cpd 122-2) l-(3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3-methoxypyrazolo[l,5- a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5-d][l,4]diazepin-6-yl)piperidin- l-yl)prop-2-en-l-one, isomer 2

[0409] A 8 mL vial was charged with (2S)-2-(5-fluoropyridin-2-yl)-2-({3-methoxy-6-[3-methyl-6- (piperidin-3-yl)-4H,5H,7H,8H-pyrazolo[l,5-d][l,4]diazepin-2-yl]pyrazolo[l,5- a]pyridin-4- yl}oxy)ethanol (50 mg, 0.093 mmol, 1.00 equiv), DCM (2 mL, 0.024 mmol) and N,N- Diisopropylethylamine (0.07 mL, 0.390 mmol, 4.19 equiv). The prop-2-enoyl prop2-enoate (11.77 mg, 0.093 mmol, 1.00 equiv) was added and stirred for 30 min at room temperature. The reaction was quenched with water (20 mL) and extracted with DCM (3x10 mL). The combined organic layers were washed with brine (3x5 mL), dried over anhydrous Na2SO4. After filtration, the filtrate was concentrated under reduced pressure. The Confidential crude product (50 mg) was purified by Prep-HPLC with the following conditions: Column: XBridge Prep OBD C18 Column, 30*150 mm, 5pm; Mobile Phase A: Water (lOmmol / L NH4HCO3), Mobile Phase B: ACN; Flow rate: 60 mL / min mL / min; Gradient (B%): 27% B to 40% B in lOmin; Wave Length: 254nm / 220nm to afford the hybrid. Then the hybrid was separated by Prep-Chiral-HPLC with the following conditions: Column: (R, R)-WHELK-O1, 5 pm, 250 mm x 21.2 mm; Mobile Phase A: MtBE (with 0.5% NH3 (2 M in MeOH)), Mobile Phase B: MeOH: DCM = 1: 1; Flow rate: 20 mL / min; Gradient (B%): isocratic 30% B; Wave Length: 254 / 220 nm to afford l-(3-(2-(4-((S)-l-(5-fluoropyridin-2-yl)-2-hydroxyethoxy)-3- methoxypyrazolo[l,5-a]pyridin-6-yl)-3-methyl-4,5,7,8-tetrahydro-6H-pyrazolo[l,5- d][l,4]diazepin-6-yl)piperidin-l-yl)prop-2-en-l-one, isomer 1 (the first elution, 7.9 mg, 14.35%yield, 99.2%purity) as a white solid, and isomer 2 (the second elution, 7.9 mg, 14.35%yield, 99.3%purity) as a white solid.

[0410] Table 2 Experimental characterization of compounds according to an embodiment of the invention

[0411] Biological Examples

[0412] Overview The FGFR3 phosphorylation assay aims at measuring the reduction of FGFR3 autophosphorylation, a marker for FGFR.3 activity, upon treatment with potential FGFR3 inhibitors. 293T cells were transfected with a doxycycline inducible construct encoding the FGFR3 S249C+V555M double mutant fused to a C-terminal FLAG-tag. After exposing the transfected cells for two hours to potential FGFR3 inhibitors, phospho-FGFR (pFGFR) and total FGFR (tFGFR) levels were measured in a high-content microscopy assay by means of an anti- phospho-FGFR antibody and an anti-FLAG antibody.

[0413] Protocol

[0414] Fourteen million 293T cells (ATCC, Cat. No. CRL-3216) were transiently transfected using 75 pL JetPEI (Polyplus, Cat. No. 101000020), 2.5 pg of a FGFR3 S249C+V555M FLAG construct and 22.5 pg pBluescript DNA. The transfected cells were seeded in a T175 culture flask in culture medium consisting of Dulbecco's Modified Eagle's Medium (DMEM, Gibco, Cat. No. 41966-029) complemented with 10% heat inactivated (HI) fetal bovine serum (FBS), 2 mM L-glutamine and 1% penicillin / streptomycin (P / S) and incubated overnight (O / N) at 37°C, 5% CO2.

[0415] Assay plates (384-well format) were coated with 30 pL of a 50 pg / mL poly-D-lysine solution per well, followed by a brief centrifugation at 1,000 RPM and incubated for 1 hour (h) at 37°C, 5% CO2. The coating solution was then removed by tapping the plates on sterile tissues and subsequently the plates were washed lx with PBS and dried for Ih at room temperature (RT).

[0416] The transfected 293T cells were harvested by trypsinization and resuspended in culture medium (DMEM supplemented with 10% FBS, 1% Pen / strep, 2 mM L-Glutamine and 100 ng / ml doxycycline (Sigma, Cat. No. D3072)). 45 pl cell suspension, containing 20,000 transfected cells, was seeded into each well of the assay plates and the plates were incubated O / N at 37°C, 5% CO2.

[0417] The test compounds were prepared in compound dilution plates as a 10-point 1 / 5 dilution series in 100% DMSO starting from 10 mM. Control samples were added to the compound dilution plates: 100% DMSO was added to the negative control wells and 1 mM G1756296 to the positive control wells.

[0418] Compounds and controls were diluted 1 / 33 in culture medium supplemented with 100 ng / ml doxycycline and 5 pL of the dilution was transferred to the assay plates containing the cells. This resulted in a final concentration of 0.3% DMSO for the negative control wells, 3 pM G1756296 for the positive control wells and a 10-point 1 / 5 dilution series starting at a final concentration of 30 pM for the test compounds. The plates were briefly centrifuged at 1000 rpm and incubated at 37°C, 5% CO2 for 2h.

[0419] Cell fixation was performed by addition of 2.7 pL of a 37% paraformaldehyde (PFA) solution to each well of the assay plates still containing the medium (final PFA concentration: 2%), followed by a 15 min incubation at RT. Afterwards the plates were washed 3 times with PBS using a BlueWasher (BlueCatBio) device to remove the PFA. After the last wash the PBS was kept on the plates and plates were either stored at 4°C until further staining or stained immediately.

