WO2023138412A1

WO2023138412A1 - Fused pyrimidin-2-amine compounds as cdk20 inhibitors

Info

Publication number: WO2023138412A1
Application number: PCT/CN2023/071020
Authority: WO
Inventors: Feng Ren; Xiao DING; Yingtao LIU; Min Zheng; Wei Zhu
Original assignee: InSilico Medicine IP Ltd
Current assignee: InSilico Medicine IP Ltd
Priority date: 2022-01-20
Filing date: 2023-01-06
Publication date: 2023-07-27
Anticipated expiration: 2024-07-20

Abstract

The present disclosure relates to compounds of Formula (I) or pharmaceutically acceptable salts, solvate, stereoisomer, or isotopic variant thereof useful as CDK20 inhibitors, pharmaceutical compositions comprising the same, and use thereof in the treatment of CDK20-associated diseases or conditions such as hepatocellular carcinoma (HCC), colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, renal cell carcinoma, etc.

Description

FUSED PYRIMIDIN-2-AMINE COMPOUNDS AS CDK20 INHIBITORS

CROSS-REFERENCE TO RELATED APPLICATION (S)

The present application claims priority to PCT Application No. PCT/CN2022/072865 filed on January 20, 2022, entitled “FUSED PYRIMIDIN-2-AMINE COMPOUNDS AS CDK20 INHIBITORS” , the disclosure of which is hereby incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present disclosure relates to novel fused pyrimidin-2-amine compounds or pharmaceutically acceptable salts, solvate, stereoisomer, or isotopic variant thereof, which are useful as CDK20 inhibitors. The present disclosure further relates to pharmaceutical compositions comprising one or more of such compounds or pharmaceutically acceptable salts, solvate, stereoisomer, or isotopic variant thereof as an active ingredient, and use of such compounds or pharmaceutically acceptable salts, solvate, stereoisomer, or isotopic variant thereof in the treatment of CDK20-associated diseases or conditions, such as hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, renal cell carcinoma, etc.

BACKGROUND

Cyclin-dependent kinase 20 (CDK20) , also known as cell cycle-related kinase (CCRK) , is a newly identified member of the cyclin-dependent kinase (CDK) family with increased attraction in recent years due to its functions (both cell cycle-dependent and -independent) in a variety of human tissues ¹. CDK20 is widely expressed at a comparable translational level in many human tissues including brain, lung, liver, pancreas, and gastrointestinal tract ². More importantly, increasing preclinical evidence has shown that CDK20 is overexpressed in many tumor cell lines as well as tumor samples from patients with different types of cancer such as hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma ^1, 3–6, indicating its potential role in cancer development, which was proven in numerous basic and preclinical studies by several different research groups. In vitro studies showed that androgen receptor (AR) , CDK20, and β-catenin constitute a positive feedback circuit to promote cell cycle progression in HCC cells, and CDK20 overexpression frequently correlates with ectopic expression of AR and β-catenin in primary HCC tissue samples, and with the tumor staging and poor overall survival of patients ⁴. In lung cancer cells, CDK20 competes with NRF2 for KEAP1 binding, which prevents degradation of NRF2 and enhances its transcriptional activity, and therefore lowering the cellular reactive oxygen species (ROS) level. Moreover, CDK20 depletion in lung cancer cells demonstrates impaired cell proliferation, defective G2/M arrest, and increased radiochemosensitivity ⁵. In addition to its pro-tumorigenic role through modulation of cell cycle and oncogenic signaling, CDK20 is also involved in immunosuppression in certain types of tumors. Zhou et al. reported that, by activating the EZH2-NF-κB pathway, CDK20 expressed in HCC cells increased IL-6 production and induced immunosuppressive MDSC expansion from human peripheral blood mononuclear cells; inhibition of tumorous CDK20 increased IFN-γ ⁺TNF-α ⁺CD8 ⁺ T cell infiltration and upregulated PD-L1 expression level in tumors, providing a greater chance of combination therapy with PD-L1 blockade to eradicate HCC tumors ⁷.

There is currently no CDK20-specific inhibitor publicly available, and no development of such a reagent has been reported. A major reason for this is perhaps the lack of a 3D structure for the CDK20 protein (with or without its cyclin partner) which may be useful for the design of selective inhibitors. There remains a need to develop new compounds that effectively act as CDK20 inhibitors.

SUMMARY OF THE INVENTION

Disclosed herein are novel compounds as CDK20 inhibitors. As a result, the compounds of the present disclosure are particularly useful in the modulation of CDK20 and thus in the treatment of CDK20-associated diseases and conditions.

In one aspect, the present disclosure is directed to a compound of Formula (I)

or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof, wherein X ¹, X ², X ³, X ⁴, ring A, R ², R ³, R ⁴, and n are described herein.

In another aspect, the present disclosure is directed to a pharmaceutical composition for treating a CDK20-associated disease or condition, which comprises the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as provided herein, and a pharmaceutically acceptable carrier or excipient.

In a further aspect, the present disclosure is directed to a method of treating a CDK20-associated disease or condition in a subject in need thereof, which comprises administering to the subject a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as provided herein.

In a further aspect, the present disclosure is directed to the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as provided herein for use in the treatment of a CDK20-associated disease or condition.

In a further aspect, the present disclosure is directed to use of the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as provided herein in the manufacture of a medicament for treating a CDK20-associated disease or condition.

In a further aspect, the present disclosure is directed to a kit for treating a CDK20-associated disease or condition, which comprises a compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as provided herein, a container, and optionally a package insert or label indicating a treatment.

DESCRIPTION OF THE DRAWINGS

The summary, as well as the following detailed description, may be further understood when read in conjunction with the appended drawings. For the purpose of illustrating the present disclosure, there are shown in the drawings exemplary embodiments of the present disclosure; however, the present disclosure is not limited to the specific disclosure of the drawings.

In the drawings:

Figure 1 shows predicted binding pose for an exemplary compound of the present disclosure (Compound I-001) with CDK20;

Figure 2 shows assay results of an exemplary compound of the present disclosure (Compound I-001) .

DETAILED DESCRIPTION OF THE INVENTION

Reference will now be made in detail to certain embodiments, examples of which are illustrated in the accompanying detailed description. While enumerated embodiments will be described, it shall be understood that they are not intended to limit the present disclosure to those embodiments. On the contrary, the present disclosure is intended to cover all alternatives, modifications, and equivalents, which may be included within the scope of the present disclosure as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present disclosure. The present disclosure is in no way limited to the methods and materials as described. In the event that one or more of the incorporated literatures and similar materials differs from or contradicts this disclosure, including but not limited to defined terms, term usage, described techniques, or the like, this disclosure controls.

It is appreciated that certain features of the present disclosure, which are, for clarity, described in the context of separate embodiments, can also be provided in combination in a single embodiment. Conversely, various features of the present disclosure, which are, for brevity, described in the context of a single embodiment, can also be provided separately or in any suitable sub-combination.

The following embodiments are provided herein.

