NITROGEN-CONTAINING FUSED BICYCLIC COMPOUNDS AND THEIR USE AS UBIQUITIN-SPECIFIC-PROCESSING PROTEASE 1 (USP1) INHIBITORS

The present disclosure provides compounds having Formula (I): (I) and the pharmaceutically acceptable salts and solvates thereof, wherein X1, X2, X3, X4, X5, X6, X7, X8, R1, R2, R6, R6′, R7, and R7′ are defined as set forth in the specification. The present disclosure is also directed to the use of compounds of Formula (I) to inhibit a USP1 protein and/or to treat a disorder responsive to the inhibition of USP1 proteins and USP1 activity. Compounds of the present disclosure are especially useful for treating cancer.

REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

The content of the electronically submitted sequence listing (Name: 4195_021PC01-sequence-listing.txt; Size: 7,204 Bytes; and Date of Creation: Jun. 1, 2021) is herein incorporated by reference in its entirety.

BACKGROUND OF THE DISCLOSURE

Field of the Invention

The present disclosure provides nitrogen-containing fused bicyclic compounds as ubiquitin-specific-processing protease 1 (USP1) inhibitors, and therapeutic methods of treating conditions and diseases wherein inhibition of USP1 provides a benefit. In particular, the present disclosure provides methods of treating cancer by administering a USP1 inhibitor.

Background

Ubiquitin is a small (76 amino acid) protein that is post-transcriptionally attached to target proteins. The consequence of ubiquitination is determined by the number and linkage topology of ubiquitin molecules conjugated to the target protein. For example, proteins exhibiting lysine 48-linked poly-ubiquitin chains are generally targeted to the proteasome for degradation, while mono-ubiquitination or poly-ubiquitin chains linked through other lysines regulate non-proteolytic functions, such as cell cycle regulation, DNA damage repair, transcription, and endocytosis. Ubiquitination is a reversible process, and enzymes called deubiquitinases remove ubiquitin from target proteins.

USP1 is a deubiquitinase that plays a role in DNA damage repair. USP1 interacts with UAF1 (USP1-associated factor 1) to form a complex that is required for the deubiquitinase activity. The USP1/UAF1 complex deubiquitinates mono-ubiquitinated PCNA (proliferating cell nuclear antigen) and mono-ubiquitinated FANCD2 (Fanconi anemia group complementation group D2), which are proteins that play important functions in translesion synthesis (TLS) and the Fanconi anemia (FA) pathway, respectively. The USP1/UAF1 complex also deubiquitinates Fanconi anemia complementation group I (FANCI). These two pathways are essential for repair of DNA damage induced by DNA cross-linking agents, such as cisplatin and mitomycin C (MMC).

Safe and effective treatments targeting deubiquitinases are unknown, not yet commercially available, or have not yet been clinically developed.

BRIEF SUMMARY OF THE DISCLOSURE

In one aspect, the present disclosure relates to compounds, or a pharmaceutically acceptable salt or solvate thereof, having Formula I (also referred to herein as

Compounds of the Disclosure):

X1is selected from N and CR5;

X2is selected from N and CH;

each of X5, X6, X7, and X8are independently selected from N and CR8;

R3and R3′are taken together with the atom to which they are attached to form a carbonyl or an optionally substituted cycloalkyl;

R7and R7′are taken together with the atom to which they are attached to form a carbonyl or an optionally substituted cycloalkyl; and

R8and R4or R4′are taken together with the atoms to which they are attached to form an optionally substituted cycloalkyl.

R3and R3′are taken together with the atom to which they are attached to form a carbonyl or an optionally substituted (C3-C8) cycloalkyl.

R7and R7′are taken together with the atom to which they are attached to form a carbonyl or an optionally substituted (C3-C8) cycloalkyl.

R8and R4or R4′are taken together with the atoms to which they are attached to form an optionally substituted (C3-C8) cycloalkyl.

In some embodiments,

is independently selected from

In some embodiments,

is independently selected from

In some embodiments, one of X5, X6, X7, and X8is N.

In some embodiments, two of X5, X6, X7, and X8are N.

In some embodiments, R8is selected from hydrogen and fluoro.

In some embodiments, two of the optional substituents on R1are taken together with the carbon or nitrogen atoms to which they are attached to form an optionally substituted cycloalkyl, optionally substituted heterocyclo, optionally substituted aryl, or optionally substituted heteroaryl group.

In some embodiments, R1is an optionally substituted 5- or 6-membered nitrogen-containing heteroaryl, wherein nitrogen is the only heteroatom.

In some embodiments, R1is an optionally substituted pyrimidin-5-yl.

In some embodiments, R1is an optionally substituted pyrimidin-5-yl, and wherein the pyrimidin-5-yl is optionally substituted at the 4-position, optionally substituted at the 6-position, optionally disubstituted at the 4- and 6-positions, or optionally trisubstituted at the 2-, 4-, and 6-positions.

In some embodiments, R1is an optionally substituted pyrazol-5-yl.

In some embodiments, R1is an optionally substituted pyrazol-5-yl, wherein the pyrazol-5-yl is optionally substituted at the 1-position, optionally substituted at the 4-position, or optionally disubstituted at the 1- and 4-positions.

In some embodiments, R1is an optionally substituted pyrid-3-yl or optionally substituted pyrid-4-yl.

In some embodiments, R1is an optionally substituted pyrid-3-yl or optionally substituted pyrid-4-yl, wherein the pyrid-3-yl is optionally substituted at the 2-position, optionally substituted at the 4-position, or optionally disubstituted at the 2- and 4-positions; and wherein the pyrid-4-yl is optionally substituted at the 3-position, optionally substituted at the 5-position, or optionally disubstituted at the 3- and 5-positions.

In some embodiments, R1is an optionally substituted phenyl.

In some embodiments, R1is an optionally substituted phenyl, wherein the phenyl is optionally substituted at the 2-position, optionally substituted at the 6-position, optionally disubstituted at the 2- and 6-positions, or optionally disubstituted at the 2- and 3-positions.

In some embodiments, R1is independently selected from:

In some embodiments, two of the optional substituents on R2are taken together with the carbon or nitrogen atoms to which they are attached to form an optionally substituted cycloalkyl, optionally substituted heterocyclo, optionally substituted aryl, or optionally substituted heteroaryl group.

In some embodiments, R2is an optionally substituted 5- or 6-membered nitrogen-containing heteroaryl, wherein nitrogen is the only heteroatom.

In some embodiments, R2is an optionally substituted imidazolyl.

In some embodiments, R2is an optionally substituted pyridyl.

In some embodiments, R2is an optionally substituted pyrazolyl.

In some embodiments, R2is an optionally substituted pyridazinyl.

In some embodiments, R2is an optionally substituted pyrimidinyl.

In some embodiments, R2is an optionally substituted triazinyl.

In some embodiments, R2is an optionally substituted pyrazinyl.

In some embodiments, R2is an optionally substituted triazolyl.

In some embodiments, R2is independently selected from:

In some embodiments,

is independently selected from

In some embodiments, the compound has Formula II, Formula III, Formula IV, Formula V, Formula VI, or Formula VII:

R1is selected from

R2is selected from

X9is selected from N and CH;

X10is selected from N and CH;

X11is selected from N and CR11;

X12is selected from N and CR12;

X13is selected from N and CR13;

X14is selected from N and CR14;

X15is selected from N and CR15;

X16is selected from N and CR16;

X17is selected from N and CR17;

X18is selected from N and CR18;

X19is selected from N and CR19;

X20is selected from N and CR20,

wherein two of R11, R12, R13, R14, and R15or two of R16, R17, R18, R19, and R20are taken together with the atom to which they are attached to form an optionally substituted cycloalkyl, optionally substituted heterocyclo, optionally substituted heteroaryl, or optionally substituted aryl.

In some embodiments, the Compound of the Disclosure has Formula VIII, or Formula IX

R4and R4′are independently selected from hydrogen and methyl; and

R8is selected from hydrogen and fluoro.

In some embodiments, the Compound of the Disclosure has Formula X, or Formula XI

R4and R4′are independently selected from hydrogen and methyl; and

R8is selected from hydrogen and fluoro.

In some embodiments, the Compound of the Disclosure has Formula XII, or Formula XIII

R4and R4′are independently selected from hydrogen and methyl; and

R8is selected from hydrogen and fluoro.

In some embodiments, the Compound of the Disclosure has Formula XIV, or Formula XV

R4and R4′are independently selected from hydrogen and methyl; and

R8is selected from hydrogen and fluoro.

In some embodiments, certain Compounds of the Disclosure exhibit favorable solubility, e.g., as measured by an ADME solubility assay as disclosed herein, compared to compounds disclosed in the art as USP1 inhibitors.

In some embodiments, certain Compounds of the Disclosure exhibit favorable metabolic stability, e.g., as measured by liver microsome and hepatocyte metabolic stability assays as disclosed herein, compared to compounds disclosed in the art as USP1 inhibitors.

In other embodiments, certain Compounds of the Disclosure exhibit favorable duration of action and oral exposure in vivo.

In some embodiments, the Compound of the Disclosure is one of the specific compounds listed in the detailed description, or a pharmaceutically acceptable salt or solvate thereof.

In some embodiments, the Compound of the Disclosure inhibits a USP1 protein.

In some embodiments, the Compound of the Disclosure inhibits a USP1 protein with an IC50value of less than about 1 μM in a Ub-Rho deubiquitinating assay.

In some embodiments, the Ub-Rho deubiquitinating assay is the assay disclosed in Example 394.

In one aspect, the present disclosure relates to a method of treating cancer in a patient comprising administering to the patient a therapeutically effective amount of a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof.

In one aspect, the present disclosure relates to a pharmaceutical composition comprising a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure relates to a pharmaceutical composition for use in treatment of cancer.

In some embodiments, the present disclosure relates to a Compound of the Disclosure for use in treatment of cancer.

In some embodiments, the present disclosure relates to a use of a Compound of the Disclosure for the manufacture of a medicament for treatment of cancer.

In one aspect, the present disclosure relates to a kit comprising a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising a Compound of the Disclosure, and instructions for administering the compound, or a pharmaceutically acceptable salt or solvate thereof, or the pharmaceutical composition to a patient having cancer.

In one aspect, the present disclosure relates to a method of treating cancer in a patient comprising administering to the patient a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising a Compound of the Disclosure.

In some embodiments, the cancer is selected from the group consisting of a hematological cancer, a lymphatic cancer, and a DNA damage repair pathway deficient cancer, and a homologous-recombination deficient cancer.

In some embodiments, the cancer comprises cancer cells with a mutation in a gene encoding p53. In some embodiments, the mutation in a gene encoding p53 is a germline mutation. In some embodiments, the mutation in a gene encoding p53 is a somatic mutation. In some embodiments, the cancer comprises cancer cells with a loss of function mutation in a gene encoding p53.

In some embodiments, the cancer is selected from the group consisting of lung cancer, non-small cell lung cancer (NSCLC), colon cancer, bladder cancer, osteosarcoma, ovarian cancer, and breast cancer.

In some embodiments, the cancer is non-small cell lung cancer (NSCLC).

In some embodiments, the cancer is colon cancer.

In some embodiments, the cancer is bladder cancer.

In some embodiments, the cancer is ovarian cancer or breast cancer.

In some embodiments, the cancer is ovarian cancer.

In some embodiments, the cancer is breast cancer.

In some embodiments, the cancer is triple negative breast cancer.

In some embodiments, the cancer comprises cancer cells with elevated levels of RAD18.

In some embodiments, the elevated levels of RAD18 are elevated RAD18 protein levels.

In some embodiments, the elevated levels of RAD18 are elevated RAD18 mRNA levels.

In some embodiments, the elevated levels of RAD18 have been detected prior to the administration.

In another aspect, the present disclosure relates to a method that further comprises detecting RAD18 levels in a cancer sample obtained from the subject.

In some embodiments, the cancer is selected from the group consisting of bone cancer, including osteosarcoma and chondrosarcoma; brain cancer, including glioma, glioblastoma, astrocytoma, medulloblastoma, and meningioma; soft tissue cancer, including rhabdoid and sarcoma; kidney cancer; bladder cancer; skin cancer, including melanoma; and lung cancer, including non-small cell lung cancer; colon cancer, uterine cancer; nervous system cancer; head and neck cancer; pancreatic cancer; and cervical cancer.

In some embodiments, the cancer is a DNA damage repair pathway deficient cancer.

In some embodiments, the cancer comprises cancer cells with a mutation in a gene encoding p53. In some embodiments, the mutation in a gene encoding p53 is a germline mutation. In some embodiments, the mutation in a gene encoding p53 is a somatic mutation. In some embodiments, the cancer comprises cancer cells with a loss of function mutation in a gene encoding p53.

In some embodiments, the cancer is a BRCA1 mutant cancer. In some embodiments, the BRCA1 mutation is a germline mutation. In some embodiments, the BRCA1 mutation is a somatic mutation. In some embodiments, the BRCA1 mutation leads to BRCA1 deficiency.

In some embodiments, the cancer is a BRCA2 mutant cancer. In some embodiments, the BRCA2 mutation is a germline mutation. In some embodiments, the BRCA2 mutation is a somatic mutation. In some embodiments, the BRCA2 mutation leads to BRCA2 deficiency.

In some embodiments, the cancer is a BRCA1 mutant cancer and a BRCA2 mutant cancer.

In some embodiments, the cancer is a BRCA1 deficient cancer.

In some embodiments, the cancer is a BRCA2 deficient cancer.

In some embodiments, the cancer is a BRCA1 deficient cancer and a BRCA2 deficient cancer.

In some embodiments, the BRCA1 or BRCA2 mutant cancer is a BRCA1 or BRCA2 deficient cancer.

In some embodiments, the cancer is a Poly (ADP-ribose) polymerase (“PARP”) inhibitor refractory or resistant cancer. In some embodiments, the cancer is a PARP inhibitor resistant or refractory BRCA1, BRCA2, or BRCA1 and BRCA2 mutant cancer. In some embodiments, the cancer is a PARP inhibitor resistant or refractory BRCA1, BRCA2, or BRCA1 and BRCA2-deficient cancer.

In some embodiments, the cancer has a mutation in the gene encoding ataxia telangiectasia mutated (ATM) protein kinase. In some embodiments the ATM mutation is a germline mutation. In some embodiments the ATM mutation is a somatic mutation. In some embodiments the cancer is an ATM-deficient cancer.

In some embodiments the cancer has a mutation in the gene encoding at least two of p53, BRCA1, BRCA2, and ATM.

In another aspect, the present disclosure relates to a method of treating a USP1 protein mediated disorder comprising administering to a patient in need thereof a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising a Compound of the Disclosure, in an effective amount to treat the USP1 protein mediated disorder.

In some embodiments, the USP1 protein comprises the amino acid sequence of SEQ ID NO:1.

In another aspect, the present disclosure relates to a method of inhibiting a USP1 protein comprising contacting a USP1 protein with a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition comprising a Compound of the Disclosure.

In some embodiments, the contacting occurs in vitro.

In some embodiments, the contacting occurs in vivo.

In some embodiments, the USP1 protein comprises the amino acid sequence

DETAILED DESCRIPTION OF THE DISCLOSURE

One aspect of the present disclosure is based on the use of Compounds of the Disclosure as inhibitors of a ubiquitin-specific-processing protease 1 (USP1) protein. In view of this property, the Compounds of the Disclosure are useful for inhibiting a USP1 protein and for treating diseases, disorders, or conditions, e.g., cancer, that are responsive to inhibition of a USP1 protein.

In some embodiments, certain Compounds of the Disclosure exhibit improved solubility, e.g., as measured by an ADME solubility assay as disclosed herein.

In some embodiments, certain Compounds of the Disclosure exhibit improved metabolic stability, e.g., as measured by liver microsome metabolic stability assays as disclosed herein.

In other embodiments, certain Compounds of the Disclosure exhibit improved duration of action and oral exposure in vivo.

In one embodiment, Compounds of the Disclosure are compounds having Formula I:

X1is selected from N and CR5;

X2is selected from N and CH;

each of X5, X6, X7, and X8are independently selected from N and CR8;

R3and R3′are taken together with the atom to which they are attached to form a carbonyl or an optionally substituted cycloalkyl;

R7and R7′are taken together with the atom to which they are attached to form a carbonyl or an optionally substituted cycloalkyl; and

R8and R4or R4′are taken together with the atoms to which they are attached to form an optionally substituted cycloalkyl.

R3and R3′are taken together with the atom to which they are attached to form a carbonyl or an optionally substituted (C3-C8) cycloalkyl.

R7and R7′are taken together with the atom to which they are attached to form a carbonyl or an optionally substituted (C3-C8) cycloalkyl.

R8and R4or R4′are taken together with the atoms to which they are attached to form an optionally substituted (C3-C8) cycloalkyl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I. wherein

is independently selected from

In another embodiment,

In another embodiment,

In another embodiment,

In another embodiment,

In another embodiment,

In another embodiment,

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein

is independently selected from

In another embodiment,

In another embodiment,

In another embodiment,

In another embodiment,

In another embodiment,

In another embodiment,

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein X2is CH.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein X2is N.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein X3is NR3.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein X3is O.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein X3is CR3R3′.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein one of X5, X6, X7, and X8is N. In another embodiment, X5is N. In another embodiment, X6is N. In another embodiment, X7is N. In another embodiment, X8is N.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein two of X5, X6, X7, and X8is N. In another embodiment, X5and X6are N. In another embodiment, X5and X7are N. In another embodiment, X6and X8are N. In another embodiment, X6and X7are N.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein X6is CR8.

In another embodiment, a Compound of the Disclosure is a compound having Formula I, wherein two of the optional substituents on R1are taken together with the carbon or nitrogen atoms to which they are attached to form an optionally substituted cycloalkyl, optionally substituted heterocyclo, optionally substituted aryl, or optionally substituted heteroaryl group.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein the optional substituents on R1are independently selected from hydrogen, halo, nitro, cyano, hydroxy, and amino.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R1is an optionally substituted 5- or 6-membered nitrogen-containing heteroaryl, wherein nitrogen is the only heteroatom.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R1is an optionally substituted pyrimidin-5-yl. In another embodiment, the pyrimidin-5-yl is optionally substituted at the 4-position. In another embodiment, the pyrimidin-5-yl is optionally substituted at the 6-position. In another embodiment, the pyrimidin-5-yl is optionally disubstituted at the 4- and 6-positions. In another embodiment, the pyrimidin-5-yl is optionally trisubstituted at the 2-, 4-, and 6-positions.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R1is an optionally substituted pyrazol-5-yl. In another embodiment, the pyrazol-5-yl is optionally substituted at the 1-position. In another embodiment, the pyrazol-5-yl is optionally substituted at the 4-position. In another embodiment, the pyrazol-5-yl is optionally disubstituted at the 1- and 4-positions.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R1is an optionally substituted pyrid-3-yl. In another embodiment, the pyrid-3-yl is optionally substituted at the 2-position. In another embodiment, the pyrid-3-yl is optionally substituted at the 4-position. In another embodiment, the pyrid-3-yl is optionally disubstituted at the 2- and 4-positions.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R1is and optionally substituted pyrid-4-yl. In another embodiment, the pyrid-4-yl is optionally substituted at the 3-position. In another embodiment, the pyrid-4-yl is optionally substituted at the 5-position. In another embodiment, the pyrid-4-yl is optionally disubstituted at the 3- and 5-positions.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R1is an optionally substituted phenyl. In another embodiment, the phenyl is optionally substituted at the 2-position. In another embodiment, the phenyl is optionally substituted at the 6-position. In another embodiment, the phenyl is optionally disubstituted at the 2- and 6-positions. In another embodiment, the phenyl is optionally disubstituted at the 2- and 3-positions.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R1is selected from optionally substituted C6aryl and optionally substituted 5- or 6-membered nitrogen-containing heteroaryl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R1is independently selected from:

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein two of the optional substituents on R2are taken together with the carbon or nitrogen atoms to which they are attached to form an optionally substituted cycloalkyl, optionally substituted heterocyclo, optionally substituted aryl, or optionally substituted heteroaryl group.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein the optional substituents on R2are independently selected from hydrogen, halo, nitro, cyano, hydroxy, and amino.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is an optionally substituted 5- or 6-membered nitrogen-containing heteroaryl. In another embodiment, R2is an optionally substituted 5- or 6-membered nitrogen-containing heteroaryl, wherein nitrogen is the only heteroatom.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is an optionally substituted imidazolyl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is an optionally substituted pyridyl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is an optionally substituted pyrazolyl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is an optionally substituted pyridazinyl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is an optionally substituted pyrimidinyl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is an optionally substituted triazinyl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is an optionally substituted pyrazinyl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is an optionally substituted triazolyl.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R2is independently selected from:

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein

is independently selected from

In one embodiment, a Compound of the Disclosure is a compound having Formula II, Formula III, Formula IV, Formula V, Formula VI, or Formula VII:

R1is selected from

R2is selected from

X9is selected from N and CH;

X10is selected from N and CH;

X11is selected from N and CR″;

X12is selected from N and CR12;

X13is selected from N and CR13;

X14is selected from N and CR14;

X15is selected from N and CR15;

X16is selected from N and CR16;

X17is selected from N and CR17;

X18is selected from N and CRIB;

X19is selected from N and CR19;

X20is selected from N and CR20,

wherein two of R11, R12, R13, R14, and R15or two of R16, R17, R18, R19, and R20are taken together with the atom to which they are attached to form an optionally substituted cycloalkyl, optionally substituted heterocyclo, optionally substituted heteroaryl, or optionally substituted aryl.

In another embodiment, a Compound of the Disclosure is a compound having Formula II:

In another embodiment, a Compound of the Disclosure is a compound having Formula III:

In another embodiment, a Compound of the Disclosure is a compound having Formula IV:

In another embodiment, a Compound of the Disclosure is a compound having Formula V:

In another embodiment, a Compound of the Disclosure is a compound having Formula VI:

In another embodiment, a Compound of the Disclosure is a compound having Formula VII:

In another embodiment, a Compound of the Disclosure is a compound having Formula II, Formula III, Formula IV, Formula V, Formula VI, or Formula VII, wherein R1is

In another embodiment, a Compound of the Disclosure is a compound having Formula II, Formula III, Formula IV, Formula V, Formula VI, or Formula VII, wherein R1is

In another embodiment, a Compound of the Disclosure is a compound having Formula II, Formula III, Formula IV, Formula V, Formula VI, or Formula VII, wherein R2is

In another embodiment, a Compound of the Disclosure is a compound having Formula II, Formula III, Formula IV, Formula V, Formula VI, or Formula VII, wherein R2is

In one embodiment, a Compound of the Disclosure is a compound having Formula VIII, or Formula IX

wherein R4and R4′are independently selected from hydrogen and methyl; and R8is selected from hydrogen and fluoro.

In another embodiment, a Compound of the Disclosure is a compound having Formula VIII,

In another embodiment, a Compound of the Disclosure is a compound having Formula IX,

In one embodiment, a Compound of the Disclosure is a compound having Formula X, or Formula XI,

wherein R4and R4′are independently selected from hydrogen and methyl; and R8is selected from hydrogen and fluoro.

In another embodiment, a Compound of the Disclosure is a compound having Formula X,

In another embodiment, a Compound of the Disclosure is a compound having Formula XI,

In one embodiment, a Compound of the Disclosure is a compound having Formula XII, or Formula XIII

wherein R4and R4′are independently selected from hydrogen and methyl; and R8is selected from hydrogen and fluoro.

In another embodiment, a Compound of the Disclosure is a compound having Formula XII,

In another embodiment, a Compound of the Disclosure is a compound having Formula XIII,

In one embodiment, a Compound of the Disclosure is a compound having Formula XIV, or Formula XV

wherein R4and R4′are independently selected from hydrogen and methyl; and R8is selected from hydrogen and fluoro.

In another embodiment, a Compound of the Disclosure is a compound having Formula XIV,

In another embodiment, a Compound of the Disclosure is a compound having Formula XV,

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R8is selected from hydrogen and fluoro. In another embodiment, R8is hydrogen. In another embodiment, R8is fluoro.

In one embodiment, a Compound of the Disclosure is a compound having Formula I, wherein R8is selected from cyano, chloro, methoxy, methoxyethoxy, hydroxyethoxy, and difluoromethoxy.

In one embodiment, Compounds of the Disclosure are compounds selected from the group consisting of:

2-(4-cyclopropyl-6-(difluoromethoxy)pyrimidin-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-methyl-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one; and

(4S)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(4-(1-(1-fluoropropan-2-yl)-4-(trifluoromethyl)-1H-imidazol-2-yl)phenoxy)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridine, or a pharmaceutically acceptable salt or solvate, e.g., hydrate, of any of the above.

Definitions

For the purpose of the present disclosure, the term “alkyl” as used by itself or as part of another group refers to a straight- or branched-chain aliphatic hydrocarbon containing one to twelve carbon atoms (i.e., C1-12alkyl) or the number of carbon atoms designated (i.e., a Ci alkyl such as methyl, a C2alkyl such as ethyl, a C3alkyl such as propyl or isopropyl, etc.). The alkyl group can be suitably chosen from a straight chain C1-10alkyl group, a branched chain C3-10alkyl group, a straight chain C1-6alkyl group, a branched chain C3-6alkyl group, a straight chain C1-4alkyl group, a branched chain C3-4alkyl group, a straight or branched chain C3-4alkyl group. The alkyl group can be partially or completely deuterated, i.e., one or more hydrogen atoms of the alkyl group are replaced with deuterium atoms. Non-limiting exemplary C1-10alkyl groups include methyl (including —CD3), ethyl, propyl, isopropyl, butyl, sec-butyl, tent-butyl, iso-butyl, 3-pentyl, hexyl, heptyl, octyl, nonyl, and decyl. Non-limiting exemplary C1-4alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tent-butyl, and iso-butyl.