[0420] PBS was removed using a BlueWasher device and 25 pL of blocking buffer (PBS containing 0.2% Triton X-100, 2% heat inactivated FBS, 3% bovine serum albumin and 1% milk) was added to each well. Plates were incubated for Ih at RT. Next, immunostaining was performed by incubating the wells for 2 h with 25 pL of 1 / 200 diluted anti-pFGFR antibody (Cell Signaling Technology, Cat. No. 52928) and 1 / 1,000 diluted anti-FLAG antibody (Sigma, Cat. No. F1804) at RT.

[0421] The plates were washed 2x with PBS-T and lx with PBS and 25 pL of a solution containing secondary antibodies and nucleic acid stain was added. This solution consisted of PBS, 1 / 500 diluted Alex Fluor 647 conjugated goat anti-rabbit IgG (ThermoFisher Scientific, Cat. No. A- 21245), 1 / 500 diluted Alexa Fluor 488 conjugate goat anti-mouse IgG (ThermoFisher Scientific, Cat. No. A28175) and 1 / 1000 diluted Hoechst 33342 (ThermoFisher Scientific, H3570). Plates were incubated for Ih at RT.

[0422] After two wash steps with PBS-T and 1 wash step with PBS, plates were imaged on an automated high-content microscope (e.g., Operetta, Revvity).

[0423] Biological data of the compounds

[0424] A: IC5o= <25 nM

[0425] B: IC5o= >25 to 250 nM

[0426] C: IC50— >250 to 1000 nM

[0427] D: IC5o= >1000 nM to 10000 nM

[0428] Table 3 Biological data of compounds according to an embodiment of the invention

[0429] Protocol for FGFR1

[0430] The same protocol as described above for FGFR3 inhibition was used, replacing the doublemutant FGFR3 (S249C+V555M) by the FLAG-tagged full-length FGFR1.

[0431] Protocol for FGFR2

[0432] The same protocol as described above for FGFR.3 inhibition was used, replacing the doublemutant FGFR3 (S249C+V555M) by the FLAG-tagged full-length FGFR2. Protocol for FGFR4

[0433] The same protocol as described above for FGFR3 inhibition was used, replacing the doublemutant FGFR3 (S249C+V555M) by the FLAG-tagged full-length FGFR4

[0434] Certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3 over FGFR1. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, are at least about 3 fold (e.g. at least about 4-, 5- , 6-, 7-, 8-, 9-, 10-, 15-, 20-, 30-, 40-, or more) more selective for FGFR3 than for FGFR1. Certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3 over FGFR2. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, are at least about 3 fold (e.g. at least about 4-, 5- , 6-, 7-, 8-, 9-, 10-, 15-, or more) more selective for FGFR3 than for FGFR2.

[0435] Certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, selectively target FGFR3 over FGFR4. For example, certain compounds of formula (I), or a pharmaceutically acceptable salt thereof, are at least about 3 fold (e.g. at least about 4-, 5- , 6-, 7-, 8-, 9-, 10-, 15-, or more) more selective for FGFR3 than for FGFR4.

[0436] Pharmacokinetic, ADME and Toxicity Assays

[0437] Thermodynamic solubility

[0438] The thermodynamic solubility is determined in Fasted State Simulated Gastric Fluid (FaSSGF, pH 1.6), Fed State Simulated Intestine Fluid (FeSSIF v2, pH 5.8), Fasted State Simulated Intestine Fluid (FaSSIF v2, pH 6.5) and phosphate buffer (pH7.4).

[0439] Approximately 1 mg of each compound is weighed into a 12 mL clear glass screw cap vial and 1 mL of medium is added (final concentration about Img / mL). The sample is placed in a thermodynamic water bath at 37 °C and 120 rpm of orbital shaking for 24 hours.

[0440] For HPLC / DAD analysis the sample is filtered through a 0.2 pm PTFE filter mounted onto syringe and then analysed neat and accurately diluted 10 and 100 times.

[0441] The same batch of test compound is used as a standard, assigning the purity of 100%, in order to prepare the calibration curve. Test compound stock solution is prepared at a concentration of 1 mg / mL in DMSO and diluted to cover the calibration range from 0.0001 - 0.1 mg / mL.

[0442] The samples are analysed on HPLC system with a flow rate of 0.5 mL / min. Solvent A is 10 mM ammonium carbonate at pHlO and solvent B is acetonitrile. The sample is going through a XBridge C18 2.7pM (2.1 x 50mm) column, from Waters. The solvent gradient has a total run time of 12 min and ranges from 10% B to 90% B.

[0443] Peak areas are analysed and are plotted against the standard curve to obtain the solubility of the compound.

[0444] Solubility values are reported in pM or pg / mL.

[0445] Aqueous Solubility This protocol describes determination of kinetic solubility by turbidimetry.

[0446] Turbidimetric solubility allows rapid determination of solubility using small amounts of compound. Briefly, compound's DMSO solutions (prepared at specific concentrations) are spiked into aqueous buffer and turbidity is measured using VIS spectrophotometry at 620 nm. Sulfaphenazole and o-naphtoflavone are used as assay controls.

[0447] 10 mM DMSO stock solution is diluted in DMSO (in 96-well V-bottom polypropylene microplates) to a concentration of 1 mM.

[0448] 3 pl of compound's / control's dilution in DMSO in triplicates are added to appropriate wells containing 297 pl of 100 mM PBS solution to the final concentration of 10 pM (1% DMSO; done in 96-well flat bottom microplates).

[0449] The plate is incubated by gently shaking (200-300 rpm) for 45 minutes at 37°C and left at room temperature (without shaking) for 15 minutes (total incubation time = Ih).

[0450] The plate absorbance at 620nm is read on a Microplate reader (Tecan, Infinite F500 or Spark) Calculation of solubility range data and resulting curves is made by Excel tools software.