Embodiment 1. A compound of Formula (I)

or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof, wherein

each of X ¹, X ², X ³, and X ⁴ is independently N or CR ^A, in which R ^A is H or R ¹, with the proviso that no more than two of X ¹, X ², X ³, and X ⁴ are N;

ring A is a 5-or 6-membered aromatic or heteroaromatic ring which contains 0, 1, or 2 heteroatoms selected from the group consisting of N, O, and S, optionally further fused to one or two cyclic rings independently selected from -cycloalkyl, -heterocyclyl, -aryl, -heteroaryl ring;

R ¹ is H, halo, -CN, -NO ₂, -alkyl, -haloalkyl, -alkenyl, -alkynyl, - (CH ₂) _p-aryl, - (CH ₂) _p-heteroaryl, -cycloalkyl, -heterocyclyl, -aryl, -heteroaryl, -OH, -OR ^C, -SR ^C, -COR ^C, -CO ₂R ^C, - CONR ^CR ^D, -S (O) R ^C, -SO ₂R ^C, -SO ₃R ^C, -SO ₂NR ^CR ^D, -P (O) (OR ^C) ₂, or -NR ^CR ^D, in which each of R ^C and R ^D is independently H, -alkyl, -haloalkyl, -alkenyl, -alkynyl, - (CH ₂) _p-aryl, - (CH ₂) _p-heteroaryl, -cycloalkyl, -heterocyclyl, -aryl, or -heteroaryl, or R ^C and R ^D together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said heteroaryl, said heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic, or spiro bicyclic, contains 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by alkyl or oxo, and p is 1, 2, or 3;

R ² is H, halo, -C _1-6alkyl, -C _3-6cycloalkyl, or -C _1-6haloalkyl;

R ³ is R ^E, -NR ^ER ^B, or -CONR ^ER ^B, wherein each of R ^E and R ^B is independently H, -C _1-6alkyl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, -C _1-6alkylene-NR ^FR ^G, -C _1-6alkylene-C _3-6cycloalkyl, or -C _1- ₆alkylene-C _3-6heterocyclyl, or R ^E and R ^B together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said C _3-6heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic, or spiro bicyclic, contains 1 or 2 heteroatoms selected from the group consisting of N, O and S, and is unsubstituted or substituted by -C _1- ₆alkyl or oxo, and each of R ^F and R ^G is independently H or -C _1-6alkyl;

R ⁴ is independently H, -C _1-6alkyl, -OC _1-6alkyl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, or - (CH ₂) _q-C _3-6heterocyclyl, in which said C _3-6heterocyclyl is monocyclic, fused bicyclic, or spiro bicyclic, contains 1 or 2 heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by -C _1-6alkyl or oxo, and q is 1, 2, or 3; and

n is 0, 1, 2, 3, or 4.

Embodiment 2. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to embodiment 1, wherein each of X ¹, X ², X ³, and X ⁴ is independently CR ^A, in which R ^A has the same meaning as defined above.

Embodiment 3. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to embodiment 1, wherein one of X ¹, X ², X ³, and X ⁴ is N, and the remainder of X ¹, X ², X ³, and X ⁴ are independently CR ^A, in which R ^A has the same meaning as defined above.

Embodiment 4. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein ring A is a 5-or 6-membered aromatic or heteroaromatic ring which contains 0, 1, or 2 heteroatoms independently selected from N and S, optionally fused to a 5-or 6-membered -cycloalkyl or heterocyclyl ring.

Embodiment 5. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein ring A is a pyrrole, imidazole, benzene, pyridine, pyrazole, thiazole, isothizole, or 1, 2, 3, 4-tetrahydroisoquinoline ring.

Embodiment 6. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein ring A is a pyrrole ring.

Embodiment 7. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein

is a group represented by the following formula:

Embodiment 8. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein

is a group represented by the following formula:

Embodiment 9. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein n is 0, 1, or 2.

Embodiment 10. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein n is 1, and

is a group represented by the following formula:

Embodiment 11. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ¹ is independently H, halo, -CN, -NO ₂, -C _1-6alkyl, -C _1-6haloalkyl, -C _2- ₆alkenyl, -C _2-6alkynyl, - (CH ₂) _p-aryl, - (CH ₂) _p-heteroaryl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, aryl, heteroaryl, -OH, -OR ^C, -SR ^C, -COR ^C, -CO ₂R ^C, -CONR ^CR ^D, -S (O) R ^C, -SO ₂R ^C, -SO ₃R ^C, -SO ₂NR ^CR ^D, -P (O) (OR ^C) ₂, or -NR ^CR ^D, in which each of R ^C and R ^D is independently H, -C _1-6alkyl, -C _2-6alkenyl, -C _2-6alkynyl, - (CH ₂) _p-aryl, - (CH ₂) _p-heteroaryl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, aryl, or heteroaryl, or R ^C and R ^D together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said heteroaryl, said C _3- ₆heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic or spiro bicyclic, contains 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by C _1-6alkyl or oxo, and p is 1, 2, or 3.

Embodiment 12. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ¹ is independently H, halo, -CN, -C _1-6alkyl, -C _1-6haloalkyl, - (CH ₂) _p- (5-to 10-membered aryl) , - (CH ₂) _p- (5-to 10-membered heteroaryl) , -C _3-6cycloalkyl, -C _3- ₆heterocyclyl, 5-to 10-membered aryl, 5-to 10-membered heteroaryl, -OR ^C, -SR ^C, -COR ^C, -CO ₂R ^C, -CONR ^CR ^D, -S (O) R ^C, -SO ₂R ^C, -SO ₂NR ^CR ^D, or -NR ^CR ^D, in which each of R ^C and R ^D is independently H or -C _1-6alkyl, or R ^C and R ^D together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said 5-to 10-membered heteroaryl, said C _3-6heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic or spiro bicyclic, contains 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by C _1-6alkyl or oxo, and p is 1 or 2.

Embodiment 13. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ¹ is independently H, halo, -CN, -C _1-3alkyl, -C _1-3haloalkyl, -CH ₂-phenyl, -CH ₂-pyridinyl, -O-phenyl, -O-pyridinyl, -OC _1-3alkyl, 2-oxopyrrolidin-1-yl, -CONHC _1-3alkyl, -SO ₂C _1-3alkyl, imidazolopyrimidinyl, tetrahydroimidazolopyridinyl, pyrazolyl, -C _3-6cycloalkyl, or -C _3-6heterocycyl, in which said C _3-6heterocycyl is monocyclic, fused bicyclic or spiro bicyclic, contains 1, 2, or 3 heteroatoms selected from N, O, and S, and is unsubstituted or further substituted by C _1-6alkyl or oxo.

Embodiment 14. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ¹ is independently H, -Br, -CN, -CH ₃, -CF ₃, pyridin-2-ylmethyl, pyridin-2-yloxy, -OCH ₃, 2-oxopyrrolidin-1-yl, -CONHCH ₃, -SO ₂CH ₃, imidazo [1, 2-a] pyrimidin-3-yl, 5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyridin-3-yl, pyrazol-4-yl, 3-methyl-pyrazol-4yl, cyclopentyl, or oxetan-3-yl.

Embodiment 15. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ² is H, -C _1-6alkyl, or -C _3-6cycloalkyl.

Embodiment 16. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ³ is R ^E, -NHR ^E, -NR ^E’R ^B’, -CONHR ^E, or -CONR ^E’R ^B’; wherein R ^E is H, -C _1-6alkyl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, -C _1-6alkylene-NHC _1-3alkyl, -C _1-6alkylene-N (C _1- ₃alkyl) ₂, -C _1-6alkylene-C _3-6cycloalkyl, or -C _1-6alkylene-C _3-6heterocyclyl, and R ^E’a nd R ^B’ together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said C _3-6heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic, or spiro bicyclic, contains 1 or 2 N heteroatoms, and is unsubstituted or substituted by -C _1-6alkyl.

Embodiment 17. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ³ is a group represented by the following formula:

Embodiment 18. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ³ is a group represented by the following formula:

Embodiment 19. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ⁴ is H, -C _1-6alkyl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, or - (CH ₂) _q-C _3- ₆heterocyclyl, in which said C _3-6heterocyclyl is monocyclic, fused bicyclic, or spiro bicyclic, contains 1 or 2 heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by -C _1-6alkyl, and q is 1, 2, or 3.

Embodiment 20. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein R ⁴ is H, (1-methylpiperidin-4-yl) methyl, or (2, 6-diazaspiro [3.3] heptan-2-yl) ethyl.

Embodiment 21. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of the preceding embodiments, wherein the isotopic variant is a deuterated variant.

Embodiment 22. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to embodiment 1, wherein the compound wherein the compound is one of the compounds in Table 1.

Table 1

Embodiment 23. A pharmaceutical composition for treating a CDK20-associated disease or condition, which comprises the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of embodiments 1 to 22, and a pharmaceutically acceptable carrier or excipient.

Embodiment 24. The pharmaceutical composition according to embodiment 23, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.

Embodiment 25. The pharmaceutical composition according to embodiment 24, which further comprises a second therapeutic agent useful for treating said disease or condition.

Embodiment 26. A method of treating a CDK20-associated disease or condition in a subject in need thereof, which comprises administering to the subject a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of embodiments 1 to 22.

