Compositions and methods for treating kit and pdgfra mediated diseases

Abstract

The present disclosure provides compounds of Formula (I-0), pharmaceutically acceptable salts thereof, and / or solvates of any of the foregoing, for use in the treatment of diseases and disorders associated with mutant KIT and PDGFRa, and exhibit advantageous non-brain penetration properties for the treatment of diseases and disorders associated with mutant KIT and PDGFRa. The present disclosure also provides methods for the treatment of gastrointestinal stromal tumors and systemic mastocytosis.

Description

[0001] This application claims priority to U.S. Provisional Application No. 63 / 091,486, filed October 14, 2020. The entire contents of the aforementioned application are incorporated herein by reference.

[0002] This disclosure relates to novel compounds and their use as selective inhibitors of activated KIT and PDGFRα mutant protein kinases. The compounds disclosed herein can be used, for example, in pharmaceutical compositions for treating chronic conditions. The KIT receptor belongs to the class III receptor tyrosine kinase family, which also includes the structurally associated protein PDGFRα. Normally, stem cell factors bind to and activate KIT by inducing dimerization and autophosphorylation, thereby initiating downstream signaling. However, in some tumor types, somatic activation mutations in KIT drive ligand-independent constitutive oncogenic activity, including acute myeloid leukemia, melanoma, intracranial germ cell tumors, mediastinal B-cell lymphoma, seminoma, and gastrointestinal stromal tumors. Mutant KIT is also known to play a role in mast cell activation, which is common and likely essential for maintenance. Mast cell activation syndrome occurs when mast cells are pathologically overproduced or if their activation is disproportionate to perceived homeostatic threats. Mast cell activation syndrome refers to a group of symptoms caused by multiple factors, presenting as episodic, multisystemic symptoms resulting from the release of mast cell mediators. One type of mast cell activation syndrome is mastocytosis. The World Health Organization (WHO) classifies mastocytosis into seven different categories: cutaneous mastocytosis, indolent systemic mastocytosis (ISM), smoldering systemic mastocytosis (SSM), mastocytosis associated with hematologic malignancies (SM-AHN), aggressive systemic mastocytosis (ASM), mast cell leukemia (MCL), and mast cell sarcoma.

[0003] Systemic mastocytosis (SM) is a clonal disease of mast cells characterized by an increased mast cell burden, with focal and / or diffuse infiltration of tumor mast cells in the skin, bone marrow, spleen, liver, gastrointestinal tract, and other organs, and increased release of mast cell mediators. SM includes five subtypes of mastocytosis: indolent SM (ISM), smoldering SM (SSM), SM associated with non-MC lineage hematologic malignancies (SM-AHN), aggressive SM (ASM), and MC leukemia (MCL). The latter three subtypes are associated with reduced overall survival and are grouped as advanced SM (AdvSM). ISM is a chronic disease associated with normal or near-normal life expectancy, while SSM has an intermediate prognosis. ISM and SSM are combined as non-advSM.

[0004] In all subtypes of SM and in most patients with the disease, tumor mast cells exhibit a mutation at exon 17, D816, of KIT, leading to ligand-independent activation of KIT kinase activity. Differentiation and survival of wild-type mast cells require KIT activity; therefore, constitutive activation of KIT via the D816V mutation is considered a pathogenic driver of SM. Specifically, the KIT D816V mutation is found in 90% to 98% of SM patients, and rare KIT D816Y, D816F, and D816H variants have been identified. Based on these findings, KIT D816V is considered a major therapeutic target for SM.

[0005] Chronic indolent spondylitis (ISM) and supra-spondylitis (SSM) are characterized by severe symptoms, including itching, flushing, GI cramps, diarrhea, allergic reactions, bone pain, and osteoporosis. These symptoms can lead to severe debility and negatively impact quality of life. Currently, there are no approved treatments for ISM or SSM. Therefore, discovering new treatments targeting ISM or SSM would be beneficial.

[0006] Compounds exhibiting mutagenic KIT and PDGFRα inhibitory activity have been described in WO2015 / 057873, CN108191874, and WO2019 / 034128. The chemical structures of compounds known in the art differ from those of the compounds disclosed herein.

[0007] Furthermore, although compounds with mutagenic KIT and PDGFRα inhibitory activities have been disclosed in the art, the properties of these known compounds are completely different from those of the compounds disclosed herein.

[0008] The purpose of this disclosure is to provide novel compounds with highly selective and potent activity against mutant KIT and PDGFRα kinases for the safe and effective treatment of chronic conditions such as ISM and SSM, as well as other diseases mediated by mutant KIT or PDGFRα. Any new therapy should be well-tolerated when treating these conditions, especially chronic ones such as ISM and SSM. In particular, there is a need for novel compounds that target mutant KIT and PDGFRα kinases and reduce the levels of adverse CNS side effects associated with other known KIT and PDGFRα inhibitors.

[0009] The inventors have discovered novel compounds with high selectivity and potency against mutant KIT and PDGFRα kinases, while simultaneously possessing other desired properties, such as little or no penetration into the CNS, low unbound concentrations in the brain, and high levels or active transport out of the brain, i.e., high efflux rates from the CNS. Given this desired balance of properties, the compounds of this disclosure are particularly suitable for peripheral therapy, especially chronic peripheral therapy, while reducing or minimizing side effects in the CNS.

[0010] Therefore, the compounds disclosed herein are intended to provide a treatment with the desired efficacy, safety, and pharmaceutical properties for treating KIT and PDGFRA-mediated diseases. More specifically, compared to compounds known in the art that have mutagenic KIT and PDGFRα inhibitory activity, the compounds disclosed herein exhibit a range of advantageous properties, including reduced brain penetration levels, while maintaining efficacy and other desired pharmaceutical properties.

[0011] Abbreviations and Definitions

[0012] The following abbreviations and terms have their designated meanings throughout the text:

[0013] The term “KIT” refers to human tyrosine kinase, which may be referred to as mast / stem cell growth factor receptor (SCFR), proto-oncogene c-KIT, tyrosine-protein kinase Kit, or CD117. As used herein, the term “KIT nucleotide” includes the KIT gene, KIT mRNA, KIT cDNA and their amplified products, mutants, variants, and fragments. “KIT gene” is used to refer to a gene encoding a polypeptide with KIT kinase activity, for example, whose sequence is located between nucleotides 55,524,085 and 55,606,881 on chromosome 4 of the reference human genome hg19. “KIT transcript” refers to the transcript of the KIT gene, one example having the sequence of the NCBI reference sequence NM_000222.2. The term “KIT protein” refers to a polypeptide sequence produced by the translation of a KIT nucleotide or a portion thereof.

[0014] The term “PDGFRA” refers to human tyrosine kinase, also known as platelet-derived growth factor α. As used herein, the term “PDGFRA nucleotide” includes the PDGFRA gene, PDGFRA mRNA, KIT cDNA and their amplification products, mutants, variants, and fragments. “PDGFRA gene” is used to refer to a gene encoding a polypeptide with PDGFRA kinase activity, for example, whose sequence is located between nucleotides 54,229,089 and 54,298,247 on chromosome 4 of reference Homo sapiens annotated version 109, GRCh38.p12. “PDGFRA transcript” refers to the transcript of the PDGFRA gene, one instance of which has the sequence of NCBI reference sequence NM_006206.6. The terms “PDGFRA protein” or “PDGFRα” refer to the polypeptide sequence produced by the translation of PDGFRA nucleotides or portions thereof.

[0015] As used herein, "malignant disease" refers to a disease in which abnormal cell division is uncontrolled and can invade nearby tissues. Malignant cells can also spread to other parts of the body via the blood or lymphatic system. Non-limiting examples of malignant diseases are carcinoma, sarcoma, leukemia, and lymphoma. Cancer is a non-limiting example of a malignant disease. In some implementations, systemic mastocytosis is a non-limiting example of a malignant disease.

[0016] Non-limiting examples of cancer include gastrointestinal tumors (GIST), AML (acute myeloid leukemia), melanoma, seminoma, intracranial germ cell tumors, and mediastinal B-cell lymphoma.

[0017] As used herein, "eosinophilic disorder" refers to a condition in which an elevated number of eosinophils are found in various parts of the body and / or there is an elevated ratio of low-density to normal eosinophils (e.g., greater than 30%). The eosinophilic disorder described herein is characterized by an excess of eosinophils (eosinophilia). This increase in eosinophil count causes inflammation of tissues and leads to organ damage. The heart, lungs, skin, and nervous system are most commonly affected, but any organ can be damaged.

[0018] Diagnosing eosinophilia based on the location of elevated eosinophil levels:

[0019] Eosinophilic pneumonia (lung)

[0020] Eosinophilic cardiomyopathy (heart)

[0021] Eosinophilic esophagitis (esophageal EoE)

[0022] Eosinophilic gastritis (Gastric EG)

[0023] Eosinophilic gastroenteritis (stomach and small intestine - E GE)

[0024] Eosinophilic enteritis (small intestine)

[0025] Eosinophilic colitis (colon-EC)

[0026] Eosinophilia (blood and any organ - HES)

[0027] As used herein, the terms "subject" or "patient" refer to an organism treated by the methods of this disclosure. Such organisms include, but are not limited to, mammals (e.g., rodents, apes, equines, bovines, suidae, canines, felines, etc.), and in some embodiments, humans.

[0028] As used herein, the phrase “therapeutic effective amount” refers to an amount of active agent sufficient to achieve a beneficial or desired outcome. Therapeutic effective amounts may be administered in one or more applications, by application or by dose, and are not intended to be limited to a particular formulation or route of administration.

[0029] As used in this article, the phrase “the weight of its pharmaceutically acceptable salt eq” for a particular compound includes the weight of both the compound and the associated salt.

[0030] As used in this article, the phrase “its pharmaceutically acceptable salt”, when used in the context of an active agent distributed in salt form, refers to any pharmaceutically acceptable salt form of the active agent.

[0031] As used in this article, the term “treatment” includes any effect that results in the improvement or amelioration of symptoms of a condition, disease, disorder, etc., such as reducing, lowering, regulating, improving, or eliminating them.

[0032] While the active agent can be administered alone, in some embodiments, it can be administered as a pharmaceutical formulation in combination with one or more pharmaceutically acceptable excipients or carriers. For example, the active agent can be formulated for administration in any convenient manner for human or veterinary use. In some embodiments, the compound contained in the pharmaceutical formulation may be active itself, or may be a prodrug, such as one capable of being converted to the active compound in a physiological environment.

[0033] As used herein, the phrase “pharmaceutically acceptable” means those compounds, materials, compositions, and / or dosage forms that are suitable for contact with human and animal tissues within the bounds of reasonable medical judgment and without excessive toxicity, irritation, allergic reactions, or other problems or complications, and that are commensurate with a reasonable benefit / risk ratio.

[0034] As used in this paper, when an expression (e.g., m, n, etc.) appears more than once in any structure, the definition of each expression is intended to distinguish it from its definition elsewhere in the same structure.

[0035] Certain compounds disclosed herein may exist in specific geometric or stereoisomeric forms. This invention covers all such compounds, including cis and trans isomers, R- and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures thereof, and other mixtures thereof, as within the scope of this invention. Additional asymmetric carbon atoms may be present in the substituents. All such isomers and mixtures thereof are included in this disclosure.

[0036] For example, if a specific enantiomer of the compound of this disclosure is required, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary agent, followed by separation of the resulting diastereomer mixture and cleavage of the auxiliary group to obtain the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), a diastereomer salt is formed with a suitable optically active acid or base, followed by resolution of the diastereomer formed by fractional crystallization or chromatographic methods well known in the art, and then recovery of the pure enantiomer.

[0037] Unless otherwise stated, when a disclosed compound is named or described by a structure without specifying stereochemistry and has one or more chiral centers, it should be understood to represent all possible stereoisomers of the compound and mixtures of their enantiomers.

[0038] The “enantiomer excess” or “% enantiomer excess” of the composition can be calculated using the equation shown below. In the example shown below, the composition contains 90% of one enantiomer (e.g., the S enantiomer) and 10% of another enantiomer (i.e., the R enantiomer).

[0039] ee = (90-10) / 100 = 80%.

[0040] Therefore, a composition containing 90% of one enantiomer and 10% of the other enantiomer is considered to have an 80% enantiomer excess.

[0041] The compounds or compositions described herein may contain at least 50%, 75%, 90%, 95%, or 99% enantiomer excess in one form of the compound, for example, the S-enantiomer. That is, such compounds or compositions contain an enantiomer excess of more than the R-enantiomer of the S-enantiomer.

[0042] In one embodiment, the compounds described herein may also contain non-natural proportions of deuterium at one or more atoms constituting such compounds, and all tautomer forms of the compounds described herein are intended to be included within the scope of this disclosure.

[0043] The compounds disclosed herein can be used as free bases or salts. Representative salts include hydrobromide, hydrochloride, sulfate, hydrogen sulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthalate, methanesulfonate, glucono-p-oxophosphate, lacturonate, and laurylsulfonate, etc. (See, for example, Berge et al., (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

[0044] Some of the compounds disclosed herein can exist in both non-solventized and solvated forms (including hydrated forms). As used herein, the terms "hydrate" or "hydrated" refer to compounds formed by the combination of water with a parent compound.

[0045] Generally, the solvated form is equivalent to the unsolvated form and is covered within the scope of this disclosure. Some of the compounds disclosed herein may exist in various crystalline or amorphous forms. Generally, all physical forms are equivalent for the intended use of this disclosure and are intended to be covered within the scope of this disclosure.

[0046] This disclosure provides compounds of formula (I-0):

[0047]

[0048] Its pharmaceutically acceptable salt or stereoisomer, and / or solvate of any of the foregoing, wherein:

[0049] Selected from single and double bonds;

[0050] Selected from single and double bonds;

[0051] Z is selected from CH and NH;

[0052] Y is selected from C and N;

[0053] X1 is selected from CH, C, and N;

[0054] X2 is selected from CH, C, and N;

[0055] The condition is that when X1 and X2 are both N, then Y is not N and Z is not CH;

[0056] A is

[0057] R1 is selected from hydrogen and methyl;

[0058] R2 is selected from hydrogen and methyl, or

[0059] R1 and R2 together form a cyclopropyl group;

[0060] R3 is selected from hydrogen and methyl;

[0061] R4 is selected from hydrogen and methyl, or

[0062] R3 and R4 together form a cyclopropyl group;

[0063] R5 is selected from hydrogen and methyl;

[0064] R6 is selected from hydrogen and methyl, or

[0065] R5 and R6 together form a cyclopropyl group, or

[0066] One of R2 or R4 together with R6 forms a cyclobutyl group;

[0067] R7 is hydrogen, or one of R2, R4 or R6 together with R7 to form a ring selected from oxetane, tetrahydrofuran and tetrahydropyran, wherein the tetrahydrofuran or tetrahydropyran is optionally substituted with a hydroxyl group;

[0068] m is 0 or 1;

[0069] n is 0 or 1; and

[0070] B is selected from OH and NH2, provided that the compound is not...

[0071] Alternatively, R7 is hydrogen, or one of R2, R4, or R6 together with R7 forms a ring selected from tetrahydrofuran and tetrahydropyran, wherein the tetrahydrofuran or tetrahydropyran is optionally substituted with a hydroxyl group.

[0072] The non-restrictive embodiments disclosed herein include:

[0073] Implementation Plan 1. Compounds of formula (I):

[0074]

[0075] Its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein:

[0076] Selected from single and double bonds;

[0077] Selected from single and double bonds;

[0078] Z is selected from CH and NH;

[0079] Y is selected from C and N;

[0080] X1 is selected from CH, C, and N;

[0081] X2 is selected from CH, C, and N;

[0082] The condition is that when X1 and X2 are both N, then Y is not N and Z is not CH;

[0083] A is

[0084] R1 is selected from hydrogen and methyl;

[0085] R2 is selected from hydrogen and methyl, or

[0086] R1 and R2 together form a cyclopropyl group;

[0087] R3 is selected from hydrogen and methyl;

[0088] R4 is selected from hydrogen and methyl, or

[0089] R3 and R4 together form a cyclopropyl group;

[0090] R5 is selected from hydrogen and methyl;

[0091] R6 is selected from hydrogen and methyl, or

[0092] R5 and R6 together form a cyclopropyl group, or

[0093] One of R2 or R4 together with R6 forms a cyclobutyl group;

[0094] R7 is hydrogen, or one of R2, R4 or R6 together with R7 to form a ring selected from oxetane, tetrahydrofuran and tetrahydropyran, wherein the tetrahydrofuran or tetrahydropyran is optionally substituted with a hydroxyl group;

[0095] m is 0 or 1;

[0096] n is 0 or 1; and

[0097] B is selected from OH and NH2.

[0098] Alternatively, R7 is hydrogen, or one of R2, R4, or R6 together with R7 forms a ring selected from tetrahydrofuran and tetrahydropyran, wherein the tetrahydrofuran or tetrahydropyran is optionally substituted with a hydroxyl group.

[0099] In one implementation, the compound of formula (I) is not The S-isomer.

[0100] In some implementations of Implementation Scheme 1, R1 and R2 do not exist when m is 0. In some implementations of Implementation Scheme 1, R3 and R4 do not exist when n is 0. In some implementations of Implementation Scheme 1, m + n = 1 or m and n cannot both be 0.

[0101] It should be noted that in this disclosure, when any two R groups (e.g., R1 and R2) together form a ring structure (e.g., cyclopropyl), it is intended to include the insertion of carbon and / or oxygen atoms into the same ring structure.