[0451] The absorbance is proportionally increased with concentration of insoluble particles. Compound / control samples are compared to a solvent control in aqueous buffer (DMSO 1% final concentration), where significant increase of sample absorbance is considered when its absorbance is 3-fold standard deviation of average DMSO absorbance.

[0452] Results are accepted depending on solubility values obtained for assay controls: o-naphtoflavone: <10 pM sulfaphenazole: >10pM

[0453] Results are expressed as an average of three replicas (<10 pM or >10 pM) for one concentration set-up.

[0454] Plasma Protein Binding (Equilibrium Dialysis)

[0455] The binding of the test compound to plasma proteins is tested by equilibrium dialysis using a Teflon unit from HTDialysis, performed on a Freedom EVO liquid handling unit.

[0456] Prior to the start of the experiment, dialysis membranes (Dialysis membranes MW cut-off 12- 14kDa - HTDialysis - Cat. No. 1101) are soaked in deionised water for 60 minutes, transferred and left overnight in 20% ethanol. On the day of experiment, compound and control stock solutions are diluted, spiked into plasma in order to achieve a final concentration of 5 pM (0.5% DMSO). Immediately after assembly of the Teflon unit, a volume of 100 pl of plasma (spiked with compound / control) is placed on one side of the well and 100 pl of blank PPB buffer (60 mM Na2HPO4, 14 mM KH2PO4, 70 mM NaCI) is added to the other side, respectively, with each compound assayed in duplicate. The test compound is incubated for 4 hours at 37°C with gentle shaking. Thereafter, an aliquot is taken from each side of the well and matrix matched (mix of equal volumes of spiked plasma with blank PPB buffer and samples from buffer compartment with blank plasma). In order to estimate the recovery an aliquot is also taken at t=0 min from spiked plasma solutions and matrix matched as described above.

[0457] Matrix matched samples are further mixed with six volumes of STOP solution (acetonitrile: methanol, 2: 1 v / v with an internal standard (IS)). After brief mixing and centrifugation (at 4500 rpm for 30 min, at +4°C), the supernatant is transferred into new plates and analysed and quantified by LC-MS / MS.

[0458] The percentage bound (PPB) is determined using the following equation:

[0459] PPB = [( Cplasma ~ Cbuffer) / Cplasma ]*100

[0460] Cpiasma = Peak area of the compound in the plasma I Peak area of the IS in the plasma Cbuffer = Peak area of the compound in the buffer / Peak area of the IS in the buffer

[0461] The recovery is a control, it allows to be sure that the compound has not a non-specific binding to the plates or it is not stable in the plasma in these conditions.

[0462] % recovery = [(Buffer+Plasma) / Recovery]*100

[0463] Buffer = (ratio of the peak area of the compound / peak area of IS) in Buffer compartment after 4h.

[0464] Plasma = (ratio of the peak area of the compound / peak area of IS) in Plasma compartment after 4h.

[0465] Recovery = ratio at TO of the peak area of the compound in the well recovery I peak area of the IS in the well recovery.

[0466] Aldehyde oxidase stability

[0467] An in vitro metabolic stability in human and rat liver S9 (human: 20-donor pool, mixed gender (Corning, Cat. No.452961) and rat: pooled, male Sprague-Dawley (Corning, Cat. No. 452591)) is done in order to investigate the test compound as substrate of aldehyde oxidase. A 10 mM DMSO stock solution of the test compound is first diluted in DMSO (40 fold) to obtain a 250 pM working solution. Incubation mixtures are prepared by adding liver S9 suspension (final protein concentration of 2 mg / mL) to 50mM potassium phosphate buffer, pH 7.4. Hydralazine (Sigma, Cat.No.H1753) (selective inhibitor of aldehyde oxidase; final concentration of 100 pM) or miliQ water are added for incubations with and without addition of selective inhibitor, respectively. After pre-warming for 5 min at 37°C, the reaction is initiated by addition of test compound to the incubation mixture (final concentration in the incubation mixture of 1 pM, 0.4%DMSO). After 0, 3, 6, 12, 18 and 30 minutes of incubation, the reaction (aliquot of 100 pL) is terminated with 300 pL of acetonitrile: methanol (2: 1) mixture with 1% acetic acid and the analytical internal standard. Samples are mixed, centrifuged and the supernatant analysed by LC-MS / MS. Phthalazine (Aldrich, Cat. No. P38706) is included as a positive control.

[0468] S9 stability, expressed as the percentage of remaining parent compound, is calculated from the peak area ratio of compound and internal standard following different incubation times compared to the same ratio at the t=0 min (100%). The half-life (tl / 2) is calculated in GraphPadPrism software from % remaining vs. time regression using non-linear regression fit (one phase exponential decay) with following constrain parameters: Span = 100, Plateau=0, K=no constraint).

[0469] In vitro intrinsic clearance (CLint) is calculated from the half-life value using following equation: CLint[pl / min / mg] = (In2 / tl / 2)*(ml per incubation / mg S9 protein) .

[0470] Liver microsomal stability

[0471] The test compound is diluted from DMSO stock solution to obtain a final concentration of IpM in the final incubation mix (final DMSO 0.03%). A NADPH generating system (Cofactor solution) is prepared by adding to a 50mM PBS pH 7.4 buffer, NADP (Sigma, N0505), G6P (Sigma, G6526), MgChxSH?© (Sigma, M2670) and G6P DH (Sigma, G637) to obtain in the final incubation mix NADP at 0.5mM , G6P at 5mM , MgChx6H2O at 0.5mM and G6P DH at 1.5 U / mL. Liver microsomes (Corning; Discovery Life Sciences) solution is prepared from 20mg / mL stock concentration to obtain a final concentration in the incubation mix of 0.5mg / mL.