Embodiment 27. The method according to embodiment 26, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.

Embodiment 28. The method according to embodiment 27, wherein the CDK20-associated disease or condition is hepatocellular carcinoma (HCC) .

Embodiment 29. A compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of embodiments 1 to 22 for use in the treatment of a CDK20-associated disease or condition.

Embodiment 30. The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to embodiment 29, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.

Embodiment 31. Use of a compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of embodiments 1 to 22 in the manufacture of a medicament for treating a CDK20-associated disease or condition.

Embodiment 32. The use according to embodiment 31, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.

Embodiment 33. A kit for treating a CDK20-associated disease or condition, which comprises a compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof according to any one of embodiments 1 to 22, a container, and optionally a package insert or label indicating a treatment.

Embodiment 34. The kit according to embodiment 33, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.

Embodiment 35. The kit according to embodiment 34, which further comprises a second therapeutical agent useful for treating said disease or disorder.

DEFINITIONS

The terms used herein but not defined have their ordinary meaning and the meaning of such terms is independent at each occurrence thereof. Nevertheless, unless otherwise stated, the following definitions apply throughout the specification and claims.

As used herein, the singular forms “a” , “an” , and “the” include plural referents unless expressly stated to the contrary.

As used herein, the terms “comprise” and “include” are intended to specify the presence of stated features, integers, components, or steps, but they do not preclude the presence or addition of one or more other features, integers, components, steps, or groups thereof.

Definitions of specific functional groups and chemical terms are described in more detail below. For purpose of this disclosure, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 ^th Edition, inside cover, and specific functional groups are generally defined as described therein. Additionally, general principles of organic chemistry, as well as specific functional moieties and reactivity, are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March, March’s Advanced Organic Chemistry, 5 ^th Edition, John Wiley &Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modem Methods of Organic Synthesis, 3 ^rd Edition, Cambridge University Press, Cambridge, 1987.

All ranges cited herein are inclusive, unless expressly stated to the contrary.

When a range of values is listed, it is intended to encompass each value and sub-range within the range. For example, “C _1-6” is intended to encompass, C ₁, C ₂, C ₃, C ₄, C ₅, C ₆, C _1-6, C _1- ₅, C _1-4, C _1-3, C _1-2, C _2-6, C _2-5, C _2-4, C _2-3, C _3-6, C _3-5, C _3-4, C _4-6, C _4-5, and C _5-6.

When any variable occurs more than one time in any constituent or in Formula (I) or in any other formula depicting and describing the compounds of the present disclosure, its definition at each occurrence is independent of its definition at every other occurrence. Also, combinations of substituents and/or variables are permissible only if such combinations result in stable compounds.

As used herein, the term “alkyl” refers to a linear or branched chain saturated hydrocarbon group. The term “C _i-j alkyl” refers to an alkyl having i to j carbon atoms. Alkyl groups may contain 1 to 10 carbon atoms, unless otherwise stated. In certain embodiments, alkyl groups contain 1 to 6 carbon atoms, such as, 1 to 5 carbon atoms, 1 to 4 carbon atoms, 1 to 3 carbon atoms, or 1 to 2 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-and iso-propyl, n-, sec-, iso-, and tert-butyl, neopentyl, and the like. As used herein, the term “alkylene” refers to a divalent substituent that is a monovalent alkyl having one hydrogen atom replaced with a valency.

As used herein, the term “alkenyl” refers to a linear or branched-chain hydrocarbon radical having at least one carbon-carbon double bond, and includes radicals having “cis” and “trans” orientations, or alternatively, “E” and “Z” orientations. Alkenyl groups may contain 2 to 10 carbon atoms, unless otherwise stated. In certain embodiments, alkenyl groups may contain 2 to 6 carbon atoms, such as 2 to 5 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms. In certain embodiments, alkenyl groups contain 2 carbon atoms. Non-limiting examples of alkenyl groups include ethylenyl (vinyl) , propenyl, butenyl, pentenyl, 1-methyl-2-buten-1-yl, 5-hexenyl, etc.

As used herein, the term “alkynyl” refers to a linear or branched hydrocarbon radical having at least one carbon-carbon triple bond. Alkynyl groups may contain 2 to 10 carbon atoms, unless otherwise stated. In certain embodiments, alkynyl groups contain 2 to 8 carbon atoms, 2 to 6 carbon atoms, 2 to 4 carbon atoms, 2 to 3 carbon atoms. In certain embodiments, alkynyl groups contain 2 carbon atoms. Non-limiting examples of alkynyl groups include ethynyl, 1-propynyl, 2-propynyl, etc.

As used herein, the term “cycloalkyl” refers to a monovalent non-aromatic, saturated monocyclic and polycyclic ring system, in which all the ring atoms are carbons and which contains at least three ring forming carbon atoms. Cycloalkyl groups may contain 3 to 10 ring forming carbon atoms, unless otherwise stated. In certain embodiments, cycloalkyl groups may contain 3 to 8 ring forming carbon atoms, 3 to 6 ring forming carbon atoms (i.e., C ₃- ₆cycloalkyl) , etc. Particularly, cycloalkyl groups may be monocyclic or bicyclic. Alternatively, bicyclic cycloalkyl groups may include fused, spiro, and bridged cycloalkyl structures. Also included are rings in which 1, 2, or 3 heteroatoms replace carbons. Such groups are termed as “heterocyclyl” or “heterocyclic ring” , which refers to a cycloalkyl group further bearing at least one heteroatom selected from N, O, and S as ring forming atoms. Non-limiting examples of heterocyclyl groups include oxiranyl, pyrrolidinyl, piperidyl, tetrahydropyranyl, piperazinyl, pyrrolidinyl, and morpholinyl. Heterocyclyl groups may be described by using a carbon count. For example, C _3- ₉heterocyclyl refers to a heterocyclyl group which contains three to nine ring forming carbon atoms, and may further contain at least one heteroatom, such as 1, 2, or 3 heteroatoms as ring forming atoms. For example, C _3-6heterocyclyl refers to a heterocyclyl group which contains three to six ring forming carbon atoms, and may further contain at least one heteroatom, such as 1, 2, or 3 heteroatoms as ring forming atoms. Non-limiting examples of C _3-6heterocyclyl groups include

etc. In certain embodiments, heterocyclyl groups or heterocyclic rings contain 1 or 2 heteroatoms, as ring forming atoms. In certain embodiments, heterocyclyl group may be monocyclic or bicyclic, such as fused bicyclic and spiro bicyclic. In the context of the present disclosure, the terms “heterocyclyl” and “heterocyclic ring” may be used interchangeably.

As used herein, the term “aryl” or “aromatic ring” refers to a mono-, bicyclic, or multicyclic carbocyclic ring system having at least one aromatic rings. Aryl groups may be 6-to 10-membered, unless otherwise stated. In certain embodiments, aryl groups may contain 6 ring forming carbon atoms. All atoms within a carbocyclic aryl group are carbon atoms. Non-limiting examples of aryl groups include phenyl, naphthyl, 1, 2-dihydronaphthyl, 1, 2, 3, 4-tetrahydronaphthyl, fluorenyl, indanyl, indenyl, etc. In the context of the present disclosure, the terms “aryl” and “aromatic ring” may be used interchangeably.