[0102] Implementation Plan 2. Compound of embodiment 1 of formula (II):

[0103]

[0104] Its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing. The definitions of variables A and B are provided in equation (I-0) or equation (I).

[0105] Implementation Plan 3. Compound of embodiment 1 of formula (III):

[0106]

[0107] Its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein:

[0108] X1 is selected from CH, C, and N;

[0109] X2 is selected from CH, C, and N; and

[0110] The condition is that only one of X1 and X2 is N.

[0111] The definitions of variables A and B are provided in equation (I-0) or equation (I).

[0112] In one implementation, the compound of formula (III) is not The S-isomer.

[0113] Implementation Plan 4. The compound described in Implementation Scheme 3, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein: X1 is N and It is a single bond, and X2 is C and It is a double bond.

[0114] Implementation Plan 5. The compound described in Implementation Scheme 3, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein: X1 is C and It is a double bond, and X2 is N and It is a single key.

[0115] Implementation Plan 6. The compound of any one of embodiments 1-5, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein:

[0116] A is

[0117] R3 is selected from hydrogen and methyl;

[0118] R4 is selected from hydrogen and methyl, or R3 and R4 together form a cyclopropyl group;

[0119] R5 is selected from hydrogen and methyl; or

[0120] R5 and R6 together form a cyclopropyl group, and

[0121] R7 is hydrogen.

[0122] Implementation Plan 7.The compound of any one of embodiments 1-5, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein:

[0123] A is

[0124] w is 1 or 2;

[0125] t is 1 or 2; and

[0126] s is 0 or 1.

[0127] Implementation Plan 8. The compound of any one of embodiments 1-7, its pharmaceutically acceptable salt, and / or the solvate of any of the foregoing, wherein B is NH2.

[0128] Implementation Plan 9. The compound of any one of embodiments 1-7, its pharmaceutically acceptable salt, and / or the solvate of any of the foregoing, wherein B is OH.

[0129] Implementation Plan 10. The compound of any one of embodiments 1-9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound has K p <0.4.

[0130] In some embodiments of implementation 10, the compound has a K content measured according to the procedure described in biological example 3. p <0.4. In some embodiments of embodiment 10, the compound, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing are selected from compounds 1 and 2.

[0131] Implementation Plan 11. The compound of any one of embodiments 1-9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound has K p ≤0.30.

[0132] In some embodiments of implementation 11, the compound has a K content measured according to the procedure described in biological example 3. p ≤0.30. In some embodiments of embodiment 11, the compound, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing are selected from compounds 1 and 2.

[0133] Implementation Plan 12. The compound of any one of embodiments 1-9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound has K p ≤0.20.

[0134] In some embodiments of implementation 12, the compound has a K content measured according to the procedure described in biological example 3. p ≤0.20. In some embodiments of embodiment 12, the compound, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing are selected from compounds 1 and 2.

[0135] Implementation Plan 13. The compound of any one of embodiments 1-9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound has K p ≤0.10.

[0136] In some embodiments of implementation 13, the compound has a K content measured according to the procedure described in biological example 3. p ≤0.10. In some embodiments of embodiment 12, the compound, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing are selected from compound 2.

[0137] Implementation Plan 14. The compound of any one of embodiments 1-13, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound has K in homogenized rat brain p，uu ≤0.2.

[0138] In some embodiments of implementation 14, the compound has K in homogenized rat brain measured according to the procedure described in biological example 3. p，uu ≤0.2. In some embodiments of embodiment 14, the compound, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing are selected from compounds 1 and 2.

[0139] Implementation Plan 14-1. The compound of any one of embodiments 1-13, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound has K in homogenized rat brain p，uu <0.1.

[0140] In some embodiments of implementation 14-1, the compound has a Kc in homogenized rat brain measured according to the procedure described in biological example 3. p，uu <0.1. In some embodiments of embodiment 14-1, the compound, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing are selected from compounds 1 and 2.

[0141] Implementation Plan 14-2. The compound of any one of embodiments 1-13, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound has K in homogenized rat brainp，uu ≤0.05.

[0142] In some embodiments of Implementation 14-2, the compound has a K0 in homogenized rat brain measured according to the procedure described in Biological Example 3. p，uu ≤0.05. In some embodiments of embodiments 14-2, the compound, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing are selected from compounds 1 and 2.

[0143] Implementation plan 14-3. The compound of any one of embodiments 1-13, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound has K in rat brain slices p，uu ≤0.02.

[0144] In some embodiments of embodiments 14-3, the compound has a K0 in homogenized rat brain measured according to the procedure described in biological example 3. p，uu ≤0.02. In some embodiments of embodiments 14-3, the compound, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing are selected from compounds 1 and 2.

[0145] Implementation Plan 15. The compound of any one of embodiments 1-14, a pharmaceutically acceptable salt thereof, and / or a solvate of any of the foregoing, wherein said compound has an unbound clearance rate of <900 mL / min / kg in rats (Cl u ).

[0146] Implementation Plan 16. The compound of any one of embodiments 1-14, a pharmaceutically acceptable salt thereof, and / or a solvate of any of the foregoing, wherein the compound has an unbound clearance rate of <750 mL / min / kg in rats (Cl u ).

[0147] Implementation Plan 17. The compound of any one of embodiments 1-14, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound has an IC50 against <10 μM CYP3A4. 50 .

[0148] Implementation Plan 18. The compound of any one of embodiments 1-5, 8 and 9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein A is selected from...

[0149] Implementation Plan 19.The compound of any one of embodiments 1-5, 8 and 9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein A is selected from...

[0150] Implementation Plan 20. The compound of any one of embodiments 1-5, 8 and 9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein A is selected from...

[0151] Implementation Plan 21. The compound of any one of embodiments 1-5, 8 and 9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein A is selected from...

[0152] Implementation Plan 22. The compound of any one of embodiments 1-5, 8 and 9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein A is selected from...

[0153] Implementation Plan 23. The compound of any one of embodiments 1-5, 8 and 9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein A is selected from...

[0154] Implementation Plan 24. The compound of any one of embodiments 1-5, 8 and 9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein A is selected from...

[0155] Implementation Plan 25. The compound of any one of embodiments 1-5, 8 and 9, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing, wherein the compound is any one of the compounds listed in Table 1.

[0156] In some embodiments of embodiment 25, the compound is any one of compounds 1-7, 13, 14, 14-A, 28, 33, and 38. In some embodiments of embodiment 25, the compound is any one of compounds 1-7, 13, 14, 28, 33, and 38. In some embodiments of embodiment 25, the compound is any one of compounds 1-6. In some embodiments of embodiment 25, the compound is any one of compounds 1 and 2.

[0157] Implementation Plan 26. A pharmaceutical composition comprising:

[0158] The compound according to any one of embodiments 1-25, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing; and

[0159] Pharmaceutically acceptable excipients.

[0160] Implementation Plan 27. A method for treating a disease or condition in a patient in need, wherein the method comprises administering to the patient a compound according to any one of embodiments 1-25, a pharmaceutically acceptable salt thereof, and / or a solvate of any of the foregoing, wherein the disease or condition is selected from: systemic mastocytosis, gastrointestinal stromal tumor, acute myeloid leukemia, melanoma, seminoma, intracranial germ cell tumor, mediastinal B-cell lymphoma, Ewing's sarcoma, diffuse large B-cell lymphoma, dysgerminoma, myelodysplastic syndrome, nasal NK / T-cell lymphoma, chronic myelomonocytic leukemia, and brain cancer.

[0161] Implementation Plan 28. A method for treating a disease or condition mediated by a mutant KIT or PDGFRα in a patient in need, wherein the method comprises administering to the patient a compound according to any one of embodiments 1-25, a pharmaceutically acceptable salt thereof, and / or a solvate of any of the foregoing.

[0162] Implementation Plan 29. According to the method described in implementation scheme 28, the disease or condition is selected from: systemic mastocytosis, gastrointestinal stromal tumor, acute myeloid leukemia, melanoma, seminoma, intracranial germ cell tumor, mediastinal B-cell lymphoma, Ewing's sarcoma, diffuse large B-cell lymphoma, dysgerminoma, myelodysplastic syndrome, nasal NK / T-cell lymphoma, chronic myelomonocytic leukemia, and brain cancer.

[0163] Implementation Plan 30. The compound according to any one of embodiments 1-25, its pharmaceutically acceptable salt, and / or a solvate of any of the foregoing are used as a medicament for treating a disease or condition in a patient in need, wherein the disease or condition is selected from: systemic mastocytosis, gastrointestinal stromal tumor, acute myeloid leukemia, melanoma, seminoma, intracranial germ cell tumor, mediastinal B-cell lymphoma, Ewing's sarcoma, diffuse large B-cell lymphoma, dysgerminoma, myelodysplastic syndrome, nasal NK / T-cell lymphoma, chronic myelomonocytic leukemia, and brain cancer.

[0164] Implementation Plan 31. The compound, a pharmaceutically acceptable salt thereof, and / or a solvate thereof, according to any one of embodiments 1-25, is used as a medicament for treating diseases or conditions mediated by mutant KIT or PDGFRα in patients in need.

[0165] Implementation Plan 32. According to the compound of embodiment 31, the disease or condition is selected from: systemic mastocytosis, gastrointestinal stromal tumor, acute myeloid leukemia, melanoma, seminoma, intracranial germ cell tumor, mediastinal B-cell lymphoma, Ewing's sarcoma, diffuse large B-cell lymphoma, dysgerminoma, myelodysplastic syndrome, nasal NK / T-cell lymphoma, chronic myelomonocytic leukemia, and brain cancer.

[0166] Implementation Plan 33. A method for treating eosinophilia, comprising administering to a subject in need a therapeutically effective amount of any one of embodiments 1-25, a pharmaceutically acceptable salt thereof, and / or a solvate of any of the foregoing.

[0167] Implementation Plan 34. According to the method of embodiment 33, the eosinophilic disease is selected from: hypereosinophilic syndrome, eosinophilic granuloma, eosinophilic gastroenteritis, eosinophilic leukemia, eosinophilic granuloma and Kimura disease.

[0168] Implementation Plan 35. According to the method of embodiment 33, the eosinophilic disorder is a hypereosinophilic syndrome.

[0169] Implementation Plan 36. According to the method described in embodiment 33, the eosinophilic disease is eosinophilic leukemia.

[0170] Implementation Plan 37. According to the method of embodiment 36, the eosinophilic disease is chronic eosinophilic leukemia.

[0171] Implementation Plan 38. The method according to any one of embodiments 33-27, wherein the eosinophilia is refractory to imatinib, sunitinib and / or regorafenib.

[0172] Implementation Plan 39. The compound, a pharmaceutically acceptable salt thereof, and / or a solvate thereof, according to any one of embodiments 1-25, is used as a drug for treating eosinophilia.

[0173] Implementation Plan 40. According to the compound method of embodiment 39, the eosinophilic disease is selected from: hypereosinophilic syndrome, eosinophilic granuloma, eosinophilic gastroenteritis, eosinophilic leukemia, eosinophilic granuloma and Kimura disease.

[0174] Implementation Plan 41.According to the compound of embodiment 39, the eosinophilic disorder is hypereosinophilic syndrome.

[0175] Implementation Plan 42. According to the compound of embodiment 39, the eosinophilic disease is eosinophilic leukemia.

[0176] Implementation Plan 43. According to the compound of embodiment 42, the eosinophilic disease is chronic eosinophilic leukemia.

[0177] Implementation Plan 44. The method according to any one of embodiments 39-43, wherein the eosinophilia is refractory to imatinib, sunitinib and / or regorafenib.

[0178] Implementation Plan 45. A method for treating mast cell syndrome, comprising administering to a subject in need a therapeutically effective amount of a compound according to any one of embodiments 1-25, a pharmaceutically acceptable salt thereof, and / or a solvate of any of the foregoing.

[0179] Implementation Plan 46. According to the method of embodiment 45, the mast cell syndrome is mediated by a mutant KIT or PDGFRα.

[0180] Implementation Plan 46-1. According to the method of embodiment 45, the mast cell syndrome is mediated by wild-type KIT or PDGFRα.

[0181] Implementation Plan 47. According to the method of any one of embodiments 46, the mast cell syndrome is selected from: mast cell activation syndrome (MCAS) and hereditary alpha trypsinemia (HAT).

[0182] Implementation Plan 48. According to the method of embodiment 47, the MCAS is selected from: monoclonal mast cell activation syndrome (MMAS), secondary MCAS, and idiopathic MCAS.

[0183] Implementation Plan 48-1. According to the method described in implementation scheme 27, the disease or condition is systemic mastocytosis.

[0184] Implementation Plan 49. According to any one of the embodiments 48, the systemic mastocytosis is selected from: indolent systemic mastocytosis and smoldering systemic mastocytosis.

[0185] It should be noted that in this disclosure, any of the above-defined implementation schemes is intended to include corresponding sub-implementations. For example, when referring to implementation scheme 14, it is intended to include implementation scheme 14, implementation scheme 14-1, implementation scheme 14-2, implementation scheme 14-3, and the specific implementation schemes described therein.

[0186] Table 1 lists the compounds prepared by the synthetic methods described herein.

[0187]

[0188]

[0189]

[0190]

[0191]

[0192]

[0193]

[0194]

[0195]

[0196]

[0197]

[0198]

[0199]

[0200]

[0201]

[0202]

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213]

[0214]

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221] The compounds disclosed herein are selective KIT inhibitors. In some embodiments, the compounds disclosed herein are selective D816V KIT inhibitors. The compounds disclosed herein are selective PDGFRα inhibitors. In some embodiments, the compounds disclosed herein are selective PDGFRα exon 18 inhibitors. In some embodiments, the compounds disclosed herein are selective PDGFRα D842V inhibitors. As used herein, "selective KIT inhibitor" or "selective PDGFRα inhibitor" means a compound, its pharmaceutically acceptable salt, or a solvation of any of the foregoing, that selectively inhibits KIT protein kinase or PDGFRα protein kinase relative to another protein kinase, and exhibits at least 2-fold selectivity to KIT protein kinase or PDGFRα protein kinase relative to another kinase. For example, selective KIT inhibitors or selective PDGFRα inhibitors exhibit at least 9-fold, 10-fold, at least 15-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 60-fold, at least 70-fold, at least 80-fold, at least 90-fold, at least 100-fold, at least 125-fold, at least 150-fold, at least 175-fold, or at least 200-fold selectivity relative to another kinase (e.g., VEGFR2 (vascular endothelial growth factor receptor 2), SRC (non-receptor protein tyrosine kinase), and FLT3 (fmS-like tyrosine kinase 3)). In some embodiments, selective KIT or selective PDGFRα inhibitors exhibit selectivity relative to PDGRFβ, CSF1R (colony-stimulating factor receptor 1), and FLT3. In some embodiments, selective KIT or selective PDGFRα inhibitors exhibit selectivity relative to LCK (lymphocyte-specific protein kinase), ABL (nucleoprotein tyrosine kinase), NIMA-associated kinase 5 (NEK5), and ROCK1 (rho-associated coil-and-coil persistent protein kinase-1). In some embodiments, selectivity to KIT or PDGFRα protein kinase relative to another kinase is measured in cellular assays (e.g., cytometry). In some embodiments, selectivity to KIT or PDGFRα protein kinase relative to another kinase is measured in biochemical assays (e.g., biochemical assays).

[0222] The compounds disclosed herein are selective relative to ion channels. In some embodiments, selective KIT or selective PDGFRα inhibitors have limited potential to inhibit human voltage-gated sodium channels (hNav1.2).

[0223] The compounds disclosed herein are selective for mutant KIT relative to wild-type KIT. In some embodiments, the compounds disclosed herein are selective for exon 17 mutant KIT relative to wild-type KIT.

[0224] The compounds disclosed herein can be used to treat diseases or conditions in humans or non-humans associated with mutant KIT or mutant PDGFRA activity. In some embodiments, the compounds disclosed herein are used as pharmaceuticals. In some embodiments, the compounds disclosed herein are used in treatment. In some embodiments, the compounds disclosed herein are used in the preparation of pharmaceuticals. In some embodiments, this disclosure provides methods for treating KIT-driven malignancies, including mastocytosis (SM), GIST (gastrointestinal stromal tumor), AML (acute myeloid leukemia), melanoma, seminoma, intracranial germ cell tumor, and / or mediastinal B-cell lymphoma. Furthermore, mutations in KIT are associated with Ewing's sarcoma, DLBCL (diffuse large B-cell lymphoma), dysgerminoma, MDS (myelodysplastic syndrome), NKTCL (nasal NK / T-cell lymphoma), CMML (chronic myelomonocytic leukemia), and brain cancer. In some embodiments, this disclosure provides methods for treating Ewing's sarcoma, DLBCL, NKTCL, CMML, and / or brain cancer. KIT mutations have also been found in thyroid cancer, colorectal cancer, endometrial cancer, bladder cancer, NSCLC, and breast cancer (AACR Project GENIE). In some embodiments, the compounds disclosed herein can be used to treat mast cell activation syndrome (MCAS). The compounds disclosed herein can be used to treat systemic mastocytosis. The compounds disclosed herein can be used to treat advanced systemic mastocytosis. The compounds disclosed herein can be used to treat indolent SM and smoldering SM. The compounds disclosed herein can be used to treat GIST.