For the purpose of the present disclosure, the term “optionally substituted alkyl” as used by itself or as part of another group means that the alkyl as defined above is either unsubstituted or substituted with one, two, or three substituents independently chosen from halo, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyl, aralkyloxy, alkylthio, carboxamido, sulfonamido, alkylcarbonyl, arylcarbonyl, alkyl sulfonyl, aryl sulfonyl, carboxy, carboxyalkyl, alkoxyalkyl, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (cyano)alkyl, (carboxamido)alkyl, mercaptoalkyl, (heterocyclo)alkyl, or (heteroaryl)alkyl. The alkyl can be an optionally substituted C1-4alkyl. The optionally substituted alkyl can be substituted with two substituents, or one substituent. Non-limiting exemplary optionally substituted alkyl groups include —CH2CH2NO2, —CH2CH2CO2H, —CH2CH2SO2CH3, —CH2CH2COPh, and —CH2C6H11.

For the purpose of the present disclosure, the term “alkylene” or “alkylenyl” refers to a divalent alkyl radical. Any of the above mentioned monovalent alkyl groups may be an alkylene by abstraction of a second hydrogen atom from the alkyl. The alkylene group may also be a C1-C6alkylene or a C1-C4alkylene. Non-limiting exemplary alkylene groups include, —CH2—, —CH(CH3)—, —C(CH3)2—, —CH2CH2—, —CH2CH(CH3)—, —CH2C(CH3)2—, —CH2CH2CH2—, and —CH2CH2CH2CH2—.

For the purpose of the present disclosure, the term “cycloalkyl” as used by itself or as part of another group refers to saturated and partially unsaturated (containing one or two double bonds) cyclic aliphatic hydrocarbons containing one to three rings having from three to twelve carbon atoms (i.e., C3-12cycloalkyl) or the number of carbons designated. The cycloalkyl group can have two rings, or one ring. The cycloalkyl group can be chosen from a C3-8cycloalkyl group and a C3-6cycloalkyl group. The cycloalkyl group can contain one or more carbon-to-carbon double bonds or one carbon-to-carbon double bond. Non-limiting exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, decalin, adamantyl, cyclohexenyl, and spiro[3.3]heptane.

For the purpose of the present disclosure, the term “optionally substituted cycloalkyl” as used by itself or as part of another group means that the cycloalkyl as defined above is either unsubstituted or substituted with one, two, or three substituents independently chosen from halo, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyl, aralkyloxy, alkylthio, carboxamido, sulfonamido, alkylcarbonyl, arylcarbonyl, alkyl sulfonyl, aryl sulfonyl, carboxy, carboxyalkyl, alkyl, optionally substituted cycloalkyl, alkenyl, alkynyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted heterocyclo, alkoxyalkyl, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (cyano)alkyl, (carboxamido)alkyl, mercaptoalkyl, (heterocyclo)alkyl, or (heteroaryl)alkyl. The optionally substituted cycloalkyl can be substituted with two substituents or one substituent.

For the purpose of the present disclosure, the term “cycloalkyloxy” as used by itself or as part of another group refers to a cycloalkyl group attached to a terminal oxygen atom. A non-limiting exemplary of a cycloalkyloxy group is:

For the purpose of the present disclosure, the term “alkenyl” as used by itself or as part of another group refers to an alkyl group as defined above containing one, two or three carbon-to-carbon double bonds. The alkenyl group can be chosen from a C2-6alkenyl group and a C2-4alkenyl group. Non-limiting exemplary alkenyl groups include ethenyl, propenyl, isopropenyl, butenyl, sec-butenyl, pentenyl, and hexenyl.

For the purpose of the present disclosure, the term “optionally substituted alkenyl” as used herein by itself or as part of another group means the alkenyl as defined above is either unsubstituted or substituted with one, two or three substituents independently chosen from halo, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido, sulfonamido, alkylcarbonyl, arylcarbonyl, alkyl sulfonyl, aryl sulfonyl, carboxy, carboxyalkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclo.

For the purpose of the present disclosure, the term “alkynyl” as used by itself or as part of another group refers to an alkyl group as defined above containing one to three carbon-to-carbon triple bonds. The alkynyl can have one carbon-to-carbon triple bond. The alkynyl group can be chosen from a C2-6alkynyl group and a C2-4alkynyl group. Non-limiting exemplary alkynyl groups include ethynyl, propynyl, butynyl, 2-butynyl, pentynyl, and hexynyl groups.

For the purpose of the present disclosure, the term “optionally substituted alkynyl” as used herein by itself or as part of another group means the alkynyl as defined above is either unsubstituted or substituted with one, two or three substituents independently chosen from halo, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyloxy, alkylthio, carboxamido, sulfonamido, alkylcarbonyl, arylcarbonyl, alkyl sulfonyl, aryl sulfonyl, carboxy, carboxyalkyl, cycloalkyl, aryl, heteroaryl, or heterocyclo.

For the purpose of the present disclosure, the term “haloalkyl” as used by itself or as part of another group refers to an alkyl group substituted by one or more fluorine, chlorine, bromine and/or iodine atoms. The alkyl group can be substituted by one, two, or three fluorine and/or chlorine atoms. The haloalkyl group can be chosen from a C1-4haloalkyl group. Non-limiting exemplary haloalkyl groups include fluoromethyl, difluoromethyl, trifluoromethyl, pentafluoroethyl, 1,1-difluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, 3,3,3-trifluoropropyl, 4,4,4-trifluorobutyl, and trichloromethyl groups.

For the purpose of the present disclosure, the term “hydroxyalkyl” as used by itself or as part of another group refers to an alkyl group substituted with one or more, e.g., one, two, or three, hydroxy groups. The hydroxyalkyl group can be chosen from a monohydroxyalkyl group, i.e., substituted with one hydroxy group, a dihydroxyalkyl group, i.e., substituted with two hydroxy groups, and a C1-4hydroxyalkyl group. Non-limiting exemplary hydroxyalkyl groups include hydroxymethyl, hydroxyethyl, hydroxypropyl and hydroxybutyl groups, such as 1-hydroxyethyl, 2-hydroxyethyl, 1,2-dihydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 3-hydroxybutyl, 4-hydroxybutyl, 2-hydroxy-1-methylpropyl, and 1,3-dihydroxyprop-2-yl.

For the purpose of the present disclosure, the term “alkoxy” as used by itself or as part of another group refers to an optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclo, optionally substituted alkenyl or optionally substituted alkynyl attached to a terminal oxygen atom. The alkoxy group can be chosen from a C1-4alkoxy group and a C1-4alkyl attached to a terminal oxygen atom, e.g., methoxy, ethoxy, and tert-butoxy.

For the purpose of the present disclosure, the term “alkylthio” as used by itself or as part of another group refers to a sulfur atom substituted by an optionally substituted alkyl group. The alkylthio group can be chosen from a C1-4alkylthio group. Non-limiting exemplary alkylthio groups include —SCH3(i.e., methylthio), and —SCH2CH3.

For the purpose of the present disclosure, the term “alkoxyalkyl” as used by itself or as part of another group refers to an alkyl group substituted with an alkoxy group. Non-limiting exemplary alkoxyalkyl groups include methoxymethyl, methoxyethyl, methoxypropyl, methoxybutyl, ethoxymethyl, ethoxyethyl, ethoxypropyl, ethoxybutyl, propoxymethyl, iso-propoxymethyl, propoxyethyl, propoxypropyl, butoxymethyl, tert-butoxymethyl, isobutoxymethyl, sec-butoxymethyl, and pentyloxymethyl.

For the purpose of the present disclosure, the term “halo” as used by itself or as part of another group refers to a halogen atom. Non-limiting exemplary halo groups include fluoro, chloro, bromo, and iodo.

For the purpose of the present disclosure, the term “haloalkoxy” as used by itself or as part of another group refers to a haloalkyl attached to a terminal oxygen atom. Non-limiting exemplary haloalkoxy groups include fluoromethoxy, difluoromethoxy, trifluoromethoxy, and 2,2,2-trifluoroethoxy.

For the purpose of the present disclosure, the term “heteroalkyl” as used by itself or part of another group refers to a stable straight or branched chain hydrocarbon radical containing 1 to 10 carbon atoms and at least two heteroatoms, which can be the same or different, selected from O, N, or S, wherein: 1) the nitrogen atom(s) and sulfur atom(s) can optionally be oxidized; and/or 2) the nitrogen atom(s) can optionally be quaternized. The heteroatoms can be placed at any interior position of the heteroalkyl group or at a position at which the heteroalkyl group is attached to the remainder of the molecule. The heteroalkyl group can contain two oxygen atoms, one oxygen and one nitrogen atom, or two nitrogen atoms. Non-limiting exemplary heteroalkyl groups include —CH2OCH—2CH2OCH3, —OCH2CH2OCH2CH2OCH3, —CH2NHCH2CH2OCH2, —OCH2CH2NH2, —NHCH2CH2N(H)CH3, —NHCH2CH2OCH3and —OCH2CH2OCH3.

For the purpose of the present disclosure, the term “aryl” as used by itself or as part of another group refers to a monocyclic or bicyclic aromatic ring system having from six to fourteen carbon atoms (i.e., C6-14aryl). The aryl group can be chosen from a C6-14aryl group and a C6-10aryl group. Non-limiting exemplary aryl groups include phenyl (abbreviated as “Ph”), naphthyl, phenanthryl, anthracyl, indenyl, azulenyl, biphenyl, biphenylenyl, and fluorenyl groups. The aryl group can be chosen from phenyl or naphthyl. The aryl group can be phenyl.

For the purpose of the present disclosure, the term “aryloxy” as used by itself or as part of another group refers to an optionally substituted aryl attached to a terminal oxygen atom. A non-limiting exemplary aryloxy group is PhO—.

For the purpose of the present disclosure, the term “heteroaryloxy” as used by itself or as part of another group refers to an optionally substituted heteroaryl attached to a terminal oxygen atom.

For the purpose of the present disclosure, the term “aralkyloxy” or “arylalkyloxy” as used by itself or as part of another group refers to an aralkyl group attached to a terminal oxygen atom. A non-limiting exemplary aralkyloxy group is PhCH2O—.

For the purpose of the present disclosure, the term “heterocycle” or “heterocyclo” as used by itself or as part of another group refers to saturated and partially unsaturated (e.g., containing one or two double bonds) cyclic groups containing one, two, or three rings having from three to fourteen ring members (i.e., a 3- to 14-membered heterocyclo) and at least one heteroatom. The heterocyclo group can be chosen from a C3-14heterocyclo group and a C3-8heterocyclo group. Each heteroatom is independently selected from the group consisting of oxygen, sulfur, including sulfoxide and sulfone, and/or nitrogen atoms, which can be quaternized. The term “heterocyclo” is meant to include cyclic ureido groups such as imidazolidinyl-2-one, cyclic amide groups such as β-lactam, γ-lactam, δ-lactam and c-lactam, and cyclic carbamate groups such as oxazolidinyl-2-one. The term “heterocyclo” is also meant to include groups having fused optionally substituted aryl groups, e.g., indolinyl, indolinyl-2-one, benzo[d]oxazolyl-2(3H)-one. The term “heterocyclo” is also meant to include groups having fused optionally substituted heteroaryl groups, e.g., 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazine. The heterocyclo group can be chosen from a 4-, 5-, 6-, 7- or 8-membered cyclic group containing one ring and one or two oxygen and/or nitrogen atoms, a 5- or 6-membered cyclic group containing one ring and one or two nitrogen atoms, an 8-, 9-, 10-, 11-, or 12-membered cyclic group containing two rings and one or two nitrogen atoms. The heterocyclo can be optionally linked to the rest of the molecule through a carbon or nitrogen atom. Non-limiting exemplary heterocyclo groups include 2-oxopyrrolidin-3-yl, 2-imidazolidinone, piperidinyl, morpholinyl, piperazinyl, pyrrolidinyl, azetidinyl, 8-azabicyclo[3.2.1]octane (nortropane), 6-azaspiro[2.5]octane, 6-azaspiro[3.4]octane, indolinyl, indolinyl-2-one, 1,3-dihydro-2H-benzo[d]imidazol-2-one.

For the purpose of the present disclosure, the term “optionally substituted heterocyclo” as used herein by itself or part of another group means the heterocyclo as defined above is either unsubstituted or substituted with one to four substituents independently selected from halo, nitro, cyano, hydroxy, amino, alkylamino, dialkylamino, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyl aralkyloxy, alkylthio, carboxamido, sulfonamido, alkylcarbonyl, arylcarbonyl, alkyl sulfonyl, aryl sulfonyl, carboxy, carboxyalkyl, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclo, alkoxyalkyl, (amino)alkyl, hydroxyalkylamino, (alkylamino)alkyl, (dialkylamino)alkyl, (cyano)alkyl, (carboxamido)alkyl, mercaptoalkyl, (heterocyclo)alkyl, and (heteroaryl)alkyl. Substitution may occur on any available carbon or nitrogen atom, and may form a spirocycle.

For the purpose of the present disclosure, the term “heterocyclooxy” as used by itself or as part of another group refers to a heterocyclo group attached to a terminal oxygen atom.

For the purpose of the present disclosure, the term “amino” as used by itself or as part of another group refers to —NH2.

For the purpose of the present disclosure, the term “alkylamino” as used by itself or as part of another group refers to —NHR21, wherein R21is C1-6alkyl. R21can be C1-4alkyl. Non-limiting exemplary alkylamino groups include —N(H)CH3and —N(H)CH2CH3.

For the purpose of the present disclosure, the term “dialkylamino” as used by itself or as part of another group refers to —NR22aR22b, wherein R22aand R22bare each independently C1-6alkyl. R22aand R22bcan each independently be C1-4alkyl. Non-limiting exemplary dialkylamino groups include —N(CH3)2and —N(CH3)CH2CH(CH3)2.

For the purpose of the present disclosure, the term “hydroxyalkylamino” as used by itself or as part of another group refers to —NHR23, wherein R23is hydroxyalkyl.

For the purpose of the present disclosure, the term “cycloalkylamino” as used by itself or as part of another group refers to —NR24aR24b, wherein R24ais optionally substituted cycloalkyl and R24bis hydrogen or C1-4alkyl.

For the purpose of the present disclosure, the term “aralkylamino” as used by itself or as part of another group refers to —NR25aR25b, wherein R25ais aralkyl and R25bis hydrogen or C1-4alkyl. Non-limiting exemplary aralkylamino groups include —N(H)CH2Ph and —N(CH3)CH2Ph.

For the purpose of the present disclosure, the term “(amino)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with an amino group. The alkyl can be a C1-4alkyl. Non-limiting exemplary (amino)alkyl groups include —CH2NH2, —C(NH2)(H)CH3, —CH2CH2NH2, —CH2C(NH2)(H)CH3, —CH2CH2CH2NH2, —CH2CH2CH2CH2NH2, and —CH2C(CH3)2CH2NH2

For the purpose of the present disclosure, the term “(alkylamino)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with an alkylamino group. The alkyl can be a C1-4alkyl. A non-limiting exemplary (alkylamino)alkyl group is —CH2CH2N(H)CH3.

For the purpose of the present disclosure, the term “(dialkylamino)alkyl” as used by itself or as part of another group refers to an alkyl group substituted by a dialkylamino group. The alkyl can be a C1-4alkyl. Non-limiting exemplary (dialkylamino)alkyl groups are —CH2CH2N(CH3)2.

For the purpose of the present disclosure, the term “(cycloalkylamino)alkyl” as used by itself or as part of another group refers to an alkyl group substituted by a cycloalkylamino group. The alkyl can be a C1-4alkyl. Non-limiting exemplary (cycloalkylamino)alkyl groups include —CH2N(H)cyclopropyl, —CH2N(H)cyclobutyl, and —CH2N(H)cyclohexyl.

For the purpose of the present disclosure, the term “(aralkylamino)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with an aralkylamino group. The alkyl can be a C1-4alkyl. A non-limiting exemplary (aralkylamino)alkyl group is —CH2CH2CH2N(H)CH2Ph.

For the purpose of the present disclosure, the term “(cyano)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with one or more cyano, e.g., —CN, groups. The alkyl can be a C1-4alkyl. Non-limiting exemplary (cyano)alkyl groups include —CH2CH2CN, —CH2CH2CH2CN, and —CH2CH2CH2CH2CN.

For the purpose of the present disclosure, the term “(amino)(hydroxy)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with one amino, alkylamino, or dialkylamino group and one hydroxy group. The alkyl is a C1-6alkyl or a C1-4alkyl.

For the purpose of the present disclosure, the term “(amino)(aryl)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with one amino, alkylamino, or dialkylamino group and one optionally substituted aryl group. The alkyl can be a C1-6alkyl. The optionally substituted aryl group can be an optionally substituted phenyl.

For the purpose of the present disclosure, the term “(cycloalkyl)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with one optionally substituted cycloalkyl group. The alkyl can be a C1-4alkyl or a C3-6cycloalkyl. The optionally substituted cycloalkyl group can be substituted with an amino or (amino)alkyl group.

For the purpose of the present disclosure, the term “(cycloalkyl)alkyloxy” as used by itself or as part of another group refers to a (cycloalkyl)alkyl group attached to a terminal oxygen atom. The alkyloxy can be a C1-4alkyloxy or a C3-6cycloalkyloxy.

For the purpose of the present disclosure, the term “(hydroxy)(aryl)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with one hydroxy group and one optionally substituted aryl group. The alkyl can be a C1-6alkyl. The optionally substituted aryl group can be an optionally substituted phenyl. Non-limiting exemplary (hydroxy)(aryl)alkyl groups include:

For the purpose of the present disclosure, the term “carboxamido” as used by itself or as part of another group refers to a radical of formula —C(═O)NR26aR26b, wherein R26aand R26bare each independently hydrogen, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl, or R26aand R26btaken together with the nitrogen to which they are attached from a 3- to 8-membered heterocyclo group. R26aand R26bcan each independently be hydrogen or optionally substituted alkyl. Non-limiting exemplary carboxamido groups include —CONH2, —CON(H)CH3, —CON(CH3)2, and —CON(H)Ph.

For the purpose of the present disclosure, the term “(carboxamido)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with a carboxamido group. Non-limiting exemplary (carboxamido)alkyl groups include —CH2CONH2, —C(H)CH3—CONH2, and —CH2CON(H)CH3.

For the purpose of the present disclosure, the term “sulfonamido” as used by itself or as part of another group refers to a radical of the formula —SO2NR27aR27b, wherein R27aand R27bare each independently hydrogen, optionally substituted alkyl, or optionally substituted aryl, or R27aand R27btaken together with the nitrogen to which they are attached from a 3- to 8-membered heterocyclo group. Non-limiting exemplary sulfonamido groups include —SO2NH2, —SO2N(H)CH3, and —SO2N(H)Ph.

For the purpose of the present disclosure, the term “alkylcarbonyl” as used by itself or as part of another group refers to a carbonyl group, i.e., —C(═O)—, substituted by an alkyl group. A non-limiting exemplary alkylcarbonyl group is —COCH3.

For the purpose of the present disclosure, the term “arylcarbonyl” as used by itself or as part of another group refers to a carbonyl group, i.e., —C(═O)—, substituted by an optionally substituted aryl group. A non-limiting exemplary arylcarbonyl group is -COPh.

For the purpose of the present disclosure, the term “alkylsulfonyl” as used by itself or as part of another group refers to a sulfonyl group, i.e., —SO2—, substituted by any of the above-mentioned optionally substituted alkyl groups. Non-limiting exemplary alkylsulfonyl groups are —SO2CH3(i.e., methylsulfonyl) and —SO2CH2CH3(i.e., ethyl sulfonyl).

For the purpose of the present disclosure, the term “arylsulfonyl” as used by itself or as part of another group refers to a sulfonyl group, i.e., —SO2—, substituted by any of the above-mentioned optionally substituted aryl groups. A non-limiting exemplary arylsulfonyl group is —SO2Ph.

For the purpose of the present disclosure, the term “mercaptoalkyl” as used by itself or as part of another group refers to any of the above-mentioned alkyl groups substituted by a —SH group.

For the purpose of the present disclosure, the term “carboxy” as used by itself or as part of another group refers to a radical of the formula —COOH.

For the purpose of the present disclosure, the term “carboxyalkyl” as used by itself or as part of another group refers to any of the above-mentioned alkyl groups substituted with a —COOH. A non-limiting exemplary carboxyalkyl group is —CH2CO2H.

For the purpose of the present disclosure, the term “alkoxycarbonyl” as used by itself or as part of another group refers to a carbonyl group, i.e., —C(═O)—, substituted by an alkoxy group. Non-limiting exemplary alkoxycarbonyl groups are —CO2Me and —CO2Et.

For the purpose of the present disclosure, the term “aralkyl” or “arylalkyl” as used by itself or as part of another group refers to an alkyl group substituted with one, two, or three optionally substituted aryl groups. The aralkyl group can be a C1-4alkyl substituted with one optionally substituted aryl group. Non-limiting exemplary aralkyl groups include benzyl, phenethyl, —CHPh2, —CH2(4-OH—Ph), and —CH(4-F—Ph)2.

For the purpose of the present disclosure, the term “(heterocyclo)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with one, two, or three optionally substituted heterocyclo groups. The (heterocyclo)alkyl can be a C1-4alkyl substituted with one optionally substituted heterocyclo group. The heterocyclo can be linked to the alkyl group through a carbon or nitrogen atom. Non-limiting exemplary (heterocyclo)alkyl groups include:

For the purpose of the present disclosure, the term “heterocycloalkyloxy” or “(heterocyclo)alkyloxy” as used by itself or as part of another group refers to a heterocycloalkyl group attached to a terminal oxygen atom. A non-limiting exemplary heterocycloalkyloxy is:

For the purpose of the present disclosure, the term “heteroaralkyl” or “(heteroaryl)alkyl” as used by itself or as part of another group refers to an alkyl group substituted with one, two, or three optionally substituted heteroaryl groups. The (heteroaryl)alkyl group can be a C1-4alkyl substituted with one optionally substituted heteroaryl group. Non-limiting exemplary (heteroaryl)alkyl groups include:

For the purpose of the present disclosure, the term “heteroaralkyloxy” or “heteroarylalkyloxy” as used by itself or as part of another group refers to a heteroaralkyl group attached to a terminal oxygen atom. A non-limiting exemplary of a heteroaralkyloxy group is:

For the purpose of the present disclosure, the term “alkylcarbonylamino” as used by itself or as part of another group refers to an alkylcarbonyl group attached to an amino. A non-limiting exemplary alkylcarbonylamino group is —NHCOCH3.

The present disclosure encompasses any of the Compounds of the Disclosure being isotopically-labelled (i.e., radiolabeled) by having one or more atoms replaced by an atom having a different atomic mass or mass number. Examples of isotopes that can be incorporated into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine and chlorine, such as2H (or deuterium (D)),3H,11C,13C,14C,15N,18O,17O,31P,32P,35S,18F, and36Cl, respectively, e.g.,3H,11C, and14C. The present disclosure also provides a composition wherein substantially all of the atoms at a position within the Compound of the Disclosure are replaced by an atom having a different atomic mass or mass number. The present disclosure also provides a composition wherein a portion of the atoms at a position within the Compound of the disclosure are replaced, i.e., the Compound of the Disclosure is enriched at a position with an atom having a different atomic mass or mass number. In one embodiment, the present disclosure provides a composition wherein a Compound of the Disclosure has from 1 to 8 hydrogens replaced with deuterium. Isotopically-labelled Compounds of the Disclosure can be prepared by methods known in the art.

Compounds of the Disclosure may contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms. The present disclosure is meant to encompass the use of all such possible forms, as well as their racemic and resolved forms and mixtures thereof. The individual enantiomers can be separated according to methods known in the art in view of the present disclosure. When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that they include both E and Z geometric isomers. All tautomers are intended to be encompassed by the present disclosure as well.

As used herein, the term “stereoisomers” is a general term for all isomers of individual molecules that differ only in the orientation of their atoms in space. It includes enantiomers and isomers of compounds with more than one chiral center that are not mirror images of one another (diastereomers).

The term “chiral center” or “asymmetric carbon atom” refers to a carbon atom to which four different groups are attached.

The terms “enantiomer” and “enantiomeric” refer to a molecule that cannot be superimposed on its mirror image and hence is optically active wherein the enantiomer rotates the plane of polarized light in one direction and its mirror image compound rotates the plane of polarized light in the opposite direction.

The term “racemic” refers to a mixture of equal parts of enantiomers and which mixture is optically inactive.

The term “absolute configuration” refers to the spatial arrangement of the atoms of a chiral molecular entity (or group) and its stereochemical description, e.g., R or S.

The stereochemical terms and conventions used in the specification are meant to be consistent with those described inPure&Appl. Chem68:2193 (1996), unless otherwise indicated.

The term “enantiomeric excess” or “ee” refers to a measure for how much of one enantiomer is present compared to the other. For a mixture of R and S enantiomers, the percent enantiomeric excess is defined as |R−S|*100, where R and S are the respective mole or weight fractions of enantiomers in a mixture such that R+S=1. With knowledge of the optical rotation of a chiral substance, the percent enantiomeric excess is defined as ([α]obs/[α]max)*100, where [α]obsis the optical rotation of the mixture of enantiomers and [α]maxis the optical rotation of the pure enantiomer. Determination of enantiomeric excess is possible using a variety of analytical techniques, including NMR spectroscopy, chiral column chromatography or optical polarimetry.