As used herein, the term “heteroaryl” or “heteroaromatic ring” refers to a monocyclic ring system, or a fused or bridged bicyclic ring system, in which the ring system contains one, two, three, or four heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur; and at least one of the rings is an aromatic ring. Heteroaryl groups may be 5-to 10-membered, unless otherwise stated. In certain embodiments, heteroaryl groups may be 5-to 6-membered. In certain embodiments, heteroaryl groups may contain one, two, or three heteroatoms. In certain embodiments, heteroaryl groups may contain one or two heteroatoms. In certain embodiments, heteroraryl groups may further substituted by alkyl or oxo. Non-limiting examples of heteroaryl groups include benzimidazolyl, benzotriazolyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, furyl, imidazolyl, indolyl, isoindazolyl, isoquinolinyl, isothiazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, purinyl, pyrrolyl, pyridinyl, pyrazinyl, pyrimidinyl, qunazolinyl, quinolinyl, thiadiazolyl, thiazolyl, thienyl, triazolyl, tetrazolyl, dihydroindolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, pyrrolopyridinyl, pyrrolopyrimidinyl, pyrrolopyrazinyl, pyrrolopyridazinyl, pyrazolopyridinyl, pyrazolopyrimidinyl, pyrazolopyrazinyl, pyrazolopyridazinyl, imidazolopyridinyl, imidazolopyrimidinyl, imidazolopyrazinyl, and imidazolopyridazinyl, etc. Heteroaryl groups include at least one ring having at least one heteroatom as described above and at least one aromatic ring. For example, a ring having at least one heteroatom may be fused to one, two, or three carbocyclic rings, e.g., an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring, or another monocyclic heterocyclic ring. Non-limiting examples of fused heteroaryl groups include 1, 2, 3, 5, 8, 8a-hexahydroindolizine, 2, 3-dihydrobenzofuran, 2, 3-dihydroindole, 2, 3-dihydrobenzothiophene, 1, 2, 3, 4-tetrahydroisoquinoline, and 5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyridine, etc. In the context of the present disclosure, the terms “heteroaryl” and “heteroaromatic ring” may be used interchangeably.

As used herein, the term “oxo” refers to a divalent oxygen atom and the structure of oxo may be shown as =O.

As used herein, the term “halogen” or “halo” refers to fluoride, chloride, bromide, and iodide. In certain embodiments, non-limiting examples of halo include fluoride, chloride and bromide, and more particularly fluoride and bromide.

As used herein, the term “haloalkyl” refers to an alkyl group substituted with one or more halogen atoms selected from the group consisting of fluoride, chloride, bromide, and iodide. Non-limiting examples of haloalkyl groups include -CH ₂F, -CHF ₂, -CF ₃, -CH ₂CF ₃, -CF ₂CF ₃, etc.

As used herein, the term “substituted” , when refers to a chemical group, means that the chemical group has one or more hydrogen atoms that is/are removed and replaced by substituents. The term “substituent” as used herein has the ordinary meaning known in the art and refers to a chemical moiety that is covalently attached to, or if appropriate, fused to, a parent group. It is to be understood that substitution at a given atom is limited by valency. It is understood that the substituent can be further substituted.

When a moiety is noted as being “optionally” substituted in Formula (I) or any embodiment thereof, it means that Formula (I) or the embodiment thereof encompasses both compounds that are substituted with the noted substituent (s) on the moiety and compounds that do not contain the noted substituent (s) on the moiety (i.e., wherein the moiety is unsubstituted) .

The compounds as provided herein are described with reference to both generic formulas and specific compounds. In addition, the compounds of the present disclosure may exist in a number of different forms or derivatives, all within the scope of the disclosure. These include, for example, pharmaceutically acceptable salts, tautomers, stereoisomers, racemic mixtures, regioisomers, prodrugs, solvated forms, different crystal forms or polymorphs, and active metabolites, etc.

As used herein, the term “pharmaceutically acceptable salt” , unless otherwise stated, includes salts that retain the biological effectiveness of the free acid/base form of the specified compound and that are not biologically or otherwise undesirable. Contemplated pharmaceutically acceptable salt forms include, but are not limited to, mono, bis, tris, tetrakis, and so on. Pharmaceutically acceptable salts are non-toxic in the amounts and concentrations at which they are administered. The preparation of such salts can facilitate the pharmacological use by altering the physical characteristics of a compound without preventing it from exerting its physiological effect. Useful alterations in physical properties may include, for example, increasing the solubility to facilitate administering higher concentrations of the drug.

Pharmaceutically acceptable salts of the compounds of Formula (I) include acid addition and base salts. Suitable acid addition salts can be formed from acids which form non-toxic salts. Non-limiting examples may include the acetate, adipate, aspartate, benzoate, besylate, bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate, citrate, cyclamate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate, maleate, malonate, mesylate, methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate/hydrogen phosphate/dihydrogen phosphate, pyroglutamate, saccharate, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, 1, 5-naphathalenedisulfonic acid and xinafoate salts. Suitable base salts are formed from bases which form non-toxic salts. Non-limiting examples may include the aluminium, arginine, benzathine, calcium, choline, diethylamine, bis (2-hydroxyethyl) amine (diolamine) , glycine, lysine, magnesium, meglumine, 2-aminoethanol (olamine) , potassium, sodium, 2-Amino-2- (hydroxymethyl) propane-1, 3-diol (tris or tromethamine) and zinc salts. Hemisalts of acids and bases may also be formed, for example, hemisulfate and hemicalcium salts. For a review on suitable salts, see, Stahl and Wermuth, Handbook of Pharmaceutical Salts: Properties, Selection, and Use (Wiley-VCH, 2002) .

Pharmaceutically acceptable salts of the compound of Formula (I) may be prepared by one or more of three methods: (i) by reacting the compound of Formula (I) with the desired acid or base; (ii) by removing an acid-or base-labile protecting group from a suitable precursor of the compound of Formula (I) or by ring-opening a suitable cyclic precursor, for example, a lactone or lactam, using the desired acid or base; or (iii) by converting one salt of the compound of Formula (I) to another by a reaction with an appropriate acid or base or by means of a suitable ion exchange column. The three reactions may be typically carried out in solution. The resulting salt may precipitate out and be collected by filtration or may be recovered by evaporation of the solvent. The degree of ionization in the resulting salt may vary from completely ionized to almost non-ionized.

The compound of Formula (I) and pharmaceutically acceptable salts thereof may exist in unsolvated and solvated forms. As used herein, the term “solvate” refers to a molecular complex comprising the compound of Formula (I) , or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable solvent molecules. For example, the term “hydrate” is employed when said solvent is water.

The compounds of Formula (I) may have one or more chiral (asymmetric) centers. The present disclosure encompasses all stereoisomeric forms of the compounds of Formula (I) . Centers of asymmetry that are present in the compounds of Formula (I) can all independently of one another have (R) or (S) configuration. When bonds to a chiral carbon are depicted as straight lines in the structural formulas of the present disclosure, or when a compound name is recited without an (R) or (S) chiral designation for a chiral carbon, it is understood that both the (R) and (S) configurations of each such chiral carbon and hence each enantiomer or diastereomer and mixtures thereof are embraced within the formula or by the name. The production of specific stereoisomers or mixtures thereof may be identified in the Examples where such stereoisomers or mixtures were obtained, but this in no way limits the inclusion of all stereoisomers and mixtures thereof from being within the scope of the disclosure.

The present disclosure includes all possible enantiomers and diastereomers and mixtures of two or more stereoisomers, for example mixtures of enantiomers and/or diastereomers, in all ratios. Thus, enantiomers are a subject of the present disclosure in enantiomerically pure form, both as levorotatory and as dextrorotatory antipodes, in the form of racemates and in the form of mixtures of the two enantiomers in all ratios. In the case of a cis/trans isomerism, the present disclosure includes both the cis form and the trans form as well as mixtures of these forms in all ratios. The preparation of individual stereoisomers can be carried out, if desired, by separation of a mixture by customary methods, for example by chromatography or crystallization, by use of stereochemically uniform starting materials for the synthesis or by stereoselective synthesis. Optionally, a derivatization can be carried out before separation of stereoisomers. The separation of a mixture of stereoisomers can be carried out in an intermediate step during the synthesis of a compound of Formula (I) , or it can be done on a final racemic product. Absolute stereochemistry may be determined by X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing a stereogenic center of known configuration. Alternatively, absolute stereochemistry may be determined by Vibrational Circular Dichroism (VCD) spectroscopy analysis.