[0225] The compounds disclosed herein can be used to treat diseases or conditions associated with KIT mutations in exons 9, 11, 14, 17, and / or 18 of the KIT gene sequence. The compounds disclosed herein can be used to treat diseases or conditions associated with PDGFRA mutations in exons 12, 14, and / or 18 of the PDGFRA gene sequence. In some embodiments, methods are provided herein for treating diseases or conditions associated with at least one KIT mutation in exons 9, 11, 14, 17, and / or 18 of the KIT gene sequence. In some embodiments, methods are provided for treating diseases or conditions associated with at least one PDGFRA mutation in exons 12, 14, and / or 18 of the PDGFRA gene sequence.

[0226] The compounds disclosed herein are active against one or more KIT protein kinases that have mutations in exon 17 of the KIT gene sequence (e.g., KIT protein mutations D816V, D816Y, D816F, D816K, D816H, D816A, D816G, D816E, D816I, D816F, D820A, D820E, D820G, D820Y, N822K, N822H, V560G, Y823D, and A829P), and have much less activity against wild-type KIT protein kinases. In some embodiments, this document provides methods for treating diseases or conditions associated with at least one KIT mutation, such as those selected from D816V, D816Y, D816F, D816K, D816H, D816A, D816G, D816E, D816I, D816F, D820A, D820E, D820G, D820Y, N822K, N822H, V560G, Y823D, and A829P. In some embodiments, this document provides methods for treating diseases or conditions associated with at least one KIT mutation, such as those selected from C809, C809G, D816H, D820A, D820G, N822H, N822K, and Y823D.

[0227] The compounds disclosed herein are active against one or more KIT protein kinases (e.g., KIT protein mutations del557-559insF, V559G / D) that have mutations in exon 11 of the KIT gene sequence. In some implementations, this document provides methods for treating diseases or conditions associated with at least one KIT mutation, such as those selected from L576P, V559D, V560D, V560G, W557G, Del 554-558EVQWK, del557-559insF, Del EVQWK554-558, DelEVQWKVVEEIN GNNYVYI554-571, Del KPMYEVQWK550-558, Del KPMYEVQW550-557FL, DelKV558-559, Del KV558-559N, Del MYEVQW552-557, Del PMYE551-554, Del VV559-560, DelWKVVE557-561, Del WK557-558, Del WKVV557-560C, Del WKVV557-560F, DelYEVQWK553-558 and insert K558NP.

[0228] The compounds disclosed herein are active against one or more KIT protein kinases (e.g., KIT protein mutations V559D / V654A, V560G / D816V, and V560G / 822K) that have mutations in exons 11 / 13 of the KIT gene sequence. In some embodiments, methods are provided herein for treating diseases or conditions associated with one or more KIT mutations in exons 11 / 13.

[0229] The compounds disclosed herein are active against one or more KIT protein kinases with mutations in exon 9 of the KIT gene sequence. In some embodiments, methods are provided herein for treating diseases or conditions associated with at least one KIT mutation in exon 9.

[0230] In some embodiments, the compounds disclosed herein have no activity against KIT protein kinases with mutant V654A, N655T, T670I, and / or N680.

[0231] The compounds disclosed herein are active against one or more mutated PDGFRα protein kinases. In some embodiments, methods are provided herein for treating diseases or conditions associated with at least one PDGFRA mutation (e.g., PDGFRα protein mutation V561D, Del RV560-561, Del RVIES560-564, Ins ER561-562, SPDGHE566-571R, SPDGHE566-571K, or Ins YDSRW582-586) in exon 12 of the PDGFRA gene sequence. In some embodiments, methods are provided herein for treating diseases or conditions associated with at least one PDGFRA mutation (e.g., PDGFRα protein mutation N659K) in exon 14 of the PDGFRA gene sequence. In some embodiments, this document provides for the treatment of patients with at least one PDGFRA mutation in exon 18 of the PDGFRA gene sequence (e.g., PDGFRα protein mutations D842V, D842Y, D842I, DI842-843IM, D846Y, Y849C, Del D842, Del I843, Del RD841-842, DelDIM842-845, Del DIMH842-845, Del IMHD843-846, Del...). Methods related to diseases or conditions (e.g., MHDS844-847, RD841-842KI, DIMH842-845A, DIMH842-845V, DIMHD842-846E, DIMHD842-846S, DIMHD842-846N, DIMHD842-846G, IMDHS843-847T, IMDHS8843-847M, or HDSN845-848P).

[0232] The compounds disclosed herein are active against one or more PDGFRα protein kinases (e.g., protein mutations PDGFRαD842V, PDGFRαD842I, or PDGFRαD842Y) having a mutated exon 18 in the PDGFRA gene sequence. In some embodiments, methods are provided herein for treating diseases or conditions associated with at least one PDGFRA mutation in exon 18 (e.g., the protein mutation PDGFRαD842V).

[0233] The compounds disclosed herein can be used to treat eosinophilic disorders. In some embodiments, eosinophilic disorders are mediated by mutant KIT or PDGFRα. In some embodiments, eosinophilic disorders are mediated by wild-type KIT or PDGFRα. In some embodiments, a method for treating eosinophilic disorders is provided herein, comprising administering to a subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof and / or a solvate of any of the foregoing. In one embodiment, eosinophilic disorders are selected from: hypereosinophilic syndrome, eosinophilic granulomatosis, eosinophilic gastroenteritis, eosinophilic leukemia, eosinophilic granulomatosis, and Kimura disease.

[0234] In some implementations, eosinophilic disorders are selected from: hypereosinophilic syndrome, eosinophilia, eosinophilic gastroenteritis, eosinophilic leukemia, eosinophilic granuloma, and Kimura disease. Other eosinophilic disorders include eosinophilic esophagitis, eosinophilic gastroenteritis, eosinophilic fasciitis, and Churg-Strauss syndrome.

[0235] In one embodiment, eosinophilia is hypereosinophilic syndrome. In a specific embodiment, eosinophilic syndrome is idiopathic hypereosinophilic syndrome. In one embodiment, eosinophilia is eosinophilic leukemia. In a specific embodiment, eosinophilic leukemia is chronic eosinophilic leukemia. In another embodiment, eosinophilia is refractory to imatinib, sunitinib, and / or regorafenib. In a specific embodiment, eosinophilia is refractory to imatinib treatment.

[0236] The compounds disclosed herein can be used to reduce the number of eosinophils in subjects in need. In some embodiments, a method for reducing the number of eosinophils in subjects in need is provided herein, comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof and / or a solvate of any of the foregoing.

[0237] In one embodiment, the disclosed method reduces the number of eosinophils in the blood, bone marrow, gastrointestinal tract (e.g., esophagus, stomach, small intestine, and colon), or lungs. In another embodiment, the disclosed method reduces the number of eosinophils in the blood. In yet another embodiment, the disclosed method reduces the number of eosinophils in the lungs. In still another embodiment, the disclosed method reduces the number of eosinophil precursor cells.

[0238] In another embodiment, the disclosed method reduces the number of eosinophils (after application) by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%. In one specific embodiment, the method disclosed herein reduces the number of eosinophils to below the detection limit.

[0239] In another embodiment, the disclosed method reduces the number of eosinophil precursors (after administration) by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, or at least about 99%. In one specific embodiment, the method disclosed herein reduces the number of eosinophil precursors to below the detection limit.

[0240] The compounds disclosed herein can be used to treat mast cell syndrome. Mast cell syndrome is subdivided into two groups of conditions: (1) cutaneous mast cell syndrome (CM), which describes a form limited to the skin; and (2) systemic mast cell syndrome (SM), which describes a form involving mast cell infiltration of extracutaneous organs, with or without involvement of the skin. SM is further subdivided into five forms: indolent (ISM); smoldering (SSM); aggressive (ASM); SM associated with non-mast cell lineage diseases of the blood (SM-AHNMD); and mast cell leukemia (MCL).

[0241] The diagnosis of SM is partly based on histological and cytological studies of bone marrow infiltration showing mast cell infiltration with typically atypical morphology, which usually aberrantly expresses non-mast cell markers (CD25 and / or CD2). The diagnosis of SM can be confirmed when bone marrow mast cell infiltration occurs in one of the following contexts: (1) abnormal mast cell morphology (fusiform cells); (2) serum trypsin levels above 20 ng / mL; or (3) presence of mutations in activating KIT proteins, such as exon 17 mutations, such as D816 mutations (e.g., D816V).

[0242] Activating mutations at the D816 position have been found in the vast majority (90-98%) of mastocytosis cases, with the most common mutations being D816V, D816H, and D816Y. The D816V mutation has been found in the activation loop of the protein kinase domain and leads to constitutive activation of KIT kinase.

[0243] No medications are approved for non-late forms of systemic mastocytosis, ISM, or SSM. Current treatments for these chronic conditions include non-specific symptom-targeting therapies with varying degrees of efficacy that do not affect MC burden. Cytoreductive therapies (e.g., cladribine and interferon-alpha) are occasionally used to treat refractory symptoms. Based on current treatment regimens, there is an unmet medical need among patients with moderate to severe ISM and SSM whose symptoms cannot be adequately treated with existing symptom-targeting therapies.

[0244] The compounds disclosed herein can be used to treat ISM or SSM. In some embodiments, patients with ISM or SSM have symptoms that are not adequately controlled by at least one, at least two, or at least three symptomatic treatments. Symptoms can be assessed using patient-reported outcomes (PRO) tools, such as the Indolent Systemic Mastocytosis-Symptom Assessment Form (ISM-SAF) (I ISPOR Europe 2019, Copenhagen Denmark, 2-6 Nov 2019). The compounds disclosed herein can be used to improve symptoms associated with ISM or SSM, such as reducing or eliminating itching, flushing, headache, and / or GI events, such as vomiting, diarrhea, and abdominal pain. Improvement in symptoms can be assessed using the ISM-SAF.

[0245] The compounds disclosed herein may be used to treat other mast cell syndromes, such as mast cell activation syndrome (MCAS) and hereditary alpha trypsinemia (HAT) (Picard Clin. Ther. 2013, May 35(5)548; Akin J. Allergy Clin. Immuno. 140(2)34962). The compounds disclosed herein may be used to treat mast cell syndromes associated with KIT and PDGFRα mutations. The compounds disclosed herein may be used to treat mast cell diseases associated with wild-type KIT and PDGFRα.

[0246] The disclosed compounds can be used to treat mast cell activation syndrome (MCAS), an immune disorder in which mast cells inappropriately and excessively release chemical mediators, causing a range of chronic symptoms, sometimes including anaphylactic or near-anaphylactic reactions. Unlike mastocytosis, where patients have an abnormally high number of mast cells, MCAS patients have a normal number of mast cells, but these cells cannot function properly and are defined as "overreacting." Types of MCAS include primary MCAS (monoclonal mast cell activation syndrome (MMAS)), secondary MCAS (MCAS caused by another disease), and idiopathic MCAS (MCAS excluding primary or secondary MCAS).

[0247] The compounds disclosed herein can be used to treat hereditary alpha trypsinemia (HAT) (overexpression of TPSAB1, which leads to elevated trypsin levels).

[0248] Other mast cell disorders include mast cell-mediated asthma, allergic reactions (including idiopathic, IgE- and non-IgE-mediated), urticaria (including idiopathic and chronic), atopic dermatitis, swelling (angioedema), irritable bowel syndrome, mast cell gastroenteritis, mast cell colitis, pruritus, chronic pruritus, and pruritus secondary to chronic renal failure and mast cell-related cardiac, vascular, intestinal, brain, kidney, liver, pancreas, muscle, bone, and skin conditions. In some implementations, mast cell disorders are not associated with mutant KIT or mutant PDGFRα.

[0249] KIT and PDGFRA mutations have been extensively investigated in GIST. The compounds disclosed herein may be used to treat GIST associated with KIT mutations. The compounds disclosed herein may be used to treat unresectable or metastatic GIST. Nearly 80% of metastatic GISTs have primary activating mutations in either the extracellular region (exon 9) or the juxtamembrane (JM) domain (exon 11) of the KIT gene sequence. Many mutant KIT tumors respond to targeted therapies such as imatinib (a selective tyrosine kinase inhibitor that specifically inhibits BCR-ABL, KIT, and PDGFRA proteins). However, most GIST patients eventually relapse due to secondary mutations in KIT that significantly reduce the binding affinity of imatinib. These resistance mutations always occur within the adenosine 5-triphosphate (ATP)-binding pocket (exons 13 and 14) or activation loop (exons 17 and 18) of the kinase gene. None of the currently approved agents for GIST are selectively targeted agents. Imatinib is currently approved for the treatment of GIST; followed by the use of a multi-kinase inhibitor. In many cases, these multi-kinase inhibitors (e.g., sunitinib, regorafenib, and midostaurin) only weakly inhibit imatinib resistance mutations and / or multi-kinase inhibitors are limited by more complex safety profiles and small therapeutic windows. In some embodiments, the compounds disclosed herein can be used to treat GIST in patients who have already been treated with imatinib. The compounds disclosed herein can be used as first-line (1L), second-line (2L), third-line (3L), or fourth-line (4L) therapy for the treatment of GIST.

[0250] The compounds disclosed herein can be used to treat GIST when a specific mutation in KIT is absent or present. In some embodiments, the compounds disclosed herein are able to treat GIST when a specific mutation in KIT is absent. In some embodiments, the compounds disclosed herein are not able to treat GIST when a specific mutation in KIT is present. In some embodiments, the compounds disclosed herein do not provide clinical benefit in patients carrying KIT ATP-binding pocket mutations (KIT protein mutations V654A, N655T, and / or T670I).

[0251] The compounds disclosed herein may be used to treat GIST associated with PDGFRA mutations. In 5-6% of patients with unresectable metastatic GIST, an activation loop mutation occurs as a primary mutation in exon 18 of the PDGFRA gene sequence located at amino acid position 842 of the protein.

[0252] The compounds disclosed herein can also be used to treat AML. AML patients often carry KIT mutations, most of which are located at position D816 of the KIT protein.

[0253] In some embodiments, the disclosed compounds are administered to a subject in need. In some embodiments, the disclosed compounds are administered as a pharmaceutical formulation, wherein the compounds are combined with one or more pharmaceutically acceptable excipients or carriers. Thus, in some embodiments, compositions are disclosed herein comprising at least one entity of a compound selected from Formula I and a pharmaceutically acceptable salt thereof and / or a solvate of any of the foregoing, and optionally further comprising at least one pharmaceutically acceptable excipient.

[0254] The compounds disclosed herein can be formulated for administration in any convenient manner for human or veterinary use. In some embodiments, the compounds contained in the pharmaceutical composition may be active themselves, or may be prodrugs, for example, capable of being converted into active compounds in a physiological environment.

[0255] As used herein, the phrase “pharmaceutically acceptable” means those compounds, materials, compositions, and / or dosage forms that are suitable for contact with human and animal tissues within the bounds of reasonable medical judgment and without excessive toxicity, irritation, allergic reactions, or other problems or complications, and that are commensurate with a reasonable benefit / risk ratio.

[0256] Examples of pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, etc. (10) Corn oil and soybean oil; (11) Diols, such as propylene glycol; (12) Polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (13) Esters, such as ethyl oleate and ethyl laurate; (14) Agar; (15) Buffers, such as magnesium hydroxide and aluminum hydroxide; (16) Alginate; (17) Pyrogen-free water; (18) Isotonic saline; (19) Ringer's solution; (20) Ethanol; (21) Phosphate buffer; (22) Cyclodextrin, such as... (22) Other non-toxic and compatible substances used in pharmaceutical preparations.

[0257] Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, etc.; (2) oil-soluble antioxidants, such as palmitic acid ascorbate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol, etc.; and (3) metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, etc.

[0258] Solid dosage forms (e.g., capsules, tablets, pills, sugar-coated pills, powders, granules, etc.) may include one or more pharmaceutically acceptable carriers, such as sodium citrate or calcium hydrogen phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and / or gum arabic; (3) humectants, such as glycerin. (4) Disintegrants, such as agar, calcium carbonate, potato or cassava starch, alginic acid, certain silicates and sodium carbonate; (5) Solution blockers, such as paraffin; (6) Absorption promoters, such as quaternary ammonium compounds; (7) Wetting agents, such as cetyl alcohol and glyceryl monostearate; (8) Absorbents, such as kaolin and bentonite clay; (9) Lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium laurate sulfate and mixtures thereof; and (10) Colorants.

[0259] Liquid dosage forms may include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active ingredient, liquid dosage forms may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol, oils (e.g., cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters and mixtures thereof of sorbitol.

[0260] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, tragacanth gum and mixtures thereof.

[0261] In addition to active compounds, ointments, pastes, creams and gels may also contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silicic acid, talc, zinc oxide or mixtures thereof.

[0262] In addition to the active compound, powders and aerosols may contain excipients such as lactose, talc, silica, aluminum hydroxide, calcium silicate, and polyamide powders or mixtures thereof. Aerosols may also contain conventional propellants such as chlorofluorocarbons and volatile unsubstituted hydrocarbons such as butane and propane.

[0263] Non-limiting examples of dosage forms for topical or transdermal application of the compounds of this disclosure include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalers. The active compounds may be mixed under sterile conditions with pharmaceutically acceptable carriers and with any preservatives, buffers, or propellants that may be required.

[0264] When the compounds of this disclosure are administered to humans and animals as pharmaceuticals, the compounds may be given on their own or as pharmaceutical compositions comprising, for example, 0.1 to 99.5% (e.g., 0.5 to 90%) of an active ingredient in combination with a pharmaceutically acceptable carrier.

[0265] The formulation can be administered topically, orally, transdermally, rectally, vaginally, parenterally, intranasally, intrapulmonaryly, intraocularly, intravenously, intramuscularly, intra-arterially, intrathecally, intracapsularly, intradermally, intraperitoneally, subcutaneously, subepidermally, or by inhalation.