The terms “enantiomerically pure” or “enantiopure” refer to a sample of a chiral substance all of whose molecules (within the limits of detection) have the same chirality sense.

The terms “enantiomerically enriched” or “enantioenriched” refer to a sample of a chiral substance whose enantiomeric ratio is greater than 50:50. Enantiomerically enriched compounds may be enantiomerically pure.

It is understood that embodiments of the invention described herein include “consisting” and/or “consisting essentially of” embodiments. As used herein, the singular form “a,” “an,” and “the” includes plural references unless indicated otherwise. Use of the term “or” herein is not meant to imply that alternatives are mutually exclusive.

In this application, the use of “or” means “and/or” unless expressly stated or understood by one skilled in the art. In the context of a multiple dependent claim, the use of “or” refers back to more than one preceding independent or dependent claim.

The term “about,” as used herein, includes the recited number ±10%. Thus, “about 10” means 9 to 11. As is understood by one skilled in the art, reference to “about” a value or parameter herein includes (and describes) instances that are directed to that value or parameter per se. For example, description referring to “about X” includes description of “X.”

The present disclosure encompasses the preparation and use of salts of the Compounds of the Disclosure, including non-toxic pharmaceutically acceptable salts. Examples of pharmaceutically acceptable addition salts include inorganic and organic acid addition salts and basic salts. The pharmaceutically acceptable salts include, but are not limited to, metal salts such as sodium salt, potassium salt, cesium salt and the like; alkaline earth metals such as calcium salt, magnesium salt and the like; organic amine salts such as triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N′-dibenzylethylenediamine salt and the like; inorganic acid salts such as hydrochloride, hydrobromide, phosphate, sulphate and the like; organic acid salts such as citrate, lactate, tartrate, maleate, fumarate, mandelate, acetate, dichloroacetate, trifluoroacetate, oxalate, formate and the like; sulfonates such as methanesulfonate, benzenesulfonate, p-toluenesulfonate and the like; and amino acid salts such as arginate, asparginate, glutamate and the like. The term “pharmaceutically acceptable salt” as used herein, refers to any salt, e.g., obtained by reaction with an acid or a base, of a Compound of the Disclosure that is physiologically tolerated in the target patient (e.g., a mammal, e.g., a human).

Acid addition salts can be formed by mixing a solution of the particular Compound of the Disclosure with a solution of a pharmaceutically acceptable non-toxic acid such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid, phosphoric acid, oxalic acid, dichloroacetic acid, or the like. Basic salts can be formed by mixing a solution of the compound of the present disclosure with a solution of a pharmaceutically acceptable non-toxic base such as sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate and the like.

The present disclosure encompasses the preparation and use of solvates of Compounds of the Disclosure. Solvates typically do not significantly alter the physiological activity or toxicity of the compounds, and as such may function as pharmacological equivalents. The term “solvate” as used herein is a combination, physical association and/or solvation of a compound of the present disclosure with a solvent molecule such as, e.g. a disolvate, monosolvate or hemisolvate, where the ratio of solvent molecule to compound of the present disclosure is about 2:1, about 1:1 or about 1:2, respectively. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. In certain instances, the solvate can be isolated, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid. Thus, “solvate” encompasses both solution-phase and isolatable solvates. Compounds of the Disclosure can be present as solvated forms with a pharmaceutically acceptable solvent, such as water, methanol, ethanol, and the like, and it is intended that the disclosure includes both solvated and unsolvated forms of Compounds of the Disclosure. One type of solvate is a hydrate. A “hydrate” relates to a particular subgroup of solvates where the solvent molecule is water. Solvates typically can function as pharmacological equivalents. Preparation of solvates is known in the art. See, for example, M. Caira et al,J. Pharmaceut. Sci.,93(3):601-611 (2004), which describes the preparation of solvates of fluconazole with ethyl acetate and with water. Similar preparation of solvates, hemisolvates, hydrates, and the like are described by E.C. van Tonder et al.,AAPS Pharm. Sci. Tech.,5(1):Article 12 (2004), and A. L. Bingham et al.,Chem. Commun.603-604 (2001). A typical, non-limiting, process of preparing a solvate would involve dissolving a Compound of the Disclosure in a desired solvent (organic, water, or a mixture thereof) at temperatures above 20° C. to about 25° C., then cooling the solution at a rate sufficient to form crystals, and isolating the crystals by known methods, e.g., filtration. Analytical techniques such as infrared spectroscopy can be used to confirm the presence of the solvent in a crystal of the solvate.

Since Compounds of the Disclosure are inhibitors of USP1 proteins, the present disclosure provides a method for inhibiting a USP1 protein comprising contacting a USP1 protein or a composition comprising a USP1 protein with one or more Compounds of the Disclosure.

Since Compounds of the Disclosure are inhibitors of USP1 proteins, a number of diseases, conditions, or disorders mediated by USP1 proteins can be treated by employing these compounds. The present disclosure is thus directed generally to a method for treating a disease, condition, or disorder responsive to the inhibition of USP1 proteins in an animal suffering from, or at risk of suffering from, the disorder, the method comprising administering to the animal an effective amount of one or more Compounds of the Disclosure.

The present disclosure is further directed to a method of inhibiting USP1 proteins in an animal in need thereof, the method comprising administering to the animal a therapeutically effective amount of at least one Compound of the Disclosure.

As used herein, “treatment” is an approach for obtaining beneficial or desired clinical results. “Treatment” as used herein, covers any administration or application of a therapeutic for disease in a mammal, including a human. For purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, any one or more of: alleviation of one or more symptoms, diminishment of extent of disease, preventing or delaying spread (for example, metastasis) of disease, preventing or delaying recurrence of disease, delay or slowing of disease progression, amelioration of the disease state, inhibiting the disease or progression of the disease, inhibiting or slowing the disease or its progression, arresting its development, and remission (whether partial or total). Also encompassed by “treatment” is a reduction of pathological consequence of a proliferative disease. The methods provided herein contemplate any one or more of these aspects of treatment. In-line with the above, the term treatment does not require one-hundred percent removal of all aspects of the disorder.

In the context of cancer, the term “treating” includes, but is not limited to, inhibiting growth of cancer cells, inhibiting replication of cancer cells, lessening of overall tumor burden, and delaying, halting, or slowing tumor growth, progression, or metastasis.

As used herein, “delaying” means to defer, hinder, slow, retard, stabilize, suppress and/or postpone development or progression of the disease (such as cancer). This delay can be of varying lengths of time, depending on the history of the disease and/or individual being treated.

A “therapeutically effective amount” of a substance can vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the substance to elicit a desired response in the individual. A therapeutically effective amount is also one in which any toxic or detrimental effects of the substance are outweighed by the therapeutically beneficial effects. A therapeutically effective amount can be delivered in one or more administrations. A therapeutically effective amount refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic effect.

The terms “administer,” “administering,” “administration,” and the like refer to methods that can be used to enable delivery of the therapeutic agent to the desired site of biological action. Administration techniques that can be employed with the agents and methods described herein are found in e.g., Goodman and Gilman,The Pharmacological Basis of Therapeutics,current ed.; Pergamon; and Remington's,Pharmaceutical Sciences(current edition), Mack Publishing Co., Easton, Pa.

The terms “pharmaceutical formulation” and “pharmaceutical composition” refer to a preparation which is in such form as to permit the biological activity of the active ingredient(s) to be effective, and which contains no additional components which are unacceptably toxic to a subject to which the formulation would be administered. Such formulations may be sterile.

A “pharmaceutically acceptable carrier” refers to a non-toxic solid, semisolid, or liquid filler, diluent, encapsulating material, formulation auxiliary, or carrier conventional in the art for use with a therapeutic agent that together comprise a “pharmaceutical composition” for administration to a subject. A pharmaceutically acceptable carrier is non-toxic to recipients at the dosages and concentrations employed and is compatible with other ingredients of the formulation. The pharmaceutically acceptable carrier is appropriate for the formulation employed.

A “sterile” formulation is aseptic or essentially free from living microorganisms and their spores.

The term “container” means any receptacle and closure therefore suitable for storing, shipping, dispensing, and/or handling a pharmaceutical product.

The term “insert” or “package insert” means information accompanying a pharmaceutical product that provides a description of how to administer the product, along with the safety and efficacy data required to allow the physician, pharmacist, and patient to make an informed decision regarding use of the product. The package insert generally is regarded as the “label” for a pharmaceutical product.

The term “disease” or “condition” or “disorder” as used herein refers to a condition where treatment is needed and/or desired and denotes disturbances and/or anomalies that as a rule are regarded as being pathological conditions or functions, and that can manifest themselves in the form of particular signs, symptoms, and/or malfunctions. As demonstrated below, Compounds of the Disclosure inhibit USP1 proteins and can be used in treating diseases and conditions such as proliferative diseases, wherein inhibition of USP1 proteins provides a benefit.

The terms “polypeptide” and “protein” are used interchangeably to refer to a polymer of amino acid residues and are not limited to a minimum length. Such polymers of amino acid residues may contain natural or non-natural amino acid residues, and include, but are not limited to, peptides, oligopeptides, dimers, trimers, and multimers of amino acid residues. Both full-length proteins and fragments thereof are encompassed by the definition. The terms also include post-expression modifications of the polypeptide, for example, glycosylation, sialylation, acetylation, phosphorylation, and the like. Furthermore, for purposes of the present disclosure, a “polypeptide” refers to a protein which includes modifications, such as deletions, additions, and substitutions (generally conservative in nature), to the native sequence, as long as the protein maintains the desired activity. These modifications may be deliberate, as through site-directed mutagenesis, or may be accidental, such as through mutations of hosts which produce the proteins or errors due to PCR amplification.

“USP1” and “ubiquitin-specific-processing protease 1” as used herein refer to any native polypeptide or USP1-encoding polynucleotide. The term “USP1” encompasses “full-length,” unprocessed USP1 polypeptide as well as any forms of USP1 that result from processing within the cell (e.g., removal of the signal peptide). The term also encompasses naturally occurring variants of USP1, e.g., those encoded by splice variants and allelic variants. The USP1 polypeptides described herein can be isolated from a variety of sources, such as from human tissue types or from another source, or prepared by recombinant or synthetic methods. Human USP1 sequences are known and include, for example, the sequences publicly available as UniProt No. O94782 (including isoforms). As used herein, the term “human USP1 protein” refers to USP1 protein comprising the amino acid sequence as set forth in SEQ ID NO:1 in U.S. provisional patent application No. 62/857,986 filed Jun. 6, 2019.

USP1 is a deubiquitinating enzyme that acts as part of a complex with UAF1. USP1's “deubiquitinase activity” includes its ability to deubiquitinate as part of the USP1-UAF1 complex.

The term “specifically binds” to a protein or domain of a protein is a term that is well understood in the art, and methods to determine such specific binding are also well known in the art. A molecule is said to exhibit “specific binding” or “preferential binding” if it reacts or associates more frequently, more rapidly, with greater duration and/or with greater affinity with a particular protein or domain of a protein than it does with alternative proteins or domains. It should be understood that a molecule that specifically or preferentially binds to a first protein or domain may or may not specifically or preferentially bind to a second protein or domain. As such, “specific binding” or “preferential binding” does not necessarily require (although it can include) exclusive binding. Generally, but not necessarily, reference to binding means preferential binding. For example, a USP1 inhibitor that specifically binds to USP1, UAF1, and/or the USP1-UAF1 complex may not bind to other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) or may bind to other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) with a reduced affinity as compared to binding to USP1.

The terms “reduction” or “reduce” or “inhibition” or “inhibit” refer to a decrease or cessation of any phenotypic characteristic or to the decrease or cessation in the incidence, degree, or likelihood of that characteristic. To “reduce” or “inhibit” is to decrease, reduce or arrest an activity, function, and/or amount as compared to a reference. In some embodiments, by “reduce” or “inhibit” is meant the ability to cause an overall decrease of 20% or greater. In some embodiments, by “reduce” or “inhibit” is meant the ability to cause an overall decrease of 50% or greater. In some embodiments, by “reduce” or “inhibit” is meant the ability to cause an overall decrease of 75%, 85%, 90%, 95%, or greater. In some embodiments, the amount noted above is inhibited or decreased over a period of time, relative to a control over the same period of time.

In some embodiments inhibiting USP1 proteins is the inhibition of one or more activities or functions of USP1 proteins. It should be appreciated that the activity or function of the one or more USP1 proteins may be inhibited in vitro or in vivo. Non-limiting examples of activities and functions of USP1 include deubiquitinase activity, and formation of a complex with UAF1 and are described herein. Examplary levels of inhibition of the activity of one or more USP1 proteins include at least 10% inhibiton, at least 20% inhibition, at least 30% inhibition, at least 40% inhibition, at least 50% inhibition, at least 60% inhibition, at least 70% inhibition, at least 80% inhibition, at least 90% inhibition, and up to 100% inhibition.

The terms “individual” or “subject” are used interchangeably herein to refer to an animal; for example, a mammal, such as a human. In some instances, methods of treating mammals, including, but not limited to, humans, rodents, simians, felines, canines, equines, bovines, porcines, ovines, caprines, mammalian laboratory animals, mammalian farm animals, mammalian sport animals, and mammalian pets, are provided. In some examples, an “individual” or “subject” refers to an individual or subject in need of treatment for a disease or disorder. In some instances, the subject to receive the treatment can be a patient, designating the fact that the subject has been identified as having a disorder of relevance to the treatment, or being at particular risk of contracting the disorder.

As used herein, the terms “cancer” and “tumor” refer to or describe the physiological condition in mammals in which a population of cells are characterized by unregulated cell growth. The terms encompass solid and hematological/lymphatic cancers. Examples of cancer include but are not limited to, DNA damage repair pathway deficient cancers. Additional examples of cancer include, but are not limited to, ovarian cancer, breast cancer (including triple negative breast cancer), non-small cell lung cancer (NSCLC), and osteosarcoma. The cancer can be BRCA1 or BRCA2 wildtype. The cancer can also be BRCA1 or BRCA2 mutant. The cancer can further be a PARP inhibitor resistant or refractory cancer, or a PARP inhibitor resistant or refractory BRCA1 or BRCA2-mutant cancer.

As used herein, the term “loss of function” mutation refers to a mutation that that results in the absence of a gene, decreased expression of a gene, or the production of a gene product (e.g. protein) having decreased activity or no activity. Loss of function mutations include for example, missense mutations, nucleotide insertions, nucleotide deletions, and gene deletions. Loss of function mutations also include dominant negative mutations. Thus, cancer cells with a loss of function mutation in a gene encoding p53 include cancer cells that contain missense mutations in a gene encoding p53 as well as cancer cells that lack a gene encoding p53.

In various embodiments, the Compounds of the Disclosure are USP1 inhibitors that reduce the level of USP1 protein and/or inhibit or reduce at least one biological activity of USP1 protein.

In some embodiments, the Compounds of the Disclosure specifically bind to USP1 protein. In some embodiments, the Compounds of the Disclosure specifically bind to USP1 protein in a USP1-UAF1 complex. In some embodiments, the Compounds of the Disclosure specifically bind to USP1 mRNA. In some embodiments, the Compounds of the Disclosure specifically bind to USP1 protein (alone or in a USP1-UAF1 complex) or USP1 mRNA. In some embodiments, the Compounds of the Disclosure specifically bind to UAF1 (alone or in a USP1-UAF1 complex) and inhibit or reduces formation or activity of the USP1-UAF1 complex.

In some embodiments, the Compounds of the Disclosure decrease the formation of the USP1-UAF1 complex. In some embodiments, the Compounds of the Disclosure decrease the activity of the USP1-UAF1 complex. In some embodiments, the Compounds of the Disclosure decrease the deubiquitinase activity of USP1. In some embodiments, the Compounds of the Disclosure increase mono-ubiquitinated PCNA. In some embodiments, the Compounds of the Disclosure increase mono-ubiquitinated FANCD2. In some embodiments, the Compounds of the Disclosure increase mono-ubiquitinated FANCI.

In some embodiments, the Compounds of the Disclosure do not bind to other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) or bind deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) with at least 5-fold, at least 10-fold, at least 20-fold, or at least 100-fold reduced affinity compared to the affinity for USP1 (i.e., the Ku of the USP1 inhibitor for other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) is at least 5-fold, at least 10-fold, at least 20-fold, or at least 100-fold higher than the KDfor USP1).

In some embodiments, the Compounds of the Disclosure inhibit USP1 deubiquitinase activity with an IC50 of less than about 50 nM, between about 50 nM and about 200 nM, between about 200 nM and about 2 pM, or greater than 2 pM, e.g., as measured using the assay disclosed in US Patent Application Publication No. 2017/0145012 or IC50 of 50 nM to 1000 nM, e.g., as measured using the assay disclosed in Liang et al.,Nat Chem Biol10: 289-304 (2014). In some embodiments, the Compounds of the Disclosure inhibit USP1 deubiquitinase activity with an IC50as measured using the assay disclosed in Chen, et al.,Chem Biol.,18(11):1390-1400 (2011). In some embodiments, the Compounds of the Disclosure do not inhibit the activity of other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) or inhibit the activity of other deubiquitinases, other USP proteins, or other UAF1 complexes (e.g., USP46-UAF1) with at least 5-fold, at least 10-fold, at least 20-fold, or at least 100-fold higher IC50 compared to the IC50 for inhibition of USP1 deubiquitinase activity.

Exemplary Assays

Any suitable assay in the art can be used to determine an activity, detect an outcome or effect, determine efficacy, etc. Certain non-limiting exemplary assays that can be used in the methods provided herein are described.

In some instances, a method of determining whether a Compound of the Disclosure inhibits USP1 deubiquitinase activity measure a change in mass upon di-ubiquitin cleavage of deubiquitinase binding. For example, ubiquitin aldehyde and ubiquitin vinyl sulfone form covalent irreversible linkages to deubiquitinases that result in observable mass changes to the deubiquitinases. Similarly, cleavage of di-ubiquitins results in an observable mass change.

In some instances, a method of determining whether a Compound of the Disclosure inhibits USP1 deubiquitinase activity involves an increase in luminescence or fluorescence upon cleavage, e.g., that can be monitored on a plate reader. Such assays can use ubiquitin linked to a flurophore through a linker linkage, such as ubiquitin-7-amino-4-methylcoumarin (Ub-AMC) or ubiquitin-Rhodamine110. Such assays can also use a di-ubiquitin containing an isopeptide linkage. Exemplary di-ubiquitins can comprise a flurophore on one ubiquitin and a quencher on the other ubiquitin such that fluorescence increases with then di-ubiquitin is cleaved. Such assays can also use enzyme coupled systems wherein ubiquitin is coupled to an enzyme that is only active in producing a fluorescence enzyme product when released from the ubiquitin.

Exemplary Deubiquitination Assay for USP1/UAF1 Activity and Inhibitor Testing.

Deubiquitinase activity can be measured using ubiquitin-rhodamine 110 as a substrate. Cleavage of the amide bond between rhodamine and the c-terminal glycine of ubiquitin yields an increase in fluorescence signal. The assay can be conducted in 20 ul total volume of assay buffer (50 mM Tris-HCl, pH 7.8, 0.5 mM EDTA, 0.01% Bovine Serum Albumin, 1 mM DTT, 0.01% Tween-20), and 0.05 nM USP1/UAF1 enzyme. Reaction can be initiated by addition of 150 nM Ubiquitin-rhodamine (Boston Biochem) substrate.

Compounds of the Disclosure can be dissolved in DMSO and tested in dose response format, beginning at 10 μM.

Compounds of the Disclosure can be added to enzyme/assay buffer mix and incubated 10 min. Substrate mix can be added, and reaction mix can be read in kinetic mode for 30 min at Ex480/Em540 and IC50response curves can be plotted. See, e.g., Chen, et al.,Chem Biol.,18(11):1390-1400 (2011).

Exemplary Deubiquitination Assay for ES2 Ub-PCNA IF Activity and Inhibitor Testing.

USP1 is a deubiquitinating enzyme that removes ubiquitin from mono-ubiquitinated Proliferating Cell Nuclear Antigen (Ub-PCNA). The levels of Ub-PCNA in the nucleus of cells can be used to assess the activity of USP1. An immunofluorescence assay can be established to monitor Ub-PCNA levels in the ovarian cancer cell line, ES2.

An assay can be performed by first plating 5000 ES2 cells per well in black 96 well plates (Corning #3904) and then incubating overnight at 37° C. and 5% CO2. Compounds, resuspended in DMSO, can be added to the cells, to a final DMSO concentration of 0.3%. Plates can be incubated at 37° C. and 5% CO2for 3 hours.

Cells can be then fixed and stained by first removing the media from each plate and fixing the cells with −20° C. methanol for 5 minutes at room temperature. Fixation plates can be washed with Tris buffered saline with tween (Boston Bioproducts #IBB-855) 5 times for 5 minutes each. Plates can be blocked for 1 hour with 50 μl of Odyssey blocking buffer (Licor #327-50000) at room temperature with rocking. Block can be removed from all wells and 50 μl of primary antibody was added to each well. Ub-PCNA ab (Cell Signaling Technology #13439) can be diluted at 1:400 in Odyssey blocking buffer. Plates can be sealed and incubated overnight at 4° C. Primary antibodies can be removed from the plates and plates can be washed with Tris buffered saline with tween 5 times for 5 minutes. Plates can be stained for 1 hour at room temperature with rocking with 50 μl of secondary antibodies diluted 1:10,000 in Odyssey blocking buffer at room temperature (anti-rabbit Alexa 488). Antibodies can be removed from the plates and plates can be washed with Tris buffered saline with tween 3 times for 5 minutes. Plates can be washed once with DAPI (Chemometec #910-3012) stain diluted 1:5000 (Stock solution of 500 μg/ml) in tris buffered saline with tween for 5 minutes. DAPI stain can be removed and plates can be washed one additional time for 5 minutes with tris buffered saline with tween. Wash can be removed from the plate and 100 μl tris buffered saline with tween can be added back to all wells, Plates can be sealed with Foil seals or black plate seals and plates can be stored at 4° C. until they can be imaged.

Imaging of the plates can be performed by first locating the nuclei using the DAPI stain. A mask can be created by drawing a circle slightly smaller than each nucleus. Ub-PCNA intensities can be measured for each individual nucleus counted. A histogram of all Ub-PCNA nuclear intensities measured in untreated wells can be generated and a 95% cutoff can be established. This 95% cutoff can be used to determine the number of Positive cells that have Ub-PCNA values higher than the 95% cutoff. The number of positive cells can be expressed as a percentage by dividing by the total number of cells in the well. This percent positive value can be used to graph all data and to determine AC50 values.

Methods of Use

In some embodiments, the Compounds of the Disclosure can be used to inhibit the activity of a USP1 protein. For example, in some embodiments, a method of inhibiting a USP1 protein comprises contacting the USP1 protein with a Compound of the Disclosure. The contacting can occur in vitro or in vivo.

In some embodiments, the Compounds of the Disclosure can be used to treat a “USP1 protein mediated disorder.” A USP1 protein mediated disorder is any pathological condition in which a USP1 protein is known to play a role. In some embodiments, a USP1 protein mediated disorder is a proliferative disease such as cancer.

Various methods of treating diseases and disorders with the Compounds of the Disclosure are provided herein. Exemplary diseases and disorders that may be treated with the Compounds of the Disclosure include, but are not limited to, cancer.

In some embodiments, methods of treating cancer with Compounds of the Disclosure are provided. Such methods comprise administering to a subject with cancer a therapeutically effective amount of a Compound of the Disclosure.

In some embodiments, the cancer to be treated with a Compound of the Disclosure is selected from a hematological cancer, a lymphatic cancer, and a DNA damage repair pathway deficient cancer. In some embodiments, the cancer to be treated with a Compound of the Disclosure is a cancer that comprises cancer cells with a mutation in a gene encoding p53. In some embodiments, the cancer to be treated with a Compound of the Disclosure is a cancer that comprises cancer cells with a loss of function mutation in a gene encoding p53. In some embodiments, the cancer to be treated with a Compound of the Disclosure is a cancer that comprises cancer cells with a mutation in a gene encoding BRCA1. In some embodiments, the cancer to be treated with a Compound of the Disclosure is a cancer that comprises cancer cells with a mutation in a gene encoding BRCA2. In some embodiments, the cancer to be treated with a Compound of the Disclosure is a cancer that comprises cancer cells with a loss of function mutation in a gene encoding ATM.

In some embodiments, the cancer to be treated with a Compound of the Disclosure is selected from non-small cell lung cancer (NSCLC), osteosarcoma, ovarian cancer, and breast cancer. In some embodiments, the cancer is ovarian cancer or breast cancer. In some embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is a triple negative breast cancer.

In some embodiments, the cancer to be treated with a Compound of the Disclosure is selected from the group consisting of bone cancer, including osteosarcoma and chondrosarcoma; brain cancer, including glioma, glioblastoma, astrocytoma, medulloblastoma, and meningioma; soft tissue cancer, including rhabdoid and sarcoma; kidney cancer; bladder cancer; skin cancer, including melanoma; and lung cancer, including non-small cell lung cancer; colon cancer, uterine cancer; nervous system cancer; head and neck cancer; pancreatic cancer; and cervical cancer.

Various methods of treating cancer with a Compound of the Disclosure are provided herein. In some embodiments, a therapeutically effective amount of Compound of the Disclosure is administered to a subject with cancer, wherein the cancer comprises cancer cells with elevated levels of RAD18. In some embodiments, the elevated levels of RAD18 are elevated RAD18 protein levels. In some embodiments, the elevated levels of RAD18 are elevated RAD18 mRNA levels. In some embodiments, elevated levels of RAD18 (e.g., RAD18 protein and/or RAD18 mRNA) have been detected (e.g., in a cancer sample obtained from the subject) prior to the administration. That is, in some embodiments, a subject's cancer has been tested for RAD18 protein or mRNA prior to beginning treatment with a USP1 inhibitor.