Unless otherwise stated, the structures depicted herein are also meant to include the compounds that differ only in the presence of one or more isotopically enriched atoms, in other words, the compounds wherein one or more atoms are replaced by atoms having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number which predominates in nature. Such compounds are referred to as a “isotopic variant” . The present disclosure is intended to include all pharmaceutically acceptable isotopic variants of the compounds of Formula (I) . Examples of isotopes suitable for inclusion in the compounds of the present disclosure include, but not limited to, isotopes of hydrogen, such as ²H and ³H; carbon, such as ¹¹C, ¹³C and ¹⁴C; chlorine, such as ³⁶Cl; fluorine, such as ¹⁸F; iodine, such as ¹²³I and ¹²⁵I; nitrogen, such as ¹³N and ¹⁵N; oxygen, such as ¹⁵O, ¹⁷O and ¹⁸O; phosphorus, such as ³²P; and sulfur, such as ³⁵S. Certain isotopic variants of the compounds of Formula (I) , for example those incorporating a radioactive isotope, may be useful in drug and/or substrate tissue distribution studies. Particularly, compounds having the depicted structures that differ only in the replacement with heavier isotopes, such as the replacement of hydrogen by deuterium ( ²H) , can afford certain therapeutic advantages, for example, resulting from greater metabolic stability, increased in vivo half-life, or reduced dosage requirements and, hence, may be utilized in some particular circumstances. Isotopic variants of compounds of Formula (I) can generally be prepared by conventional techniques known to one skilled in the art or by processes analogous to those described in the accompanying examples and synthesis using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.

Pharmaceutically acceptable solvates in accordance with the present disclosure may include those wherein the solvent of crystallization may be isotopically substituted, e.g., D ₂O, d ₆-acetone, d ₆-DMSO.

One way of carrying out the present disclosure is to administer a compound of Formula (I) in the form of a prodrug. Thus, certain derivatives of a compound of Formula (I) which may have little or no pharmacological activity themselves can, when administered into or onto the body, be converted into a compound of Formula (I) having the desired activity, for example by hydrolytic cleavage, particularly hydrolytic cleavage promoted by an esterase or peptidase enzyme. Such derivatives are referred to as “prodrugs” . Further information on the use of prodrugs may be found in, e.g., T. Higuchi and W. Stella, “Pro-drugs as Novel Delivery Systems” , Vol. 14, ACS Symposium Series, and E. B. Roche (Ed. ) , “Bioreversible Carriers in Drug Design” , Pergamon Press, 1987, American Pharmaceutical Association. Reference can also be made to Nature Reviews/Drug Discovery, 2008, 7, 355, and Current Opinion in Drug Discovery and Development, 2007, 10, 550.

Prodrugs in accordance with the present disclosure can, for example, be produced by replacing appropriate functionalities present in the compounds of Formula (I) with certain moieties known to those skilled in the art as “pro-moieties” as described, for example, in H. Bundgaard, “Design of Prodrugs” , Elsevier, 1985, and Y.M. Choi-Sledeski and C.G. Wermuth, “Designing Prodrugs and Bioprecursors” , Practice of Medicinal Chemistry, 4 ^th Edition, Chapter 28, 657-696, Elsevier, 2015. Thus, a prodrug in accordance with the present disclosure may include, but not limited to, (a) an ester or amide derivative of a carboxylic acid in a compound of Formula (I) , if any; (b) an amide, imine, carbamate or amine derivative of an amino group in a compound of Formula (I) ; (c) an oxime or imine derivative of a carbonyl group in a compound of Formula (I) , if any; or (d) a methyl, primary alcohol or aldehyde group that can be metabolically oxidized to a carboxylic acid in a compound of Formula (I) , if any.

References to compounds of Formula (I) are taken to include the compounds themselves and prodrugs thereof. The present disclosure includes such compounds of Formula (I) as well as pharmaceutically acceptable salts of such compounds and pharmaceutically acceptable solvates of said compounds and salts.

ADMINISTRATION AND DOSING

The compounds of the present disclosure may be administered in an amount effective to treat the diseases or conditions as described herein. The compounds of the present disclosure can be administered as compound per se, or alternatively, as a pharmaceutically acceptable salt. For administration and dosing purposes, the compound of the present disclosure per se or pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof will simply be referred to as the compounds of the disclosure.

The compounds of the disclosure are administered by any suitable route in the form of a pharmaceutical composition adapted to such a route, and in a dose effective for the treatment intended. The compounds of the disclosure may be administered orally, rectally, vaginally, parenterally, or topically.

As used herein, the terms “administration” and “administer” refer to absorbing, ingesting, injecting, inhaling, implanting, or otherwise introducing the compound of the disclosure, or a pharmaceutical composition thereof. The terms “treatment” and “treat” refer to reversing, alleviating, delaying the onset of, or inhibiting the progress of a “pathological condition” (e.g., a disease, disorder, or condition, or one or more signs or symptoms thereof) described herein. In certain embodiments, treatment may be administered after one or more signs or symptoms of a disease or condition have developed or have been observed. In other embodiments, treatment may be administered in the absence of signs or symptoms of the disease or condition. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and/or in light of genetic or other susceptibility factors) . Treatment may also be continued after symptoms have resolved, for example, to delay or prevent recurrence. As used herein, the terms “disease” , “disorder” , “condition” , and “pathological condition” are used interchangeably.

Dosage levels for administration can be determined by those skilled in the art by routine experimentation. The dosage regimen for the compounds of the disclosure and/or compositions comprising said compounds is based on a variety of factors, including the type, age, weight, sex, and medical condition of the patient; the severity of the condition; the route of administration; and the activity of the particular compound employed. Thus, the dosage regimen may vary widely. It is not uncommon that the administration of the compounds of the disclosure will be repeated a plurality of times in a day.

In certain embodiments, the compound of the disclosure may be administered in combination with a second therapeutical agent. In certain embodiments, non-limiting examples of second therapeutical agents may include an anti-cancer agent. In certain embodiments, non-limiting examples of second therapeutical agents may include an additional CDK20 inhibitor. The second therapeutical agent can be administered before, after, or at the same time that the compound of the present disclosure is administered.

PHARMACEUTICAL COMPOSITIONS

In some aspect, the present disclosure is directed to a pharmaceutical composition comprising the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as provided herein, and at least one pharmaceutically acceptable carrier or excipient.

As used herein, the term “pharmaceutically acceptable carrier or excipient” refers to a carrier or excipient which is useful for preparing a pharmaceutical composition that is generally safe, non-toxic, and neither biologically nor otherwise undesirable, and includes carrier or excipient that is acceptable for veterinary use as well as human pharmaceutical use. A “pharmaceutically acceptable carrier or excipient” as used herein includes both one and more than one such carrier or excipient. The particular carrier or excipient used will depend upon the means and purpose for which the compounds of the disclosure is being applied. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C, et al., Ansel’s Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams &Wilkins, 2004; Gennaro, Alfonso R., et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams &Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more of buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents, and other known additives to provide an elegant presentation of the drug (i.e., the compound or pharmaceutical composition as provided herein) or aid in the manufacturing of the pharmaceutical product (i.e., medicament) .

The compositions of the present disclosure may be in a variety of forms. These include, for example, liquid, semi-solid and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions) , dispersions or suspensions, tablets, pills, powders, liposomes, suppositories, etc. The form depends on the intended mode of administration and therapeutic application.

Pharmaceutical compositions of the present disclosure may be prepared by any of the well-known techniques of pharmacy, such as effective formulation and administration procedures. The above considerations in regard to effective formulations and administration procedures are well known in the art, and are described in standard textbooks. Formulation of pharmaceutical products is discussed in, e.g., Hoover, John E., Remington’s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania, 1975; Liberman, et al., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, N.Y., 1980; and Kibbe, et al., Eds., Handbook of Pharmaceutical Excipients, 3 ^rd Edition, American Pharmaceutical Association, Washington, 1999.

In a further aspect, the present disclosure relates to a kit for treating a CDK20-associated disease or condition, which comprises a compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as provided herein, a container, and optionally a package insert or label indicating a treatment.

METHODS OF TREATMENT

In a further aspect, the present disclosure is directed to a method of treating a CDK20-associated disease or condition in a subject in need thereof, which comprises administering to the subject a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as provided herein, owning to the CDK20 inhibitor activity of the compound of the present disclosure.

As used herein, the term “subject in need thereof” is a subject having a CDK20-associated disease or condition, or a subject having an increased risk of developing CDK20-associated disease or condition relative to the population at large. In certain embodiments, the subject is a warm-blooded animal. In certain embodiments, the warm-blooded animal is a mammal. In certain embodiments, the warm-blooded animal is a human.