[0266] Furthermore, the compounds of this disclosure can be administered alone or in combination with other compounds (including other KIT or PDGFRα modulating compounds) or other therapeutic agents. In some embodiments, the compounds of this disclosure can be administered in combination with ripretinib. In some embodiments, the compounds of this disclosure can be administered in combination with one or more compounds selected from imatinib, sunitinib, regorafenib, cabozantinib, clarnone, midotulin, bentuximab, and mastitinib for the treatment of the diseases or conditions disclosed herein.

[0267] The compounds disclosed herein can be administered to patients who have previously been treated with one or more other compounds. The compounds disclosed herein can be used as first-line (1L), second-line (2L), third-line (3L), or fourth-line (4L) therapy.

[0268] In some embodiments, the compounds disclosed herein are administered after prior treatment with imatinib.

[0269] The compounds disclosed herein can be administered to patients who have not previously been treated with midostaurin. In some embodiments, the compounds disclosed herein can be administered to patients who have previously been treated with midostaurin. Example

[0270] definition

[0271]

[0272]

[0273]

[0274] The methods for preparing the compounds of this disclosure can be carried out in suitable solvents, which can be readily selected by those skilled in the art of organic synthesis. At the temperature at which the reaction proceeds, for example, in the range of the solvent's freezing temperature to its boiling temperature, a suitable solvent can be substantially unreactive with the starting materials (reactants), intermediates, or products. A given reaction can be carried out in one solvent or a mixture of more than one solvent. Depending on the specific reaction step, those skilled in the art can select a suitable solvent for that particular reaction step.

[0275] The preparation of the compounds disclosed herein may involve the protection and deprotection of various chemical groups. Those skilled in the art can readily determine the need for protection and deprotection, as well as the selection of appropriate protecting groups. Chemical reactions of protecting groups can be found, for example, in Wuts and Greene, Protective Groups in Organic Synthesis, 5th ed., John Wiley & Sons: New Jersey, (2014), the entirety of which is incorporated herein by reference.

[0276] The reaction can be monitored using any suitable method known in the art. For example, product formation can be detected by spectroscopic means such as nuclear magnetic resonance (NMR) spectroscopy (e.g. 1 H or 13 C) Monitoring by infrared (IR) spectroscopy, spectrophotometry (e.g., UV-Vis), mass spectrometry (MS), or chromatographic methods such as high-performance liquid chromatography (HPLC) or thin-layer chromatography (TLC). Analytical instruments and methods for compound characterization:

[0277] LC-MS: Unless otherwise specified, all liquid chromatography-mass spectrometry (LC-MS) data (analytical sample purity and identity) were obtained using an Agilent Modlel 6120 mass spectrometer with an Agilent Modlel-1260LC system, utilizing ES-API ionization with an Agilent Poroshel 120 (EC-C18, 2.7 μM particle size, 3.0 × 50 mm) reversed-phase column at 22.4 °C. The mobile phase consisted of a mixture of 0.1% formic acid in H₂O and 0.1% formic acid in acetonitrile. A constant gradient was used from 95% aqueous / 5% organic to 5% aqueous / 95% organic mobile phase over 4 minutes. The flow rate was kept constant at 1 mL / min.

[0278] Silica gel chromatography: in Teledyne Isco Rf unit or Biota Silica gel chromatography was performed on an Isolera Four unit.

[0279] Proton NMR: Unless otherwise specified, all 1H NMR spectra were obtained using a Varian 400MHz Unity Inova 400MHz NMR instrument (acquisition time = 3.5 seconds with a 1-second delay; 16 to 64 scans). For characterization, all protons were reported in DMSO-d6 solvent at parts per million (ppm) relative to residual DMSO (2.50 ppm).

[0280] Those skilled in the art will recognize that variations in gradient, column length, and flow rate are possible, and that some conditions may be more suitable for compound characterization than others, depending on the type of chemical being analyzed.

[0281] Example

[0282] Synthesis and preparation

[0283] Preparation of intermediates

[0284] Preparation 1: N-((S)-1-(2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)-1-(4-fluorophenyl)ethyl)-2-methylpropane-2-sulfinamide (I-1)

[0285]

[0286] Step 1: Synthesis of ethyl 2-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-carboxylate (iii): ethyl 2-chloropyrimidin-5-carboxylate (5.00 g, 26.8 mmol, 1.00 eq), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-3,6-dihydropyridin-1(2H)-carboxylate tert-butyl ester (9.11 g, 29 mmol, 29 mmol, 1.00 eq), and ethyl 2-chloropyrimidin-5-carboxylate (5.00 g, 26.8 mmol, 1.00 eq) were prepared. A mixture of Cs₂CO₃ (17.5 g, 53.6 mmol, 2.00 eq) in Pd(dppf)Cl₂·CH₂Cl₂ (2.63 g, 3.22 mmol, 0.12 eq), dioxane (150 mL), and H₂O (15.0 mL) was degassed and purged three times with N₂(g), and then stirred at 60 °C for 12 h under N₂(g) atmosphere. The reaction mixture was filtered, diluted with H₂O (300 mL), and extracted with EtOAc (400 mL × 3). The combined organic layers were washed with brine (500 mL × 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give the residue. The residue was purified by column chromatography (silica gel, petroleum ether: EtOAc = 50:1 to 10:1, plate 1, Rf = 0.71) to give the title compound (4.90 g, 54.9% yield). LCMS: RT=0.977min, m / z=278.2(M-56+H)+.

[0287] Step 2: Synthesis of 2-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidine-5-carboxylic acid (iv): A mixture of NaOH (803 mg, 20.1 mmol, 2.23 eq) in 20.1 mL of H₂O was added to a solution of ethyl 2-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidine-5-carboxylic acid (3.00 g, 9.00 mmol, 1.00 eq) in 30.0 mL of THF. The mixture was then stirred at 25 °C for 2 hours. The mixture was concentrated under vacuum to remove THF, then the pH was adjusted to 2-3 with 1 M HCl and filtered. The filter cake was concentrated under vacuum to give the title compound (2.49 g, 90.6% yield) as a pale yellow solid, which was used in the next step without further purification. 1H NMR (400MHz, d6-DMSO) δ9.11 (s, 2H), 7.28 (br s, 1H), 4.12 (br s, 3H), 3.52-3.55 (br t, J = 5.2Hz, 2H), 2.63 (s, 2H), 1.43 (s, 9H).

[0288] Step 3: Synthesis of tert-butyl 4-(5-(methoxy(methyl)carbamoyl)pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid (v): HATU (6.20 g, 16.3 mmol, 2.00 eq), DIPEA (5.27 g, 40.8 mmol, 7.10 mL, 5.00 eq), and N,O-dimethylhydroxylamine hydrochloride (1.19 g, 12.2 mmol, 1.50 eq) were added to a solution of 2-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridine-4-yl)pyrimidin-5-carboxylic acid (2.49 g, 8.16 mmol, 1.00 eq) in DMF (50.0 mL). The mixture was then stirred at 25 °C for 9 h. The mixture was diluted with EtOAc (50.0 mL), washed with water (100 mL × 2), and the aqueous layer was extracted with EtOAc (50.0 mL × 2). The organic layers were then combined and washed successively with 1 M HCl (40.0 mL), saturated NaHCO3 (40.0 mL), and brine (50.0 mL). The organic layers were dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by column chromatography (SiO2, petroleum ether:EtOAc = 5:1 to 10:3, petroleum ether:EtOAc = 1:1, Rf = 0.60) to give the title compound (1.89 g, 66.5% yield) as a pale yellow solid: LCMS: RT = 0.888 min, m / z = 293.3 (M-56+H)+.1H NMR: (400 MHz, d6-DMSO) δ 9.00 (s, 2H), 7.30 (br s, 1H), 4.13 (br s, 2H), 3.60 (s, 3H), 3.54 (br t, J = 5.6 Hz, 2H), 3.31 (s, 3H), 2.69 (s, 1H), 2.63 (br d, J = 1.6 Hz, 2H), 1.43 (s, 9H).

[0289] Step 4: Synthesis of N-methoxy-N-methyl-2-(1,2,3,6-tetrahydropyridin-4-yl)pyrimidine-5-carboxamide trifluoroacetate (vi): TFA (7.48 g, 65.6 mmol, 4.86 mL, 12.1 eq) was added to a solution of tert-butyl 4-(5-(methoxy(methyl)carbamoyl)pyrimidine-2-yl)-3,6-dihydropyridin-1(2H)-carboxylic acid (1.89 g, 5.42 mmol, 1.00 eq) in DCM (18.9 mL). The mixture was stirred at 25 °C for 0.5 h and then concentrated under vacuum to give the title compound (1.97 g, crude) as a yellow oil, which was used in the next step without further purification.

[0290] Step 5: Synthesis of 2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-N-methoxy-N-methylpyrimidine-5-carboxamide (viii): DIPEA (4.22 g, 32.6 mmol, 5.68 mL, 6.00 eq) was added to a solution of N-methoxy-N-methyl-2-(1,2,3,6-tetrahydropyridin-4-yl)pyrimidine-5-carboxamide trifluoroacetate (1.97 g, 5.44 mmol, 1.00 eq) in DCM (20.0 mL), and the resulting mixture was stirred at 25 °C for 5 min. 6-Bromo-4-chloropyrrolo[2,1-f][1,2,4]triazine (1.33 g, 5.71 mmol, 1.05 eq) was added, and the mixture was stirred at 25 °C for another 2 hours. The reaction mixture was concentrated under vacuum, diluted with isopropanol (20.0 mL) and water (5.00 mL), stirred for 16 h, and filtered. The filter cake was washed with isopropanol (10.0 mL × 3), then with petroleum ether (20.0 mL × 2), and concentrated under vacuum to give the title compound (1.99 g, 82.4% yield) as a yellow solid, which was used in the next step without further purification: LCMS: RT = 0.897 min, m / z = 446.2 (M + H) + 1H NMR: (400 MHz, d6-DMSO) δ 9.03 (s, 2H), 7.96 (d, J = 1.6 Hz, 1H), 7.93 (s, 1H), 7.40 (br s, 1H), 7.24 (d, J = 1.6 Hz, 1H), 4.75 (br s, 2H), 4.14 (t, J = 5.6 Hz, 2H), 3.61 (s, 3H), 3.32 (s, 3H), 2.83 (br s, 2H).

[0291] Step 6: Synthesis of (2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)(4-fluorophenyl)methyl ketone (ix): Under N2 (g), magnesium (4-fluorophenyl) bromide (1.00 M solution in THF, 28.1 mL, 7.00 eq) was added dropwise to a cooled solution (0 °C) of 2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)-N-methoxy-N-methylpyrimidin-5-carboxamide (1.78 g, 4.01 mmol, 1.00 eq) at THF (110 mL). The resulting mixture was stirred at 25 °C for 4 h. The mixture was slowly poured into a saturated NH4Cl aqueous solution (50.0 mL), and then diluted with EtOAc (50.0 mL). The aqueous layer was extracted with EtOAc (30.0 mL × 3), the organic layers were combined and washed with brine (50.0 mL), dried over Na2SO4 and concentrated. The crude product was ground with isopropanol (30.0 mL) at 25 °C for 3 h with stirring for 12 h. The mixture was filtered with isopropanol (3.00 mL × 3), and the filter cake was washed with petroleum ether (3.00 mL) and dried under vacuum to give the title compound (1.36 g, 69.6% yield, 98.3% purity) as a pale yellow solid: LCMS: RT = 1.024 mins, m / z = 479.2 (M + H) + 1H NMR (400 MHz, d6-DMSO) δ 9.09 (s, 2H), 7.92–7.98 (m, 4H), 7.49 (br s, 1H), 7.43 (t, J = 8.8 Hz, 2H), 7.25 (d, J = 1.6 Hz, 1H), 4.78 (br s, 2H), 4.16 (t, J = 5.6 Hz, 2H), 2.87 (br s, 2H).

[0292] Step 7: Synthesis of (S,E)-N-((2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazine-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)(4-fluorophenyl)methylene)-2-methylpropane-2-sulfinamide (x): (2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazine) was added to THF (60.0 mL). (S)-2-methylpropane-2-sulfinamide (1.05 g, 8.70 mmol, 4.00 eq) and Ti(OEt)4 (22.0 g, 96.5 mmol, 20.0 mL, 44.4 eq) were added to a solution of (-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)(4-fluorophenyl)methyl ketone (1.06 g, 2.17 mmol, 1.00 eq). The mixture was then stirred at 90 °C for 14 hours. The reaction mixture was cooled to room temperature, quenched with water (100 mL), and filtered. The filter cake was washed with EtOAc (30.0 mL × 3), and the filtrate was washed with brine (50.0 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (silica gel, petroleum ether: EtOAc = 3:1 to 0:1) to give the title compound (1.07 g, 82.1% yield) as a pale yellow solid: LCMS: RT = 2.749 mins, m / z = 584.3 (M+H)+.1H NMR: (400 MHz, d6-DMSO) δ 8.23 ​​(s, 2H), 7.97 (d, J = 1.6 Hz, 1H), 7.95 (s, 1H), 7.58–7.82 (m, 2H), 7.43 (br s, 1H), 7.37 (br t, J = 8.8 Hz, 2H), 7.26 (d, J = 1.2 Hz, 1H), 4.78 (br s, 2H), 4.17 (br t, J = 5.6 Hz, 2H), 2.86 (br s, 2H), 2.86 (br t, J = 5.6 Hz, 2H). s,2H),1.26(s,9H).

[0293] Step 8: Synthesis of N-((S)-1-(2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)-1-(4-fluorophenyl)ethyl)-2-methylpropane-2-sulfinamide (I-1): Under N2 (g), (S,E)-N-((2-(1-) (6-bromopyrrolo[2,1-f][1,2,4]triazin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)(4-fluorophenyl)methylene)-2-methylpropane-2-sulfinamide (600 mg, 1.00 mmol, 1.00 eq) was cooled (0 °C) and MeMgBr (3.00 M solution in diethyl ether, 3.34 mL, 10.0 eq) was added. The mixture was then stirred at 25 °C for 1 h. The mixture was carefully quenched with saturated NH4Cl aqueous solution (30.0 mL) and extracted with EtOAc (20.0 mL × 3). The organic extract was washed with brine (20.0 mL), dried over Na2SO4, and concentrated under vacuum. The residue was purified by preparative TLC (SiO2, petroleum ether: EtOAc = 1:1, Rf = 0.35, 0.30) to give the title compound I-1 (632 mg, 89.3% yield, 84.7% purity) as a yellow solid: LCMS: RT = 1.024 mins, m / z = 598.2 (M+H)+.

[0294] In some preparations, I-1 is mixed with 4M HCl in MeOH (7 vol.) and dioxane (6.0 eq) and heated to 40 °C. After the reaction is complete, the mixture is cooled to room temperature and charged with MTBE (10 vol.). The mixture is filtered, washed with MTBE, and dried under vacuum to give amine (I-1a):

[0295]

[0296] The solution was purged with N2(g) for 10 minutes with KOAc (4.0 mmol) in amine I-1a, 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborane) (3.1 mmol), Pd(dppf)Cl2 (200 μmol), dppf (300 μmol), and 1,4-dioxane (30 mL), and stirred at 80 °C for 16 hours. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH = 15 / 1) to give compound (I-1A):

[0297]

[0298] In some preparations, I-1A was mixed with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborphane-2-yl)-1H-pyrazole (6.0 mmol), Pd(dppf)Cl2 (605 μmol), and K2CO3 (18.2 mmol) in DMF / H2O (40 mL / 10 mL), purged with N2(g) for 10 min, and stirred at N2 and 70 °C for 16 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH = 10 / 1) to give compound (I-1B):

[0299]

[0300] The corresponding (S)-hydroxy intermediates are: (I-8), (I-8A), and (I-8B):

[0301]

[0302] It was prepared at 0°C to room temperature by treating intermediate ix (obtained from step 6 above) in THF with methyl magnesiumgrinard. After the reaction was complete, the mixture was quenched in NH4Cl solution and extracted with EA. The combined organic layers were washed with H2O and brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography to obtain the racemic product. The enantiomers were separated by chiral HPLC to give (R)- and (S)-I-8. (S)-hydroxy compounds I-8A and I-8B were prepared by replacing I-1a with I-8 in the method of preparation 1.

[0303] Preparation 2: (S)-1-(4-fluorophenyl)-1-(2-(4-(6-iodo-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)pyrimidin-5-yl)ethane-1-amine hydrochloride (I-2).

[0304]

[0305] Add DIEA (1.66 g, 12.9 mmol, 2.24 mL, 4.00 eq) and 4-chloro-6-iodo-7H-pyrrolo[2,3-d]pyrimidine (900 mg, 3.22 mmol, 1.00 eq) to a suspension of compound I-5 (see Preparation 6) (1.32 g, 3.22 mmol, 1.00 eq) in DMF (23.2 mL). Stir the mixture at 40 °C for 60 h. Add the mixture dropwise to H2O (150 mL) and filter the resulting suspension. Wash the filter cake with H2O (20.0 mL × 3). The crude product was purified by preparative HPLC (column: Phenomenex Synergi Max-RP 250 x 50 mm x 10 mm; mobile phase: [water (0.05% HCl) - CH3CN]; B%: 10%-35%, 20 min) to give the title compound (1.20 g, 64.0% yield, 99.8% purity) as a yellow solid: LCMS: RT = 0.882 min, m / z = 528.3 (M-16)+.