[0472] Test compounds are incubated at 37°C with and without cofactors and reaction is started by adding cofactors or buffer (for negative control) to incubation mix. Time points are taken at 0, 10, 25 and 40'. At the desired incubation time point, a STOP solution (acetonitrile: methanol, 2:1 v / v) is added at a ratio 3: 1 to the incubation mix. Samples are then centrifuged (at 4500 rpm, at 4°C, for 30 min). The supernatant is then analyzed using a LC-MS / MS method. Metabolic stability, expressed as the percentage of remaining parent compound, is calculated from the ratio of peak area of the remaining compound and peak area of the internal standard after different time of incubation compared to the same ratio at the t=0 min (100%):

[0473] % remaining = (peak area ratios of test compound vs IS at appointed time) / (peak area ratios of test compound vs IS at Omin) x 100

[0474] The half-life (ti / 2) is calculated in GraphPad Prism software from % remaining vs. time regression using non-linear regression fit (one phase exponential decay with following constrain parameters: Span = 100, Plateau=0, K=no constraint).

[0475] In vitro intrinsic clearance (CLint) is calculated from half-life using following equation:

[0476] CLint = [pl / min / mg] = 0.693 / ti / 2 / min x (mL of incubation / mg protein) x 1000

[0477] Hepatocyte stability

[0478] The test compound is diluted from DMSO stock solution to obtain a final concentration of IpM in the final incubation mix. The test compound is incubated in dog, monkey, human, mouse and rat hepatocytes (BioIVT, mouse Cat. No. M005052, rat Cat. No. M00005 , dog, Cat. No. M00205 , monkey Cat. No. M00305, human Cat. No. X008001) (final concentration: 0.5x106 cells / ml), resuspended in Krebs-Henseleit modified buffer for 3 hours (except in human - 90 minutes) at 37°. Aliquots are taken at different time points and the reaction is terminated by addition of 3 volumes of STOP solution (acetonitrile: methanol = 2: 1 with an internal standard). Aliquots are then centrifuged (at 4500 rpm, at 4°C, for 30 min). The supernatant is then analysed using a LC-MS / MS method.

[0479] Metabolic stability, expressed as the percentage of remaining parent compound, is calculated from the ratio of peak area of the remaining compound and peak area of the internal standard after different time of incubation compared to the same ratio at the t=0 min (100%).

[0480] The half-life (tl / 2) is calculated in GraphPadPrism software from % remaining vs. time regression using non-linear regression fit (one phase exponential decay) with following constrain parameters: Span = 100, Plateau=0, K=no constraint).

[0481] In vitro intrinsic clearance (CLint) is calculated from half-life using the following equation:

[0482] CL]nt[pl / min / 106cellsj = (In2 / tl / 2)*(ml per incubation / #cells per incubation)

[0483] Caco-2 Permeability Bi-directional Caco-2 assays are performed as described below. Cells are seeded at 1 x 105 cells / cm2 in 96-well HTS Transwell plates (Corning). Permeability assays are performed with the cells at days 21-25 post-seeding.

[0484] This assay is performed in both the apical (A) to basolateral (B) A-B or B-A direction. Compounds and the references (Propranolol, Labetalol, Ranitidine, Colchicine) are prepared at 10 pM in HBSS-MES (pH 6.5) or HBSS-HEPES (pH 7.4) with a final DMSO concentration of 1 % (v / v). The working solution is then centrifuged and the supernatant is added to the donor side.

[0485] The assay plate is incubated at 37 °C for 60 min or 40 min for the A-B or B-A assay, respectively. Samples are aliquoted from the donor side at time zero and the end point, and from the receiver side at the end point.

[0486] In addition, Lucifer yellow (LY), a membrane integrity marker, is co-incubated with the test compound at the start of the experiment to assess integrity of the cell layers. As LY cannot freely permeate lipophilic barriers, a high Papp of the Lucifer yellow (measured by fluorescence) indicates poor formation of the cell monolayer.

[0487] Moreover, the Transepithelial electrical resistance (TEER) of each Multiscreen™ well is also evaluated prior to the initiation of the assay so as to evaluate monolayer integrity.

[0488] Reference compounds namely, Propranolol (highly permeable), Labetalol (moderately permeable), Ranitidine (poorly permeable), and Colchicine (P-glycoprotein substrate) are included in this assay. Samples are analysed by LC / MS-MS.

[0489] Apparent permeability (Papp) values are calculated from the relationship:

[0490] Papp=[COmpOUnd] acceptor final X Vacceptor / ([COmpOUnd]donor initial X Vdonor) / Tine X Vdonor / SUCfdCO area x 60 x 10~6cm / s

[0491] V = chamber volume

[0492] Tine = incubation time.

[0493] Surface area = 0.33cm2

[0494] The Efflux ratios, as an indication of active efflux from the apical cell surface, are calculated using the ratio of Papp B>A / Papp A>B.

[0495] The following assay acceptance criteria are used: If at least 3 out of 4 control molecules are in these ranges determined by the historical data, the assay is validated.

[0496] Lucifer yellow permeability: <0.5 10-6 cm / s

[0497] Colchicine: Papp (A>B) value < 1 (xlO-6 cm / s) with Efflux ratio >4

[0498] Labetalol: Papp (A>B) value < 6 (xlO-6 cm / s) Propranolol: Papp (A>B) value < 25 (xlO-6 cm / s)

[0499] Ranitidine: Papp (A>B) value < 2 (xlO-6 cm / s)

[0500] MDCKII-MDR1 Permeability

[0501] MDCKII-MDR1 cells are Madin-Darby canine kidney epithelial cells, over-expressing human multi-drug resistance (MDR1) gene, coding for P-glycoprotein (P-gp). Cells are obtained from Netherlands Cancer Institute and used after a 3-4 day cell culture in 24-well Millicell cell culture insert plates (Millipore, PSRP010R5). Bi-directional MDCKII-MDR1 permeability assay is performed as described below.