Furthermore, in certain embodiments, the CDK20-associated disease or condition is selected from a group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma. In certain embodiments, the CDK20-associated disease or condition is hepatocellular carcinoma (HCC) .

The method of treating a CDK20-associated disease or condition as described herein may be used as a monotherapy. As used herein, the term “monotherapy” refers to the administration of a single active or therapeutic compound to a subject in need thereof. In certain embodiments, monotherapy will involve administration of a therapeutically effective amount of one of the compounds of the present disclosure or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof, to a subject in need of such treatment.

Depending upon the particular disease or condition to be treated, the method of treating a CDK20-associated disease or condition described herein may involve, in addition to administration of the compound of Formula (I) , combination therapy of one or more additional therapeutic agent (s) , for example, a second therapeutic agent which is an anti-cancer agent. As used herein, the term “combination therapy” refers to the administration of a combination of multiple active therapeutic agents. In certain embodiments, the compound of the present disclosure or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof may be administered simultaneously, separately or sequentially to treatment with the one or more additional therapeutic agent (s) . For example, the additional therapeutic agent (s) may be administered separately from the compound of the present disclosure, as part of a multiple dosage regimen. Alternatively, the additional therapeutic agent (s) may be part of a single dosage form, mixed with the compound of the present disclosure in a single composition.

SYNTHESIS

The compounds of the present disclosure may be prepared by the general and specific methods described below, using the common general knowledge of those skilled in the art of synthetic organic chemistry. Such common general knowledge can be found in standard reference books, e.g., Barton and Ollis (Ed. ) , Comprehensive Organic Chemistry, Elsevier; Richard Larock, Comprehensive Organic Transformations: A Guide to Functional Group Preparations, John Wiley and Sons; and Compendium of Organic Synthetic Methods, Vol. I-XII, Wiley-Interscience. The starting materials used herein are commercially available or may be prepared by routine methods known in the art.

The Schemes described hereinafter are intended to provide a general description of the methodology employed in the preparation of the compounds of the present disclosure. Some of the compounds of the present disclosure may contain single or multiple chiral centers with the stereochemical designation (R) or (S) . It will be apparent to those skilled in the art that all of the synthetic transformations can be conducted in a similar manner no whether the materials are enantioenriched or racemic. Moreover, the resolution to the desired optically active material may take place at any desired point in the procedure using well known methods such as those described herein and in the chemistry literature.

EXAMPLES

In order that the disclosure may be more fully understood, the following examples are set forth. The examples described herein are offered to illustrate the compounds, methods and compositions provided herein and are not to be construed in any way as limiting their scope.

During synthetic procedures, it may be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This may be achieved by means of conventional protecting groups, such as those described in T.W. Greene and P.G.M. Wutts, Protective Groups in Organic Synthesis, 4 ^th Edition, John Wiley and Sons. The protective groups are optionally removed at a convenient subsequent stage using methods well known in the art.

The compounds of the present disclosure can be readily prepared according to the following reaction schemes and examples, or modifications thereof, using readily available starting materials, reagents, and conventional synthesis procedures. In these reactions, it is also possible to make use of variants which are themselves known to those skilled in the art, but are not mentioned in greater detail. Furthermore, other methods for preparing the compounds of the present disclosure will be readily apparent to those skilled in the art in light of the reaction schemes and examples as described herein. Unless otherwise indicated, all variables are as defined above. In general chemical procedures, all reagents and materials may be purchased from commercial vendors or may be readily prepared by those skilled in the art.

TARGET SELECTION AND IDENTIFICATION

Primary liver cancer is the sixth most frequently occurring cancer and the third most common cause of cancer mortality worldwide according to GLOBOCAN 2020 update released by International Agency for Research on Cancer (IARC) . ⁸ Hepatocellular carcinoma (HCC) is the dominant type of liver cancer, accounting for approximately 75%of the total patient population. The incidence rate of liver cancer is very close to its mortality rate due to very poor prognosis in all regions around the world. PD-L1 inhibitor atezolizumab in combo with bevacizumab has become the new standard-of-care (SoC) first-line treatment for advanced HCC after demonstrating a 42%reduction in the risk of death and a 41%reduction in the risk of disease worsening or death over the previous SoC Nexavar, but there’s still a huge unmet medical need for HCC patients.

PandaOmics is one of automated drug discovery AI engines to accelerate and optimize key steps of the early stages of drug discovery. This biocomputational platform combines bioinformatics methods for data analysis, visualization and interpretation with advanced multimodal deep learning approaches for target identification. PandaOmics therapeutic target and biomarker identification system is based on the combination of multiple scores derived from text and OMICs data associating genes with a disease of interest. Text evidence prioritization (Text, Financial and KOL (Key Opinion Leader) score families) singles out the genes, extensively mentioned across scientific literature and grant description. OMICs-based scores, in contrast, explore the molecular connection of genes with diseases based on differential expression, gene variants, interactome topology, signaling pathways activation analysis using iPanda algorithm ⁹, knockout\overexpression experiments and more. This kind of approach allows users to unveil the hidden hypotheses that might not be obvious over common general knowledge or simple bioinformatics analysis. AI tools are extremely helpful for efficient target hypothesis generation. The overall scoring approach results in the ranked list of target hypotheses for a given disease which can be subsequently filtered according to their novelty, accessibility by small molecules and antibodies, safety, tissue specificity, crystal structure availability and major biological structures.

Another unique feature of the PandaOmics platform is the ability to combine the data from different experiments into a single Meta-analysis and leverage the insights from all the datasets together for the precise target prioritization. During this study we’ve created a Meta-analysis for each of the diseases of interest composed from 10 datasets for HCC (1133 disease samples and 674 healthy controls) . After obtaining the ranked list of target hypotheses we applied PandaOmics filters in order to get the list of the most promising targets that satisfy First-in-class (see Methods section) scenarios and share the current unavailability of crystal structure but have structure folds predicted by AlphaFold. The final list of top-20 targets was then manually curated to nominate the most promising candidates. For hepatocellular carcinoma CDK20 was chosen since it had the highest scores aligned with the First-in-class scenario. The proposed therapeutic target CDK20 was passed to Chemistry42 platform for the automated generation of small molecule inhibitors.

Seven compounds shown in Table 1 are selected from Chemistry42 for synthesis and assessing the binding abilities towards CDK20. The results suggested one compound 001 (also referred to as “compound I-001” ) demonstrated a Kd value of 8.9 ± 1.6 μM (n = 4) in CDK20 kinase binding assay. As shown in Fig. 1, we also proposed the binding mode for I-001: four hydrogen bond interactions are represented as dash lines. Besides the two hydrogen bonds formed with the hinge residue Met84, I-001 also interacts with residue Leu85 via amide -NH group and residue Ile10 in P-loop via pyrrole -NH group. Alternatively, amide -NH group or pyrrole -NH group may form hydrogen bond interactions with the two acid centers Asp87 and Glu90 in the solvation region. Based on the predicted binding pose, hit expansion upon I-001 is ongoing to further improve the enzymatic activity.

Table 2

More specific compounds that exhibited CDK20 kinase binding abilities are listed in Table 3.

Table 3

METHODS AND EXPERIMENTS

Target ID and Target proposal:

PandaOmics platform was used to conduct a hypothesis generation for Hepatocellular carcinoma, limiting the target list to the proteins whose structures were predicted by AlphaFold2. Hepatocellular carcinoma Meta-analysis combined the data from ten experiments: GSE36376 ³⁰ (Cho et al. 2020) , GSE107170 ^30, 31 (Cho et al. 2020; Diaz et al. 2018) , GSE102079 ³², GSE45267 ^32, 33, GSE133039 ³⁴, GSE104766 ³⁵, GSE77314 ^35, 36, GSE60502 ³⁷, E-MTAB-5905 ³⁸ and TCGA-LIHC ³⁹, resulting in 1133 disease samples and 674 healthy controls.