In some embodiments, such methods comprise (a) identifying a cancer in a subject as a USP1 inhibitor-sensitive cancer and then (b) administering a therapeutically effective amount of a Compound of the Disclosure to the subject.

In some embodiments, such methods comprise (a) detecting levels of RAD18 (e.g., RAD18 protein and/or RAD18 mRNA) in cancer cells (e.g., in a cancer sample obtained from the subject) and then (b) administering a therapeutically effective amount of a Compound of the Disclosure to a subject having a cancer comprising cells with elevated levels of RAD18.

In some embodiments, such methods comprise administering to a subject with triple negative breast cancer a therapeutically effective amount of a Compound of the Disclosure.

In some embodiments, a Compound of the Disclosure is used to treat a cancer, wherein the cancer is a homologous-recombination deficient cancer. In some embodiments, a Compound of the Disclosure is used to treat a cancer, wherein the cancer comprises cancer cells with a mutation in a gene encoding p53. In some embodiments, a Compound of the Disclosure is used to treat a cancer, wherein the cancer comprises cancer cells with a loss of function mutation in a gene encoding p53. In some embodiments, a Compound of the Disclosure is used to treat a cancer that does not have a defect in the homologous recombination pathway.

In some embodiments, a Compound of the Disclosure is used to treat a cancer, wherein the cancer is a BRCA1 mutant cancer. In some embodiments, a Compound of the Disclosure is used to treat a cancer, wherein the cancer is a BRCA2 mutant cancer. In some embodiments, a Compound of the Disclosure is used to treat a cancer, wherein the cancer is a BRCA1 mutant cancer and a BRCA2 mutant cancer. In some embodiments, the cancer is not a BRCA1 mutant cancer or a BRCA2 mutant cancer. In some embodiments, the cancer is a BRCA1 deficient cancer. In some embodiments, the cancer is a BRCA2 deficient cancer. In some embodiments, the cancer is a BRCA1 deficient cancer and a BRCA2 mutant cancer.

In some embodiments, a Compound of the Disclosure is used to treat a cancer, wherein the cancer is an ATM mutant cancer. In some embodiments, the cancer is not an ATM mutant cancer. In some embodiments, the cancer is an ATM deficient cancer.

In some embodiments, a Compound of the Disclosure is used to treat a cancer, wherein the cancer is a PARP inhibitor resistant or refractory cancer. In some embodiments, a Compound of the Disclosure is used to treat a cancer, wherein the cancer is a PARP inhibitor resistant or refractory BRCA1-deficient cancer.

In some embodiments, the cancer is a BRCA1 and/or BRCA2 mutant cancer, wherein the cancer comprises cells with elevated levels of RAD18, e.g., wherein the elevated levels of RAD18 are at least as high as the RAD18 protein and/or mRNA levels in ES2 cells or wherein the elevated levels of RAD18 are higher than the RAD18 protein and/or mRNA levels in HEP3B217 cells. In some embodiments, a triple negative breast cancer is a BRCA1 and/or BRCA2 mutant cancer.

In some instances, the cancer is a solid cancer. In some instances, the cancer is a hematological/lymphatic cancer. In some instances, the cancer is a DNA damage repair pathway deficient cancer. In some instances, the cancer is a homolgous-recombination deficient cancer. In some instances, the cancer comprises cancer cells with a mutation in a gene encoding p53. In some instances, the cancer comprises cancer cells with a loss of function mutation in a gene encoding p53. In some instances, the cancer is selected from the group consisting of non-small cell lung cancer (NSCLC), osteosarcoma, ovarian cancer, and breast cancer (including triple negative breast cancer). In some instances, the cancer is ovarian cancer or breast cancer (including triple negative breast cancer). In some instances, the cancer is ovarian cancer. In some instances, the cancer is breast cancer (including triple negative breast cancer.)

In some embodiments, a Compound of the Disclosure is used in combination with one or more additional therapeutic agents to treat cancer. It has been reported that p53 status determines PARP inhibitor sensitization (Sa et al. Genome Biology, (2019) 20:253) and that BRCA1/2 status predicts the efficacy of PARP inhibitors in the clinic (Audeh et al. Lancet (2010) 376 (9737), 245-51). As shown below, p53 mutant cancers and BRCA mutant cancers have increased sensitivity to USP1 inhibitors. Accordingly, in some embodiments, a Compound of the Disclosure is used in combination with a PARP inhibitor to treat cancer.

In some embodiments, provided herein are Compounds of the Disclosure for use as a medicament or for use in preparing a medicament, e.g., for the treatment of cancer. In some embodiments, provided herein are Compounds of the Disclosure for use in a method for the treatment of cancer.

Pharmaceutical Compositions

Compounds of the Disclosure can be administered to a mammal in the form of a raw chemical without any other components present, or Compounds of the Disclosure can also be administered to a mammal as part of a pharmaceutical composition containing the compound combined with a suitable pharmaceutically acceptable carrier (see, for example, Gennaro, Remington: The Science and Practice of Pharmacy with Facts and Comparisons: Drugfacts Plus, 20th ed. (2003); Ansel et al., Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th ed., Lippencott Williams and Wilkins (2004); Kibbe et al., Handbook of Pharmaceutical Excipients, 3rd ed., Pharmaceutical Press (2000)). Such a carrier can be selected from pharmaceutically acceptable excipients and auxiliaries. The term “pharmaceutically acceptable carrier” or “pharmaceutically acceptable vehicle” encompasses any of the standard pharmaceutical carriers, solvents, surfactants, or vehicles. Standard pharmaceutical carriers and their formulations are described in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 19th ed. 1995.

A pharmaceutical composition of the present disclosure may be prepared as liquid suspensions or solutions using a liquid, such as an oil, water, an alcohol, and combinations of these.

A pharmaceutical composition of the present disclosure may be prepared as a sterile injectable, which may be aqueous or oleaginous suspensions. These suspensions may be formulated according to techniques known in the art.

A pharmaceutical composition of the present disclosure may be orally administered in any orally acceptable dosage form including capsules, tablets, aqueous suspensions or solutions.

A pharmaceutical composition of the present disclosure may be administered in the form of suppositories for rectal administration.

A pharmaceutical composition of the present disclosure may also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Topical application for the lower intestinal tract may be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used. For topical applications, the pharmaceutical compositions may be formulated in a suitable ointment, lotion, or cream containing the active component suspended or dissolved in one or more carriers.

A pharmaceutical composition of the present disclosure may also be administered ophthalmically and formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic uses, the pharmaceutical compositions may be formulated in an ointment such as petrolatum.

The pharmaceutical compositions to be used for in vivo administration can be sterile. This is readily accomplished by filtration through, e.g., sterile filtration membranes.

Pharmaceutical compositions within the scope of the present disclosure include all compositions where a Compound of the Disclosure is combined with one or more pharmaceutically acceptable carriers. In one embodiment, the Compound of the Disclosure is present in the composition in an amount that is effective to achieve its intended therapeutic purpose.

A pharmaceutical composition of the present disclosure can be administered to any patient that may experience the beneficial effects of a Compound of the Disclosure. Foremost among such patients are mammals, e.g., humans and companion animals, although the disclosure is not intended to be so limited. In one embodiment, the patient is a human. In another embodiment, a pharmaceutical compositions of the present disclosure can be administered to a patient having PARP inhibitor resistant or refractory cancer. In another embodiment, a pharmaceutical compositions of the present disclosure can be administered to a patient having PARP inhibitor resistant or refractory BRCA1-deficient cancer. In another embodiment, a pharmaceutical compositions of the present disclosure can be administered to a patient in combination with a PARP inhibitor.

In another embodiment, the present disclosure provides kits which comprise a Compound of the Disclosure (or a composition comprising a Compound of the Disclosure) packaged in a manner that facilitates their use to practice methods of the present disclosure. In one embodiment, the kit includes a Compound of the Disclosure (or a composition comprising a Compound of the Disclosure) packaged in a container, such as a sealed bottle or vessel, with a label affixed to the container or included in the kit that describes use of the compound or composition to practice the method of the disclosure. In one embodiment, the compound or composition is packaged in a unit dosage form. The kit further can include a device suitable for administering the composition according to the intended route of administration. In some embodiments, the present disclosure provides a kit which comprise a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, and instructions for administering the compound, or a pharmaceutically acceptable salt or solvate thereof, to a patient having cancer.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula I, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula II, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula III, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula IV, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula V, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula VI, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula VII, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula VIII, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula IX, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula X, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula XI, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula XII, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula XIII, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula XIV, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a compound having Formula XV, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier, wherein the compound binds to a protein encoded by the USP1 gene.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is for use in treating cancer.

In some embodiments, the present disclosure provides a pharmaceutical composition comprising a Compound of the Disclosure, or a pharmaceutically acceptable salt or solvate thereof, and a pharmaceutically acceptable carrier, wherein the pharmaceutical composition is for the manufacture of a medicament for treatment of cancer.

EXAMPLES

General Synthetic Methods

Compounds of the Disclosure are prepared using methods known to those skilled in the art in view of this disclosure, or by the illustrative methods shown in the General Schemes below. In any of the General Schemes, suitable protecting groups can be employed in the synthesis. (See, Wuts, P. G. M.; Greene, T. W., “Greene's Protective Groups in Organic Synthesis”, 4th Ed., J. Wiley & Sons, NY, 2007).

Unless otherwise noted, all reagents were used without further purification.1H-NMR spectra were obtained in DMSO-d6 or CD3OD at room temperature on a Bruker AVANCE 400 MHz spectrometer. When more than one conformer was detected, the chemical shifts for the most abundant one is reported. Chemical shifts of1H NMR spectra were recorded in parts per million (ppm) on the 6 scale from an internal standard of residual solvent. Splitting patterns are designed as s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br, broad. LC-MS and prep-HPLC conditions are described below:

LCMS Agilent 5 Minute Method

Prepartive HPLC Generic Methods

Chiral Preparative HPLC Generic Methods

Compound Synthesis: The compounds of the invention may be prepared by methods well known to those skilled in the art, and as described in the synthetic experimental procedures shown below.

To a solution of 4-cyclopropyl-6-methoxypyrimidine (1.6 g, 10.7 mmol) in EtOH (10 mL) was added Br2(1.87 g, 11.7 mmol) at −20° C. The resulting mixture was slowly warmed to room temperature and stirred at the same temperature for 16 h. Solvent was removed under reduced pressure and the residue dissolved in EtOAc and washed with saturated Na2CO3, water and brine. The organic layer was dried over Na2SO4and concentrated to give 5-bromo-4-cyclopropyl-6-methoxypyrimidine (2.3 g).1H NMR (400 MHz, Methanol-d4) δ 8.40 (s, 1H), 4.03 (s, 3H). MS (ESI) m/z 229.0, 231.0 [M+H]+.

To a solution of 1-isopropyl-4-methyl-1H-pyrazole (3 g, 24.2 mmol) in dry THF (10 mL) at 0° C. was added n-BuLi (2.5 M in hexane, 11.6 mL, 29.0 mmol) dropwise. After addition, the solution was stirred at room temperature for 1 h before cooling to −78° C. 2-Isopropoxy-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (5.4 g, 29 mol) was added dropwise and the resulting solution was warmed to room temperature and stirred for 2 h then quenched with saturated NH4Cl solution. The mixture was diluted with EtOAc, washed with water and brine, dried with Na2SO4and concentrated under vacuum to give 1-isopropyl-4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.5 g). MS (ESI) m/z 251.2 [M+H]+.

To a solution of 1-isopropyl-4-methyl-1H-pyrazole (2 g, 16.1 mmol) in dry THF (15 mL) was added n-BuLi (2.5 M in hexane, 9.7 mL, 24.2 mmol) dropwise over 10 min at −78° C. The resulting solution was stirred at −78° C. for 40 min then triisopropyl borate (9.1 g, 48.3 mmol) was added dropwise over 5 min. The final solution was stirred at −78° C. for 30 min then allowed to warm to room temperature and quenched with saturated NH4Cl solution. The reaction mixture was directly purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to give (1-isopropyl-4-methyl-1H-pyrazol-5-yl)boronic acid (420 mg). MS (ESI) m/z 169.1 [M+H]+.

1-isopropyl-4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was prepared from 4-chloro-1H-pyrazole following step 1 and step 2 of Intermediate 2. MS (ESI) m/z 271.2 [M+H]+.

To a solution of 4-methoxy-3-nitro-2-(prop-1-en-2-yl)pyridine (2.5 g, 12.8 mmol) in MeOH (25 mL) was added 5% Pd/C (1 g). The resulting mixture was stirred vigorously under an atmosphere of H2at room temperature for 2.5 h. The mixture was filtered through celite and concentrated under vacuum to give 2-isopropyl-4-methoxypyridin amine (1.98 g). MS (ESI) m/z 167.2 [M+H]+.

To the solution of 2-isopropyl-4-methoxypyridin-3-amine in HBr aqueous solution (4M, 15 mL) at 0° C. was added NaNO2(368 mg, 5.3 mmol) in H2O (5 mL) dropwise. After addition, the solution was stirred at 0° C. for 1 h then added dropwise into a stirred solution of CuBr2in H2O (10 mL). The final mixture was then stirred at room temperature for 1 h before 4M NaOH aqueous solution was added to adjust the pH to 8-9. The resulting mixture was extracted with EtOAc, dried with Na2SO4, concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to give 3-bromo-2-isopropyl-4-methoxypyridine (630 mg). MS (ESI) m/z 230.1, 232.1 [M+H]+.

4-chloro-1-ethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was prepared from 4-chloro-1H-pyrazole following step 1 and step 2 of Intermediate 2. MS (ESI) m/z 257.1 [M+H]+.

4-chloro-1-cyclobutyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was prepared from 4-chloro-1H-pyrazole following step 1 and step 2 of Intermediate 2. MS (ESI) m/z 283.1 [M+H]+.

4-fluoro-1-isopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was prepared from 4-fluoro-1H-pyrazole following step 1 and step 2 of Intermediate 2. MS (ESI) m/z 255.2 [M+H]+.

To a stirred mixture of magnesium powder (2.5 g, 104 mmol) in dry THF (40 mL) was added drowise bromocyclobutane (9.45 g, 70 mmol) at room temperature to keep the mixture slightly boiling. After the magnesium powder was consumed, ZnCl2(14 g, 104 mmol), 4-chloro-6-methoxypyrimidine (1.5 g, 10.4 mmol) and Pd (dppf)C12(510 mg, 0.7 mmol) were added. The resulting mixture was heated at 70° C. for 12 h then quenched with water (1 mL) and filtered through celite. The filtrate was concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to give 4-cyclobutyl-6-methoxypyrimidine (1.3 g). MS (ESI) m/z 165.1 [M+H]+.

To a solution of 7-bromoindoline (800 mg, 4 mmol) in THF (8 mL) was added NaH (323 mg, 60% in mineral oil, 8 mmol) at 0° C. The mixture was stirred for 30 min then MeI (574 mg, 4 mmol) was added. The resulting mixture was stirred at room temperature for 5 h then quenched with water (20 mL) and extracted with EtOAc (20 mL×3). The combined organic phases were dried over Na2SO4and purified by silica gel chromatography (eluting with 1/30 EtOAc/PE) to afford 7-bromo-1-methylindoline (410 mg). MS (ESI) m/z 212.0, 214.0 [M+H]+.

To a solution of 7-bromo-1-methylindoline (180 mg, 0.85 mmol) and triisopropyl borate (207.5 mg, 1.1 mmol) in dry THF (3 mL) at −78° C. was added n-BuLi (2.5 M in hexane, 0.44 mL, 1.1 mmol) dropwise. The resulting solution was allowed to warm to room temperature and stirred for 4 h then quenched with saturated NH4Cl solution. Solvent was removed under vacuum and the crude was directly purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to give (1-methylindolin-7-yl)boronic acid (80 mg). MS (ESI) m/z 178.1 [M+H]+.

4-methoxy-6-(prop-1-en-2-yl)pyrimidine was prepared from 4-chloro-6-methoxypyrimidine and 4,4,5,5-tetramethyl-2-(prop-1-en-2-yl)-1,3,2-dioxaborolane according to step 1 of Intermediate 1. MS (ESI) m/z 151.1 [M+H]+.

4-chloro-1-(tetrahydrofuran-3-yl)-1H-pyrazole was prepared from 4-chloro-1H-pyrazole according to step 1 of Intermediate 2. MS (ESI) m/z 173.0 [M+H]+.

To a solution of 4-chloro-1-(tetrahydrofuran-3-yl)-1H-pyrazole (50 mg, 0.29 mmol) in dry THF (5 mL) at −78° C. was added n-BuLi (2.5 M in hexane, 0.175 mL, 0.44 mmol) dropwise. The solution was stirred at −78° C. for 30 min then a solution of I2 (111 mg, 0.44 mmol) in dry THF (2 mL) was added. The resulting solution was stirred at −78° C. for another 30 min then allowed to warm slowly to room temperature and quenched with saturated NH4Cl solution. The mixture was diluted with EtOAc, washed with water and brine, dried with Na2SO4, concentrated under vacuum and purified by silica gel chromatography (eluting with 1/7 EtOAc/PE) to afford 4-chloro-5-iodo-1-(tetrahydrofuran-3-yl)-1H-pyrazole (63 mg). MS (ESI) m/z 298.9 [M+H]+.

methyl 2-(4-chloro-1H-pyrazol-1-yl)propanoate was prepared from 4-chloro-1H-pyrazole according to step 1 of Intermediate 2. MS (ESI) m/z 189.0 [M+H]+.

To a solution of methyl 2-(4-chloro-1H-pyrazol-1-yl)propanoate(6.3 g, 33.5 mmol) in dry THF (200 mL) was added LiAlH4(1.3 g, 33.5 mmol) at 0° C. After addition, the reaction mixture was stirred at room temperature for 30 min. The reaction mixture was quenched with saturated aqueous Na2CO3, diluted with DCM (50 mL), filtered through celite and concentrated under vacuo to give 2-(4-chloro-1H-pyrazol-1-yl)propan-1-ol (3.6 g). The crude material was used in the next step without further purification. MS (ESI) m/z 161.3 [M+H]+.

To a solution of 2-(4-chloro-1H-pyrazol-1-yl)propan-1-ol (3.2 g, 20 mmol) in dry DMF (80 mL) was added NaH (60% in mineral oil, 962 mg, 24 mmol) in portions at 0° C. The reaction mixture was stirred for 30 min then iodomethane (8.5 g, 60 mmol) was added. The reaction was stirred at room temperature for 2 h then quenched with water (100 mL). The mixture was extracted with DCM (50 mL×3) and the combined organic layers dried over Na2SO4, concentrated under vacuum and purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to give 4-chloro-1-(1-methoxypropan-2-yl)-1H-pyrazole(1.8 g). MS (ESI) m/z 175.1 [M+H]+.

1-isopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was prepared from 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole according to step 1 of Intermediate 2. MS (ESI) m/z 237.2 [M+H]+.

To a solution of 1-isopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (5.61 g, 23.8 mmol) in THF (50 mL) at 0° C. was added NaOH (2.5 M aqueous solution) and H2O2(30%, 5.3 mL). The resulting solution was allowed to warm to room temperature and stirred for 3 h. The pH was adjusted to 2 with 1.0 M HCl and the resulting solution extracted with EtOAc (50 mL×3), dried over Na2SO4, concentrated under vacuum to afford 1-isopropyl-1H-pyrazol-4-ol (4.0 g). MS (ESI) m/z 127.1 [M+H]+.

To a solution of 1-isopropyl-1H-pyrazol-4-ol (1.0 g, 7.94 mmol, 1.0 eq) in MeCN (20 mL) was added TMSCHN2(2 M in hexanes, 15.8 mL, 31.8 mmol). The resulting solution was stirred at room temperature for 24 h. The reaction solution was concentrated under vacuum and purified by silica gel chromatography (eluting with 1/7 EtOAc/PE) to afford 1-isopropyl-4-methoxy-1H-pyrazole (415.9 mg). MS (ESI) m/z 141.2 [M+H]+.

5-iodo-1-isopropyl-4-methoxy-1H-pyrazole was prepared from 1-isopropyl-4-methoxy-1H-pyrazole according to step 2 of Intermediate 14. MS (ESI) m/z 267.0 [M+H]+.

A mixture of ethyl 5-bromo-1-isopropyl-1H-pyrazole-4-carboxylate (593 mg, 2.27 mmol) and NaOH (276 mg, 6.9 mmol) in EtOH (5 mL) and (5 mL) was heated at 60° C. for 1 h. EtOH was removed under reduced pressure and the pH of the aqueous solution was adjusted to 3 with 2M HCl. The mixture was extracted with EtOAc (20 mL×3), dried over Na2SO4, concentrated under vacuum to afford 5-bromo-1-isopropyl-1H-pyrazole-4-carboxylic acid (650 mg), which was used in next step without further purification. MS (ESI) m/z 233.0, 235.0 [M+H]+.

To a solution of 5-bromo-1-isopropyl-1H-pyrazole-4-carboxamide (160 mg, 0.69 mmol) in DMF (3 mL) was added POCl3(317.4 mg, 2.07 mmol) slowly at room temperature. The resulting mixture was stirred at room temperature for 2 h then quenched with saturated NH4Cl (20 mL) and extracted with EtOAc (20 mL×2). The combine organic phases were washed with brine (20 mL×2), dried over Na2SO4and concentrated under vacuum to afford 5-bromo-1-isopropyl-1H-pyrazole-4-carbonitrile (156 mg) without further purification. MS (ESI) m/z 255.0, 257.0 [M+H]+.

1-isopropyl-4-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was prepared from 1-isopropyl-4-methoxy-1H-pyrazole (prepared from step 3 of Intermediate 16) following step 2 of Intermediate 2. MS (ESI) m/z 267.2 [M+H]+.

1-methyl-4-(prop-1-en-2-yl)-1H-pyrazole was prepared from 4-iodo-1-methyl-1H-pyrazole according to step 1 of intermediate 7. MS (ESI) m/z 123.1 [M+H]+.

4-isopropyl-1-methyl-1H-pyrazole was prepared from 1-methyl-4-(prop-1-en-2-yl)-1H-pyrazole according to step 2 of intermediate 7. MS (ESI) m/z 125.1 [M+H]+.

5-iodo-4-isopropyl-1-methyl-1H-pyrazole was prepared from 4-isopropyl-1-methyl-1H-pyrazole according to step 2 of Intermediate 14. MS (ESI) m/z 251.0 [M+H]+.

To a solution of tert-butyl 2-(4-chloro-1H-pyrazol-1-yl)acetate (1.0 g, 4.63 mmol) in THF (10 mL) was added LiHMDS (1 M in THF, 4.6 mL) at −78° C. The mixture was stirred at −78° C. for 30 min then 1,3,2-dioxathiolane 2,2-dioxide (574 mg, 4.63 mmol) was added. The reaction was allowed to warm to room temperature and stirred for 2 h. The mixture was quenched with water, concentrated and purified with a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to afford tert-butyl 1-(4-chloro-1H-pyrazol-1-yl)cyclopropane-1-carboxylate (171 mg). MS (ESI) m/z 243.1 [M+H]+.

4-isopropyl-6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine was prepared from 4-isopropyl-6-methoxypyrimidine following step 2 and step 3 of Intermediate 1. MS (ESI) m/z 279.2 [M+H]+.

A mixture of 4-cyclopropyl-1((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole (1.0 g, 4.2 mmol) in HCl-EtOAc solution (4M, 10 mL) was heated at 40° C. for 4 h. Solvent was removed under vacuum to give crude 4-cyclopropyl-1H-pyrazole (450 mg) which was used directly in next step without further purification. MS (ESI) m/z 109.1 [M+H]+.

To a solution of 2,3-dichloro-4-iodopyridine (10 g, 36.6 mmol) in MeOH (100 mL) was added sodium methoxide (1.98 g, 36.6 mmol) slowly at room temperature. The resulting mixture was heated at 70° C. overnight. The mixture was concentrated and purified with a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to afford 2,3-dichloro-4-methoxypyridine (1.82 g). MS (ESI) m/z 178.0 [M+H]+.

(R)-4-chloro-5-iodo-1-(1-methoxypropan-2-yl)-1H-pyrazole was prepared from (R)-4-chloro-1-(1-methoxypropan-2-yl)-1H-pyrazole according to step 2 of Intermediate 14. MS (ESI) m/z 301.0 [M+H]+.

To a solution of 1-(2-((tert-butyldimethylsilyl)oxy)ethyl)-4-chloro-1H-pyrazole (3.8 g, 14.57 mmol) in MeCN (50 mL) was added NBS (2.6g, 14.57 mmol) and TFA (1.7 g, 14.57 mmol). The mixture was stirred at room temperature for 12 h. The resulting mixture was concentrated under vacuum and purified by a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) and prep-HPLC (condition 2) to afford 2-(5-bromo-4-chloro-1H-pyrazol-1-yl)ethan-1-ol (400 mg). MS (ESI) m/z 224.9, 226.9 [M+H]+.

To a solution of 4-cyclopropyl-6-(difluoromethoxy)pyrimidine (430 mg, 2.3 mmol) in DMF (4 mL) was added Br2(1.8 g, 11.6 mol) dropwise at 0° C. The resulting mixture was heated at 50° C. for 2 h. The mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL×2). The combine organic phases were washed with brine, dried over Na2SO4, concentrated under vacuum and purified by silica gel chromatography (eluting with 1/40 EtOAc/PE) to 5-bromo-4-cyclopropyl-6-(difluoromethoxy)pyrimidine (275 mg). MS (ESI) m/z 265.0, 267.0 [M+H]+.