[0502] 3x105 cells / mL (1.2x105 cells / well) are seeded in plating medium consisting of DMEM + 1% Glutamax-100 + 1% Antibiotic / Antimycotic + 10% FBS (Biowest, S1810). Cells are left in CO2 incubator for 3-4 days. The medium is changed 24h after seeding and on the day of experiment.

[0503] Test and reference compounds (amprenavir and propranolol) are prepared in Dulbecco's phosphate buffer saline (D-PBS, pH7.4) and added to either the apical (400pL) or basolateral (800pL) chambers of the Millicell cell culture insert plates assembly at a final concentration of 10 pM (0.5 pM in case of amprenavir) with a final DMSO concentration of 1%. lOOpM Lucifer Yellow (Sigma) is added to the all donor buffer solutions, in order to assess integrity of the cell monolayers by monitoring Lucifer Yellow permeation. Lucifer yellow is a fluorescent marker for the paracellular pathway and it is used as an internal control in every monolayer to verify tight junction integrity during the assay.

[0504] After a 1 h incubation at 37°C while shaking at an orbital shaker at 150rpm, 75pL aliquots are taken from both apical (A) and basal (B) chambers and added to 225pL acetonitrile:water solution (2: 1) containing analytical internal standard (10 ng / mL warfarin) in a 96 well plate. Aliquoting is also performed at the beginning of the experiment from donor solutions to obtain initial (Co) concentration.

[0505] Concentration of compound in the samples is measured by high performance liquid- ch romatography / mass spectroscopy (LC-MS / MS).

[0506] Lucifer yellow is measured with a Fluoroscan Ascent FL Thermo Scientific (Ex 485nm and Em 530nm) in a 96 well plate containing 150pL of liquid from all receiver wells (basolateral or apical side).

[0507] The apparent permeability coefficient (Papp) is calculated according to the following equation: Papp = (dQ / dT)*(l / CO)*(l / A) where dQ / dT = permeability rate

[0508] CO = initial concentration in donor compartment

[0509] A = surface area of the cell monolayer (0.11 cm2)

[0510] "Concentration" is the ratio between compound and internal standard peak areas. The Papp value has a dimension of a rate (xlO-6 cm / sec).

[0511] Permeability values are classified as follows: low permeability: <2x10-6 cm / sec, moderate permeability: 2-10x10-6 cm / sec, high permeability: >10x10-6 cm / sec

[0512] The Efflux ratio, as an indication of active efflux from the apical cell surface, is calculated using following equation: Efflux ratio = Papp (B2A) / Papp (A2B)

[0513] A compound is considered to be a possible P-gp substrate when the efflux ratio is > 2.

[0514] Mass balance (MB) is calculated from equation: MB = (Md+Mr) / Mo where Md = mass of drug in donor compartment at time 60 min Mr = mass of drug in receiver compartment at time 60 min Mo = mass of drug in donor compartment at time 0 min

[0515] Mass of drug = Analyte / IS ratio * volume of compartment (75 pL or 250 pL)

[0516] Mass balance results should be evaluated as follows: Mass balance between 80-120%: acceptable, Mass balance between 50-80% and 120-135% should be interpreted with caution, Mass balance <50% and >135% should be discarded

[0517] Results are accepted based on obtained data: Lucifer yellow: all Papp values should be <3x10-6 cm / sec, Amprenavir: low permeability (PappA2B <2x10-6 cm / sec), efflux ratio >2, Diclofenac: high permeability (PappA2B >10x10-6 cm / sec), efflux ratio <2

[0518] Glutathione stability assay

[0519] Solutions of test compound (10 pM, 1% final DMSO concentration) are incubated in 50 mM PBS buffer only to evaluate the buffer stability of the test compound and in 50 mM PBS buffer containing 2 mM reduced L-glutathione to determine the reactivity of the test compound. The incubation is performed in duplicate for 5h at 37°C and pH 7.4. Omeprazole, Afatinib and Ibrutinib are used as positive controls. At 0, 30, 60, 180 and 300 min, aliquots are withdrawn and immediately quenched with ice-cold organic solvent (acetonitrile:methanol (2: 1)) containing diclofenac and warfarin sodium as internal standard. Samples are then centrifuged to remove precipitated proteins, diluted with H2O MilliQ:STOP solution (if needed) and the (diluted) supernatant is analysed by Liquid Chromatography-Tandem Mass Spectrometry (LC- MS / MS). GSH reactivity, expressed as the percentage of remaining parent compound, is calculated from the ratio of peak area of the remaining compound and peak area of the internal standard after different times of incubation compared to the same ratio at the t=0 min (100%). peak area ratios of test compound vs. IS at appointed time

[0520] % remaining = - - - - - - - x 100. peak area ratios of test compound vs. IS at 0 min

[0521] The half-life (ti / 2) is calculated in GraphPad Prism software from % remaining vs. time regression using non-linear regression fit (one phase exponential decay with following constrain parameters: Span=100, Plateau=0, K=no constraint). The half-life and % remaining after 5 h is reported for the test compound and the reference compounds. For omeprazole, the % remaining at 5h should be < 50% and the half-life < 5h. For Afatinib, the % remaining at 5h should be < 35% and the half-life < 5h. For Ibrutinib, the % remaining at 5h should be > 80% and the half-life > 5h. The % remaining in the buffer stability assay condition should be >80%.

[0522] Aurora B kinase inhibition assay

[0523] Aurora-B (h) is incubated with the compound of interest dissolved in DMSO, 8 mM MOPS pH 7.0, 0.2 mM EDTA, 30 pM AKRRRLSSLRA, 10 mM Magnesium acetate and [y-33P]-ATP (specific activity and concentration as required). The reaction is initiated by the addition of the Mg / ATP mix. After incubation for 40 minutes at room temperature, the reaction is stopped by the addition of phosphoric acid to a concentration of 0.5%. An aliquot of the reaction is then spotted onto a filter and washed four times for 4 minutes in 0.425% phosphoric acid and once in methanol prior to drying and scintillation counting. The biochemical kinase activity assays are performed with the gold standard radioisotope-based technology.