[0306] In some preparations, a mixture of I-2 (2.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborane) (3.1 mmol), Pd(dppf)Cl2 (200 μmol), dppf (300 μmol), and KOAc (4.0 mmol) in 1,4-dioxane (30 mL) was purged with N2 (g) for 10 min and stirred at 80 °C for 16 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH = 15 / 1) to give compound (I-2A):

[0307]

[0308] In some preparations, I-2 was mixed with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborphane-2-yl)-1H-pyrazole (6.0 mmol), Pd(dppf)Cl2 (605 μmol), and K2CO3 (18.2 mmol) in DMF / H2O (40 mL / 10 mL), and purged with N2(g) for 10 min, then stirred at N2 and 70 °C for 16 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH = 10 / 1) to give compound (I-2B):

[0309]

[0310] The corresponding (S)-hydroxy intermediates are: (I-7), (I-7A), and (I-7B):

[0311]

[0312] It is prepared by replacing I-5 with intermediate I-6 (see Preparation 6) in Preparation 2.

[0313] Preparation 3: 1-(2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)-1-(4-fluorophenyl)ethane-1-ol (1k):

[0314]

[0315]

[0316] Step 1: 22 mmol of 6-bromopyrrolo[2,1-f][1,2,4]triazine-4(3H)-one 1a was dissolved in 100 mL of phosphorus oxychloride and reacted at 130 °C for 3 hours. The reaction solution was concentrated under reduced pressure. A saturated aqueous solution of sodium bicarbonate (100 mL) was added to the resulting residue and extracted with DCM. The organic phases were combined and washed with saturated NaCl solution (100 mL). The organic phases were dried over anhydrous Na₂SO₄ and concentrated under reduced pressure to give 6-bromo-4-chloropyrrolo[2,1-f][1,2,4]triazine 1b.

[0317] Step 2: Under an argon atmosphere, ethyl 2-chloropyrimidine-5-carboxylate 1c (10 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate tert-butyl 1d (11 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (1.2 mmol), and cesium carbonate (20 mmol) were dissolved in 1,4-dioxane / H₂O (66 mL, V / V = 10 / 1) and reacted at 60 °C for 5 hours. The reaction solution was diluted with EA (150 mL) and washed sequentially with water (30 mL × 2) and saturated NaCl aqueous solution (30 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by rapid column chromatography on silica gel to give ethyl 2-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidine-5-carboxylic acid ester 1e.

[0318] Step 3: Ethyl 2-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidine-5-carboxylic acid 1e (5.4 mmol) was dissolved in THF (15 mL), and NaOH aqueous solution (10 mL, 1 M) was added dropwise. The reaction was allowed to proceed for 5 h at room temperature. The reaction solution was concentrated under reduced pressure to remove THF, H2O (20 mL) was added, and the solution was adjusted to approximately pH 2-3 with 1 M HCl aqueous solution. A large amount of white solid product precipitated, which was filtered and dried to obtain crude 2-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidine-5-carboxylic acid 1f.

[0319] Step 4: 2-(1-(tert-butoxycarbonyl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidine-5-carboxylic acid 1f (4.86 mmol) was dissolved in DCM (50 mL), and N,N-diisopropylethylamine (24 mmol), 2-(7-azabenzotriazole)tetramethylurea hexafluorophosphate (9.7 mmol), and N,O-dimethylhydroxylamine hydrochloride (7.3 mmol) were added sequentially, and the reaction was carried out at room temperature for 6 hours. The reaction solution was diluted by adding DCM (150 mL), and washed sequentially with H2O (20 mL × 2), 1M HCl aqueous solution (20 mL), saturated sodium bicarbonate aqueous solution (20 mL), and saturated NaCl aqueous solution (20 mL). The organic phase was dried over anhydrous Na2SO4 and concentrated under reduced pressure. The residue was purified by rapid column chromatography on silica gel to obtain 1 g of tert-butyl 4-(5-(methoxy(methyl)carbamoyl)pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid.

[0320] Step 5: Under an argon atmosphere, 1 g (3.4 mmol) of 4-(5-(methoxy(methyl)carbamoyl)pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester was dissolved in 15 mL of THF. The reaction solution was cooled to 0 °C in an ice-water bath, and 14 mL (1 M / THF) of 4-fluorophenyl magnesium bromide was added dropwise, and the reaction was allowed to proceed at room temperature for 4 hours. The reaction was quenched by adding saturated ammonium chloride aqueous solution. The reaction solution was diluted with EA (100 mL) and washed successively with H2O (20 mL × 2) and saturated NaCl solution (20 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by rapid column chromatography on silica gel to give 1 h of 4-(5-(4-fluorobenzoyl)pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester.

[0321] Step 6: Dissolve 2.4 mmol of 4-(5-(4-fluorobenzoyl)pyrimidin-2-yl)-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester in DCM (10 mL), add trifluoroacetic acid (2 mL), and react at room temperature for 2 h. Concentrate the reaction solution under reduced pressure to obtain crude (4-fluorophenyl)(2-(1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl) methyl ketone, which is used directly in the next step.

[0322] Step 7: Dissolve (4-fluorophenyl)(2-(1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl) methyl ketone 1i (2.4 mmol) in DCM (15 mL). Add DIPEA (9.5 mmol) dropwise and stir for 5 minutes at room temperature. Then add 6-bromo-4-chloropyrrolo[2,1-f][1,2,4]triazine 1b (2.8 mmol) and react at room temperature for 12 hours. Concentrate the reaction solution under reduced pressure to remove DCM. Dilute the residue by adding EA (70 mL) and wash successively with H2O (10 mL), 1M HCl aqueous solution (10 mL), and saturated NaCl aqueous solution (20 mL). Dry the organic phase with anhydrous sodium sulfate and concentrate under reduced pressure. The residue was purified by rapid column chromatography on silica gel to obtain (2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazine-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)(4-fluorophenyl)methyl ketone 1j.

[0323] Step 8: Under an argon atmosphere, 0.4 mmol of (2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazine-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)(4-fluorophenyl)methyl ketone was dissolved in 10 mL of THF. The reaction solution was cooled to 0 °C in an ice-water bath, and 4.2 mL of methyl magnesium bromide (1 M / THF) was added dropwise, and the reaction was allowed to proceed at room temperature for 3 hours. The reaction was quenched at 0 °C by adding 10 mL of saturated ammonium chloride aqueous solution. The reaction solution was distilled under reduced pressure to remove THF. The reaction solution was diluted by adding 50 mL of EA to the residue. The aqueous layer was separated, and the organic phase was washed successively with 10 mL × 2 and 20 mL of saturated NaCl aqueous solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by rapid column chromatography on silica gel to give 1-(2-(1-(6-bromopyrrolo[2,1-f][1,2,4]triazine-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)-1-(4-fluorophenyl)ethane-1-ol (1k).

[0324] In some preparations, a mixture of KOAc (4.0 mmol) in 1k (2.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborane) (3.1 mmol), Pd(dppf)Cl2 (200 μmol), dppf (300 μmol), and 1,4-dioxane (30 mL) was purged with N2 (g) for 10 min and stirred at 80 °C for 16 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH) to give compound (1k-A):

[0325]

[0326] In some preparations, 1k-A was mixed with 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborphane-2-yl)-1H-pyrazole (6.0 mmol), Pd(dppf)Cl2 (605 μmol), and K2CO3 (18.2 mmol) in DMF / H2O (40 mL / 10 mL), and purged with N2(g) for 10 min, then stirred at N2 and 70 °C for 16 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH) to give compound (1k-B):

[0327]

[0328] The corresponding (S)-hydroxy intermediates are: (I-9), (I-9A), and (I-9B):

[0329]

[0330] It was prepared by treating intermediate 1j (obtained from step 7 above) with (S)-2-methylpropane-2-sulfinamide (0.908 mmol) and ethyl orthotitanate (0.715 mmol), and stirring in THF (3.2 mL) at 70 °C until the reaction was confirmed to be complete by TLC. Upon reaching room temperature, water was added, and the product was extracted into EA. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, concentrated under vacuum, and loaded onto a Celite plate. The residue was purified by MPLC (0-10% MeOH-EtOAc) to obtain the sulfonamide. The resulting sulfonamide was dissolved in THF and cooled to 0 °C. Methyl magnesium bromide (3 M solution in diethyl ether, 1.5 mmol) was added, and the mixture was stirred at 0 °C until complete. Additional methyl magnesium bromide was added if necessary. Saturated ammonium chloride was added, and the product was extracted into EA. The combined organic extracts were washed with brine, dried over Na₂SO₄, filtered, and concentrated under vacuum while being loaded onto a Celite plate. The residue was purified by MPLC (0–10% MeOH-EtOAc) to give methylsulfinamide, which was deprotected by stirring in 4M HCl in 1,4-dioxane (1.5 mL) / MeOH (1.5 mL) at room temperature. The solvent was removed under vacuum, and the residue was ground in EAc to give the racemic mixture. The enantiomers were separated by chiral HPLC to give (R)- and (S)-I-9.

[0331] Preparation 4: (R)-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborphanecyclopentan-2-yl)-1H-pyrazol-1-yl)propane-2-ol (I-3)

[0332]

[0333] Triethylamine (4.38 g, 43.3 mmol, 6.03 mL, 4.20 eq) was added to a solution of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1H-pyrazole (2.00 g, 10.3 mmol, 1.00 eq) and (R)-2-methylethylene oxide (1.50 g, 25.8 mmol, 1.81 mL, 2.50 eq) in acetonitrile (6.70 mL). The mixture was stirred at 100 °C for 12 h, then cooled and concentrated under vacuum. The residue was purified by column chromatography (SiO2, petroleum ether: EA = 2:1) to give the title compound (2.05 g, 78.9% yield) as a white oil. LCMS: RT=0.759min, m / z=253.2(M+H)+.1H NMR: (400MHz, d6-DMSO)δ7.85(s1H),7.56(s,1H),7.88(d,J=4.8Hz 1H),4.00-4.02(m,2H),3.93-3.96(m,1H),1.24(s,12H),1.00-1.01(m,3H).

[0334] Preparation 5: (S)-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazol-1-yl)propane-2-ol (I-4)

[0335]

[0336] Triethylamine (4.38 g, 43.3 mmol, 6.03 mL, 4.20 eq) was added to a solution of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (2.00 g, 10.3 mmol, 1.00 eq) and (S)-2-methylethylene oxide (1.80 g, 30.9 mmol, 2.17 mL, 3.00 eq) in acetonitrile (6.70 mL). The mixture was stirred at 100 °C for 12 hours, then cooled and concentrated under vacuum. The residue was purified by column chromatography (SiO2, petroleum ether: EtOAc = 2:1) to give the title compound (2.13 g, 72.4% yield, 88.4% purity) as a white oil: LCMS: RT = 0.767 min, m / z = 253.2 (M+H) + 1H NMR (400 MHz, d6-DMSO) δ 7.85 (s 1H), 7.56 (s, 1H), 7.88 (d, J = 4.8 Hz 1H), 4.00–4.02 (m, 2H), 3.93–3.96 (m, 1H), 1.24 (s, 12H), 1.01 (d, J = 6.0 Hz, 3H).

[0337] Preparation 6: (S)-1-(4-fluorophenyl)-1-(2-(piperazin-1-yl)pyrimidin-5-yl)ethane-1-amine hydrochloride (I-5A)

[0338]

[0339] Step 1: Synthesis of (S,Z)-4-(5-(((tert-butylsulfinyl)imino)(4-fluorophenyl)methyl-pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester

[0340] 20.0 g (1.0 eq) of 4-(5-(4-fluorobenzoyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester, (S)-(-)-2-methyl-2-propanesulfinamide (9.43 g, 1.5 eq) and LiOH (0.64 g, 0.5 eq) were added to a reaction vessel containing toluene (160 mL). Titanium isopropoxide (IV) (18.42 g, 1.25 eq) was added to the mixture, and the reaction was stirred at 50–60 °C for 1 h. The reactants were then distilled to remove 80 mL, while another 80 mL of toluene was added at 40–60 °C. The reaction mixture was cooled to 20–30 °C, and then a sodium citrate solution (80 mL, 30% w / w citric acid, pH 3–4) was added. The mixture was stirred at 45–55 °C for 1.5 h, and then the phases were separated. The organic phase was washed with potassium bicarbonate (40 mL, 25% w / w aqueous solution) and distilled to remove 40 mL. The product solution was diluted with tetrahydrofuran (30 mL) and then used directly as a solution for the next step (approximately 15% w / w (S,Z)-4-(5-(((tert-butylsulfinyl)imino)(4-fluorophenyl)methyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester).

[0341] Step 2: Synthesize 4-(5-((S)-1-(((S)-tert-butylsulfinyl)amino)-1-(4-fluorophenyl)methyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester by isolation

[0342] At 10°C, over 2–3 hours, 27.8 g of methyl magnesium chloride (22% w / w, 2.0 eq in THF) was added to a reaction solution (120 g, corresponding to 20 g of input substance) of (S,Z)-4-(5-(((tert-butylsulfinyl)imino)(4-4-fluorophenyl)methyl)-pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester in toluene / THF. The reaction mixture was allowed to be stirred for 1.5 hours until the reaction was complete. The reaction mixture was quenched by adding methanol (40 mL), followed by water (10 mL). The mixture was distilled to remove 100–110 mL of distillate, and then washed with ammonium chloride (80 mL, 20% w / w in water). The organic phase was washed with water (80 mL), diluted with toluene (60 mL), and distilled to remove 60–80 mL of distillate. At 50-60°C, 80 mL of n-heptane was added to a solution of 4-(5-((S)-1-(((S)-tert-butylsulfinyl)amino)-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester, and the solution was cooled to 42°C. At this point, 25-50 mg of seed crystals were added. The solution was kept for 30 min, and then cooled to 0-10°C for 30 min. The solid was separated by filtration, washed with a mixture of n-heptane and toluene (1:1, 30 mL), and then washed with n-heptane (30 mL). The product was dried to give 9 g (40-45%) of crude 4-(5-((S)-1-(((S)-tert-butylsulfinyl)amino)-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (96.4-97.2% de).

[0343] Step 3: Recrystallization of 4-(5-((S)-1-(((S)-tert-butylsulfinyl)amino)-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester.

[0344] 10.0 g of 4-(5-((S)-1-(((S)-tert-butylsulfinyl)amino)-1-(4-fluorophenyl)ethyl)pyrimidin-2-ylS,Z)piperazine-1-carboxylic acid tert-butyl ester was dissolved in isopropanol (100 mL) and heated to 40-60 °C, then passed through a filter washed / rinsed with isopropanol (20 mL). The resulting solution was vacuum distilled at 40-60 °C to remove 60-70 mL of distillate. The mixture was diluted with water (45 mL) at 50-60 °C and then cooled to 40 °C, at which point seed crystals (25-50 mg) were added. The mixture was further cooled to 20-25 °C and water (20 mL) was added. The solid was separated by filtration, washed with an isopropanol / water mixture (1:1, 20 mL), and the slurry was washed with isopropanol / water (1:2, 30 mL). Drying yielded 8.5 g (85%) of the product 4-(5-((S)-1-(((S)-tert-butylsulfinyl)amino)-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)piperazine-1-carboxylic acid tert-butyl ester (>99.8% de).

[0345] Step 4: Synthesis of (S)-1-(4-fluorophenyl)-1-(2-(piperazin-1-yl)pyrimidin-5-yl)ethane-1-amine hydrochloride (I-5A)

[0346] 4-(5-((S)-1-(((S)-tert-butylsulfinyl)amino)-1-(4-fluorophenyl)ethyl)pyrimidin-2-ylS,Z)piperazine-1-carboxylic acid tert-butyl ester (7) (9.0 g) was heated to 45-55 °C for 1 h in acetonitrile (40 mL) containing hydrochloric acid (33%, 8.14 g, 4.1 eq) to give (S)-1-(4-fluorophenyl)-1-(2-(piperazin-1-yl)pyrimidin-5-yl)ethane-1-amine hydrochloride (I-5A). Preparation of (S)-1-(4-fluorophenyl)-1-(2-(piperazin-1-yl)pyrimidin-5-yl)ethanol (I-6):

[0347] Its R-enantiomer is described in WO2015 / 057873.

[0348] Preparation of compounds

[0349] Example 1: Synthesis of (S)-2-(4-(4-(4-(5-(1-amino-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)pyrrolo[2,1-f][1,2,4]triazine-6-yl)-1H-pyrazol-1-yl)ethane-1-ol (1)

[0350]

[0351] Step 1: Synthesis of N-((S)-1-(4-fluorophenyl)-1-(2-(1-(6-(1-(2-hydroxyethyl)-1H-pyrazol-4-yl)pyrrolo[2,1-f][1,2,4]triazin-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)ethyl)-2-methylpropane-2-sulfinamide (i): Under N2 (g), I-1 (300 mg, 424 μL) was added to DMF (5.00 mL) to form 2-methylpropane-2-sulfinamide (i). Pd(dppf)Cl2·CH2Cl2 (52.0 mg, 63.7 μmol, 0.15 eq) was added to a solution of 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazol-1-yl)ethane-1-ol (142 mg, 594 mmol, 1.40 eq) and K2CO3 (117 mg, 849 μmol, 2.00 eq). The mixture was then stirred at 90 °C for 2 hours. The mixture was diluted with EtOAc (50.0 mL) and then washed successively with water (10.0 mL) and brine (35.0 mL × 3). The aqueous layer was extracted with EtOAc (20.0 mL × 2), and the combined organic layers were dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC (column: 3-Phenomenex LunaC18 75 x 30 mm x 3 mm; mobile phase [water (0.05% HCl) - CH3CN]; B%: 29%-49%, 6.5 min). The fractions containing the product were combined and the solution was saturated with NaHCO3 aqueous solution, and the pH was adjusted to 9. Extraction was performed with EtOAc (20.0 mL × 3), followed by concentration under vacuum to give the title compound as a pale yellow solid (66.0 mg, 105 μmol, 24.7% yield): LCMS: RT = 0.877 min, m / z = 630.3 (M+H)+.