A solution of 2-bromo-4-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (344 mg, 1.0 mmol) in HCl EtOAc solution (4M, 10 mL) was stirred at room temperature for 2 h. The resulting mixture was concentrated and dried under vacuum to afford 2-bromo-4-(trifluoromethyl)-1H-imidazole (160 mg). MS (ESI) m/z 215.0, 217.0 [M+H]+.

To a mixture of 2-bromo-4-(trifluoromethyl)-1H-imidazole (363 mg, 1.7 mmol) and Cs2CO3(1.7 g, 5.1 mmol) in MeCN (20mL) was added iodoethane (312 mg, 2.0 mmol). The resulting mixture was stirred at room temperature for 2 h then filtered through celite. The filtrate was concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to afford 2-bromo-1-ethyl-4-(trifluoromethyl)-1H-imidazole (120 mg). MS (ESI) m/z 243.0, 245.0 [M+H]+.

A mixture of 3,3-dibromo-1,1,1-trifluoropropan-2-one (36 g, 0.13 mol) and NaOAc (20 g, 0.24 mol) in water (80 mL) was heated at 95° C. for 30 min. After cooling to 0° C., a cold solution of methyl 4-formylbenzoate (20 g, 0.12 mol) in a mixture of ammonium hydroxide (28%, 100 mL) and MeOH (300 mL) was added. The resulting mixture was stirred at ambient temperature for 4 h then concentrated under vacuum and purified by silica gel chromatography (eluting with 1/2 EtOAc/PE) to afford methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (30.1 g). MS (ESI) m/z 271.3 [M+H]+.

To a solution of methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (10 g, 37 mmol) in DMF (100 mL) at 0° C. was added 60% NaH (2.2 g, 60% in mineral oil, 55 mmol). The mixture was stirred at 0° C. for 30 min then MeI (7.8 g, 55 mmol) was added. After stirring at ambient temperature for 2 h, the reaction mixture was diluted with water (300 mL) and extracted with EtOAc (3×300 mL). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, concentrated under vacuum to afford methyl 4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (9.8 g), which was used directly in next step without further purification. MS (ESI) m/z 285.3 [M+H]+.

To a solution of methyl 4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (5 g , 17.6 mmol) in dry THF (50 mL) at 0° C. was added LiAlH4(3.3 g, 88 mmol). The reaction mixture was stirred at room temperature for 30 min then quenched with saturated Na2CO3(10 mL). The mixture was diluted with DCM (50 mL) and filtered through celite. The filtrate was concentrated and purified by silica gel chromatography (eluting with 1/10 MeOH/DCM) to afford (4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (3.0 g). MS (ESI) m/z 257.5 [M+H]+.

(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol was synthesized from methyl 4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate according to step 3 of Intermediate BB-1. MS (ESI) m/z 285.3 [M+H]+.

3-fluoro-4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzonitrile was prepared from 3-fluoro-4-formylbenzonitrile by following step 1 of Intermediate BB-1. MS (ESI) m/z 256.1 [M+H]+.

A mixture of 3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzonitrile (1.12 g, 4.11 mmol) and NaOH (3.32 g, 82.3 mmol) in EtOH/H2O (50 mL/25 mL) was heated at 100° C. for 3 h. The mixture was concentrated under vacuum to remove the EtOH and the pH adjusted to 3 by slow addition of 3M HCl. The resulting solution was extracted with EtOAc (3×50 mL). The organic layers were combined, dried over Na2SO4, filtered and concentrated under vacuum. The residue was dissolved in dry THF (10 mL) and cooled to 0° C., LiAlH4(312 mg, 8.21 mmol) was added slowly and the reaction mixture allowed to warm to ambient temperature. The reaction was stirred for 20 min, then quenched with 2M NaOH and diluted with DCM (20 mL). The resulting mixture was filtered through celite and the filtrate dried over Na2SO4and concentrated under vacuum to afford (3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol yl)phenyl)methanol (1.01 g). MS (ESI) m/z 275.2 [M+H]+.

(3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol was prepared from 3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzonitrile according to step 3 of Intermediate BB-4. MS (ESI) m/z 303.2 [M+H]+.

To a solution of 4-hydrazinylbenzoic acid hydrochloride (10 g, 53 mmol) in EtOH (100 mL) was slowly added 1,1,1-trifluoropentane-2,4-dione (8.2 g, 53 mmol). The solution was stirred at ambient temperature for 2 h then concentrated under vacuum and purified by silica gel chromatography (eluting with 1/10 MeOH/DCM) to afford 4-(5-methyl (trifluorometH-yl)-1H-pyrazol-1-yl)benzoic acid (4.2 g). MS (ESI) m/z 271.1 [M+H]+.

(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol was synthesized from 4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoic acid according to step 3 of Intermediate BB-1. MS (ESI) m/z 257.2 [M+H]+.

To a solution of ethyl 4-amino-3-fluorobenzoate (8.0 g, 43.7 mmol) in concentrated HCl (60 mL) was added a solution of NaNO2(3.59 g, 52.0 mmol) in H2O (6 mL) at 0° C. After stirred at the same temperature for 30 min, the mixture was added dropwise to a solution of SnCl2.2H2O (32 g, 141 mmol) in concentrated HCl (80 mL) at −20° C. The reaction was stirred at −10° C. for 30 min then solid precipitate that formed was collected by filtration. The solid was dissolved in 10% Na2CO3and extracted with EtOAc (3×100 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to afford ethyl 3-fluoro-4-hydrazinylbenzoate hydrochloride (7.2 g). MS (ESI) m/z 185.12 [M+H]+.

To a solution of ethyl 3-fluoro-4-hydrazinylbenzoate hydrochloride (5 g, 21.3 mmol) in EtOH (50 mL) was slowly added 1,1,1-trifluoropentane-2,4-dione (4 g, 26 mmol). The solution was stirred at ambient temperature for 2 h then concentrated under vacuum and purified by silica gel chromatography (eluting with 1/50 Acetone/PE) to ethyl 3-fluoro-4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (2.2 g) and ethyl 3-fluoro-4-(3-methyl-5-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (1.0 g).

(3-fluoro-4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol was prepared from ethyl 3-fluoro-4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate according to step 3 of Intermediate BB-1. MS (ESI) m/z 275.11 [M+H]+.

A solution of 4-hydrazinylbenzoic acid hydrochloride (5.0 g, 26.5 mmol) and ethyl 4,4,4-trifluoro-3-oxobutanoate (4.9 g, 26.51 mmol) in a mixture of MeOH (60 mL) and concentrated HCl (12 mL) was heated at 70° C. for 12 h. After cooling to ambient temperature, solvent was removed under reduced pressure to give methyl 4-(5-hydroxy-3-(trifluoromethyl)-1H-pyrazol-1-yl) benzoate (7.5 g). The crude product was used in next step directly without further purification. MS (ESI) m/z 287.1 [M+H]+.

To a solution of methyl 4-(5-hydroxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (1.1 g 3.85 mmol) in DMF at 0° C. (5 mL) was added NaH (230.8 mg, 60% in mineral oil, 5.77 mmol). the reaction was stirred at 0° C. for 30 min then methyl iodide (546.7 mg, 3.85 mmol) was added. The reaction was allowed to warm to ambient temperature and stirred 1 h then quenched with ice water, extracted with EtOAc (200 mL) and concentrated to give a crude product. The crude was purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to afford methyl 4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (715 mg). MS (ESI) m/z 301.1 [M+H]+.

(4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)methanol was synthesized from methyl 4-(5-methoxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate according to step 3 of Intermediate BB-1. MS (ESI) m/z 273.1 [M+H]+.

A mixture of 4-bromobenzimidamide hydrochloride (26.2 g, 111.3 mmol), 80% hydrazine monohydrate (30 mL) in EtOH (150 mL) was stirred at room temperature for 3 h. The reaction mixture was diluted with H2O (200 mL) and extracted with EtOAc (3×150 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered, concentrated under vacuum to give 4-bromobenzimidohydrazide (21.9 g), which was used directly in next step without further purification. MS (ESI) m/z 214.0, 216.0 [M+H]+.

A mixture of 3-(4-bromophenyl)-5-(trifluoromethyl)-1H-1,2,4-triazole (13.5 g, 46.4 mmol) in DMF (50 mL) at 0° C. was treated with portion-wise with NaH (2.78 g, 60% in mineral oil, 69.6 mmol). The resulting mixture was stirred at room temperature for 30 min. lodomethane (13.2 g, 92.8 mmol) was added and the resulting solution stirred for 1 h at room temperature. The reaction mixture was poured into ice water (50 mL), extracted with EtOAc (3×150 mL). The organic layers were combined, dried over anhydrous Na2SO4, filtered, concentrated and purified by column chromatography (eluting with 0 to 33.3% EtOAc/PE) to give 5-(4-bromophenyl)-1-methyl-3-(trifluoromethyl)-1H-1,2,4-triazole (4.3 g) and 3-(4-bromophenyl)-1-methyl-5-(trifluoromethyl)-1H-1,2,4-triazole (4.7 g). MS (ESI) m/z 306.0, 307.9 [M+H]+.

(4-(1-methyl-3-(trifluoromethyl)-1H-1,2,4-triazol-5-yl)phenyl)methanol was synthesized from ethyl 4-(1-methyl-3-(trifluoromethyl)-1H-1,2,4-triazol-5-yl)benzoate according to step 3 of Intermediate BB-1. MS (ESI) m/z 258.2 [M+H]+.

To a solution of ethyl 1-(4-bromophenyl)-5-hydroxy-1H-pyrazole-3-carboxylate (18.9 g, 60.8 mmol) in DMF (150 mL) at 0° C. was added NaH (3.45 g, 60% in mineral oil, 86.2 mmol). The resulting solution was stirred for at 0° C. for 30 min before MeI (12.2 g, 86.2 mmol) was added. The resulting solution was stirred at room temperature for 12 h then quenched with H2O and extracted with EtOAc (2×200 mL). The organic phase was dried over Na2SO4, filtered, concentrated and purified by silica gel chromatography (eluting with 1/5 EtOAc/PE) to afford ethyl 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carboxylate (7.1 g). MS (ESI) m/z 325.0, 227.0 [M+H]+.

A mixture of ethyl 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carboxylate (7.1 g, 21.8 mmol) in 7 M NH3in MeOH (70 mL) was heated at 100° C. in a sealed tube for 12 h. The solvent was removed under vacuum to afford of 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carboxamide (6.4 g). MS (ESI) m/z 296.0, 297.1 [M+H]+.

ethyl 4-(3-cyano-5-methoxy-1H-pyrazol-1-yl)benzoate was prepared from 1-(4-bromophenyl)-5-methoxy-1H-pyrazole-3-carbonitrile according to step 4 of Intermediate BB-11. MS (ESI) m/z 272.1 [M+H]+.

1-(4-(hydroxymethyl)phenyl)-5-methoxy-1H-pyrazole-3-carbonitrile was prepared from ethyl 4-(3-cyano-5-methoxy-1H-pyrazol-1-yl)benzoate according to step 3 of Intermediate BB-1. MS (ESI) m/z 230.1 [M+H]+.

1-ethyl-2-(4-nitrophenyl)-4-(trifluoromethyl)-1H-imidazole was prepared from 4-nitrobenzaldehyde following step 1 and step 2 of intermediate BB-1. MS (ESI) m/z 286.1 [M+H]+.

To a solution of 4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)aniline (2.85 g, 10 mmol) in MeOH was added Pd/C (10%, 285 mg). The mixture was evacuated and backfilled with hydrogen three times and then charged with hydrogen. The resulting mixture was stirred for 16 h at ambient temperature, then was filtered through celite and concentrated under vacuum to give 4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)aniline (2.35 g). MS (ESI) m/z 256.1 [M+H]+.

To a solution of 3-bromo-1-methyl-1H-pyrazole (4.0 g, 24.8 mmol) in mixture of dioxane (24 mL) and H2O (6 mL) was added (4-(methoxycarbonyl)phenyl)boronic acid (8.9 g, 49.6 mmol), Pd(dppf)Cl2(906 mg, 1.24 mmol) and K2CO3(7.0 g, 49.6 mmol). The mixture was heated at reflux for 16 h under N2. After cooling to ambient temperature, the mixture was filtered through celite and the filtrate concentrated under vacuum. The residue was purified by silica gel chromatography (eluting with 1/10 EtOAc/PE) to afford methyl 4-(1,4-dimethyl-1H-pyrazol-3-yl)benzoate (3.2 g). MS (ESI) m/z 217.1 [M+H]+.

(4-(1-methyl-1H-pyrazol-3-yl)phenyl)methanol was prepared from methyl 4-(1-methyl-1H-pyrazol-3-yl)benzoate according to step 3 of Intermediate BB-1. MS (ESI) m/z 189.1 [M+H]+.

To a solution of 3-iodo-1,4-dimethyl-1H-pyrazole (1.4 g, 6.3 mmol) in a mixture of dioxane (8 mL) and H2O (2 mL) was added (4-(methoxycarbonyl)phenyl)boronic acid (2.3 g, 13 mmol), Pd(dppf)Cl2(230 mg, 0.3 mmol) and K2CO3(1.73 g, 12.6 mmol). The mixture was heated at 80° C. for 12 h under N2. After cooling to ambient temperature, the mixture was filtered through celite and the filtrate concentrated under vacuum. The residue was purified by silica gel chromatography (eluting with 1/10 EtOAc/PE) to afford methyl 4-(1,4-dimethyl-1H-pyrazol-3-yl)benzoate (1.2 g). MS (ESI) m/z 231.1 [M+H]+.

(4-(1,4-dimethyl-1H-pyrazol-3-yl)phenyl)methanol was prepared from methyl 4-(1,4-dimethyl-1H-pyrazol-3-yl)benzoate according to step 3 of Intermediate BB-1. MS (ESI) m/z 203.1 [M+H]+.

A mixture of methyl 4-(5-(bromomethyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (570 mg, 1.5 mmol) and MeONa (170.1 mg, 3.1 mmol) in MeOH (8.5 mL) was stirred at 90° C. for 0.5 h. The reaction mixture was diluted with water (20 mL), extracted with DCM (20 mL×3), the combined organic layer was dried over Na2SO4, concentrated under vacuum to give crude product (500 mg), which was used directly in next step without further purification. MS (ESI) m/z 315.1 [M+H]+.

A solution of methyl 4-(5-(bromomethyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (1g, 2.75 mmol) in ethanolic methylamine solution (2M, 10 mL) was stirred at room temperature overnight. Solvent was removed and the crude was purified by silica gel chromatography (eluting with 1/4 EtOAc/PE) to afford methyl 4-(5-((methylamino)methyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (0.6 g). MS (ESI) m/z 314.1 [M+H]+.

To a solution of methyl 4-(5-((methylamino)methyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (550 mg, 1.76 mmol) and di-tert-butyl dicarbonate (574.5 mg, 2.63 mmol) in DCM (10 mL) was added TEA (532.8 mg, 5.26 mmol). The resulting solution was stirred at room temperature for 2 h. Solvent was removed and the crude was purified by silica gel chromatography (eluting with 1/8 EtOAc/PE) to afford methyl 4-(5-(((tert-butoxycarbonyl)(methyl)amino)methyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (450 mg). MS (ESI) m/z 414.2 [M+H]+.

To a solution of methyl 4-(5-methyl-3-(4,4, 5,5-tetramethyl-1,3 ,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)benzoate (720 mg, 2.11 mmol) in THF (10 mL) was added NaOH (2 M, 1.5 ml) and H2O2(30%, 3 ml). The resulting solution was stirred at room temperature for 2 h. Solvent was removed in vacuo and the crude residue was purified by silica gel chromatography (eluting with 1/1 EtOAc/PE) to afford methyl 4-(3-hydroxy-5-methyl-1H-pyrazol-1-yl)benzoate (280 mg). MS (ESI) m/z 233.1 [M+H]+.

To a solution of methyl 4-(3-hydroxy-5-methyl-1H-pyrazol-1-yl)benzoate (450 mg, 1.94 mmol) in EtOAc (16 mL) and MeOH (1.6 mL) was added TMSCHN2(1.6 mL, 3.10 mmol) at 0° C. The resulting solution was stirred at room temperature for overnight. Solvent was removed and the crude was purified by silica gel chromatography (eluting with 1/5 EtOAc/PE) to afford methyl 4-(3-methoxy-5-methyl-1H-pyrazol-1-yl)benzoate (150 mg). MS (ESI) m/z 247.1 [M+H]+.

A mixture of 4-hydrazinylbenzoic acid hydrochloride (5.0 g, 26.5 mmol) and methyl 3-oxobutanoate (4.6 g, 39.8 mmol) in concentrated HCl (6mL) and MeOH (30 mL) was heated at 70° C. for overnight. The pH of the mixture was adjusted to 7-8 with saturated aqueous NaHCO3. The mixture was extracted with EtOAc (30 mL×3). The organic layers were combined, concentrated under vacuum and purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford methyl 4-(5-hydroxy-3-methyl-1H-pyrazol-1-yl)benzoate (4.8 g). MS (ESI) m/z 233.1 [M+H]+.

(4-(1-(2-fluoroethyl)-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol was prepared from methyl 4-(1-(2-fluoroethyl)-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate according to step 3 of Intermediate BB-1. MS (ESI) m/z 289.1 [M+H]+.

To a mixture of 3-(4-bromophenyl)-5-(trifluoromethyl)-1H-1,2,4-triazole (8.2 g, 28.1 mmol) in DMF (20 mL) at 0° C. was added portion-wise sodium hydride (2.25 g, 60% in mineral oil, 56.16 mmol). The mixture was stirred for at room temperature 30 min then iodoethane (4.85 g, 30.89 mmol) was added. The resulting solution was stirred at room temperature for 1 h then poured into ice water (50 mL), extracted with EtOAc (3×50 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, concentrated and purified by column chromatography (eluting with a gradient of 0 to 33.3% EtOAc/PE) to give 5-(4-bromophenyl)-1-ethyl-3-(trifluoromethyl)-1H-1,2,4-triazole (1.6 g) and 3-(4-bromophenyl)-1-ethyl-5-(trifluoromethyl)-1H-1,2,4-triazole (4.1 g). MS (ESI) m/z 320.0, 322.0 [M+H]+.

(4-(1-ethyl-5-(trifluoromethyl)-1H-1,2,4-triazol-3-yl)phenyl)methanol was synthesized from ethyl 4-(1-ethyl-3-(trifluoromethyl)-1H-1,2,4-triazol-5-yl)benzoate according to step 3 of Intermediate BB-1. MS (ESI) m/z 272.1 [M+H]+.

A solution of (4-nitrophenyl)hydrazine hydrochloride (332 mg, 1.23 mmol) and NaOAc (110 mg, 1.34 mmol) in H2O (2 mL) was heated at 95° C. for 30 min. After cooled to 0° C., a solution of 1,1,1-trifluoropentane-2,4-dione (200 mg, 1.3 mmol) in ammonium hydroxide solution (28%, 2 mL) and MeOH (6 mL) was added into the above reaction mixture at 0° C. The resulting reaction mixture was stirred for room temperature for 14 h. The reaction mixture was diluted with H2O, extracted with DCM. The combined organic layers were washed with water and brine, dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The crude residue was purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to afford 5-methyl-1-(4-nitrophenyl) (trifluoromethyl)-1H-pyrazole (212 mg). MS (ESI) m/z 272.1 [M+H]+.

To a solution of 5-methyl-1-(4-nitrophenyl)-3-(trifluoromethyl)-1H-pyrazole (570 mg, 2.1 mmol) in MeOH (20 mL) was added 5% Pd/C (300 mg). The reaction mixture was stirred at room temperature overnight under H2atmosphere. The reaction mixture was then filtered through celite. The filtrate was concentrated under vacuum to afford 4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)aniline (480 mg). MS (ESI) m/z 242.1 [M+H]+.

1-methyl-2-(trifluoromethyl)-1H-imidazole was prepared from 2-(trifluoromethyl)-1H-imidazole and iodomethane according to step 1 of intermediate BB-3. MS (ESI) m/z 151.0 [M+H]+.

To a stirred solution of 1-methyl-2-(trifluoromethyl)-1H-imidazole (540 mg, 3.6 mmol) in MeCN (10 mL) was added NBS (1.92 g, 10.8 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. The solvent was removed in vacuo and the crude residue was purified by column chromatography (eluting with a gradient of 0 to 33.3% EtOAc/PE) to give 4,5-dibromo-1-methyl-2-(trifluoromethyl)-1H-imidazole (390 mg). MS (ESI) m/z 306.9, 308.9, 310.9 [M+H]+.

To a stirred solution of 4,5-dibromo-1-methyl-2-(trifluoromethyl)-1H-imidazole (360 mg, 1.17 mmol) in dry THF (3 mL) was added n-BuLi (2.5 M in hexane, 0.52 mL, 1.29 mmol) dropwise at −78° C. After addition, the resulting solution was stirred at −60° C. for 1 h then warmed to room temperature and quenched with aqueous saturated NH4Cl (20 mL). The mixture was extracted with EtOAc (15 mL×3), dried over anhydrous Na2SO4, concentrated and purified by column chromatography (eluting with a gradient of 0 to 33.3% EtOAc/PE) to give 4-bromo-1-methyl-2-(trifluoromethyl)-1H-imidazole (220 mg).1H NMR (400 MHz, Chloroform-d) δ 6.97 (s, 1H), 3.77 (d, J=1.1 Hz, 3H).

1-(4-(bromomethyl)-2-fluorophenyl)-3-methyl-5-(trifluoromethyl)-1H-pyrazole was prepared from ethyl 3-fluoro-4-(3-methyl-5-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (prepared in step 2 of intermediate BB-8) according to step 3 and step 4 of intermediate BB-1. MS (ESI) m/z 337.0, 339.0 [M+H]+.

1-(4-(bromomethyl)phenyl)-5-methoxy-3-(trifluoromethyl)-1H-pyrazole was prepared from methyl 4-(5-hydroxy-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzoate (prepared from step 1 of intermediate BB-9) and iodoethane following step 2 to step 4 of intermediate BB-9. MS (ESI) m/z 349.0, 351.0 [M+H]+.

A mixture of methyl 4-(4-chloro-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)benzoate (1 g, 2.73 mmol) in HCl-EtOAc solution (4M, 20 mL) was stirred at room temperature for 12 h. The pH of the resulting mixture was adjusted to 8 with saturated Na2CO3solution, extracted with EtOAc (30 mL×3), dried over anhydrous Na2SO4and concentrated under vacuum to afford methyl 4-(4-chloro-1H-imidazol-2-yl)benzoate (640 mg). MS (ESI) m/z 237.0 [M+H]+.

To a solution of 4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-methoxybenzoic acid (prepared from 4-formyl-3-methoxybenzonitrile following Intermediate BB-4, 3.28 g, 10 mmol) in MeOH (50 mL) was added dropwise thionyl chloride (5 mL) at 0° C. After addition, the solution was heated at 70° C. for 3 h. Solvent was removed and the crude was purified on a Biotage Isolera One (C18column, eluting with 30% to 90% MeCN/H2O, containing 0.1% TFA) to afford methyl 4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-methoxybenzoate (2.9 g). MS (ESI) m/z 343.1 [M+H]+.

A solution of methyl 4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-methoxybenzoate (1.96 g, 5.73 mmol) in DCM (15 mL) was added BBr3(1 M in DCM, 23 mL) dropwise at 0° C. The resulting solution was stirred at the same temperature for 2 h. The reaction mixture was quenched with MeOH (30 mL) and concentrated under vacuum. The crude product was purified by Biotage Isolera One (C18column, eluting with 30% to 90% MeCN/H2O, containing 0.1% TFA) to afford methyl 3-hydroxy-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (1.45 g). MS (ESI) m/z 329.1 [M+H]+.

2-(4-(bromomethyl)-2-(2-((tert-butyldimethylsilyl)oxy)ethoxy)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole was prepared from methyl 3-hydroxy-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (synthesized in step 2 of intermediate BB-40) and (2-bromoethoxy)(tert-butyl)dimethylsilane according to Intermediate BB-40. MS (ESI) m/z 521.1, 523.1 [M+H]+.

To a solution of 1-methyl-2-(trifluoromethyl)-1H-imidazole (3.3 g, 22.0 mmol) in MeCN (100 mL) was added NBS (11.7 g, 66.0 mmol). The mixture was stirred at room temperature for 12 hoh, then filtered through celite. The filtrate was concentrated under vacuum. The crude product was purified by silica gel chromatography (eluting with 1/6 EtOAc/PE) to afford 4,5-dibromo-1-methyl-2-(trifluoromethyl)-1H-imidazole (4.4 g). MS (ESI) m/z 306.9 [M+H]+.

To a solution of 4,5-dibromo-1-methyl-2-(trifluoromethyl)-1H-imidazole (4.2 g, 13.6 mmol) in dry THF (40 ml) was added n-BuLi (2.5 M in THF, 6 mL) slowly at −78° C. The mixture was stirred at −78° C. for 1 h, then warmed to room temperature and quenched with saturated aqueous NH4Cl (50 mL) and extracted with EtOAc (100 mL×3). The combined organic layers were washed with brine, dried over Na2SO4, concentrated and purified by silica gel chromatography (eluting with 1/6 EtOAc/PE) to afford 4-bromo-1-methyl-2-(trifluoromethyl)-1H-imidazole (1.85 g). MS (ESI) m/z 229.0, 231.0[M+H]+.