Preparation of 4-amino-N-methyl-1H-pyrrole-2-carboxamide (INT-1)

Step 1: To a solution of methyl 4-nitro-1H-pyrrole-2-carboxylate (3.0 g, 17.6 mmol) in THF (45 mL) was added LiOH (aq., 2 M, 44.1 mL) . The mixture was stirred at 85℃ for 4 hrs. The reaction mixture was neutralized by addition of HCl (aq., 1 M) and then extracted with EtOAc (40 mL × 3) . The combined organic layers were washed with brine (50 mL × 2) , dried over anhydrous sodium sulfate, filtered, and concentrated to afford INT-1-1. ¹H NMR (400 MHz, DMSO-d ₆) δ 13.19 (s, 1H) , 13.08 -12.88 (m, 1H) , 8.01 (s, 1H) , 7.21 (s, 1H) .

Step 2: A mixture of INT-1-1 (2.96 g, 18.9 mmol) , DIPEA (9.91 mL, 56.8 mmol, T ₃P (7.84 g, 24.6 mmol, 7.33 mL) and in DCM (30 mL) , followed by the addition of MeNH ₂ (1.92 g, 28.4 mmol) . The mixture was stirred at 25℃ for 2 hrs. The reaction mixture was diluted with water (30 mL) , and then filtered to give a residue, which was triturated with CH ₃CN at 25 ℃. The mixture was filtered to afford INT-1-2. LCMS [M-H] ^-: 168.0.

Step 3: To a solution of INT-1-2 (2.0 g, 11.8 mmol) in MeOH (20 mL) was added Pd/C (0.40 g, 10 wt%) under N ₂ atmosphere. The suspension was degassed and purged with H ₂ for 3 times. The mixture was stirred under H ₂ (15 psi) at 25℃ for 12 hrs. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to afford INT-1. LCMS [M+H] ⁺: 140.1.

Preparation of tert-butyl 4- (2-chloroquinazolin-7-yl) -1H-pyrazole-1-carboxylate (INT-2)

A mixture of 7-bromo-2-chloroquinazoline (1.0 g, 4.11 mmol) , tert-butyl 4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole-1-carboxylate (1.21 g, 4.11 mmol) in THF (20 mL) was added Pd (dppf) Cl ₂ (0.30 g, 0.41 mmol) and a solution of K ₂CO ₃ (1.42 g, 10.3 mmol) in water (4.0 mL) . The result mixture was stirred at 100℃ for 1 hr under N ₂ atmosphere. The mixture was cooled to 25℃, filtered and washed with EtOAc. The filtrate was diluted with water (100 mL) and extracted with EtOAc (70 mL × 3) . The organic phases were separated and dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel chromatography to afford INT-2. ¹H NMR (400 Hz, CD ₃CN) δ 9.30 (s, 1 H) 8.72 (s, 1 H) 8.25 (s, 1 H) 8.12 -8.18 (m, 1 H) 8.04 -8.12 (m, 1 H) 7.98 (dd, J = 8.44, 1 H) 1.62 (s, 9 H) .

Preparation of 2-chloro-7- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-4-yl) quinazoline (INT-3)

To a degassed solution of 7-bromo-2-chloroquinazoline (200 mg, 0.516 mmol) in dioxane (6 mL) was added 1- (tetrahydro-2H-pyran-2-yl) -4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (215 mg, 0.77 mmol) , Pd (dppf) Cl ₂ (37.7 mg, 0.05 mmol) and a solution of K ₂CO ₃ (178 mg, 1.29 mmol) in H ₂O (2 mL) . The mixture was stirred at 90℃ for 3hrs under N ₂ atmosphere. The mixture was diluted with water and extracted with EtOAc (30 mL × 3) . The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and filtered _. The filtrate was concentrated and purified by silica gel chromatography to give INT-3. LCMS [M+H] ⁺: 315.1.

Preparation of 2-chloro-6- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-4-yl) quinazoline (INT-4)

A mixture of 6-bromo-2-chloroquinazoline (2.00 g, 8.21 mmol) , 1- (tetrahydro-2H-pyran-2-yl) -4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (2.51 g, 9.04 mmol) , Cs ₂CO ₃ (8.03 g, 24.6 mmol) , 2, 4, 6-trimethyl-1, 3, 5, 2, 4, 6-trioxatriborinane (0.10 g, 0.821 mmol) in THF (20 mL) and H ₂O (5 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 70℃ for 1 hr under N ₂ atmosphere. The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (20.0 mL × 3) . The combined organic layers were washed with brine (10 mL) and dried over anhydrous sodium sulfate. The obtained organic phase was concentrated under reduced pressure and purified by silica gel chromatography to afford INT-4. LCMS [M+H] ⁺: 315.2.

Preparation of 7- (1- (tetrahydro-2H-pyran-2-yl) -1H-pyrazol-4-yl) quinazolin-2-amine (INT-5)

A mixture of 7-bromoquinazolin-2-amine (1.0 g, 4.46 mmol) , 1- (tetrahydro-2H-pyran-2-yl) -4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (1.49 g, 5.36 mmol) , Pd(dppf) Cl ₂ (0.33 g, 0.446 mmol) , K ₃PO ₄ (2.84 g, 13.4 mmol) in H ₂O (2.50 mL) in dioxane (10.0 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 2 hrs under N ₂ atmosphere. The mixture was diluted with water (10.0 mL) and extracted with EtOAc (10 mL × 3) . The combined organic layers were washed with brine (10 mL) , dried over anhydrous sodium sulfate, and concentrated under reduced pressure to INT-5. LCMS [M+H] ⁺: 296.2.

Example 1: Preparation of N-methyl-4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxamide (I-001)

The title compound was synthesized according to the synthetic procedures:

General procedure for preparation of ethyl 4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxylate (3) : To a solution of ethyl 4-amino-1H-pyrrole-2-carboxylate (500 mg, 3.24 mmol) in DMF (15 mL) was added K ₂CO ₃ (672 mg, 4.86 mmol) , 2-chloroquinazoline (534 mg, 3.24 mmol) . The mixture was stirred at 80℃ for 16 hrs. The reaction mixture was washed with water (30 mL) and extracted with EtOAc (15 mL×3) . The combined organic layers were washed with NaCl (30 mL×2) , dried over Na ₂SO ₄, filtered and concentrated. The crude was purified by column chromatography on silico gel (PE: EA= 5: 1) to give ethyl 4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxylate (632 mg, 2.24 mmol, 69.03%yield) as a yellow solid.

General procedure for preparation of 4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxylic acid (4) : To a solution of ethyl 4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxylate (100 mg, 354 umol) in THF (2 mL) was added LiOH (1 M in water, 1.77 mL) . The mixture was stirred at 50℃ for 2 hrs. The reaction mixture was quenched by addition HCl (1 M) to neutral and then extracted with EtOAc (3 mL×3) . The combined organic layers were washed with NaCl (5 mL×2) , dried over Na ₂SO ₄, filtered and concentrated to give 4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxylic acid as yellow oil. LCMS: 253.0 [M-H] ⁺. ¹H NMR (400 MHz, DMSO-d ₆) : δ 12.14 -11.97 (m, 2H) , 11.43 (br s, 1H) , 9.67 (s, 1H) , 9.19 (s, 1H) , 7.84 (d, J = 8.00 Hz, 1H) , 7.78 -7.72 (m, 1H) , 7.58 (br d, J = 8.40 Hz, 2H) , 7.34 -7.26 (m, 1H) , 6.87 (s, 1H) .

General procedure for preparation of compound I-001: To a solution of 4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxylic acid (100 mg, 393 umol) , MeNH ₂ (2 M, 1.97 mL) in DMF (0.2 mL) was added CDI (76.5 mg, 472 umol) and DCC (122 mg, 590 umol) . The mixture was stirred at 25℃ for 12 hrs. The reaction mixture was diluted with Water 2 mL and extracted with EtOAc (3 mL×3) . The combined organic layers were washed with NaCl (5 mL×2) , dried over Na ₂SO ₄, filtered and concentrated. The crude was purified by prep-HPLC to give I-001 (35.0 mg, 124 umol, 31.7%yield) as a yellow solid. LCMS: 268.0 [M+H] ⁺. ¹H NMR: (400 MHz, CDCl ₃) : δ 9.24 -9.02 (m, 2H) , 7.85 -7.69 (m, 3H) , 7.57 -7.47 (m, 1H) , 7.43 -7.33 (m, 1H) , 6.68 (br s, 1H) , 5.94 -5.79 (m, 1H) , 3.01 (d, J = 4.80 Hz, 3H) .