[0352] Step 2: Synthesis of (S)-2-(4-(4-(4-(5-(1-amino-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)-3,6-dihydropyridin-1(2H)-yl)pyrrolo[2,1-f][1,2,4]triazin-6-yl)-1H-pyrazol-1-yl)ethane-1-ol (1): N-((S)-1-(4-fluorophenyl)-1-(2-( HCl / MeOH (4.00 M, 0.50 mL, 19.1 eq) was added to a solution of 1-(6-(1-(2-hydroxyethyl)-1H-pyrazol-4-yl)pyrrolo[2,1-f][1,2,4]triazine-4-yl)-1,2,3,6-tetrahydropyridin-4-yl)pyrimidin-5-yl)ethyl)-2-methylpropane-2-sulfinamide (66.0 mg, 105 mmol, 1.00 eq). The mixture was then stirred at 25 °C for 2 h. The pH of the mixture was adjusted to 5 and concentrated under vacuum. The residue was diluted with DMF (1.00 mL). The residue was purified by preparative HPLC (column: 3-Phenomenex Luna C18 75 x 30 mm x 3 mm; mobile phase [water (0.05% HCl)-CH3CN]; B%: 14%-34%, 6.5 min). The fractions containing the product were combined, and the pH was adjusted to 8 with a saturated aqueous solution of NaHCO3. Extraction was performed with EtOAc (20.0 mL x 3), and the combined organic layers were washed with brine (20.0 mL), dried over Na2SO4, and concentrated to give the title compound (18.0 mg, 31.7% yield) as a yellow gel: LCMS: RT = 2.178 mins, m / z = 526.5(M+H)+.1H NMR(400MHz,d6-DMSO)δ8.79(s,2H),8.08(s,1H),7.99(d,J=1.6Hz,1H),7.87(s,1H),7.85(s ,1H),7.44-7.51(m,2H),7.23-7.30(m,2H),7.08-7.16(m,2H),4.94(t,J=5.6Hz,1H),4.74(br s,2H),4.12-4.19(m,4H),3.72-3.79(m,2H),2.82(br s,2H),1.80(s,3H).

[0353] Example 2: Synthesis of (S)-2-(4-(4-(4-(5-(1-amino-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-1H-pyrazol-1-yl)ethane-1-ol hydrochloride (2)

[0354]

[0355] Add Pd(dppf)Cl2·CH2Cl2 (45.0 mg, 55.1 mmol), K2CO3 (63.1 mg, 457 mmol, 3.00 eq), and H2O (0.50 mL) to a solution of I-2 (90.0 mg, 152 mmol, 98.3% purity, 1.00 eq), 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazol-1-yl)ethane-1-ol (43.5 mg, 183 mmol, 1.20 eq) in DMF (2.00 mL). Degas the mixture three times with N2 (g) and then stir at 95 °C for 5 hours. Dilute the mixture with EtOAc (20.0 mL), wash with water (10.0 mL × 3), and wash with brine (10.0 mL). The aqueous layer was extracted with EtOAc (20 mL). The combined organic layers were dried over Na2SO4 and concentrated under vacuum. The residues were purified by preparative HPLC (column: 3-Phenomenex LunaC18 75 x 30 mm x 3 mm; mobile phase: [water (0.05% HCl)-CH3CN]; B%: 9%-29%, 7 min). The fraction containing the product was freeze-dried to give the title compound (30.3 mg, 34.4% yield) as a pale yellow solid: LCMS: RT = 0.711 min, m / z = 512.4 (M-16) + 1H NMR (400 MHz, d6-DMSO) δ 13.21–12.41 (m, 1H), 9.63–8.94 (m, 3H), 8.42 (s, 2H), 8.30 (s, 1H), 8.22 (s, 1H), 8.00 (s, 1H), 7.53–7.42 (m, 2H), 7.31 (t, J = 8.8 Hz, 2H), 7.12 (br d, J = 1.2 Hz, 1H), 4.19 (br The t, J = 5.2 Hz, 2H values ​​were 4.13–4.06 (m, 4H), 4.03–3.97 (m, 4H), 3.76 (t, J = 5.2 Hz, 2H), and 2.02 (s, 3H). Compounds 12, 13, 14, 14A, and 15 were prepared using the same procedure with appropriate intermediates instead of I-2.

[0356] Example 3: Synthesis of (S)-1-(4-fluorophenyl)-1-(2-(4-(6-(1-(oxecyclobutane-3-yl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)pyrimidin-5-yl)ethane-1-amine (3)

[0357]

[0358] The title compound was prepared as described in Example 2, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1H-pyrazole was replaced with 1-(oxecyclobutan-3-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1H-pyrazole-1-yl)ethane-1-ol was used. The crude product was purified by preparative HPLC (column: 3-Phenomenex Luna C18 75 x 30 mm x 3 mm; mobile phase [water (0.05% HCl) – CH3CN]; B%: 11%–31%, 6.5 min). The fractions containing the product were combined, the pH was adjusted to 8–9 with solid sodium carbonate, and extracted with EtOAc (20.0 mL × 3). The organic layer was washed with brine (10.0 mL), dried over Na₂SO₄, filtered, and concentrated under vacuum to give the title compound (97.5 mg, 50.8% yield) as a grayish-white solid: LCMS: EW25770-9-P1C, product: RT = 0.876 min, m / z = 524.4 (M⁻¹⁶)⁺. ¹H NMR: EW25770-9-P1A 400MHz, d6-DMSO) δ 12.0(s, 1H), 8.40(s, 2H), 8.34(s, 1H), 8.15(s, 1H), 8.09(s, 1H), 7.41–7.51(m, 2H), 7.05–7.15(m, 2H), 6.92(d, J = 1.2Hz, 1H), 5.61(q, J = 6.4Hz, 1H), 4.93–5.00(m, 2H), 4.84–4.92(m, 2H), 3.95–3.98(m, 4H), 3.87–3.88(m, 4H), 1.73(s, 3H). Compounds 28, 29, 30, 31, and 32 were prepared using the same procedure with appropriate intermediates instead of I-2.

[0359] Example 4: Synthesis of (R)-1-(4-(4-(4-(5-((S)-1-amino-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-1H-pyrazol-1-yl)propane-2-ol (4)

[0360]

[0361] The title compound was prepared as described in Example 3, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazol-1-yl)ethane-1-ol was replaced with I-3. The crude product was purified by preparative HPLC (column: 3-Phenomenex Luna C18 75 x 30 mm x 3 mm; mobile phase [water (0.05% HCl)-CH3CN]; B%: 11%-31%, 6.5 min). The fraction containing the product was adjusted to pH 8 and extracted with EtOAc (20.0 mL × 3). The combined organic extracts were washed with brine (20.0 mL), dried over Na2SO4, and concentrated to give the title compound (51.3 mg, 17.0% yield) as a grayish-white solid: LCMS: RT = 0.858 min, m / z = 526.4 (M-16) + 1H. NMR (400 MHz, d6-DMSO) δ 12.0 (s, 1H), 8.39 (s, 2H), 8.05–8.20 (m, 2H), 7.94 (s, 1H), 7.41–7.53 (m, 2H), 7.01–7.18 (m, 2H), 6.86 (d, J = 2.0 Hz, 1H), 4.97 (d, J = 4.8 Hz, 1H), 4.01–4.05 (m, 2H), 3.92–3.98 (m, 4H), 3.81–3.91 (m, 4H), 1.73 (s, 3H), 1.06 (d, J = 6.0 Hz, 3H). Compounds 7, 8, 9, 10, and 11 were prepared using the same procedure with appropriate intermediates instead of I-2.

[0362] Example 5: Synthesis of (S)-1-(4-(4-(4-(5-((S)-1-amino-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)piperazin-1-yl)-7H-pyrrolo[2,3-d]pyrimidin-6-yl)-1H-pyrazol-1-yl)propane-2-ol hydrochloride (5)

[0363]

[0364] The title compound was prepared as described in Example 2, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazol-1-yl)ethane-1-ol was replaced with I-4. The crude product was purified by preparative HPLC (column: 3-Phenomenex Luna C18 75 x 30 mm x 3 mm; mobile phase [water (0.05% HCl)-CH3CN]; B%: 10%-30%, 7 min). The fractions containing the desired product were combined and lyophilized to give the title compound (128.5 mg, 57.8% yield) as a yellow solid: LCMS: RT = 0.850 min, m / z = 526.5 (M-16) + .1H NMR: (400 MHz, d6-DMSO) δ 13.3 (br s, 1H), 9.55 (br s, 3H), 8.47(s, 2H), 8.35(s, 1H), 8.28(s, 1H), 8.04(s, 1H), 7.51-7.55(m, 2H), 7.26-7.34(m, 2H), 7.26(s, 1H), 4.15-4.17(m, 4H), 4.10-4.11(m, 2H), 4.02-4.04(m, 4H), 3.96-4.00(m, 2H), 2.03(s, 3H), 1.07(d, J = 6.0 Hz, 3H). Compounds 38, 39, 40, 41, and 42 were all prepared using the same procedure with appropriate intermediates instead of I-2.

[0365] Example 6: Synthesis of (S)-1-(4-fluorophenyl)-1-(2-(4-(6-(1-(oxecyclobutane-3-ylmethyl)-1H-pyrazol-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-yl)piperazin-1-yl)pyrimidin-5-yl)ethane-1-amine hydrochloride

[0366]

[0367] The title compound was prepared as described in Example 3, except that 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole was replaced with 1-(oxecyclobutane-3-ylmethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole-1-yl)ethane-1-ol was used. The crude product was purified by preparative HPLC (column: 3-Phenomenex Luna C18 75x30mm x 3um; mobile phase [water (0.05% HCl)-CH3CN]; B%: 10%-30%, 7 min). The fractions containing the desired product were combined and lyophilized to give the title compound (66.4 mg, 29.4% yield) as a pale yellow solid: LCMS: RT = 0.776 min, m / z = 555.5 (M+H)+.1H NMR: (400 MHz, d6-DMSO) δ 13.3 (s, 1H), 9.44 (br) Compounds 33, 34, 35, 36, and 37 were prepared using the same procedure with appropriate intermediates instead of I-2. The chromatogram values ​​were: 8.95 (s, 3H), 8.44 (s, 2H), 8.35 (s, 1H), 7.43–7.55 (m, 3H), 7.27–7.32 (m, 2H), 4.72 (dd, J = 8.0, 12.0 Hz, 2H), 4.49 (dd, J = 4.8, 11.6 Hz, 2H), 4.09–4.19 (m, 4H), 3.95–4.07 (m, 4H), 2.02 (s, 3H).

[0368] Example 7:

[0369]

[0370] A mixture of 1kB (prepared according to Preparation 3) (0.4 mmol), Cs₂CO₃ (0.8 mmol), and 2,2-dimethylethylene oxide (1.2 mmol) in NMP (5 mL) was stirred at 120 °C for 10 hours. The reaction mixture was diluted with EA, washed with H₂O and brine, and dried over Na₂SO₄. The organic layer was concentrated under vacuum, the residue was purified by preparative HPLC, and then lyophilized to give compound 69. Compounds 70, 71, 72, 73, and 74 were prepared using the same procedure, with appropriate intermediates substituted for 1kB.

[0371] Example 8:

[0372]

[0373] Step 1: At room temperature, cesium carbonate (50 mmol) and sodium iodide (16 mmol) were added to a solution of methyl 2-bromo-2-methylpropionate (x) (16 mmol) and 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (xi) (16 mmol) in NMP (20 mL). The resulting mixture was stirred at 120 °C for 8 hours. The reaction mixture was diluted with DCM and washed successively with H2O and brine. The organic layer was concentrated under vacuum, and the residue was purified by rapid column chromatography on silica gel (petroleum ether: ethyl acetate) to give compound (xii).

[0374] Step 2: Under N2(g), a mixture of methyl 2-methyl-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazol-1-yl)propionate (xii) (0.6 mmol), I-7 (0.6 mmol), Pd(dppf)Cl2 (0.12 mmol), and K2CO3 (1.8 mmol) in DMF / H2O (8 mL / 2 mL) was stirred at 70 °C for 4 hours. The solution was then diluted with DCM, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH) to give compound (xiii).

[0375] Step 3: At 0 °C, LiAlH4 (13.4 mmol) was added to a solution of methyl (S)-2-(4-(4-(5-(1-amino-1-(4-fluorophenyl)ethyl)pyrimidin-2-yl)piperazin-1-yl)pyrrolo[2,1-f][1,2,4]triazin-6-yl)-1H-pyrazol-1-yl)-2-propionate (xiii) (0.34 mmol) in THF (20 mL), and the mixture was stirred at room temperature for 6 hours. The reaction mixture was quenched with H2O (100 mL) and 10% NaOH H2O (300 mL), and then extracted with EA. The organic layer was concentrated under vacuum, and the residue was purified by preparative HPLC and then lyophilized to give compound (79). Compounds 65, 76, 77, 78 and 80 were prepared using the same procedure and with appropriate intermediates replacing I-7.

[0376] Example 9:

[0377]

[0378] Step 1: Add TsCl (33 mmol) to a solution of (S)-1-(benzyloxy)propane-2-ol (xvi) (30 mmol) and TEA (90 mmol) in DCM (80 mL). Stir the mixture at room temperature for 24 hours. Dilute the solution with DCM, wash with H2O, and wash with brine. Concentrate the organic layer and purify the residue by rapid column chromatography (petroleum ether / ethyl acetate) on silica gel to give compound (xvii).

[0379] Step 2: A mixture of (S)-1-(benzyloxy)propane-2-yl 4-toluenesulfonate (xvii) (6.2 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (xi) (6.2 mmol), and Cs₂CO₃ (13 mmol) in NMP (12 mL) was microwave-irradiated for 0.5 h at 110 °C. The solution was then diluted with EA, washed with H₂O, and washed with brine. The organic layer was concentrated, and the residue was purified by rapid column chromatography (PE / EA) on silica gel to give compound (x viii).

[0380] Step 3: Add Pd / C (800 mg) and HOAc (0.2 mL) to a solution of (R)-1-(1-(benzyloxy)propane-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (x viii) (2.3 mmol) in MeOH (20 mL). Purge the solution with H2 (g) for 5 minutes, then stir at room temperature under H2 (g) for 16 hours. Afterward, filter the mixture and concentrate the filtrate to obtain compound (xix).

[0381] Step 4: A mixture of ((R)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1H-pyrazol-1-yl)propane-1-ol (xix) (595 μmol), I-1a (595 μmol), Pd(dppf)Cl2 (60 μmol), and K2CO3 (1.8 mmol) in DMF / H2O (4 mL / 1 mL) was purged with N2 (g) for 10 min and stirred at 70 °C for 16 h under N2 (g). The mixture was extracted with EA, and the combined organic extracts were concentrated. The residues were purified by rapid column chromatography (DCM / MeOH) on silica gel. Compounds 16, 17, 19, and 20I-1a were prepared using the same procedure with appropriate intermediates substituted for I-1a.

[0382] Example 10:

[0383]

[0384] Step 1: TsCl (22 mmol) was added to a solution of (R)-1-(benzyloxy)propane-2-ol (xxii) (18 mmol) and TEA (54 mmol) in DCM (30 mL). The resulting mixture was stirred at 25 °C for 16 hours. The mixture was then concentrated under vacuum, and the residue was purified by rapid column chromatography (petroleum ether / ethyl acetate) on silica gel to give compound (xxiii).

[0385] Step 2: A mixture of (R)-1-(benzyloxy)propane-2-yl 4-toluenesulfonate (xxiii) (6.9 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (xi) (10 mmol), and Cs₂CO₃ (6.9 mmol) in NMP (50 mL) was stirred at 110 °C for 16 hours. The solution was then diluted with EA, washed with H₂O and brine, and concentrated. The residue was purified by rapid column chromatography (PE / EA) on silica gel to give compound (xxiv).

[0386] Step 3: Add Pd / C (800 mg) and HOAc (0.2 mL) to a mixture of (S)-1-(1-(benzyloxy)propane-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (xxiv) (2.6 mmol) in MeOH (20 mL). Purge the resulting mixture with H2 (g) for 5 min, then stir at room temperature in H2 (g) for 16 hours. Afterward, filter and concentrate the mixture to give compound (xxv).

[0387] Step 4: A mixture of (S)-2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazol-1-yl)propane-1-ol (xxv) (392 μmol), I-1 (261 μmol), K2CO3 (227 μmol), and Pd(dppf)Cl2 (7 μmol) in DMF / H2O (5 mL / 1 mL) was stirred at 70 °C and N2 (g) for 4 hours. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by preparative HPLC and then lyophilized to give compound (24). Compounds 22, 23, 25, 26, and 27 were prepared using the same procedure, with appropriate intermediates used instead of I-1a in step 4.