To a solution of 4-bromo-1-methyl-2-(trifluoromethyl)-1H-imidazole (1.61 g, 7.0 mmol) in THF (60 mL) was added (2-fluoro-4-(methoxycarbonyl)phenyl)boronic acid (4.2 g, 21.1 mmol), Pd(dppf)Cl2(513.0 mg, 0.7 mmol) and K3PO4(4.5 g, 21.1 mmol). The mixture was heated at 80° C. for 12 h under N2. The resulting mixture was filtered through celite and the filtrate was concentrated under vacuum. The crude product was purified by silica gel chromatography (eluting with 1/10 EtOAc/PE) to afford methyl 3-fluoro-4-(1-methyl-2-(trifluoromethyl)-1H-imidazol-4-yl)benzoate (1.9 g). MS (ESI) m/z 303.1 [M+H]+.

To a solution of 3-bromo-1-methyl-1H-pyrazole-4-carbonitrile (500 mg, 2.7 mmol) in THF (15 mL) was added (4-(hydroxymethyl)phenyl)boronic acid (1.2 g, 8.1 mmol), K3PO4(1.7 g, 8.1 mmol) and Pd(dppf)Cl2(200 mg, 0.3 mmol). The mixture was heated at 80° C. for overnight under N2. The solution was filtered through celite and the filtrate was concentrated under vacuum. The crude product was purified by silica gel chromatography (eluting with 1/2 EtOAc/PE) to afford 3-(4-(hydroxymethyl)phenyl)-1-methyl-1H-pyrazole-4-carbonitrile (475.0 mg). MS (ESI) m/z 214.1 [M+H]+.

2-(4-(bromomethyl)-2-fluoro-6-methoxyphenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole was prepared from 2-(4-bromo-2-fluoro-6-methoxyphenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole according to step 4 to step 6 of Intermediate BB-11. MS (ESI) m/z 381.0, 383.0 [M+H]+.

The solution of tert-butyl 2-bromo-4-methyl-1H-imidazole-1-carboxylate (4.0 g, 15.4 mmol) in dioxane (30 mL) was added (4-(methoxycarbonyl)phenyl)boronic acid (11.1 g 61.5 mmol), Pd(dppf)Cl2(1.1 g 1.5 mmol) and Cs2CO3(10.0 g, 30.8 mmol). The mixture was heated at 110° C. for 12 h under N2. The solution was filtered through celite and the filtrate was concentrated under vacuum. The crude product was purified by silica gel chromatography (eluting with 1/5 EtOAc/PE) to afford tert-butyl 2-(4-(methoxycarbonyl)phenyl)-4-methyl-1H-imidazole-1-carboxylate (3.5 g). MS (ESI) m/z 317.1 [M+H]+.

To a solution of tert-butyl 2-(4-(methoxycarbonyl)phenyl)-4-methyl-1H-imidazole-1-carboxylate (3.5 g, 11.2 mmol) in DCM (20 mL) was added TFA (20 mL) slowly at 0° C. The mixture was then stirred at room temperature for 2 h. The resulting mixture was concentrated under vacuum to afford methyl 4-(4-methyl-1H-imidazol-2-yl)benzoate (3.0 g). MS (ESI) m/z 217.1 [M+H]+.

To a solution of methyl 4-(4-methyl-1H-imidazol-2-yl)benzoate (3.0 g, 14 mmol) in MeCN (25 mL) was added EtI (4.4 g, 28.1 mmol) and Cs2CO3(9.1 g, 28.1 mmol). The mixture was heated at 80° C. for 2 h. The solution was filtered through celite and the filtrate was concentrated under vacuum. The crude product was directly purified by silica gel chromatography (eluting with 1:2 EtOAc/PE) to afford methyl 4-(1-ethyl-4-methyl-1H-imidazol-2-yl)benzoate (707.0 mg). MS (ESI) m/z 245.1 [M+H]+.

2-(4-(bromomethyl)phenyl)-1-ethyl-4-methyl-1H-imidazole was synthesized from methyl 4-(1-ethyl-4-methyl-1H-imidazol-2-yl)benzoate following step 3 and step 4 of intermediate BB-1. MS (ESI) m/z 279.0 [M+H]+.

A mixture of 2-(4-bromo-2-fluorophenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole (2.0 g, 6.0 mmol), tributyl(1-ethoxyvinyl)stannane (2.6 g, 7.1 mmol) and Pd(PPh3)4(347 mg, 0.3 mmol) in DMF (12 mL) was heated at 130° C. for 1 h in microwave reactor. The reaction mixture was diluted with H2O (20 mL), extracted with EtOAc (20 mL×3). The combined organic layers were washed with water and brine, concentrated under vacuum. The resulting residue was stirred in a mixture of THF (5 mL) and aqueous HCl (2 M, 15 mL) for 30 min. The solvent was removed and the residue was purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethan-1-one (1.2 g). MS (ESI) m/z 301.1 [M+H]+.

2-(1-bromo-6-fluoro-2,3-dihydro-1H-inden-5-yl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole was prepared from 6-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2,3-dihydro-1H-inden-1-one and 2-bromo-4-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole following step 3 to step 7 of Intermediate BB-39. MS (ESI) m/z 377.0 [M+H]+.

To a solution of methyl 2-(2-(4-bromophenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)propanoate (2.8 g ,7.43 mmol) in THF (10 mL) was added LiAlH4(338.7 mg, 8.91 mmol) slowly at 0° C. The reaction mixture was stirred at room temperature for 40 min then quenched with saturated Na2CO3and quickly filtered through celite. The filtrate was concentrated to give crude 2-(2-(4-bromophenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)propan-1-ol (2.8 g) which was used directly in next step without further purification. MS (ESI) m/z 349.0, 351 [M+H]+.

To a solution of 2-(2-(4-bromophenyl)-4-(trifluoromethyl)-1H-imidazol-1-yl)propan-1-ol (2.8 g, 8.0 mmol) in DCM (20 mL) was added DAST (40 mL). The mixture was stirred at room temperature for 16 h. Solvent was removed under vacuum. The resulting residue was purified by column chromatography on silica gel (eluting with 4/1 PE/EtOAc) to give 2-(4-bromophenyl)-1-(1-fluoropropan-2-yl)-4-(trifluoromethyl)-1H-imidazole (2.9 g). MS (ESI) m/z 351.0, 353.0 [M+H]+.

2-(4-acetylphenyl)-1H-imidazole-4-carbonitrile was prepared from (4-acetylphenyl)boronic acid and 2-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole-4-carbonitrile (prepared from 1H-imidazole-4-carbonitrile following step 1 and step 2 of Intermediate BB-39) following step 3 and step 4 of Intermediate BB-39. MS (ESI) m/z 212.1 [M+H]+.

To a solution of 2-(4-acetylphenyl)-1H-imidazole-4-carbonitrile (2.2 g, 10.4 mmol) in MeCN (25 mL) was added EtI (1.8 g, 11.5 mmol) and Cs2CO3(6.8 g, 20.9 mmol). The mixture was heated at 60° C. for 4 h then filtered through celite and concentrated under vacuum. The crude was purified by column chromatography on silica gel (eluting with 2/1 PE/EtOAc) to afford 2-(4-acetylphenyl)-1-ethyl-1H-imidazole-4-carbonitrile (1.0 g). MS (ESI) m/z 240.1 [M+H]+.

To a solution of 2-(4-acetylphenyl)-1-ethyl-1H-imidazole-4-carbonitrile (1.0, 4.2 mmol) in MeOH (15 mL) was added NaBH4 (318 mg, 8.4 mmol). The mixture was stirred at room temperature for 30 min. The mixture was concentrated under vacuum and purified by column chromatography on silica gel (eluting with 3/1 PE/EtOAc) to afford 1-ethyl-2-(4-(1-hydroxyethyl)phenyl)-1H-imidazole-4-carbonitrile (800 mg). MS (ESI) m/z 242.1 [M+H]+.

To a solution of 1-(4-(4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-one (2.0 g, 7.8 mmol), cyclopropylboronic acid (1.4 g 15.8 mmol) and Na2CO3(1.7 g, 15.8 mmol) in 1,2-dichloroethane (10 mL) was added a solution of Cu(OAc)2(572.0 mg, 3.2 mmol) and 2,2′-bipyridine (1.2 g, 7.8 mmol) in 1,2-dichloroethane (20 mL). The resulting mixture was heated at 70° C. overnight under an atmosphere of O2. The mixture was diluted with water (20 mL) and extracted with DCM (20 mL×2). The organic layers were washed with brine, dried over anhydrous Na2SO4, concentrated under vacuum and purified by column chromatography on silica gel (eluting with 4/1 PE/EtOAc) to afford 1-(4-(1-cyclopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-one (1.8 g). MS (ESI) m/z 295.1[M+H]+.

tert-butyl-(4-(4-cyano-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol yl)phenyl)carbamate was prepared from 1H-imidazole-4-carbonitrile following step 1 to step 3 of Intermediate BB-39.

A solution of tert-butyl-(4-(4-cyano-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)phenyl)carbamate (1.2 g, 2.8 mmol) in TBAF (1 M in THF, 10 mL) was stirred at room temperature for 18 h. The solvent was removed under vacuum and the crude purified by column chromatography on silica gel (eluting with 2/1 PE/EtOAc) to give tert-butyl (4-(4-cyano-1H-imidazol-2-yl)phenyl)carbamate (370 mg). MS (ESI) m/z 285.1 [M+H]+.

To a solution of tert-butyl-(4-(4-cyano-1-ethyl-1H-imidazol-2-yl)phenyl)carbamate (800 mg, 2.6 mmol) in DCM (10 mL) was added TFA (3 mL) at room temperature. The reaction solution was stirred at room temperature for 2 h. Solvent was removed under vacuum and the residue diluted with EtOAc, washed with 1M NaOH and brine. The organic phase was dried over Na2SO4and concentrated under vacuum to give 2-(4-aminophenyl)-1-ethyl-1H-imidazole-4-carbonitrile (250 mg). MS (ESI) m/z 213.1 [M+H]+.

1-(1-fluoropropan-2-yl)-2-(4-nitrophenyl)-4-(trifluoromethyl)-1H-imidazole was prepared from 2-(4-nitrophenyl)-4-(trifluoromethyl)-1H-imidazole following step 1 to step 3 of Intermediate BB-60. MS (ESI) m/z 318.1 [M+H]+.

4-(1-(1-fluoropropan-2-yl)-4-(trifluoromethyl)-1H-imidazol-2-yl)aniline was prepared according to step 3 of Intermediate BB-14. MS (ESI) m/z 288.1 [M+H]+.

1-ethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-(trifluoromethyl)-1H-imidazole was prepared from 2-(4-bromophenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole according to step 3 of intermediate 1. MS (ESI) m/z 367.2 [M+H]+.

To a solution of 1-ethyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-4-(trifluoromethyl)-1H-imidazole (500 mg, 1.36 mmol) in THF (10 mL) was added 30% aqueous hydrogen peroxide (0.2 mL) and aqueous NaOH (2 M, 0.3 mL) at 0° C. The resulting mixture was stirred at room temperature for 2 h then extracted with DCM (50 mL×3). The combined organic phases were dried over anhydrous Na2SO4, concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenol (278 mg). MS (ESI) m/z 257.1 [M+H]+.

To a solution of 2-(2-bromo-4-nitrophenyl)-4-(trifluoromethyl)-1H-imidazole (3.9 g, 11.6 mmol) in MeCN (100 mL) was added EtI (3.6 g, 23.2 mmol) and Cs2CO3(7.6 g, 23.2 mmol). The mixture was stirred at room temperature for 18 h then was filtered through celite. The filtrate was concentrated under vacuum. The crude product was purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 2-(2-bromo-4-nitrophenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole (1.24 g). MS (ESI) m/z 364.0, 366.0 [M+H]+.

A mixture of 2-(2-bromo-4-nitrophenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole (1.24 g, 3.4 mmol) and CuCN (922 mg, 10.2 mmol) in NMP (10 mL) was heated at 220° C. for 30 min in microwave reactor. The mixture was filtered through celite and the filtrate concentrated under vacuum. The crude was purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 2-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-5-nitrobenzonitrile (400 mg). MS (ESI) m/z 311.1 [M+H]+.

A mixture of 2-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-5-nitrobenzonitrile (630 mg, 2.0 mmol), iron power (560 mg, 10 mmol) and NH4Cl (162 mg, 3.0 mmol) in a mixture of EtOH (20 mL) and H2O (4 mL) was heated at 85° C. for 3 h. The resulting mixture was filtered through celite and the filtrate concentrated under vacuum. The crude was purified by column chromatography on silica gel (eluting with 2/1 PE/EtOAc) to give 5-amino-2-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzonitrile (160 mg). MS (ESI) m/z 281.1 [M+H]+.

1-(4-(1-bromoethyl)phenyl)-5-methoxy-3-(trifluoromethyl)-1H-pyrazole was synthesized from 1-(4-bromophenyl)-5-methoxy-3-(trifluoromethyl)-1H-pyrazole according to step 3 to step 5 of Intermediate BB-52. MS (ESI) m/z 349.0, 351.0 [M+H]+.

To a solution of 1-(4-(3-isopropoxypyridin-2-yl)phenyl)ethan-1-ol (123 mg, 0.48 mmol) in dry DCM (5 mL) was added PBr3(260 mg, 0.96 mmol) at 0° C. The resulting mixture was stirred at same temperature for 2 h then quenched with saturated aqueous NaHCO3(10 mL) and extracted with DCM (10 mL×3). The combined organic layers were dried over anhydrous Na2SO4, concentrated under vacuum and purified by silica gel chromatography (eluting with 1/5 EtOAc/PE) to afford 2-(4-(1-bromoethyl)phenyl)-3-isopropoxypyridine (86 mg). MS (ESI) m/z 320.0, 322.0 [M+H]+.

A mixture of 3-bromo-1H-pyrazol-5-amine (600 mg, 3.704 mmol) and ethyl acrylate (0.592 mL, 5.556 mmol) in pyridine (10 mL) and H2O (1 mL) and was heated at 135° C. for 24 h in a sealed tube. After cooling to ambient temperature, the mixture was poured into EtOAc (15 mL) and the precipitate that formed was collected by filtration to afford 2-bromo-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (270 mg). MS (ESI) m/z 216.0, 218.0 [M+H]+.

To a solution of 2-bromo-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (150 mg, 0.694 mmol) in DMF (5 mL) was added 2-(4-(bromomethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazole (221 mg, 0.694 mmol) and K2CO3(192 mg, 1.39 mmol). The resulting mixture was heated at 60° C. for 2 h. After cooling to ambient temperature, the mixture was poured into water (30 mL) and extracted with EtOAc (3×30 mL). The organic layers were combined, dried over anhydrous Na2SO4and concentrated under vacuum. The residue was purified by silica gel chromatography (eluting with 1/2 EtOAc/PE) to afford 2-bromo-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (302 mg). MS (ESI) m/z 454.0, 456.0 [M+H]+.

To a solution of 2-bromo-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (300 mg, 0.661 mmol) in a mixture of dioxane (10 mL) and H2O (1 mL) was added 4-cyclopropyl-6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (183 mg, 0.663 mmol), Pd(dppf)Cl2(48 mg, 0.066 mmol) and K3PO4(281 mg, 1.32 mmol). The mixture was heated at reflux for 16 h under N2. After cooling to ambient temperature, the mixture was filtered through celite and the filtrate was concentrated under vacuum. The residue was purified by prep-HPLC (condition 1) to afford Example 1 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (110 mg).

Table 1. The compounds listed in Table 1 were synthesized according to Example 1 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers were not separated.

To a solution of tert-butyl 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidine-4(5H)-carboxylate (500 mg, 1.34 mmol) in DCM (4 mL) at 0° C. was added TFA (4 mL) slowly. The resulting solution was stirred at room temperature for 3 h. The resulting solution was concentrated under vacuum, the residue obtained was dissolved in EtOAc (50 mL) and washed with 1M NaOH aqueous solution. The organic phase was concentrated under vacuum and purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine (220 mg). MS (ESI) m/z 272.9 [M+H]+.

Table 2. The compounds listed in Table 2 were synthesized according to Example 24 using the appropriate commercially available reagents and/or intermediates described above.

A solution of 2-bromo-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (300 mg, 0.66 mmol, prepared in step 2 of Example 1) in dry THF (8 mL) was cooled to −78° C. LDA (0.33 mL, 2 M) was added dropwise and the solution stirred at −78° C. for 30 min then a solution of MeI (84.3 mg, 0.59 mmol) in dry THF (0.5 mL) was added. The resulting solution was stirred at −78° C. for another 30 min then warmed to room temperature. The reaction was quenched with water, diluted with EtOAc, washed with water and brine. The organic phase was dried with Na2SO4, concentrated, and purified by silica gel chromatography (eluting with 2:1 PE/EtOAc) to afford 2-bromo-6-methyl-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (70 mg). MS (ESI) m/z 468.1, 470.1 [M+H]+.

Example 31 2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-6-methyl-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was synthesized from (1-isopropyl-4-methyl-1H-pyrazol-5-yl)boronic acid (intermediate 3) and 2-bromo-6-methyl-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one following step 3 of Example 24.

Table 3. The compounds listed in Table 3 were synthesized according to Example 31 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of 2-bromo-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (200 mg, 0.93 mmol) in MeCN (10 mL) was added NCS (148 mg, 1.11 mmol). The resulting solution was heated at 80° C. for 2 h. The reaction mixture was concentrated, diluted with EtOAc and washed with brine. The organic layers were combined, dried over anhydrous Na2SO4, concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 2-bromo-3-chloro-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (200 mg). MS (ESI) m/z 250.0, 252.0 [M+H]+.

Example 33 3-chloro-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was synthesized from 2-bromo-3-chloro-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one according to step 3 of Example 1.

Table 4. The compounds listed in Table 4 were synthesized according to Example 33 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers were not separated.

To a solution of 4-methyl-5-nitro-1H-pyrazole (350 mg, 2.75 mmol) in DMF (5 mL) was added Br2(878.9 mg, 5.5 mmol) and the resulting solution heated at 40° C. overnight. The reaction mixture was diluted with EtOAc and washed with brine. The organic layer was dried over anhydrous Na2SO4, concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 3-bromo-4-methyl-5-nitro-1H-pyrazole (453 mg). MS (ESI) m/z 204.0, 206.0 [M-H]+.

To a solution of 3-bromo-4-methyl-5-nitro-1H-pyrazole (450 mg, 2.19 mmol) in EtOH (10 mL) and H2O (2 mL) was added Fe powder (614 mg, 10.97 mmol) and NH4Cl (234 mg, 4.38 mmol). The resulting solution was heated at 90° C. for 30 min. The reaction mixture was filtered through celite and concentrated under vacuum. The crude was directly purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 3-bromo-4-methyl-1H-pyrazol-5-amine (251 mg). MS (ESI) m/z 176.0, 178.0 [M+H]+.

A mixture of 1H-1,2,4-triazole-3,5-diamine (1g, 10.1 mmol), ethyl acrylate (1.52 g, 15.15 mmol) and pyridine (15 mL) was heated at 135° C. in sealed tube for 12 h. Solvent was removed under vacuum. The residue was dissolved in EtOAc and filtered through celite. The filtrate was concentrated to give 2-amino-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one (1.4 g). MS (ESI) m/z 154.1 [M+H]+.

To a solution of 2-bromo-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one (200 mg, 0.92 mmol) in DMF (5 mL) was added 2-(4-(bromomethyl)phenyl)-1-methyl-4-(trifluoromethyl)-1H-imidazole (293 mg, 0.92 mmol) and K2CO3(381 mg, 2.76 mmol). The mixture was stirred at ambient temperature for 12 h then diluted with EtOAc and washed with water and brine. The organic layer was dried with Na2SO4, concentrated and purified by silica gel chromatography (eluting with 1/2 EtOAc/PE) to afford 2-bromo-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one (103 mg). MS (ESI) m/z 455.1, 457.1 [M+H]+.

Example 37 2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 2-bromo-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one and (1-isopropyl-4-methyl-1H-pyrazol-5-yl)boronic acid according to step 3 of Example 1.

Table 5. The compounds listed in Table 5 were synthesized according to Example 37 using the appropriate commercially available reagents and/or intermediates described above.

2-bromo-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine was synthesized form 2-bromo-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine (prepared according to step 1 of Example 24) according to step 5 of Example 24. MS (ESI) m/z 440.1, 442.1 [M+H]+.

To a solution of 3-bromo-1H-pyrazol-5-amine (3.1 g, 19.1 mmol) in dioxane (18 mL) was added 1,3-dibromobutane (4.5 g, 21 mmol) and DIPEA (8.1 g, 62.7 mmol) and the reaction heated at 120° C. for 20 h in a sealed tube. The solvent was removed under vacuum and the crude was purified by Prep-HPLC (condition 2) to afford 2-bromo-7-methyl-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine (2.03 g) and 2-bromo-5-methyl-4,5,6,7-tetrahydropyrazolo-[1,5-α]pyrimidine(503 mg).

Example 48 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-5-methyl-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine was synthesized from 2-bromo-5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine following step 2 to step 5 of Example 24.

Table 6. The compounds listed in Table 6 were synthesized according to Example 48 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of 2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (1.1 g 4.24 mmol) in dry THF (30 mL) was added LiAlH4(976 mg, 25.4 mmol) at 0° C. The reaction was stirred 0° C. for 30 min then quenched with aqueous NaOH (2M, 1 mL), filtered and washed with THF (20 mL). The combined filtrates were concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to give 2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidine (800 mg). MS (ESI) m/z 244.2 [M+H]+.

To a mixture of 1-(4-(bromomethyl)phenyl)-5-methyl-3-(trifluoromethyl)-1H-pyrazole (94 mg, 0.29 mmol) and 2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine (60 mg, 0.24 mmol) in DMF (3 mL) was added NaI (73 mg, 0.49 mmol) and DIPEA (63 mg, 0.49 mmol) at ambient temperature. The resulting solution was heated at 40° C.-60° C. for 12 h. The reaction mixture was diluted with EtOAc (30 mL), washed with water and brine. The organic phase was dried with Na2SO4, concentrated and purified by prep-HPLC (condition 2) to give Example 52 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(5-methyl-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine (31 mg).

Table 7. The compounds listed in Table 7 were synthesized according to Example 52 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers, when generated, were separated by chiral HPLC and absolute stereochemistries were arbitrarily assigned.

To a solution of methyl 4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzoate (7.0 g, 25.9 mmol) in anhydrous THF (40 mL) was added 60% NaH (1.14 g, 60% in mineral oil, 28.5 mmol) at 0° C. The reaction was stirred at 0° C. for 15 min before SEMCl (6 mL, 33.7 mmol) was added. The resulting mixture was warmed to room temperature and stirred for another 15 min then quenched with H2O (100 mL) and extracted with EtOAc (3×100 mL). The combined organic phases were dried over anhydrous Na2SO4, concentrated under vacuum and purified by silica gel chromatography (eluting with 1/40 EtOAc/PE) to afford methyl 4-(4-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)benzoate (8.4 g). MS (ESI) m/z 401.3 [M+H]+.

To a solution of methyl 4-(4-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)benzoate (7.77 g, 19.4 mmol) in anhydrous THF (60 mL) was added LiAlH4(1.47 g, 38.8 mmol) at 0° C. The reaction was stirred at room temperature for 1 h then quenched with aqueous NaOH (2M). The resulting mixture was diluted with DCM (100 mL) and filtered through celite. The filtrate was concentrated under vacuum and purified by silica gel chromatography (eluting with 1/40 MeOH/DCM) to afford (4-(4-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)phenyl)methanol (5.9 g). MS (ESI) m/z 373.3 [M+H]+.

To a solution of (4-(4-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)phenyl)methanol (3.8 g, 10.2 mmol), NaHCO3(1.71 g, 20.4 mmol) and PPh3(5.36 g, 20.4 mmol) in DCM (70 mL) was added CBr4(6.76 g, 20.4 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h then filtered through celite and the filtrate concentrated. The residue obtained was purified by silica gel chromatography (eluting with EtOAc/PE 1/10) to afford 2-(4-(bromomethyl)phenyl)-4-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazole (3.71 g). MS (ESI) m/z 435.3, 437.3 [M+H]+.

A mixture of 2-bromo-4-(4-(4-(trifluoromethyl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (2.37 g, 4.16 mmol) in HCl dioxane solution (4 M, 100 mL) was heated at 60° C. for 2 h. The solvent was removed in vacuo and the crude residue obtained was dissolved in EtOAc and washed with saturated Na2CO3solution. The organic layer was dried over anhydrous Na2SO4, concentrated under vacuum and purified by silica gel chromatography (eluting with MeOH/DCM 1/80) to afford 2-bromo-4-(4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (1.15 g). MS (ESI) m/z 440.3, 442.3 [M+H]+.

Example 69 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 2-bromo-4-(4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one according to step 3 of Example 1.

Table 8. The compounds listed in Table 8 were synthesized according to Example 69 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of 2-bromo-4-(4-(4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (600 mg, 1.36 mmol) in dry DMF (5 mL) was added NaH (60% in mineral oil, 60 mg, 1.5 mmol) at 0° C. The resulting solution was stirred at 0° C. for 30 min before iodoethane (319 mg, 2.01 mmol) was added. The resulting mixture was then heated at 45° C. for 3 h then cooled to room temperature. The reaction mixture was quenched with H2O (40 mL) and extracted with EtOAc (3×40 mL). The combined organic layers were dried over anhydrous Na2SO4, concentrated under vacuum and purified by silica gel chromatography (eluting with MeOH/DCM 1/50) to afford 2-bromo-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (540 mg). MS (ESI) m/z 468.3, 470.3 [M+H]+.