[0524] Pharmacokinetic study in rodents

[0525] Animals

[0526] Wistar-Han rats (male, 200-225g) are obtained from Janvier (France).

[0527] CD-I Mice (male, 25-30g) are obtained from Janvier (France).

[0528] Animals are acclimatized for at least 7 days before treatment and are kept on a 12 h light / dark cycle (0700 - 1900). Temperature is maintained at approximately 22°C, and food and water are provided ad libitum.

[0529] Pharmacokinetic study

[0530] Compounds are formulated in DMSO / 10% hydroxylpropyl-p-cyclodextrine (5 / 95) for the intravenous route and in a suitable solution for the oral route. Test compounds are orally dosed as a single oesophageal gavage at 5 mg / kg under a dosing volume of 10 mL / kg (group of 3 animals) and intravenously dosed as a bolus via the caudal vein at 0.5 mg / kg under a dosing volume of 5 mL / kg. (groups of 2 animals). Blood samples are collected at the retro- orbital sinus with EDTA as anti-coagulant at the following time points: 0.05, 1, 3, 6 and 24 h (intravenous route), and 0.25, 1, 3, 6, and 24 h (oral route). Whole blood samples are centrifuged at 5000 rpm for 10 min and the resulting plasma samples are stored at -20°C pending analysis.

[0531] Quantification of compound levels in plasma

[0532] Plasma concentrations of each test compound are determined by an LC-MS / MS method in which the mass spectrometer is operated in positive or negative electrospray mode.

[0533] Determination of pharmacokinetic parameters

[0534] Pharmacokinetic parameters are calculated for each individual animal and for each analyte by non-compartmental analysis.

[0535] Liability for QT prolongation

[0536] Potential for QT prolongation is assessed in the hERG manual patch clamp assay.

[0537] Manual patch clamp assay

[0538] Single CHO cells are used for recording hERG currents at near physiological temperature (34- 35°C) using the whole-cell patch-clamp technique with an Axopatch 700A amplifier / pCIamp software (Molecular Devices, Sunnyvale CA).

[0539] Electrodes (5-7 MQ resistance) are prepared from GC150-10 glass capillary tubes (Harvard Apparatus, Cambridge, UK) and filled with intracellular solution (in mM: KCI 120; Na2ATP 4; HEPES 10; EGTA 10; MgCI21.75; CaCI25.374, pH 7.2 adjusted with KOH).

[0540] The patch-clamp recording chamber is perfused with control extracellular solution (in mM: NaCI 145; KCI 4; MgCI2 1; HEPES, 10; glucose 10; CaCI22 at pH 7.4 adjusted with NaOH). With the formation of a gigaohm seal, the cell membrane is ruptured to establish the wholecell configuration. The minimum acceptable seal resistance is 1 gigaohm, and access resistance no greater than 20pF.

[0541] The cell membrane is then voltage-clamped at a holding potential of -80 mV and depolarised to +20 mV for 2 sec followed by a negative pulse at -40mV for 1.6 sec, before returning to - 80mV at a frequency of 0.1 Hz.

[0542] Once the hERG current is stable, the perfusion is switched to extracellular solution containing test compounds starting from the lowest concentration. Each concentration on the 4pt concentration-response curve is tested for 4min to achieve steady state. Automated patch clamp

[0543] Electrophysiological recordings are made from a Chinese Hamster Ovary cell line stably expressing the full-length ion channel. Single cell ionic currents are measured in whole-cell configuration at room temperature (21-23°C) using a QPatch II (Sophion Bioscience).

[0544] The internal solution for hERG contains (mM): 120 KCI, 20 KF, 10EGTA, 10 HEPES and is buffered to pH 7.3.

[0545] The external solution (HEPES-buffered saline, HBPS) contains (mM): 138 NaCI, 4.5 KCI, 1.8 CaCh, 1.0 MgCh, 10 HEPES, 10 glucose, buffered to pH 7.4.

[0546] Cells are clamped at a holding potential of -80mV before a step to +20mV for 500ms, and then -40mV for 500ms. The test pulse is applied every 10 seconds. Currents are measured from the -40mV step and referenced to the holding current.

[0547] Compounds are incubated for 120 seconds to allow stabilization of the parameters. Concentration-response curves are generated by cumulative addition of compound with concentrations low to high.

[0548] The present invention is in no way limited to the embodiments described in the examples and / or shown in the figures. On the contrary, methods according to the present invention may be realized in many different ways without departing from the scope of the invention.

[0549] Final remarks

[0550] It will be appreciated by those skilled in the art that the foregoing descriptions are exemplary and explanatory in nature, and intended to illustrate the invention and its preferred embodiments. Through routine experimentation, an artisan will recognize apparent modifications and variations that may be made without departing from the spirit of the invention. All such modifications coming within the scope of the appended claims are intended to be included therein. Thus, the invention is intended to be defined not by the above description, but by the following claims and their equivalents.

[0551] All publications, including but not limited to patents and patent applications, cited in this specification are herein incorporated by reference as if each individual publication are specifically and individually indicated to be incorporated by reference herein as though fully set forth. It should be understood that factors such as the differential cell penetration capacity of the various compounds can contribute to discrepancies between the activity of the compounds in the in vitro biochemical and cellular assays. At least some of the chemical names of compound of the invention as given and set forth in this application, may have been generated on an automated basis by use of a commercially available chemical naming software program, and have not been independently verified. Representative programs performing this function include the Lexichem naming tool sold by Open Eye Software, Inc. and the Autonom Software tool sold by MDL, Inc. and Chemdraw 20 by PerkinElmer Informatics Inc.

[0552] In the instance where the indicated chemical name and the depicted structure differ, the depicted structure will control.