[0388] Example 11:

[0389]

[0390] TsCl (2.0 mmol) and TEA (3.4 mmol) were added to a solution of trans-3-(benzyloxy)cyclobutanol (xxxi) (1.7 mmol) in DCM (20 mL). The mixture was stirred at room temperature for 16 hours. The solution was diluted with DCM, washed with H2O and brine, and then concentrated. The residue was purified by rapid column chromatography (PE / EA) on silica gel to give compound (xxxii).

[0391] Step 2: A mixture of trans-3-(benzyloxy)cyclobutyl 4-toluenesulfonate (xxxii) (0.95 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1H-pyrazole (xi) (0.95 mmol) and Cs₂CO₃ (1.9 mmol) in NMP (5 mL) was microwave-irradiated for 0.5 h at 110 °C. The solution was then diluted with EA and washed with H₂O and brine. The organic layer was concentrated under vacuum, and the residue was purified by rapid column chromatography (PE / EA) on silica gel to give the title compound (xxxiii).

[0392] Step 3: Add Pd / C (200 mg) and HOAc (5 drops) to a solution of cis-3-(benzyloxy)cyclobutyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (xxxiii) (0.54 mmol) in MeOH (5 mL). Purge the solution with H2 (g) for 5 min and stir at H2 (g) and room temperature for 16 h. Filter the mixture and evaporate the filtrate to dryness under vacuum to obtain compound (xxxiv).

[0393] Step 4: A mixture of cis-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborphane-2-yl)-1H-pyrazol-1-yl)cyclobutanol (xxxiv) (0.21 mmol), I-1a (0.21 mmol), Pd(dppf)Cl2 (0.021 μmol), and K2CO3 (0.63 mmol) in DMF / H2O (4 mL / 1 mL) was purged with N2 for 10 min and stirred at N2 (g) and 70 °C for 16 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was directly purified by rapid column chromatography (DCM / MeOH). The obtained substance was further purified by preparative HPLC and then lyophilized to give compound (46). Compounds 44, 45, 47, 48, and 49 were prepared using the same procedure, with appropriate intermediates replacing I-1a in step 4.

[0394] Example 12:

[0395]

[0396] Step 1: Add 4-methylbenzyloxycyclobutanol (xxxv) (2.8 mmol) and TEA (8.4 mmol) to a solution of cis-3-benzyloxycyclobutanol (10 mL) and 4-methylbenzenesulfonyl chloride (3.4 mmol), and stir the resulting mixture at room temperature for 16 h. Dilute the mixture with brine and extract with DCM. Concentrate the organic extract. Purify the residue directly by rapid column chromatography (PE / EA) on silica gel to give compound (xxxvi).

[0397] Step 2: A mixture of cis-toluene-4-sulfonic acid 3-benzyloxy-cyclobutyl ester (xxxvi) (1.5 mmol), 4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborhexacyclopentan-2-yl)-1H-pyrazole (xi) (2.2 mmol), and Cs₂CO₃ (4.5 mmol) in NMP (15 mL) was microwave-irradiated for 2 h at 120 °C. The solution was then diluted with EA, washed with H₂O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (PE / EA) to obtain compound (xxxvii).

[0398] Step 3: Add Pd / C (200 mg) and concentrated HCl (0.5 mL) to a solution of trans-1-(3-benzyloxy-cyclobutyl)-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborhexacyclopentan-2-yl)-1H-pyrazole (xxxvii) (1.2 mmol) in MeOH (10 mL). Stir the reaction mixture at room temperature with H2 (g) for 16 h. Filter the mixture and concentrate the filtrate to give compound (xxxviii).

[0399] Step 4: A mixture of trans-3-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborphane-2-yl)-pyrazol-1-yl]-cyclobutanol (xxxviii) (0.8 mmol), I-1a (0.8 mmol), Pd(dppf)Cl2 (0.08 mmol), and K2CO3 (2.3 mmol) in dioxane / H2O (4 mL / 1 mL) was purged with N2 (g) for 10 min and stirred at N2 (g) and 70 °C for 4 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel. The obtained substance was further purified by preparative HPLC and then lyophilized to give compound (52). Compounds 50, 51, 53, 54, and 55 were prepared using the same procedure, with appropriate intermediates replacing I-1a in step 4.

[0400] Example 13:

[0401]

[0402] Step 1: The mixture of 4-bromo-1H-pyrazole (xxxix) (55 mmol) and K₂CO₃ (110 mmol) in ethyl 2-chloroacetate (25 mL) was stirred at 80 °C for 15 h. The reaction mixture was cooled, diluted with EA, and washed with H₂O. The organic layer was evaporated, and the residue was purified by chromatography on silica gel (petroleum ether / ethyl acetate) to give compound (x1).

[0403] Step 2: Over 2 hours at 60°C, ethyl magnesium bromide solution (3M, in hexane, 30 mL, 90 mmol) was added dropwise to a solution of ethyl 2-(4-bromo-1H-pyrazol-1-yl)ethyl acetate (x1) (30 mmol) and titanium tetraisopropoxide (15 mmol) in anhydrous THF (60 mL). After stirring at the same temperature for 2 hours, the reaction mixture was diluted with EA and washed successively with 1N saturated HCl and H2O. The organic layer was evaporated, and the residue was purified by chromatography (petroleum ether / ethyl acetate) on silica gel to give compound (xli).

[0404] Step 3: At room temperature, pyridine p-toluenesulfonic acid (1.4 mmol) was added to a solution of 1-[(4-bromo-1H-pyrazol-1-yl)methyl]cyclopropane-1-ol (xli) (1.4 mmol) and 3,4-dihydro-2H-pyran (4.1 mmol) in DCM (8 mL). The mixture was stirred for 4 hours, then diluted with brine and washed with DCM. The organic layer was concentrated, and the residue was purified by chromatography on silica gel (PE / EA) to give compound (xlii).

[0405] Step 4: Under N2(g), a mixture of 1,4-dioxane (3 mL), H2O (1 mL), and DMF (0.5 mL) containing 4-bromo-1-{[1-(oxan-2-yloxy)cyclopropyl]methyl}-1H-pyrazole (xlii) (0.5 mmol), I-1A (1.1 mmol), Pd(dppf)Cl2 (106 μmol), and Na2CO3 (1.6 mmol) was stirred at 80 °C for 3 hours. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by chromatography (ethyl acetate / methanol) on silica gel to give compound (xliii).

[0406] Step 5: At room temperature, p-toluenesulfonic acid (1.0 mmol) was added to a solution of 1-(4-fluoro-phenyl)-1-{2-[4-(6-{1-[1-(tetrahydro-pyran-2-yloxy)-cyclopropylmethyl]-1H-pyrazol-4-yl}-pyrrolo[2,1-f][1,2,4]triazin-4-yl)-piperazin-1-yl]-pyrimidin-5-yl}-ethylamine (xliii) (0.32 mmol), and the resulting mixture was stirred for 2 h. The reaction mixture was concentrated, the residue was purified by preparative HPLC, and then lyophilized to give compound (58). Compounds 56, 57, 60, 61, and 62 were prepared using the same procedure with appropriate intermediates replacing I-1a.

[0407] Example 14:

[0408]

[0409] Step 1: At 0°C, NaH (30 mmol) was added to a solution of 4-bromo-1H-pyrazole (xxxix) (14 mmol) in THF (50 mL). The solution was stirred at room temperature for 1 hour, and then methyl 2,4-dibromobutyrate (xliv) (14 mmol) was added. The mixture was stirred for 16 hours and then diluted with EA. The organic layer was washed with H2O, washed with brine, and concentrated under vacuum. The residue was purified by rapid column chromatography on silica gel (petroleum ether / ethyl acetate) to give compound (xiv).

[0410] Step 2: Add NaBH4 (6.8 mmol) to a solution of methyl 1-(4-bromo-1H-pyrazolyl)cyclopropanecarboxylate (xiv) (2.3 mmol) in MeOH (15 mL), and stir the resulting mixture at 50 °C until complete. Dilute the reaction mixture with DCM, wash successively with H2O and brine, and concentrate under vacuum. Purify the residue by rapid column chromatography on silica gel (PE / EA) to give compound (xlvii).

[0411] Step 3: A mixture of (1-(4-bromo-1H-pyrazol-1-yl)cyclopropyl)methanol (xlvii) (463 μmol), I-1A (prepared as described in Preparation 1) (695 μmol), Pd(t-Bu3P)2 (93 μmol), and Cs2CO3 (1.4 mmol) in THF / H2O (8 mL / 2 mL) was purged with N2 (g) for 10 min and stirred at N2 (g) and 80 °C for 12 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography (DCM / MeOH). The obtained substance was further purified by preparative HPLC and then lyophilized to give compound (65). Compounds 63, 64, 66, 67, and 68 were prepared using the same procedure, with appropriate intermediates replacing I-1A in Step 3.

[0412] Example 15:

[0413]

[0414] Step 1: At room temperature, MsCl (25 mmol) was added to a solution of tetrahydro-furan-3-ol (xlviii) (23 mmol) and TEA (45 mmol) in DCM (20 mL). The mixture was stirred at room temperature for 16 hours. The reaction mixture was then diluted with DCM, washed successively with H2O and brine, dried over anhydrous Na2SO4, and concentrated to dryness to give compound (xlix).

[0415] Step 2: At room temperature, 4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborhexacyclopentan-2-yl)-1H-pyrazole (xi) (17 mmol) and Cs₂CO₃ (34 mmol) were added to a solution of (S)-tetrahydrofuran-3-ylmethanesulfonate (xlviii) (11 mmol) in NMP (50 mL). The mixture was stirred at 120 °C for 2 h. The solution was diluted with EA, washed successively with H₂O and brine, and concentrated under vacuum. The residue was purified by rapid column chromatography (PE / EA) to give compound (1).

[0416] Step 3: A mixture of (R)-1-(tetrahydro-furan-3-yl)-4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborphane-2-yl)-1H-pyrazole (1) (0.3 mmol), I-1a (0.3 mmol), Pd(dppf)Cl2 (0.06 mmol), and K2CO3 (0.9 mmol) in DMF (2 mL) and H2O (0.5 mL) was stirred at 80 °C until the reaction was complete under N2 (g). The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH). The obtained substance was then purified by preparative HPLC and lyophilized to give compound (83). Compounds 81, 82, 84, 85, and 86 were prepared using the same procedure, with appropriate intermediates replacing I-1a in step 3.

[0417] Example 16:

[0418]

[0419] Step 1: At room temperature, MsCl (12.5 mmol) was added to a solution of (R)-tetrahydrofuran-3-ol (1i) (11 mmol) and TEA (23 mmol) in DCM (20 mL), and the resulting mixture was stirred at room temperature for 6 hours. The reaction mixture was diluted with DCM, washed successively with H2O and brine, and concentrated to give compound (lii).

[0420] Step 2: A mixture of I-1B (0.6 mmol), (R)-tetrahydrofuran-3-ylmethanesulfonate (III) (0.9 mmol), and Cs₂CO₃ (1.9 mmol) in NMP (10 mL) was stirred at 120 °C, and the reaction was monitored by TLC. The solution was then diluted with EA, washed with H₂O and brine, and concentrated. The residue was directly purified by preparative HPLC and then lyophilized to give compound (89). Compounds 87, 88, 90, 91, and 92 were prepared using the same procedure, with appropriate intermediates used instead of I-1B in step 2.

[0421] Example 17:

[0422]

[0423] Step 1: At 0°C, MsCl (47 mmol) was added to a solution of tetrahydro-2H-pyran-4-ol (liii) (31 mmol) and TEA (94 mmol) in DCM (100 mL). The reaction mixture was stirred at room temperature for 3 h, then diluted with DCM and washed with a saturated aqueous solution of NaHCO3. The solution was dried over anhydrous Na2SO4. The solvent was removed to obtain compound (liv).

[0424] Step 2: A mixture of methyl tetrahydro-2H-pyran-4-methanesulfonate (1iv) (18 mmol), 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (21 mmol), and Cs₂CO₃ (27 mmol) in NMP (50 mL) was stirred at 80 °C for 4 h. The reaction mixture was diluted with DCM and washed with brine. The organic layer was evaporated under vacuum. The residue was purified by rapid column chromatography (PE / EA) on silica gel to give compound (1v).

[0425] Step 3: A mixture of I-1a (603 μmol), 1-(tetrahydro-2H-pyran-4-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)-1H-pyrazole (1v) (754 μmol), K2CO3 (754 μmol), and Pd(dppf)Cl2 (41 μmol) in DMF / H2O (10 mL / 2 mL) was stirred at 70 °C and N2(g) for 4 hours. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by preparative HPLC and then lyophilized to give compound (95). Compounds 93, 94, 96, 97, and 98 were prepared using the same procedure, with appropriate intermediates used instead of I-1a in step 3.

[0426] Example 18:

[0427]

[0428] Step 1: A solution of 3,6-dioxabicyclo[3.1.0]hexane (lvi) (61 mmol), 4-bromo-1H-pyrazole (xxxix) (61 mmol), and Cs₂CO₃ (121 mmol) in NMP (100 mL) was stirred at 120 °C for 16 h. The solution was cooled and diluted with DCM, then washed with H₂O and brine. The organic layer was concentrated and purified by rapid column chromatography on silica gel (PE / EA) to obtain compound (lvii).

[0429] Step 2: A mixture of racemic-trans-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol (1vii) (12 mmol), 4-nitrobenzoic acid (12 mmol), diisopropyl azodicarbonate (17 mmol), and triphenylphosphine (17 mmol) in THF (50 mL) was stirred at room temperature for 16 hours. The solution was diluted with EA and washed with H2O and brine. The organic layer was concentrated and purified by rapid column chromatography on silica gel (PE / EA) to give compound (lviii).

[0430] Step 3: The mixture of racemic-cis-4-(4-bromo-1H-pyrazole-1-yl)tetrahydrofuran-3-yl 4-nitrobenzoate (lvii) (11 mmol) and lithium hydroxide (53 mmol) in MeOH / THF / H2O (30 mL / 30 mL / 30 mL) was stirred at room temperature for 4 hours. The resulting mixture was diluted with EA, washed with H2O and brine, and concentrated under vacuum. The residue was purified by rapid column chromatography on silica gel (PE / EA) to give racemic-cis-4-(4-bromo-1H-pyrazole-1-yl)tetrahydrofuran-3-ol. This substance was chirally separated by SFC (column: AD 20*250 mm, 10 μm (Daicel); mobile phase: CO2 / MeOH (ammonia in 0.2% methanol) = 60 / 40; flow rate: 80 g / min) to give peak 1 (1x) and peak 2 (1xi). Peak 1 is arbitrarily designated as (3S,4S)-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol and peak 2 is arbitrarily designated as (3R,4R)-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol.

[0431] Step 4: A mixture of (3R,4R)-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol (1xi) (70 mg, 0.3 mmol) (peak 2 from Step 3), I-1A (0.6 mmol), Pd[(t-Bu)3P]2 (0.06 mmol), and Na2CO3 (0.9 mmol) in dioxane / H2O (8 mL / 2 mL) was stirred at 90 °C for 4 h. After cooling, the solution was diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH = 10 / 1) to give compound (101). Compounds 00, 100, 102, 103, and 104 were prepared using the same procedure, with appropriate intermediates replacing I-1A in Step 4.

[0432] Example 19:

[0433]

[0434] Step 1: Racemic-trans-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol (1.1 g) (from Step 1 of Example 18) was chirally separated via an SFC (column: AD 20*250 mm, 10 μm (Daicel); mobile phase: CO2 / MeOH (ammonia in 0.2% MeOH) = 80 / 20; flow rate: 80 g / min) to obtain peak 1 (1xiii) and peak 2 (1xiv). Peak 1 was arbitrarily designated as (3R,4S)-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol and peak 2 was arbitrarily designated as (3S,4R)-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol.

[0435] Step 2: A mixture of (3R,4S)-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol (0.3 mmol) (1xiii) (peak 1 from Step 1), I-1A (0.6 mmol), Pd[(t-Bu)3P]2 (0.06 mmol), and Na2CO3 (0.9 mmol) in dioxane / H2O (8 mL / 2 mL) was degassed with N2 and stirred at 90 °C for 4 h. The solution was then diluted with DCM, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH) to give the title compound (107). Compounds 105, 106, 108, 109, and 110 were prepared using the same procedure, with appropriate intermediates replacing I-1a in Step 2.

[0436] Example 20:

[0437]

[0438] A mixture of (3S,4R)-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol (70 mg, 0.3 mmol) (1 xiv) (peak 2 from step 1 of Example 19), I-1A (0.6 mmol), Pd[(t-Bu)3P]2 (0.06 mmol), and Na2CO3 (0.9 mmol) in dioxane / H2O (8 mL / 2 mL) was degassed with N2 and stirred at 90 °C for 4 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH) to give compound (113). Compounds 111, 112, 114, 115, and 116 were prepared using the same procedure, with appropriate intermediates substituted for I-1A.

[0439] Example 21:

[0440]

[0441] A mixture of (3S,4S)-4-(4-bromo-1H-pyrazol-1-yl)tetrahydrofuran-3-ol (1xiii) (0.22 mmol) (peak 1 from step 3 of Example 18), I-1A (0.44 mmol), Pd[(t-Bu)3P]2 (0.044 mmol), and Na2CO3 (0.66 mmol) in dioxane / H2O (8 mL / 2 mL) was stirred at 90 °C for 4 h. After cooling, the solution was diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH) to give the title compound (119). Compounds 117, 118, 120, 121, and 122 were prepared using the same procedure, with appropriate intermediates substituted for I-1A.