Example 71 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was synthesized from 2-bromo-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one according to step 3 of Example 1.

Table 9. The compounds listed in Table 9 were synthesized according to Example 71 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of 2-methoxy-6-methylbenzoic acid (5.0 g, 30 mmol) in MeOH (50 mL) was added SOC12(5 mL) dropwise at 0° C. The resulting solution was heated at 65° C. for 6 h then concentrated and purified by silica gel chromatography (eluting with 1/8 EtOAc/PE) to afford methyl 2-methoxy-6-methylbenzoate (4.1 g, 76%) as a white solid. MS (ESI) m/z 181.0 [M+H]+.

To a solution of 3-(2-methoxy-6-methylphenyl)-1H-pyrazol-5-amine (1.4 g, 7.4 mmol) in EtOH (6 mL) was added 80% hydrazine hydrate (2.82 mL) and the resulting solution heated at 100° C. for 24 h. The reaction was concentrated under reduced pressure and the residue obtained purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 3-(2-methoxy-6-methylphenyl)-1H-pyrazol-5-amine (610 mg). MS (ESI) m/z 204.7, [M+H]+.

solution of 2-bromo-6-methyl-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (223 mg, 0.48 mmol, prepared in step 1 of Example 31) in dry THF (8 mL) was cooled to −78° C., LDA (2.0 M in THF/hexane, 0.12 mL, 0.24 mmol) was added dropwise and the solution was stirred 30 min before a solution of MeI (33.9 mg, 0.24 mmol) in dry THF (0.5 mL) was added. The resulting solution was stirred at −78° C. for another 30 min then warmed to room temperature. The reaction was quenched with water, diluted with EtOAc, washed with water and brine. The organic phase was dried with Na2SO4, concentrated, and purified by silica gel chromatography (eluting with 2:1 PE/EtOAc) to afford 2-bromo-6,6-dimethyl-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (53 mg). MS (ESI) m/z 482.1, 484.1 [M+H]+.

2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 2-bromo-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (prepared in step 1 of Example 1) following step 3 of Example 1. MS (ESI) m/z 280.2 [M+H]+.

Example 77 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorobenzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was prepared 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one following step 2 of Example 1.

Table 10. The compounds listed in Table 10 were synthesized according to Example 77 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers, when generated, were separated by chiral prep-HPLC and absolute stereochemistries were assigned by comparing retention time on chiral HPLC with similar chiral compounds that were obtained via enantioselective synthesis.

Example 147 3-chloro-2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine was prepared from 3-chloro-2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine according to step 3 of Example 52.

Table 11. The compounds listed in Table 11 were synthesized according to Example 147 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of 2-bromo-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (10 g, 46 mmol) in DMF (50 mL) was added NaH (60% in mineral oil, 2.8 g, 69 mmol) at 0° C. The mixture was stirred for 30 min then 2-(Trimethylsilyl)ethoxymethyl chloride (12 mL, 69 mmol) was added. The resulting mixture was stirred at room temperature for 2 h then quenched with water (100 mL) and extracted with EtOAc (100 mL×3). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, concentrated and purified by column chromatography on silica gel (eluting with 1/5 EtOAc/PE) to afford 2-bromo-4-((2-(trimethylsilyl)ethoxy)methyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (6.1 g). MS (ESI) m/z 346.1 [M+H]+.

A mixture of 2-(4-methoxy-6-(1-methylcyclopropyl)pyrimidin-5-yl)-4-((2-(trimethylsilyl)ethoxy)methyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (350 mg, 0.82 mmol) in HCl EtOAc solution (4M, 10 mL) was stirred at room temperature for 12 h. The pH of the resulting mixture was adjusted to 8 with saturated Na2CO3solution, extracted with EtOAc (15 mL×3), dried over anhydrous Na2SO4and concentrated under vacuum to afford 2-(4-methoxy-6-(1-methylcyclopropyl)pyrimidin-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (189 mg). MS (ESI) m/z 300.2 [M+H]+.

Example 149 4-(3-fluoro-4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-2-(4-methoxy-6-(1-methylcyclopropyl)pyrimidin-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 2-(4-methoxy-6-(1-methylcyclopropyl)pyrimidin-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one following step 2 of Example 1.

Table 12. The compounds listed in Table 12 were synthesized according to Example 149 using the appropriate commercially available reagents and/or intermediates described above.

2-bromo-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 2-bromo-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one and 2-(4-(bromomethyl)phenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole according to step 2 of Example 1. MS (ESI) m/z 468.1

Example 154 2-(4-chloro-1-(oxetan-3-yl)-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one following step 3 of Example 149.

Table 13. The compounds listed in Table 13 were synthesized according to Example 154 using the appropriate commercially available reagents and/or intermediates described above.

2-bromo-4-((2-(trimethylsilyl)ethoxy)methyl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 2-bromo-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one (prepared in step 2 of Example 37) according to step 1 of Example 149. MS (ESI) m/z 347.0, 349.0 [M+H]+.

Example 164 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one following step 2 of Example 1.

Table 14. The compounds listed in Table 14 were synthesized according to Example 164 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers, when generated, were separated by chiral prep-HPLC and absolute stereochemistries were assigned by comparing retention time on chiral HPLC with similar chiral compounds that were obtained via enantioselective synthesis.

2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-4-((2-(trimethylsilyl)ethoxy)methyl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one according to step 4 of Example 149. MS (ESI) m/z 261.1 [M+H]+.

Example 221 2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl) benzyl)-4,5,6,7-tetrahydro-[1,2,4]triazolo[1,5-α]pyrimidine was synthesized from 2-(1-isopropyl-4-methyl-1H-pyrazol-5-yl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one according to step 2 and step 3 of Example 52.

Table 15. The compounds listed in Table 15 were synthesized according to Example 221 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (248 mg, 0.47 mmol) in MeCN (5 mL) was added NIS (315 mg, 1.41 mmol). The resulting solution was heated at 80° C. for 16 h. The reaction mixture was concentrated, diluted with EtOAc and washed with brine. The organic layers were combined, dried over anhydrous Na2SO4, concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 95% MeCN/H2O) to afford 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-3-iodo-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (140 mg). MS (ESI) m/z 658.1 [M+H]+.

A solution of tert-butyl ((1-(4-((2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-5-oxo-6,7-dihydropyrazolo[1,5-α]pyrimidin-4(5H)-yl)methyl)phenyl)-3-(trifluoromethyl)-1H-pyrazol-5-yl)methyl)(methyl)carbamate (50 mg, 0.077 mmol) in HCl dioxane solution (4M, 2 mL) was stirred at room temperature for 1 h before solvent was removed. The crude was purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O containing 0.1% TFA) to afford Example 224 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(5-((methylamino)methyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (24.7 mg).

Table 16. The compounds listed in Table 16 were synthesized according to Example 224 using the appropriate commercially available reagents and/or intermediates described above.

(4-(pyridin-2-ylmethoxy)phenyl)methanol was prepared from methyl 4-(pyridin-2-ylmethoxy)benzoate according to step 3 of intermediate BB-1. MS (ESI) m/z 216.1 [M+H]+.

To a solution of (4-(pyridin-2-ylmethoxy)phenyl)methanol (100 mg, 0.46 mmol), and TEA (56.5 mg, 0.56 mmol) in DCM (2 mL) was added p-toluenesulfonyl chloride (106 mg, 0.56 mmol) at room temperature. The resulting solution was stirred at room temperature for 2 h. The solution was concentrated under vacuum. The crude product was purified by silica gel chromatography (eluting with 1/20 EtOAc/PE) to afford 4-(pyridin-2-ylmethoxy)benzyl 4-methylbenzenesulfonate (103 mg). MS (ESI) m/z 370.1 [M+H]+.

To a solution of 2-(4-chloro-1H-pyrazol-1-yl)propan-1-ol (1.6 g, 10 mmol, prepared in step 3 of Intermediate 15) and TEA (3.04 g, 30 mmol) in dry DCM (20 mL) was added tert-Butyldimethylsilyl chloride (1.81 g, 12 mmol) in dry DCM (10 mL) slowly at 0° C. After addition, the resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with DCM (30 mL), washed with water and brine, dried over anhydrous Na2SO4, concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to give 1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-4-chloro-1H-pyrazole (742 mg). MS (ESI) m/z 275.1 [M+H]+.

1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-4-chloro-5-iodo-1H-pyrazole was prepared from 1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-4-chloro-1H-pyrazole according to step 2 of intermediate 14. MS (ESI) m/z 401.0 [M+H]+.

2-(1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-4-chloro-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-2-(4,4,5, 5-tetramethyl-1,3,2-dioxaborolan-2-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (synthesized in step 2 of Example 154) and 1-(1-((tert-butyldimethylsilyl)oxy)propan-2-yl)-4-chloro-5-iodo-1H-pyrazole following step 3 of Example 149. MS (ESI) m/z 662.3 [M+H]+.

4-(3-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 2-(4-(bromomethyl)-2-(2-((tert-butyldimethylsilyl)oxy)ethoxy)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole and 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one (prepared following step 2 and 3 of Example 164) following step 2 of Example 1. MS (ESI) m/z 721.3 [M+H]+.

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (200 mg, 0.71 mmol) in H2SO4(10 mL) was added fuming HNO3(1.5 mL) at 0° C. dropwise. The mixture was then stirred at room temperature for 2 h then quenched with NaHCO3and extracted with EtOAc. The organic phase was washed with brine, dried over anhydrous Na2SO4and concentrated under vacuum to give 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-3-nitro-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (200 mg) which was used directly in next step without further purification. MS (ESI) m/z 325.1 [M+H]+.

A mixture of 4-(3-nitro-1H-pyrazol-1-yl)benzonitrile (2 g, 9.3 mmol) and Pd/C (5%, 400 mg) was stirred at room temperature for 2 h under an atmosphere of H2. The reaction mixture was filtered through celite and concentrated under vacuum to afford 4-(3-amino-1H-pyrazol-1-yl)benzonitrile (1.7 g). MS (ESI) m/z 185.1 [M+H]+.

(4-(3-morpholino-1H-pyrazol-1-yl)phenyl)methanol was prepared from 4-(3-morpholino-1H-pyrazol-1-yl)benzonitrile according to step 3 of intermediate BB-4. MS (ESI) m/z 260.1 [M+H]+.

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-hydroxy-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (350 mg, 0.64 mmol), triethylamine (192 mg, 1.9 mmol) in DCM (10 mL) was added methanesulfonyl chloride (150 mg, 1.3 mmol) at 0° C. The resulting mixture was stirred at room temperature for 1 h. The mixture was concentrated under vacuum and used in next step directly without further purification. MS (ESI) m/z 626.2 [M+H]+.

A mixture of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-5-oxo-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-6-yl methanesulfonate (401 mg, 0.64 mmol) and NaN3(83 mg, 1.3 mmol) in DMSO (5 mL) was stirred at room temperature for 1 h. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (50 mL). The organic phase was washed with brine, dried over anhydrous Na2SO4, concentrated under vacuum and purified by column chromatography on silica gel (eluting with 3/1 PE/EtOAc) to afford 6-azido-2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl) benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (308 mg). MS (ESI) m/z 573.2 [M+H]+.

To a solution of 3-bromo-5-methyl-1H-pyrazole (15.2 g, 95.02 mmol) in H2O (250 mL) was added KMnO4(66.0 g, 0.42 mol) at 110° C. in 6 batches. The resulting solution was heated at 110° C. for 4.5 h. The mixture was filtered through celite while it was hot. The pH of filtrate was adjusted to 2 with 4 M aqueous HCl. The solution was extracted with EtOAc, dried over anhydrous Na2SO4and concentrated to afford 3-bromo-1H-pyrazole-5-carboxylic acid (10.2 g) which used directly in the next step. To a solution of 3-bromo-1H-pyrazole-5-carboxylic acid (10.2 g, 53.68 mmol) in EtOH (80 mL) was added SOCl2(16 mL) at 0° C. The resulting solution was heated at reflux for 2 h. The solvent was removed under vacuum and the residue was washed with saturated aqueous NaHCO3, dried over anhydrous Na2SO4and concentrated to give ethyl 3-bromo-1H-pyrazole-5-carboxylate (12.3 g) MS (ESI) m/z 219.1, 221.1 [M+H]+.

A solution of ethyl 3-bromo-1-(4-ethoxy-4-oxobutyl)-1H-pyrazole-5-carboxylate (5.01 g, 15.0 mmol) and t-BuOK (3.38 g, 30.1 mmol) in toluene (80 mL) was stirred at 110° C. for 2 h. The mixture was filtered through celite and concentrated under vacuum to afford ethyl 2-bromo-4-oxo-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridine-5-carboxylate (2.1 g) which was used directly in next step without further purification. MS (ESI) m/z 287.1, 289.1 [M+H]+.

2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-one was prepared from 2-bromo-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-one and 4-chloro-1-isopropyl-5-(4,4,5,5-tetramethyl-1,3 ,2-dioxaborolan-2-yl)-1H-pyrazole according to step 3 of Example 1. MS (ESI) m/z 279.1 [M+H]+.

To a solution of (E)-2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-N-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl) phenyl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-imine (150 mg, 0.29 mmol) in MeOH (5 mL) was added NaBH3CN (91.75 mg, 1.46 mmol). The resulting mixture was stirred at room temperature for 2 h. The reaction was quenched with water, extracted with EtOAc, dried over anhydrous Na2SO4, concentrated and purified by prep-HPLC (condition 2) to afford Example 237 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-N-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-amine (132 mg).

Table 17. The compounds listed in Table 17 were synthesized according to Example 237 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers, when generated, were separated by chiral HPLC and absolute stereochemistries were arbitrarily assigned.

Example 256 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-N-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-α]pyridin-8-amine was prepared from ethyl 3-bromo-1H-1,2,4-triazole-5-carboxylate following step 2 to step 7 of Example 237.

Table 18. The compounds listed in Table 18 were synthesized according to Example 256 using the appropriate commercially available reagents and/or intermediates described above.

To the solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-N-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl) phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-amine (prepared according to Example 237, 30 mg, 0.058 mmol) in dry DMF (2 mL) was added NaH (1.53 mg, 60% in mineral oil, 0.064 mmol) at 0° C. The mixture was stirred at room temperature for 20 min then iodomethane (8.24 mg, 0.058 mmol) was added. The resulting mixture was stirred at room temperature for 1 h then quenched with water concentrated and purified by prep-HPLC (condition 3) to give Example 258 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-N-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)-N-methyl-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-amine (11.5 mg).

Table 19. The compounds listed in Table 19 were synthesized according to Example 258 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-one (150 mg, 0.54 mmol) in MeOH (2 mL) was added NaBH4 (32.8 mg, 1.08 mmol) at 0° C. The resulting mixture was stirred at room temperature for 30 min then quenched with aqueous HCl (1 M, 0.1 mL). The reaction mixture was directly purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to afford 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-ol (121 mg). MS (ESI) m/z 281.2 [M+H]+.

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-ol (120 mg, 0.43 mmol) in dry THF (5 mL) was added dropwise DEAD (113.2 mg, 0.65 mmol) at room temperature. After addition, 4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenol (131.2 mg, 0.52 mmol) and PPh3(170.3 mg, 0.65 mmol) were added. The resulting mixture was stirred at room temperature for 2 h. The solvent was removed and the crude was purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to afford Example 261 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenoxy)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridine (28 mg).

Table 20. The compounds listed in Table 20 were synthesized according to Example 261 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers, when generated, were separated by chiral HPLC and absolute stereochemistries were arbitrarily assigned.

methyl 4′-chloro-2′-isopropyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H,2′H-[3,3′-bipyrazole]-5-carboxylate was synthesized from methyl 3-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-5-carboxylate and 4-chloro-1-isopropyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole following step 3 of Example 1. MS (ESI) m/z 399.2 [M+H]+.

To a solution of methyl 4′-chloro-2′-isopropyl-14(2-(trimethylsilyl)ethoxy)methyl)-1H,2′H-[3,3′-bipyrazole]-5-carboxylate (285 mg, 0.72 mmol) and 2-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl) acetic acid (235 mg, 0.79 mmol) in dry DMF (7 mL) was added NaHMDS (1 M in THF, 2.88 mL, 2.88 mmol) at −10° C. under an atmosphere of N2. The resulting mixture was stirred at −10° C. for 3 h then quenched with saturated aqueous NH4Cl (20 mL). The mixture was extracted with EtOAc (30 mL×3). The combined organic phase were washed with water and brine, dried over anhydrous Na2SO4, concentrated and purified by silica gel chromatography (eluting with 1/5 EtOAc/PE) to afford 1-(4′-chloro-2′-isopropyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H,2′H-[3,3′-bipyrazol]-5-yl)-2-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-one (200 mg). MS (ESI) m/z 621.2 [M+H]+.

1-(4′-chloro-2′-isopropyl-1H,2′H-[3,3′-bipyrazol]-5-yl)-2-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-one was synthesized from 1-(4′-chloro-2′-isopropyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H,2′H-[3,3′-bipyrazol]-5-yl)-2-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-one according to step 4 of Example 149. MS (ESI) m/z 491.1 [M+H]+.

1-(4′-chloro-2′-isopropyl-1H,2′H-[3,3′-bipyrazol]-5-yl)-2-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-ol was synthesized from 1-(4′-chloro-2′-isopropyl-1H,2′H-[3,3′-bipyrazol]-5-yl)-2-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-one according to step 1 of Example 261. MS (ESI) m/z 493.2 [M+H]+.

To a solution of tert-butyl (2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-6-yl) carbonate (690 mg, 1.81 mmol) in DMF (10 mL) was added 2-(4-(bromomethyl)-2-fluorophenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole (1.3 g, 3.62 mmol), NaI (543 mg, 3.62 mmol) and DIPEA (467 mg, 3.62 mmol) at room temperature. The resulting mixture was heated at 60° C. for 16 h. The mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL×3). The combined organic phase were washed with water and brine, dried over anhydrous Na2SO4, concentrated and purified by silica gel chromatography (eluting with 1/1 EtOAc/PE) to afford tert-butyl(2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorobenzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-6-yl) carbonate (770 mg). MS (ESI) m/z 652.2 [M+H]+

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (2 g, 7.17 mmol) in DMF (20 mL) was added 2-(4-(bromomethyl)-2-chlorophenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (2.37 g, 7.17 mmol) and K2CO3(2g, 14.35 mmol). The resulting mixture was heated at 60° C. for 2 h. After cooling to ambient temperature, the mixture was poured into water (150 mL) and extracted with EtOAc (100 mL×3). The organic layers were combined, dried over anhydrous Na2SO4and concentrated under vacuum. The residue was purified by silica gel chromatography (eluting with 1/2 EtOAc/PE) to afford 2-bromo-4-(4-(1-methyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (2.3 g). MS (ESI) m/z 530.2 [M+H]+.

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(3-chloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (80 mg, 0.15 mmol) in a mixture of dioxane (3 mL) and H2O (0.3 mL) was added 2-chloro-3-(difluoromethoxy)pyridine (27.2 mg, 0.15 mmol), Pd(dppf)Cl2(11 mg, 0.015 mmol) and K3PO4(96 mg, 0.45 mmol). The mixture was heated at 110° C. for 1 h in a microwave reactor. After cooling to ambient temperature, the mixture was filtered through celite and the filtrate concentrated under vacuum. The residue was purified by prep-TLC (eluting with 1/1 PE/EtOAc) to afford Example 271 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(3-chloro-4-(3-(difluoromethoxy)pyridin-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (110 mg).

Table 21. The compounds listed in Table 21 were synthesized according to Example 271 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of 2-(4-chloro-1H-pyrazol-1-yl)propan-1-ol (1.7 g, 10.62 mmol) in DCM (45 mL) was added diethylaminosulfur trifluoride (2.56 g, 15.94 mmol) at 0° C. The resulting mixture was stirred at room temperature for 4 days. The mixture was quenched with H2O, concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN in H2O) to afford 4-chloro-1-(1-fluoropropan-2-yl)-1H-pyrazole (730 mg). MS (ESI) m/z 163.2 [M+H]+.

To a solution of 2-(4-chloro-1-(1-fluoropropan-2-yl)-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-one (176 mg, 0.59 mmol) in dry toluene (4 mL) was added 4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)aniline (303.3 mg, 1.19 mmol) and titanium ethoxide (271.1 mg, 1.19 mmol) at room temperature. The resulting mixture was heated at 120° C. for 4 h in a microwave reactor. The mixture was concentrated and purified by Biotage (C18column, eluting with 10% to 90% MeCN in H2O) to afford 2-(4-chloro-1-(1-fluoropropan-2-yl)-1H-pyrazol-5-yl)-N-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-imine (125 mg). MS (ESI) m/z 534.2 [M+H]+.

To a solution of 2-(4-chloro-1-(1-fluoropropan-2-yl)-1H-pyrazol-5-yl)-N-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-imine (125 mg, 0.23 mmol) in MeOH (6 mL) was added NaBH3CN (73.8 mg, 1.17 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. The mixture was quenched with H2O, concentrated and purified by prep-HPLC (condition 3) to afford Example 323 2-(4-chloro-1-((R)-1-fluoropropan-2-yl)-1H-pyrazol-5-yl)-N-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-amine (19.6 mg) as the first eluting diastereomer (arbitrarily assigned stereochemistry). Further elution gave Example 324 2-(4-chloro-1-((S)-1-fluoropropan-2-yl)-1H-pyrazol-5-yl)-N-(4-(1-ethyl-4-(trifluoromethyl)-1 H-imidazol-2-yl)phenyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-amine (20.0 mg) as the second diastereomer with arbitrarily assigned stereochemistry.

Table 22. The compounds listed in Table 22 were synthesized in Example 323. The stereochemistries were arbitrarily assigned.

1-isopropyl-5-(5-oxo-4-((2-(trimethylsilyl)ethoxy)methyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-2-yl)-1H-pyrazole-4-carbonitrile was prepared according to step 1 of Example 266. MS (ESI) m/z 401.2 [M+H]+.

5-(4-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl)-5-oxo-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-2-yl)-1-isopropyl-1H-pyrazole-4-carbonitrile was synthesized following step 4 and step 5 of Example 154. The obtained racemate was purified by chiral chromatography (chiral preparative HPLC condition 2) to afford Example 325 (S)-5-(4-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl)-5-oxo-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-2-yl)-1-isopropyl-1H-pyrazole-4-carbonitrile as the first eluting compound. Further elution gave Example 326 (R)-5-(4-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl)-5-oxo-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-2-yl)-1-isopropyl-1H-pyrazole-4-carbonitrile. The absolute stereochemistries were assigned by comparing retention time on chiral HPLC with similar chiral compounds that were obtained via enantioselective synthesis.

Table 23. The compounds listed in Table 23 were synthesized in Example 325.

To a solution of 2-bromo-4-((2-(trimethylsilyl)ethoxy)methyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (723 mg, 2.1 mmol) in dry THF (10 mL) was added LDA (1 M in THF, 2.1 mL, 4.19 mmol) at −78° C. under an atmosphere of N2. The mixture was stirred at −78° C. for 30 min then methyl iodide (595 mg, 4.19 mmol) was added. The resulting solution was warmed to room temperature and stirred for 1 h then quenched with saturated aqueous NH4Cl (10 mL). The mixture was extracted with EtOAc (10 mL×3). The combined organic phases were washed with water and brine, dried over anhydrous Na2SO4, concentrated and purified by silica gel chromatography (eluting with 1/2 EtOAc/PE) to afford 2-bromo-6-methyl-4-((2-(trimethylsilyl)ethoxy)methyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (500 mg). MS (ESI) m/z 360.1 [M+H]+.

To a solution of 2-bromo-6-methyl-4-((2-(trimethylsilyl)ethoxy)methyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (500 mg, 1.39 mmol) in dry THF (6 mL) was added LDA (1 M in THF, 1.4 mL, 2.79 mmol) at −78° C. under an atmosphere of N2. The mixture was stirred at −78° C. for 30 min then methyl iodide (400 mg, 2.79 mmol) was added. The resulting solution was warmed to room temperature and stirred for 1 h then quenched with saturated aqueous NH4Cl (10 mL). The mixture was extracted with EtOAc (10 mL×3). The combined organic phases were washed with water and brine, dried over anhydrous Na2SO4, concentrated and purified by silica gel chromatography (eluting with 1/2 EtOAc/PE) to afford 2-bromo-6,6-dimethyl-4-((2-(trimethylsilyl)ethoxy) methyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (200 mg). MS (ESI) m/z 374.1 [M+H]+.

To a solution of 2-bromo-6,6-dimethyl-4-((2-(trimethylsilyl)ethoxy)methyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (630 mg, 1.69 mmol) in a mixture of dioxane (15 mL) and H2O (1.5 mL) was added 4-cyclopropyl-6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyrimidine (933 mg, 3.38 mmol), Pd(dppf)Cl2(124 mg, 0.17 mmol) and K3PO4(716 mg, 3.38 mmol) at room temperature. The resulting solution was stirred at 100° C. overnight under an atmosphere of N2. The mixture was filtered through celite, concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN in H2O) to afford 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-6,6-dimethyl-4-((2-(trimethylsilyl)ethoxy)methyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (720 mg). MS (ESI) m/z 444.3 [M+H]+.