Claims

CLAIMS1. A compound of Formula (I) :wherein:Xi is N or CH;X2 is N and X3 is C, or, alternatively, X3 is N and X2 is C;X4is N;Y is a bond, O or NH;Yi is CHR6, CH2-CHR6, CHR6-CH2, CF2, CH2-CF2 or CF2-CH2;Y2is CR4R5or CF2;Y3is CR3R4, CR3R4CR3R4, CH2CF2 or CF2; wherein Yi, Y2, Y3 and X4 together with the pyrazolyl ring form a 7-membered nitrogen containing ring; nd A is CN, R8R9C=CR10C(=O)-,wherein R4and R9are as defined herewith, or, alternatively, R4and R9together with the carbon atoms to which they are attached form a 3-5-membered carbocyclic ring.Y4 is a 4-10 membered monocyclic heterocycloalkyl or a 5-12 membered fused, spiro or bridged polycyclic heterocycloalkyl, said monocyclic or polycyclic heterocycloaklyl comprising a nitrogen atom, wherein the heterocycloalkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, methyl, halomethyl, hydroxyl or methoxy and wherein the nitrogen atom of the heterocycloalkyl is connected to A;R1is hydrogen, halogen, C1-5 alkyl, C1-5 alkoxy, CN, C3-10 cycloalkyl, C2-4 alkynyl, 4-10 membered heterocycloalkyl comprising 1, 2 or 3 heteroatoms independently selected from N, O and S, phenyl or a 5-6 membered heteroaryl, wherein the 3-10 membered cycloalkyl, 4-10 membered heterocycloalkyl, phenyl and 5-6 membered heteroaryl are unsubstituted or substituted with one or more substituents independently selected from halogen, methyl, halomethyl, hydroxyl or methoxy and wherein the C1-5 alkyl and C1-5 alkoxy, are unsubstitutedor independently substituted with one or more substituents independently selected from halogen, CN, hydroxyl and methoxy;R2is hydrogen, hydroxyl, Ci-4 alkyl, CN, -CONHCI-4 alkyl, or Ci-4 haloalkyl; each R3is independently selected from hydrogen, fluoro, hydroxyl, methoxy, C1-3 alkyl, cyclopropyl, or a R3is fused with R5or R6to form CH2, CH2-CH2 or CH2OCH2; each R4is independently selected from hydrogen, or C1-3 alkyl;R5is hydrogen, fluoro, hydroxyl, methoxy, C1-3 alkyl, cyclopropyl or is fused with one R3to form CH2, CH2-CH2 or CH2OCH2, wherein the C1-3 alkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, heteroaryl (which heteroaryl is preferably pyridyl), phenyl, hydroxyl, -CO2R4or C1-3 alkoxy;R6is hydrogen, fluoro, hydroxyl, methoxy, C1-3 alkyl or is fused with one R3to form CH2, CH2- CH2or CH2OCH2; each R8is independently selected from hydrogen and C1-6 alkyl; each R9is independently selected from hydrogen, CF3 and C1-6 alkyl, which alkyl is unsubstituted or substituted with one or more substituents independently selected from fluoro, chloro, bromo, hydroxyl, and C1-6 alkoxy; andR10is hydrogen, C1-3 alkyl or halogen; or R9and R10together with the carbon atoms to which they are attached form a 4-8- membered carbocyclic ring; each of R11and R12is independently hydrogen, C1-6 alkyl, or R11and R12together with the nitrogen atom to which they are attached form a 4-6 membered heterocycloalkyl optionally having an additional ring heteroatom which is O, wherein said ring is optionally substituted with halogen;R13is hydrogen, R14R15NCH2-, or Ci-3 alkyl which is unsubstituted or substituted with hydroxyl;R14and R15are each independently hydrogen or C1-6 alkyl;R16is C1-3 alkyl-R17or C3-6 cycloalkyl-R17, wherein the C1-3 alkyl or C3-6 cycloalkyl are unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, CN, Ci-4 alkyl optionally substituted with one or more OH, C3-6 cycloalkyl, OH, OMe, -NR4R8wherein R4and R8are as defined above;R17is aryl or heteroaryl comprising 1, 2 or 3 heteroatoms independently selected from N, O and S, wherein the aryl or heteroaryl are unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, CN, NH2, Ci-4 alkyl, and Ci-4 haloalkyl, Ci-4 alkoxy, and Ci-4 haloalkoxy; or a pharmaceutically acceptable salt and / or solvate thereof.

2. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to claim 1, wherein Xi is CH, X3 is N and X2 is C and Y is O.

3. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to claim 1, wherein Xi is N, X3 is N and X2 is C and Y is O.

4. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 3, wherein Yi is CHR6, Y2 is CR4R5or CF2 and Y3 is CH2CR3R4.

5. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 4, wherein B is -C(O)Y4-.

6. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 4, wherein B is -Y4-.

7. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 4, wherein B is -SO2-Y4-.

8. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 4, wherein B is -CR4R9Y4, such as -CH2Y4-, wherein R4and R9are as defined herewith, or, alternatively, R4and R9together with the carbon atoms to which they are attached form a 3-5-membered carbocyclic ring.

9. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 8, wherein Y4 is a 4-10 membered heterocycloalkyl comprising a nitrogen atom, wherein the heterocycloalkyl is unsubstituted or independently substituted with one or more substituents independently selected from halogen, methyl, halomethyl, hydroxyl or methoxy and wherein the nitrogen atom of the heterocycloalkyl is connected to A.

10. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 9 wherein R1is halogen, such as chloro.

11. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 9 wherein R1is C1-5 alkoxy, wherein the C1-5 alkoxy is unsubstituted or independently substituted with one or more substituents independently selected from halogen, CN, hydroxyl and methoxy such as halogen such as F.

12. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 9 wherein R1is -OMe or -OCHF2.

13. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 12 wherein R2is Ci-4 alkyl, such as methyl and R3, R4, R5and R6are hydrogen.no14. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 12 wherein R16is C1-3 alkyl-R17wherein the C1-3 alkyl group is unsubstituted or substituted with one or more substituents independently selected from the group consisting of halogen, CN, Ci-4 alkyl, C3-6 cycloalkyl, OH, OMe, -NR4R8wherein R4and R8are as defined above, or, alternatively, R4and R8together with the carbon atoms to which they are attached form a 4-8-membered carbocyclic ring, such as the C1-3 alkyl group is substituted by one OH.

15. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to claim 14, wherein R16is selected from C2 alkyl-R17or C3 alkyl-R17, and wherein the C2 alkyl or C3 alkyl is substituted with at least one OH.

16. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to claim 14, wherein R16is C2 alkyl-R17, and wherein the C2 alkyl is substituted with OH.

17. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 16 wherein R17is aryl or heteroaryl, such as pyrazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl and pyrazinyl such as pyridinyl.

18. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 16 wherein R17is 2-pyridinyl, optionally substituted with one or more F.

19. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 18, wherein A is CN.

20. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 18, wherein A is CH2=CR10C(=O)-.

21. The compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1 to 20, wherein the compound is selected from:Ill22. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of a compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1-21.

23. A compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1-21, or a pharmaceutical composition according to claim 22, for use in medicine.

24. A compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1-21, or a pharmaceutical composition according to claim 22, for use in the prophylaxis and / or treatment of a disease, disorder or condition that is associated with abnormal activity or expression of an FGFR enzyme, preferably FGFR3.

25. A compound, or a pharmaceutically acceptable salt and / or solvate thereof, according to any one of claims 1-21, or a pharmaceutical composition according to claim 22, for use in the prophylaxis and / or treatment of a disease, disorder or condition that is selected from metastatic tumours (such as but not limited to, melanoma, lymphoma, leukaemia, fibrosarcoma, rhabdomyosarcoma, and mastocytoma) and types of tissue carcinoma (such as but not limited to, colorectal cancer, prostate cancer, small cell lung cancer and non-small cell lung cancer, breast cancer, pancreatic cancer, bladder cancer, renal cancer, gastric cancer, glioblastoma, primary liver cancer, ovarian cancer, prostate cancer and uterine leiomyosarcoma), acute lymphoblastic leukemia, acute myeloidleukemia, adrenocortical carcinoma, anal cancer, appendix cancer, astrocytomas, atypical teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer (osteosarcoma and malignant fibrous histiocytoma), brain stem glioma, brain tumors, brain and spinal cord tumors, breast cancer, bronchial tumors, Burkitt lymphoma, cervical cancer, chronic lymphocytic leukemia, chronic myelogenous leukemia, colon cancer, colorectal cancer, craniopharyngioma, cutaneous T -Cell lymphoma, embryonal tumors, endometrial cancer, ependymoblastoma, ependymoma, esophageal cancer, ewing sarcoma family of tumors, eye cancer, retinoblastoma, gallbladder cancer, gastric (stomach) cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), gastrointestinal stromal cell tumor, germ cell tumor, glioma, hairy cell leukemia, head and neck cancer, hepatocellular (liver) cancer, hodgkin lymphoma, hypopharyngeal cancer, intraocular melanoma, islet cell tumors (endocrine pancreas), Kaposi sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, hairy cell leukemia, liver cancer, non-small cell lung cancer, small cell lung cancer, Burkitt lymphoma, cutaneous T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, lymphoma, Waldenstrom macroglobulinemia, medulloblastoma, medulloepithelioma, melanoma, mesothelioma, mouth cancer, chronic myelogenous leukemia, myeloid leukemia, multiple myeloma, asopharyngeal cancer, neuroblastoma, non-Hodgkin lymphoma, non-small cell lung cancer, oral cancer, oropharyngeal cancer, osteosarcoma, malignant fibrous histiocytoma of bone, ovarian cancer, ovarian epithelial cancer, ovarian germ cell tumor, ovarian low malignant potential tumor, pancreatic cancer, papillomatosis, parathyroid cancer, penile cancer, pharyngeal cancer, pineal parenchymal tumors of intermediate differentiation, pineoblastoma and supratentorial primitive neuroectodermal tumors, pituitary tumor, plasma cell neoplasm / multiple myeloma, pleuropulmonary blastoma, primary central nervous system lymphoma, prostate cancer,rectal cancer, renal cell (kidney) cancer, retinoblastoma, rhabdomyosarcoma, salivary gland cancer, sarcoma, Ewing sarcoma family of tumors, sarcoma, kaposi, Sezary syndrome, skin cancer, small cell Lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, stomach (gastric) cancer, supratentorial primitive neuroectodermal tumors, T - cell lymphoma, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, urethral cancer, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, Waldenstrom macroglobulinemia, Wilms tumor, a neoplastic disease of the blood and blood forming organs, including but not limited to: acute myeloid leukaemia (AML), and acute lymphoblastic leukemia (ALL) and chronic lymphoblastic leukaemia (CLL), breast cancer (e.g. invasive ductal cancer, invasive lobular cancer), lung cancer (e.g. non-small-cell lung cancer, lung adenocarcinoma, squamous cell lung cancer and small-cell lung cancer), urothelial cancer, bladder cancer (e.g. urothelial bladder cancer, non-muscle invasive bladder cancer, muscle invasive bladder cancer), upper tract cancer (e.g. urothelial upper tract cancer), urethral cancer, gastric cancer, pancreatic cancer, prostate cancer, colorectal cancer, multiple myeloma, liver cancer, melanoma (e.g. cutaneous melanoma), head and neck cancer (e.g. oral cancer), thyroid cancer, renal cancer (e.g. renal pelvis cancer), glioblastoma, endometrial cancer, cervical cancer, ovarian cancer, and testicular cancer.

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