[0442] Example 22:

[0443]

[0444] Step 1: At 0°C, add 11.4 mmol of NaBH4 to a solution of 5.7 mmol of 2-(benzyloxy)cyclobutanone in 20 mL of MeOH. The solution is then stirred at room temperature for 3 hours. The mixture is diluted with EA, washed with water and brine, and the organic layer is concentrated and purified by rapid column chromatography on silica gel (PE / EA) to give peak 1 (arbitrarily designated as cis-2-(benzyloxy)cyclobutanol) and peak 2 (arbitrarily designated as trans-2-(benzyloxy)cyclobutanol) as a colorless oil.

[0445] Step 2: At 0°C, add methanesulfonyl chloride (2.3 mmol) and triethylamine (4.6 mmol) to a solution of cis-2-(benzyloxy)cyclobutanol (1.5 mmol) in DCM (10 mL). Stir the mixture at room temperature for 3 hours. Then, dilute the solution with DCM, wash with water and brine, dry with anhydrous Na₂SO₄, and concentrate to obtain the desired compound.

[0446] Step 3: The mixture of cis-2-(benzyloxy)cyclobutylmethanesulfonate (1.2 mmol), 4-bromo-1H-pyrazole (1.2 mmol), and Cs₂CO₃ (3.5 mmol) in DMF (8 mL) was stirred at 100 °C for 16 hours. The solution was then diluted with EA, washed with water and brine, dried over anhydrous Na₂SO₄, concentrated, and purified by rapid column chromatography on silica gel (PE / EA) to obtain the desired compound. Chiral separation of trans-2-(benzyloxy)cyclobutyl)-4-bromo-1H-pyrazole: trans-2-(benzyloxy)cyclobutyl)-4-bromo-1H-pyrazole was chirally separated via SFC (column: IG 20*250mm, 10μm (Daicel; mobile phase: CO2 / MeOH (ammonia in 0.2% methanol) = 75 / 25; flow rate: 4g / min), yielding peak 1 (250mg) and peak 2 (250mg). Peak 1 was arbitrarily designated as 1-((1S,2S)-2-(benzyloxy)cyclobutyl)-4-bromo-1H-pyrazole and peak 2 was arbitrarily designated as 1-((1R,2R)-2-(benzyloxy)cyclobutyl)-4-bromo-1H-pyrazole).

[0447] Step 4: The 1-((1S,2S)-2-(benzyloxy)cyclobutyl)-4-bromo-1H-pyrazole (820 μmol) solution in TFA (2 mL) was stirred at 80 °C for 16 hours. The solution was then concentrated and purified by rapid column chromatography on silica gel (petroleum ether / ethyl acetate) to obtain the desired compound.

[0448] Step 5: A mixture of (1S,2S)-2-(4-bromo-1H-pyrazol-1-yl)cyclobutanol (556 μmol), I-1A (667 μmol), Pd(t-Bu3P)2 (99 μmol), and Cs2CO3 (1.1 mmol) in dioxane / H2O (8 mL / 2 mL) was purged with N2 for 10 min and stirred at N2 and 90 °C for 4 h. The solution was then diluted with DCM, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH). The obtained substance was further purified by preparative HPLC and then lyophilized to give compound 125. Compounds 123, 124, 126, 127, and 128 were prepared using the same procedure, with appropriate intermediates used instead of I-1A in step 5.

[0449] Example 23:

[0450]

[0451] Step 1: At 80°C, a solution of 1-((1R,2R)-2-(benzyloxy)cyclobutyl)-4-bromo-1H-pyrazole (820 μmol) (peak 2 from step 3 of Example 22) in TFA (2 mL) was stirred for 16 hours. The solution was then concentrated and purified by rapid column chromatography on silica gel (petroleum ether / ethyl acetate) to obtain the desired compound.

[0452] Step 2: A mixture of (1R,2R)-2-(4-bromo-1H-pyrazol-1-yl)cyclobutanol (556 μmol), I-1A (667 μmol), Pd(t-Bu3P)2 (99 μmol), and Cs2CO3 (1.1 mmol) in dioxane / H2O (8 mL / 2 mL) was purged with N2 (g) for 10 min and stirred at N2 (g) and 90 °C for 4 h. The solution was then diluted with EA, washed with H2O and brine, and concentrated. The residue was purified by rapid column chromatography on silica gel (DCM / MeOH). The obtained substance was further purified by preparative HPLC and then lyophilized to give compound 131. Compounds 129, 130, 132, 133, and 134 were prepared using the same procedure, with appropriate intermediates substituted for I-1A.

[0453] Biological Example 1: Biochemical Enzyme Activity Inhibition Assay

[0454] Enzymatic activities of PDGFRα and KIT were monitored using the Perkin Elmer electrophoretic mobility shift technology platform EZReader2. Fluorescently labeled substrate peptides were incubated in the presence of kinases and ATP, and in the presence of the test compound, such that each dose of the test compound resulted in phosphorylation of the peptide according to its reflectance ratio.

[0455] During the linear, steady-state phase of the kinase-catalyzed reaction, a mixture of phosphorylated (product) and unphosphorylated (substrate) peptides is passed through the microfluidic system of the PerkinElmer EZ Reader 2 under the influence of an applied potential difference. The presence of phosphate groups on the product peptide provides a difference in mass and charge between the substrate peptide and the product, leading to the separation of the substrate and product mixture in the sample (Perrin et al., Expert Opin Drug Discovery 2010, Jan 5(1):51-63).

[0456] These conjugates (λ) are detected when the product and substrate peptide mixture passes through a laser within the instrument. ex =488nm,λ em =568nm) and were resolved into separate peaks. The ratio between these peaks reflects the activity of the compound at that concentration in the pore under these conditions.

[0457] Inhibition of KIT(D816V) mutant biochemical enzyme activity

[0458] All test products were dissolved in 100% DMSO at a stock concentration of 10 mM. In 100% DMSO, all test compounds were serially diluted 100-fold, 10-fold, and 4-fold, starting at the relevant concentration (typically 1 mM). Using a TTPLabtech Mosquito nano-liter dispenser, 130 nL volumes of each concentration were transferred to the relevant wells of a 384-well assay plate (Greiner 781 201). Using Multidrop, the remaining components of the reaction were then added to 130 nL of the compound as follows:

[0459] KIT D816V assay for ATP-targeted APPKM: In each well of a 384-well plate, with or without a series of compound concentrations (final concentration of 1% DMSO), 0.3 nM of untreated enzyme was incubated with 1 μM Src tide (5-FAM-GEEPLYWSFPAKKK-NH2) and 20 μM ATP in a co-concentrated 13 μL buffer (100 mM HEPES pH 7.5, 0.015% Brij 35, 10 mM mgCl2, 1 mM DTT) at 25°C for 60 min. The reaction was terminated by adding 70 μL of stop buffer (100 mM HEPES pH 7.5, 0.015% Brij 35, 35 mM EDTA, and 0.2% Coating Reagent 3, CaliperLifesciences). The plate was read on a Caliper EZReader 2. The results obtained for these experiments with the compounds prepared according to the examples are summarized in Table 2 below. For biochemical D816V and D842V activities, the following designations are used: ≤0.30 nM = A; ≥0.31 and <1.4 nM = B; ≥1.4 nM = C; and ND = undetermined.

[0460] Table 2.

[0461]

[0462]

[0463] For reference, the chemical structure of comparison substance A is as follows:

[0464]

[0465] Inhibition of PDGFRA mutant biochemical enzyme activity

[0466] All test products were dissolved in 100% DMSO at a stock concentration of 10 mM. In 100% DMSO, all test compounds were serially diluted 100-fold, 10-fold, and 4-fold, starting at the relevant concentration (typically 1 mM). Using a TTPLabtech Mosquito nano-liter dispenser, 130 nL volumes of each concentration were transferred to the relevant wells of a 384-well assay plate (Greiner 781 201). Using Multidrop, the remaining components of the reaction were then added to 130 nL of the compound as follows:

[0467] PDGFRαD842V assay for the apparent Michaelis constant (APPKM) of ATP: In each well of a 384-well plate, 7 nM of untreated enzyme was incubated with 1 μM SKtide (5-FAM-AHA-KKKKDDIYFFFG-NH2) and 25 μM ATP in a co-concentration of 13 μL buffer (100 mM HEPES pH 7.5, 0.015% Brij 35, 10 mM MgCl2, 1 mM DTT) for 90 min at 25°C. The reaction was terminated by adding 70 μL of termination buffer (100 mM HEPES pH 7.5, 0.015% Brij 35, 35 mM EDTA, and 0.2% CoatingReagent 3, Caliper Lifesciences). The plate was read on a Caliper EZReader 2. Biological implementation Example 2: Determination of wild-type KIT activity using SCF-stimulated UT-7 cell proliferation assay.

[0468] UT-7 cells are a human megakaryocytic leukemia cell line that can grow in cultures dependent on granulocyte-macrophage colony-stimulating factor (GM-CSF) or stem cell factor (SCF). UT-7 cells respond to SCF stimulation by activating KIT receptor tyrosine kinase and subsequent downstream signaling that supports cell growth and proliferation (Kuriu et al., 1999; Komatsu et al., 1991; Sasaki et al., 1995). The ability of compounds to inhibit SCF-stimulated UT-7 cell proliferation was tested.

[0469] The inhibition of SCF-stimulated UT-7 cell proliferation was assessed using the CellTiter-Glo assay, which quantifies the amount of adenosine triphosphate (ATP) present, a reading of metabolically active cells proportional to the number of viable cells in the culture. The ability of the test compounds to inhibit SCF-stimulated UT-7 cell proliferation was determined using a 10-point dose-response curve from 25 μM to 95.4 pM.

[0470] UT-7 cells were maintained in IMDM supplemented with 10% FBS, 5 ng / mL GM-CSF, and 100 units / mL penicillin-streptomycin and grown in a humidified tissue culture incubator at 37°C. UT-7 cells were washed once with serum-free, GM-CSF-free IMDM. Cells were then resuspended in IMDM containing 4% FBS and 50 ng / mL SCF and seeded at 22 μL per well in 384-well microplates. A 10-point dose series of test compounds (25.0 μM to 95.4 pM) was then added in 3.1 μL to each well (0.25% DMSO final concentration), and the plates were incubated in a tissue culture incubator (5% CO2, 37°C) for 72 hours. After 3 days of treatment with the test compounds, 25 μL of freshly prepared CellTiter-Glo reagent was added to each well. The plate was mixed by oscillating at 300 rpm for 10 minutes at room temperature on a plate oscillator. The plate was read on an EnVision plate reader using the Ultra Sensitive Luminescence protocol for 384-well plates. Data were normalized to 0% and 100% suppression controls and read using a four-parameter logic IC. 50 Curve fitting calculation IC 50 .

[0471] Biological Example 3: Rat Brain-Plasma Ratio (K) p Brain penetration assessment

[0472] To understand brain permeability, the brain-to-plasma ratio of compounds was obtained from Sprague-Dawley (SD) rats. The homeostatic distribution of compounds between blood and brain in preclinical species such as rats is a commonly used parameter for evaluating brain permeability. p The concentration ratio of brain to blood (C) brain / C plasma The passive diffusion properties of the compound, its affinity for membrane transport proteins at the blood-brain barrier (BBB), and the difference in relative drug binding affinity between plasma proteins and brain tissue all affect K. p The brain has a K value of less than 0.1. p Brain compounds are restricted from entering the CNS, with K+ levels greater than 0.3-0.5%. p Compounds from the brain are believed to have good brain permeability and a K+ value greater than 1. p ,Brain compounds free via BBB (Expert Opin. Drug Delivery (2016) 13(01): 85-92).

[0473] Brain permeability of the compounds disclosed herein was measured in Sprague-Dawley rats (3 / compound). Animals received an IV infusion of 1 mg / kg / hr of the compound over 8 hours via jugular vein cannulation. At 24 hours, blood was collected by tail vein bleeding or cardiac puncture (under anesthesia) and centrifuged to obtain plasma samples. Brain tissue was collected and homogenized with phosphate-buffered saline (PBS). The concentrations of the compounds in plasma and brain homogenates were obtained by LC-MS / MS analysis. Table 3 below shows the Kp, brain results for compounds 1 and 2 prepared according to the examples described herein and for comparative compound A.

[0474] Compared to Comparative A (mean = 1.8), compounds 1 and 2 exhibited very low Kp values ​​(mean = 0.133 and 0.045, respectively).

[0475] Rat plasma protein binding of the test compound was evaluated in vitro using a balanced dialysis method. The test compound was evaluated in 100% plasma within a dialysis block for 5 hours at 37°C. Samples from both the donor and recipient sides were analyzed by LC-MS / MS. The plasma protein-bound and unbound fractions were calculated using the following equation.

[0476] Combining the fraction (fb) * (%) = 100 x ([donor]) 5h -[receptor] 5h ) / [donor] 5h (Equation 1)

[0477] Unbound fraction (fu), p*(%) = 100 - %bound* (Equation 2)

[0478] Among them: [donor] 5h Donor concentration was measured after 5 hours; [Receptor] 5h The receptor concentration was measured over 5 hours; fb* represents the bound fraction determined from plasma; (fu),p* represents the unbound fraction calculated from plasma. Warfarin and quinidine were used as positive controls.

[0479] Similarly, the binding of the test compound to rat brain proteins was evaluated in vitro using a balanced dialysis method. A 1 μM compound was evaluated in brain homogenate within a dialysis block for 5 hours at 37°C. Samples from both the donor and recipient sides were analyzed by LC-MS / MS. The bound and unbound portions of the brain protein were calculated using the equations (Equations 1 and 2) described above.

[0480] Unbound brain to plasma ratio (K puu ,brain)

[0481] Based on the brain and plasma concentrations obtained above and the fu, brain value obtained above, the unbound brain to plasma ratio (K) is calculated. puu (brain), as shown in Table 3.

[0482] Compared to comparison compound A (mean = 0.84), compounds 1 and 2 exhibit highly superior low K values. puu Brain (mean = 0.017 and 0.01). Unbound drug concentrations in tissues are free drugs that can exert their pharmacological effects within tissue compartments. Because 1 and 2 have very low Kc values ​​compared to comparison A. puu The brain, which means that the amounts of compounds 1 and 2 that can exert their pharmacological effects in the brain are very low compared to comparison A.

[0483] Table 3.

[0484] Example <![CDATA[K p ,brain]]> <![CDATA[K puu ]]> 1 0.133 0.017 2 0.045 0.01 Comparative object A 1.75 0.84

Claims

1. Compounds of formula (I): (I)， Or its pharmaceutically acceptable salt, wherein: Selected from single and double bonds; Selected from single and double bonds; Z is selected from CH and NH; Y is selected from C and N; X1 is selected from CH, C, and N; X2 is selected from CH, C, and N; The condition is that when X1 and X2 are both N, then Y is not N and Z is not CH; A is ; R1 is selected from hydrogen and methyl; R2 is selected from hydrogen and methyl, or R1 and R2 together form a cyclopropyl group; R3 is selected from hydrogen and methyl; R4 is selected from hydrogen and methyl, or R3 and R4 together form a cyclopropyl group; R5 is selected from hydrogen and methyl; R6 is selected from hydrogen and methyl, or R5 and R6 together form a cyclopropyl group, or One of R2 or R4 together with R6 forms a cyclobutyl group; R7 is hydrogen; m is 0 or 1; n is 0 or 1; and B is selected from OH and NH2.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound has formula (II): (II)。 3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound has formula (III): (III)。 4. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein X1 is N and It is a single bond, and X2 is C and It is a double bond.

5. The compound of claim 3, or a pharmaceutically acceptable salt thereof, wherein X1 is C and It is a double bond, and X2 is N and It is a single key.

6. The compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein: A is: ; R3 is selected from hydrogen and methyl; R4 is selected from hydrogen and methyl, or R3 and R4 together form a cyclopropyl group; R5 is selected from hydrogen and methyl; R6 is selected from hydrogen and methyl; or R5 and R6 together form a cyclopropyl group, and R7 is hydrogen.

7. The compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein B is NH2.

8. The compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein B is OH.

9. The compound of claim 7, or a pharmaceutically acceptable salt thereof, wherein A is selected from: , , , , , , , and .

10. The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein A is selected from: , , , , , , , , , , , and .

11. The compound of claim 7, or a pharmaceutically acceptable salt thereof, wherein A is selected from: , , , , , , , ,and .

12. The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein A is selected from: , , , , , and .

13. The compound of claim 7, or a pharmaceutically acceptable salt thereof, wherein A is selected from: , and .

14. The compound of claim 8, or a pharmaceutically acceptable salt thereof, wherein A is... .

15. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

16. A pharmaceutical composition comprising: The compound according to any one of claims 1-15, or a pharmaceutically acceptable salt thereof; and Pharmaceutically acceptable excipients.

17. Use of any compound or pharmaceutically acceptable salt thereof according to any one of claims 1-15 in the preparation of a medicament for treating a disease or condition in a patient in need, wherein the disease or condition is selected from: systemic mastocytosis, gastrointestinal stromal tumor, acute myeloid leukemia, melanoma, seminoma, intracranial germ cell tumor, mediastinal B-cell lymphoma, Ewing's sarcoma, diffuse large B-cell lymphoma, dysgerminoma, myelodysplastic syndrome, nasal NK / T-cell lymphoma, chronic myelomonocytic leukemia, and brain cancer.

18. The use according to claim 17, wherein the disease or condition is systemic mastocytosis.

19. The use according to claim 18, wherein the systemic mastocytosis is selected from indolent systemic mastocytosis and smoldering systemic mastocytosis.

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