A solution of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-6,6-dimethyl (trimethylsilyl)ethoxy)methyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (700 mg, 1.58 mmol) in HCl EtOAc solution (4 M, 15 mL) was stirred at room temperature for 5 h. The solvent was removed and the residue diluted with EtOAc (50 mL), washed with 1M NaOH, dried over anhydrous Na2SO4, concentrated and purified by silica gel chromatography (eluting with 1/2 EtOAc/PE) to afford 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-6,6-dimethyl-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one (180 mg). MS (ESI) m/z 314.2 [M+H]+.

To a solution of 1-isopropyl-1H-pyrazol-4-ol (3.2 g, 25.4 mmol) in MeCN (15 mL) and H2O (15 mL) was added diethyl(bromodifluoromethyl)phosphonate (8.9 mL, 50.7 mmol) and KOH (28.5 g, 508 mmol) at −10° C. The resulting solution was stirred at room temperature overnight. The mixture was extracted with EtOAc (50 mL×3) and the combined organic phases washed with brine, concentrated and purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN in H2O) to afford 4-(difluoromethoxy)-1-isopropyl-1H-pyrazole (704.4 mg). MS (ESI) m/z 177.1 [M+H]+.

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-one (200 mg, 0.72 mmol) in toluene (5 mL) was added 2-methylpropane-2-sulfinamide (260 mg, 2.15 mmol). The resulting solution was stirred at 120° C. for 3 h in a sealed tube. The solvent was removed and the crude was purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to afford ((E)-N-(2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-ylidene)-2-methylpropane-2-sulfinamide (100 mg). MS (ESI) m/z 382.1 [M+H]+.

To a solution of N-(2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-ylidene)-2-methylpropane-2-sulfinamide (100 mg, 0.26 mmol) in MeOH (5 mL) was added NaBH4 (30 mg, 0.78 mmol) at room temperature. The resulting solution was stirred at room temperature for 1 h. Solvent was removed and the crude was purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to afford N-(2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-yl)-2-methylpropane-2-sulfinamide (100 mg). MS (ESI) m/z 384.2 [M+H]+.

A mixture of N-(2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-yl)-2-methylpropane-2-sulfinamide (110 mg, 0.29 mmol) in HCl EtOAc solution (4 M, 5 mL) was stirred at room temperature for 1 h. The reaction solution was concentrated under vacuum to give crude 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-amine (80 mg) which was used directly in next step without further purification. MS (ESI) m/z 280.1 [M+H]+.

To a solution of 2-(4-(1-bromoethyl)-2-fluorophenyl)-1-ethyl-4-(trifluoromethyl)-1H-imidazole (284 mg, 0.79 mmol) in MeCN (5 mL) was added 2-bromopyrazolo[1,5-α]pyrimidin-5(4H)-one (168.3, 0.79 mmol) and Cs2CO3(514.8 mg, 1.58 mmol). The resulting mixture was heated at 80° C. for 2 h. After cooling to ambient temperature, the mixture was filtered through celite and the filtrate was concentrated under vacuum. The residue was purified by silica gel chromatography (eluting with 1/2 EtOAc/PE) to afford 2-bromo-4-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl)pyrazolo[1,5-α]pyrimidin-5(4H)-one (150 mg). MS (ESI) m/z 498.0, 500.0 [M+H]+.

To a solution of 2-bromo-4-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl)pyrazolo[1,5-α]pyrimidin-5(4H)-one (150 mg, 0.30 mmol) in a mixture of dioxane (10 mL) and H2O (1 mL) was added 4-cyclopropyl-6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine (124.2 mg, 0.45 mmol), Pd(dppf)Cl2(36.6 mg, 0.05 mmol) and K3PO4(130.2 mg, 0.60 mmol). The mixture was heated at 110° C. for 2 h under N2in a microwave reactor. After cooling to ambient temperature, the mixture was filtered through celite and the filtrate concentrated under vacuum. The residue was purified by Biotage (C18column, eluting with 10% to 95% MeCN/H2O, containing 0.1% formic acid) to afford 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)ethyl)pyrazolo[1,5-α]pyrimidin-5(4H)-one (132 mg). The racemate was further separated by chiral prep-HPLC (chiral preparative HPLC condition 2) to give Example 334 (S)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorophenyl)-ethyl)pyrazolo[1,5-α]pyrimidin-5(4H)-one (53 mg) as the first eluting compound. Further elution provided Example 333 (R)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(1-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluoro-phenyl)ethyl)pyrazolo[1,5-α]pyrimidin-5(4H)-one (55 mg). The absolute stereochemistries were assigned by comparing retention time on chiral HPLC with similar chiral compounds that were obtained via enantioselective synthetic methods.

Table 24. The compounds listed in Table 24 were synthesized according to Example 333 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers were separated by chiral prep-HPLC and absolute stereochemistries assigned by comparing retention time on chiral HPLC with similar chiral compounds that were obtained via enantioselective synthesis.

To a solution of 5-nitro-1H-1,2,4-triazole (20 g, 0.175 mol) in dry DMF (50 mL) was added Br2(18 mL) slowly over 30 min at room temperature. The resulting solution was heated to 40° C. overnight. After cooling to room temperature, the mixture was neutralized with saturated aqueous NaHCO3and extracted with EtOAc (50 mL×3). The combined organic phases were washed with water and brine, dried over anhydrous Na2SO4and concentrated under vacuum to give crude 3-bromo-5-nitro-1H-1,2,4-triazole (12 g) which was used directly in next step without further purification. MS (ESI) m/z 192.9, 194.9 [M+H]+.

A mixture of 3-bromo-5-nitro-1H-1,2,4-triazole (12 g, 62.5 mmol), iron power (21 g, 0.375 mol) and NH4Cl (26.8 g, 0.5 mol) in a mixture of EtOH (40 mL) and H2O (10 mL) was heated at 80° C. for 4 h. After cooling to room temperature, the reaction mixture was filtered through celite and the filtrate concentrated. The residue was dissolved in MeOH (50 mL) and filtered through a mixture of celite and silica gel. The filtrate was concentrated and dried under vacuum to give crude 3-bromo-1H-1,2,4-triazol-5-amine (5.5 g) which was used directly in next step without further purification. MS (ESI) m/z 163.0, 165.0 [M+H]+.

A mixture of 3-bromo-1H-1,2,4-triazol-5-amine (3 g, 18.5 mmol), ethyl (E)-3-ethoxyacrylate (2.7 g, 18.5 mmol) and K2CO3in MeCN (20 mL) was heated at 95° C. in a sealed tube for 16 h. The reaction mixture was filtered through celite. The residue was washed with MeCN and dried under vacuum to give crude 2-bromo-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one (1.2 g) which was used directly in next step without further purification. MS (ESI) m/z 214.9, 216.9 [M+H]+.

Example 337 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl) benzyl)-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one was synthesized following step 2 and step 3 of Example 1.

Table 25. The compounds listed in Table 25 were synthesized according to Example 337 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers, when generated, were separated by chiral prep-HPLC and absolute stereochemistries were assigned by comparing retention time on chiral HPLC with similar chiral compounds that were obtained via enantioselective synthesis.

Table 26. The compounds listed in Table 26 were synthesized according to Example 347 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers, when generated, were separated by chiral prep-HPLC and absolute stereochemistries stereochemistries were arbitrarily assigned.

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-one (300 mg, 1.08 mmol) in dry THF (10 mL) was added LDA (2 M in THF/hexane, 1.1 mL, 2.2 mmol) dropwise at −78° C. After stirring for 30 min, MeI (382 mg, 2.69 mmol) was added. The resulting mixture was allowed to warm to room temperature and stirred for 1 h then quenched with saturated aqueous NH4Cl and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, concentrated under vacuum and purified by prep-TLC (eluting with 1/2 EtOAc/PE) to afford 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-5,5-dimethyl-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-one (40 mg). MS (ESI) m/z 307.1 [M+H]+.

To a stirred solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorobenzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-6-ol (420 mg, 0.76 mmol) in dry DCM (20 mL) was added TEA (154 mg, 1.52 mmol), tosyl chloride (290.6 mg, 1.52 mmol) and DMAP (18.6 mg, 0.15 mmol). The resulting mixture was stirred at room temperature for 3 h then diluted with water and extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4and concentrated under vacuum to give crude 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorobenzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-6-yl 4-methylbenzenesulfonate (505 mg). MS (ESI) m/z 706.2 [M+H]+.

A mixture of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorobenzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidin-6-yl 4-methylbenzenesulfonate (500 mg, 0.71 mmol) and NaN3(92.2 mg, 1.42 mmol) in DMF (5 mL) was heated at 100° C. for 2 h. After cooling to room temperature, the resulting mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine, dried over Na2SO4, concentrated under vacuum and purified by column chromatography on silica gel (eluting with 1/1 PE/EtOAc) to give 6-azido-2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)-3-fluorobenzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrimidine (200 mg). MS (ESI) m/z 577.2 [M+H]+.

To a solution of 1-(4′-chloro-2′-isopropyl-1H,2′H-[3,3′-bipyrazol]-5-yl)-2-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-amine (901.0 mg, 1.8 mmol) in DCM (10 mL) was added TEA (554.5 mg, 5.5 mmol) and (Boc)20 (797.9 mg, 3.7 mmol). The mixture was stirred at room temperature for 1 h. The solvent was removed under vacuum and the crude product was purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O) to give tert-butyl(1-(4′-chloro-2′-isopropyl-1H,2′H-[3,3′-bipyrazol]-5-yl)-2-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)carbamate (591.0 mg). MS (ESI) m/z 592.2 [M+H]+.

To a solution of tert-butyl (1-(4′-chloro-2′-isopropyl-1H,2′H-[3,3′-bipyrazol]-5-yl)-2-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)carbamate (591.2 mg, 1.0 mmol) in DMF (5 mL) was added 1,2-dibromoethane (557.6 mg, 3.0 mmol) and K2CO3(276.0 mg 2.0 mmol). The mixture was heated in a sealed tube at 100° C. overnight. The resulting mixture was diluted with water (20 mL) and extracted with EtOAc (3×30 mL). The combined organic phases were washed with brine, dried over anhydrous Na2SO4and concentrated under vacuum to afford crude tert-butyl 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrazine-5(4H)-carboxylate (300 mg) which was used directly in next step without further purification. MS (ESI) m/z 618.2 [M+H]+.

To a solution of tert-butyl 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydropyrazolo[1,5-α]pyrazine-5(4H)-carboxylate (300 mg, 0.48 mmol) in DCM (2 mL) was added TFA (2 mL). The mixture was stirred at room temperature for 1 h then concentrated under vacuum. The crude product was purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O, containing 0.1% trifluoroacetic acid) and neutralized with 1M aqueous NaOH to give Example 359 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrazine (100 mg).

To a solution of 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrazine (100 mg, 0.19 mmol) in MeOH (3 mL) was added paraformaldehyde (90 mg, 1.0 mmol) and 1 drop of acetic acid at room temperature. The mixture was stirred at room temperature for 30 minutes then NaBH3CN (62.8 mg, 1.0 mmol) was added. The resulting mixture was stirred at room temperature for 1 h then quenched with water. The solvent was removed under vacuum and the crude product purified on a Biotage Isolera One (C18column, eluting with 10% to 90% MeCN/H2O, containing 0.1% trifluoroacetic acid) and neutralized with 1M aqueous NaOH to give Example 360 2-(4-chloro-1-isopropyl-1H-pyrazol-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-5-methyl-4,5,6,7-tetrahydropyrazolo[1,5-α]pyrazine (6.4 mg).

Table 27. The compounds listed in Table 27 were synthesized according to Example 359 and Example 360 using the appropriate commercially available reagents and/or intermediates described above.

Example 363. Enantioselective synthesis of (R)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethyl)-6,7-dihydropyrazolo[1,5-α]pyrimidin-5(4H)-one

To a solution of borane-methyl sulfide complex (2 M in THF, 0.5 mL, 1 mmol) in dry THF (1 mL) was added (S)-3,3-Diphenyl-1-methylpyrrolidino[1,2-c]-1,3,2-oxazaborole (1 M in toluene, 0.5 mL, 0.5 mmol) at −20° C. The solution was stirred at the same temperature for 30 min then 1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-one (100 mg, 0.34 mmol, prepared in Intermediate BB-54) in dry THF (1 mL) was added. The resulting mixture was stirred at −20° C. for 2 h then warmed to room temperature, quenched with water (10 mL) and extracted with EtOAc (10 mL×3). The combined organic layers were dried over anhydrous Na2SO4, concentrated under vacuum and purified by prep-TLC (eluting with 1/3 EtOAc/PE) to afford (R)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-ol (60 mg, 98% ee). The ee value was determined by chiral HPLC using a Superchiral S-OJ (0.46×15 cm) column and eluting with 90/10 hexane/ethanol containing 0.5% diethylamine at a flow of 0.9 mL/min at room temperature. MS (ESI) m/z 299.1 [M+H]+.

To a solution of (R)-1-(4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-ol (60 mg, 0.2 mmol) in dry DCM (3 mL) was added PBr3(108 mg, 0.4 mmol) at 0° C. The resulting mixture was stirred at same temperature for 2 h then quenched with saturated aqueous NaHCO3(5 mL) and extracted with DCM (5 mL×3). The combined organic layers were dried over anhydrous Na2SO4, concentrated under vacuum and purified by prep-TLC (eluting with 1/5 EtOAc/PE) to afford (S)-2-(4-(1-bromoethyl)phenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (30 mg). MS (ESI) m/z 361.0, 363.0 [M+H]+.

Example 364 2-(4-cyclopropyl-6-(difluoromethoxy)pyrimidin-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6,7-dihydro-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one was synthesized from 3-bromo-1H-1,2,4-triazol-5-amine (prepared in Example 337) and ethyl acrylate following step 1 to step 3 of Example 347.

Table 28. The compounds listed in Table 28 were synthesized according to Example 364 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of (3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)methanol (1 g, 3.3 mmol, synthesized in step 2 of Intermediate BB-6) in dry DCM (10 mL) at 0° C. was added pyridinium chlorochromate (815 mg, 3.8 mmol). The resulting mixture was stirred at room temperature for 4 h. The reaction was quenched with saturated aqueous sodium thiosulfate (10 mL) and extracted with DCM (10 mL×2). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, concentrated and purified by column chromatography on silica gel (eluting with 1/10 EtOAc/PE) to give 3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde (930 mg). MS (ESI) m/z 301.1 [M+H]+.

To a stirred solution of 3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzaldehyde (450 mg, 1.5 mmol) in dry THF (6 mL) at 0° C. was added TMSCF3(415 mg, 2.9 mmol) and TBAF (1 M in THF, 0.06 mL). The resulting mixture was stirred at room temperature for 2 h. The reaction mixture was then acidified with aqueous HCl (3 M, 15 mL) and extracted with EtOAc (30 mL×2). The combined organic phases were washed with brine, dried over anhydrous Na2SO4and concentrated to give a white solid. The solid was recrystallized in EtOAc/hexane and dried under vacuum to give 2,2,2-trifluoro-1-(3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-ol (294 mg). MS (ESI) m/z 371.0 [M+H]+.

To a solution of 2,2,2-trifluoro-1-(3-fluoro-4-(1-isopropyl-4-(trifluoromethyl)-1H-imidazol-2-yl)phenyl)ethan-1-ol (200 mg, 0.54 mmol) and triphenyl phosphite (167 mg, 0.54 mmol) at 0° C. under an atmosphere of N2was added NBS (141 mg, 0.79 mmol). The resulting mixture was stirred at room temperature for 12 h then diluted with water (20 mL) and extracted with EtOAc (25 mL×2). The combined organic phases were washed with brine, dried over anhydrous Na2SO4, concentrated and purified by column chromatography on silica gel (eluting with 1/20 EtOAc/PE) to give 2-(4-(1-bromo-2,2,2-trifluoroethyl)-2-fluorophenyl)-1-isopropyl-4-(trifluoromethyl)-1H-imidazole (178 mg). MS (ESI) m/z 433.0, 435.0 [M+H]+.

6-chloropyrimidin-4-ol (5g, 38.3 mmol)was dissolved in a solution of sodium methoxide methanol (5 M, 30 mL) and heated at 90° C. in a sealed tube. After cooling to room temperature, solvent was removed and the crude product was purified by column chromatrography on silica gel (eluting with 1/3 EtOAc/PE) to give 6-methoxypyrimidin-4-ol (3.6 g). MS (ESI) m/z 127.0 [M+H]+.

4-(1-(ethyl-d5)-4-(trifluoromethyl)-1H-imidazol-2-yl)aniline was prepared according to

Table 29. The compounds listed in Table 29 were synthesized according to Example 364 using the appropriate commercially available reagents and/or intermediates described above.

Table 30. The compounds listed in Table 30 were synthesized according to Example 347 using the appropriate commercially available reagents and/or intermediates described above. Enantiomers, when generated, were separated by chiral prep-HPLC and absolute stereochemistries were arbitrarily assigned.

Table 31. The compounds listed in Table 31 were synthesized according to Example 337 using the appropriate commercially available reagents and/or intermediates described above.

Table 32. The compounds listed in Table 32 were synthesized according to Example 237 using the appropriate commercially available reagents and/or intermediates described above.

2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4-(4-(1-ethyl-4-(trifluoromethyl)-1H-imidazol-2-yl)benzyl)-6-methyl-[1,2,4]triazolo[1,5-α]pyrimidin-5(4H)-one was prepared from 3-bromo-1H-1,2,4-triazol-5-amine and ethyl (E)-3-ethoxy-2-methylacrylate following step 3 to step 5 of Example 337.

Table 33. The compounds listed in Table 33 were synthesized according to Example 392 using the appropriate commercially available reagents and/or intermediates described above.

To a solution of borane-methyl sulfide complex (2 M in THF, 5.6 mL, 11.2 mmol) in dry THF (19 mL) at −15° C. was added (S)-3,3-Diphenyl-1-methylpyrrolidino[1,2-c]-1,3,2-oxazaborole (1 M in toluene, 5.6 mL, 5.6 mmol). The solution was stirred for 30 min then a solution of 2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-6,7-dihydropyrazolo[1,5-α]pyridin-4(5H)-one (800 mg, 3.72 mmol) in dry THF (1 mL) was added. The resulting mixture was stirred at −15° C. for 1 h then warmed to room temperature, quenched with water (50 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were dried over anhydrous Na2SO4, concentrated under vacuum and purified by column chromatrography on silica gel (eluting with 1/4 EtOAc/PE) to afford (R)-2-(4-cyclopropyl-6-methoxypyrimidin-5-yl)-4,5,6,7-tetrahydropyrazolo[1,5-α]pyridin-4-ol (580 mg). MS (ESI) m/z 287.1 [M+H]+.

Deubiquitination Assay for USP1/UAF1 and USP1 ES2 Activity and Inhibitor Testing

Certain Compounds of the Disclosure were assessed for USP1/UAF1 activity in a Ubiquitin Rhodamine assay modified from those described previously.

Deubiquitinase activity was measured using ubiquitin-rhodamine 110 as a substrate. Cleavage of the amide bond between rhodamine and the c-terminal glycine of ubiquitin yields an increase in fluorescence signal. The assay was conducted in 20 μl total volume of assay buffer (50 mM Tris-HCl, pH 7.8, 0.5 mM EDTA, 0.01% Bovine Serum Albumin, 1 mM DTT, 0.01% Tween-20), and 0.05 nM USP1/UAF1 enzyme. Reaction was initiated by addition of 150 nM Ubiquitin-rhodamine (Boston Biochem) substrate.

Compounds of the Disclosure, dissolved in DMSO were tested in dose response format, beginning at 10 04.

The example compounds listed below were added to enzyme/assay buffer mix and incubated 10 min. Substrate mix was added, and reaction mix was read in kinetic mode for 30 min at Ex480/Em540 and IC50response curves were plotted.

Data for all assay formats was calculated as percent inhibition compared with control wells. Percent inhibition was calculated using the following equation: % inhibition=100×[1−(X−min)/(max−min)], where X is the raw data readout, min is the average of the no enzyme control wells (n=32), max is the average of the DMSO control well (n=32). IC50values were calculated using the standard four parameter curve fitting algorithm in either Prism GraphPad (La Jolla, Calif.) software, or Collaborative Drug Discovery (Burlingame, Calif.) CDD Vault. See Chem. Biol. 20(1): 55-62 (Jan. 24, 2013); Bioorg. Med. Chem. Lett. 23(20): 5660-5666 (Oct. 15, 2013).

USP1 is a deubiquitinating enzyme that removes ubiquitin from mono-ubiquitinated

Proliferating Cell Nuclear Antigen (Ub-PCNA). The levels of Ub-PCNA in the nucleus of cells was used to assess the activity of USP1. An immunofluorescence assay was established to monitor Ub-PCNA levels in the ovarian cancer cell line, ES2.

This assay was performed by first plating 5000 ES2 cells per well in black 96 well plates (Corning #3904) and then incubated overnight at 37° C. and 5% CO2. Compounds, resuspended in DMSO, were added to the cells, to a final DMSO concentration of 0.3%. Plates were incubated at 37° C. and 5% CO2for 3 hours.

Cells were then fixed and stained by first removing the media from each plate and fixing the cells with −20° C. methanol for 5 minutes at room temperature. Following fixation plates were washed with Tris buffered saline with tween (Boston Bioproducts ABB-855) 5 times for 5 minutes each. Plates were blocked for 1 hour with 50μ.1 of Odyssey blocking buffer (Licor #327-50000) at room temperature with rocking. Block was removed from all wells and 50 μl of primary antibody was added to each well. Ub-PCNA ab (Cell Signaling Technology #13439) was diluted at 1:400 in Odyssey blocking buffer. Plates were sealed and incubated overnight at 4° C. Primary antibodies were removed from the plates and plates were washed with Tris buffered saline with tween 5 times for 5 minutes. Plates were stained for 1 hour at room temperature with rocking with 50 μl of secondary antibodies diluted 1:10,000 in Odyssey blocking buffer at room temperature (anti-rabbit Alexa 488). Antibodies were removed from the plates and plates were washed with Tris buffered saline with tween 3 times for 5 minutes. Plates were washed once with DAPI (Chemometec # 910-3012) stain diluted 1:5000 (Stock solution of 500 μg/ml) in tris buffered saline with tween for 5 minutes. DAPI stain was removed and plates were washed one additional time for 5 minutes with tris buffered saline with tween. Wash was removed from the plate and 100 μl tris buffered saline with tween was added back to all wells, Plates were sealed with Foil seals or black plate seals and plates were stored at 4° C. until they could be imaged.

Imaging of the plates was performed by first locating the nuclei using the DAPI stain. A mask was created by drawing a circle slightly smaller than each nucleus. Ub-PCNA intensities were measured for each individual nucleus counted. A histogram of all Ub-PCNA nuclear intensities measured in untreated wells was generated and a 95% cutoff was established. This 95% cutoff was used to determine the number of Positive cells that had Ub-PCNA values higher than the 95% cutoff. The number of positive cells was expressed as a percentage by dividing by the total number of cells in the well. This percent positive value was used to graph all data and determine AC50values.

The following Compounds of the Disclosure inhibit USP1 activity with the IC50values shown in Table 34 below.

Solubility Determination

Certain Compounds of the Disclosure were assessed for ADME solubility at pH 2.0 and pH 7.4.

Stock solutions were prepared by adding each compound to DMSO at a concentration of 10 mM. Samples were prepared by adding 50 μL of each stock solution to separate vials. The vials were loaded onto a 96-well rack and dried. 500 μL of Phosphate Buffered Saline (PBS) pH 7.4 or PBS pH 2.0 were added into each vial. The vials were then shaken at 25° C. and 1,100 rpm for 24 hours.

After 24 hours, the vials were centrifuged at 3220 G and 25° C. for 30 minutes. The supernatant fluid was analyzed by LC-MS/MS against a standard of known concentration. The solubility of each sample was then calculated using the equation below:

where DF is the dilution factor.

The following Compounds of the Disclosure have the ADME solubility values shown in

Liver Microsomal Stability

Certain Compounds of the Disclosure were assessed for ADME metabolic stability in human liver microsomes (HLM) and rat liver microsomes (RLM).

Samples were prepared by adding 222.5 μL of a master solution (100 mM phosphate buffer and 1 mg/mL liver microsomes (HLM or RLM)) and 25 μL of a 10 mM NADPH solution to incubation plates, which were then warmed for 10 min. Each compound was separately dissolved in DMSO to prepare 10 mM stock solutions, which were then diluted to 100 μM with acetonitrile. A reaction was started by adding 2.5 μL of the 100 μM solution of each compound to separate incubation plates such that the final concentration of for each compound in each plate was 1 μM.

25 μL aliquots of each sample were taken at 0.5, 5, 10, 15, 20 and 30 minutes, and the reaction was stopped by adding 5 volumes of cold acetonitrile with IS (100 nM alprazolam, 200 nM caffeine and 100 nM tolbutamide). The samples were then centrifuged at 3,220 G for 30 minutes, and 100 μL of the supernatant fluid was mixed with 100 μL of ultra-pure H2O.

The samples were then analyzed by LC-MS/MS. Peak areas were determined from extracted ion chromatograms. Slope values (k) were determined by linear regression of the natural logarithm of the remaining percentage of the compound vs. incubation time curve. The in vitro half-life (in vitro t1/2) was determined from the slope value using the following equation:

in vitrot1/2=−(0.693/k).

The in vitro half-life (min) was converted into the in vitro intrinsic clearance (in vitro CLint, in μL/min/mg protein) using the following equation:

The following Compounds of the Disclosure have the ADME metabolic stability values shown in Table 37 below.

Having now fully described this invention, it will be understood by those of ordinary skill in the art that the same can be performed within a wide and equivalent range of conditions, formulations, and other parameters without affecting the scope of the invention or any embodiment thereof.

All patents and publications cited herein are fully incorporated by reference herein in their entirety.