Example 2: Preparation of 4- ( (6-methoxyquinazolin-2-yl) amino) -N-methyl-1H-pyrrole-2-carboxamide (I-002)

Step 1: To a solution of 2, 4-dichloro-6-methoxyquinazoline (0.30 g, 1.31 mmol) in DCM (5.0 mL) was added zinc powder (256.9 mg, 3.93 mmol) and NH ₄Cl (aq., 44.1 mmol, 5.0 mL) . The mixture was stirred at 40℃ for 2 hrs. The resulting mixture was cooled to r.t. and filtered through a Celite pad. The filtrate was extracted with DCM (10 mL × 2) . The combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum to afford I-002-1. LCMS [M+H] ⁺: 195.0.

Step 2: To a solution of I-002-1 (50.0 mg, 256.9 μmol) and INT-1 (35.75 mg, 256.9 μmol) in DMSO (1 mL) was added K ₂CO ₃ (53.3 mg, 385.37 μmol) . The mixture was stirred at 100℃for 4 hrs. The mixture was concentrated and purified by prep-HPLC to afford I-002. ¹H NMR (400 MHz, CD ₃OD) δ 9.02 (s, 1H) , 7.58 (d, J = 9.2 Hz, 1H) , 7.46 (d, J = 1.7 Hz, 1H) , 7.41 (dd, J = 9.2, 2.9 Hz, 1H) , 7.21 (d, J = 2.8 Hz, 1H) , 6.89 (d, J = 1.6 Hz, 1H) , 3.90 (s, 3H) , 2.88 (s, 3H) . LCMS [M+H] ⁺: 298.2.

Example 3: Preparation of N-methyl-4- ( (6-methylquinazolin-2-yl) amino) -1H-pyrrole-2-carboxamide (I-004)

Step 1: A mixture of 6-bromo-2-chloroquinazoline (200 mg, 821 μmol) , 2, 4, 6-trimethyl-1, 3, 5, 2, 4, 6-trioxatriborinane (412 mg, 1.64 mmol, 459 μL) , Cs ₂CO ₃ (803 mg, 2.46 mmol) , Pd(PPh ₃) ₂Cl ₂ (115 mg, 164 μmol) in H ₂O (0.5 mL) and DME (2 mL) was degassed and purged with N ₂ for 3 times, and then the mixture was stirred at 90℃ for 4 hrs under N ₂ atmosphere. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (10 mL × 3) . The combined organic layers were washed with brine (10 mL) , dried over anhydrous sodium sulfate, filtered, and concentrated to afford I-004-1. LCMS [M+H] ⁺: 179.1.

Step 2: To a solution of I-004-1 (50.0 mg, 280 μmol) and INT-1 (38.9 mg, 280 μmol) in DMSO (1 mL) was added K ₂CO ₃ (58.0 mg, 420 μmol) . The mixture was stirred at 100℃ for 2 hrs. The mixture was concentrated and purified by prep-HPLC to afford I-004. ¹H NMR (400 MHz, CD ₃OD) δ 8.99 (s, 1H) , 7.64 -7.55 (m, 3H) , 7.47 (d, J = 1.6 Hz, 1H) , 6.89 (d, J = 1.6 Hz, 1H) , 2.88 (s, 3H) , 2.46 (s, 3H) . LCMS [M+H] ⁺: 282.0.

Example 4: Preparation of 4- ( (6-cyanoquinazolin-2-yl) amino) -N-methyl-1H-pyrrole-2-carboxamide (I-005)

Step 1: A mixture of INT-1 (200 mg, 1.44 mmol) and 6-bromo-2-chloroquinazoline (350 mg, 1.44 mmol) in DMSO (10 mL) was added K ₂CO ₃ (596 mg, 4.31 mmol) . The mixture was stirred at 90℃ for 5 hrs. The reaction mixture was washed by water (2 mL) and then extracted with EtOAc (2 mL × 3) . The combined organic layers were washed with brine (2 mL × 2) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel chromatography to afford I-005-1. LCMS [M+H] ⁺: 348.0.

Step 2: A mixture of I-005-1 (500 mg, 866 μmol) , Zn (CN) ₂ (254 mg, 2.17 mmol, 138 μL) , Pd ₂ (dba) ₃ (15.9 mg, 17.3 μmol) and DPPF (19.2 mg, 34.7 μmol) in DMF (15 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 90℃ for 4 hrs under N ₂ atmosphere. The reaction mixture was quenched by the addition of water (40 mL) , and then filtered to give a filter residue. The residue was purified by prep-HPLC to afford I-005. ¹H NMR (400 MHz, DMSO-d ₆) δ 11.23 (s, 1H) , 10.22 (s, 1H) , 9.22 (s, 1H) , 8.41 (s, 1H) , 8.06 -7.91 (m, 2H) , 7.67 (d, J = 8.8 Hz, 1H) , 7.47 (s, 1H) , 6.94 (s, 1H) , 2.74 (d, J = 4.5 Hz, 3H) . LCMS [M+H] ⁺: 293.2.

Example 5: Preparation of N-methyl-4- ( (7- (2-oxopyrrolidin-1-yl) quinazolin-2-yl) amino) -1H-pyrrole-2-carboxamide (I-007)

Step 1: A mixture of pyrrolidin-2-one (83.9 mg, 986 μmol) , 7-bromo-2-chloroquinazoline (200 mg, 821 μmol) , Xantphos (157 mg, 271 μmol) , Pd ₂ (dba) ₃ (120 mg, 131 μmol) and K ₃PO ₄ (976 mg, 4.60 mmol) in toluene (5.0 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 60℃ for 16 hrs under N ₂ atmosphere. The reaction mixture was diluted with water (10 mL) and extracted with DCM (5.0 mL × 3) . The combined organic layers were washed with brine (10 mL) , dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give I-007-1. LCMS [M+H] ⁺: 248.2.

Step 2: To a solution of I-007-1 (50.0 mg, 202 μmol) and INT1-1 (28.1 mg, 202 μmol) in DMSO (1 mL) was added K ₂CO ₃ (41.9 mg, 303 μmol) . The mixture was stirred at 100℃ for 3 hrs. The mixture was purified by prep-HPLC to afford I-007. ¹H NMR (400 MHz, DMSO-d ₆) δ11.10 (s, 1H) , 9.62 (s, 1H) , 9.06 (s, 1H) , 7.97 (q, J = 4.7, 3.9 Hz, 1H) , 7.78 (s, 2H) , 7.66 (s, 1H) , 7.48 (s, 1H) , 6.82 (d, J = 4.1 Hz, 1H) , 3.98 (t, J = 7.0 Hz, 2H) , 2.73 (d, J = 4.5 Hz, 3H) , 2.57 (t, J = 8.1 Hz, 2H) , 2.11 (p, J = 7.5 Hz, 2H) . LCMS [M+H] ⁺: 351.2.

Example 6: Preparation of N-methyl-2- ( (5- (methylcarbamoyl) -1H-pyrrol-3-yl) amino) quinazoline-7-carboxamide (I-008)

Step 1: To a solution of enzyme (CD-HLE-001, 200 mg) in buffer (33.0 mL, 0.1 M solution of sodium phosphate, pH = 7.0) was added methyl 2-chloroquinazoline-7-carboxylate (333 mg, 1.50 mmol) in DMSO (3.30 mL) at 20℃. The mixture was stirred at 35℃ for 24 hrs. The reaction mixture was extracted with DCM (40 mL × 3) . The combined organic layers were washed with brine (120 mL) , dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give I-008-1. LCMS [M+H] ⁺: 209.2.

Step 2: To a solution of I-008-2 (200 mg, 959 μmol) in DCM (1 mL) was added T ₃P (1.22 g, 1.92 mmol, 1.14 mL, 50.0%purity) and DIEA (501 μL 2.88 mmol, ) and methylamine hydrochloride (77.7 mg, 1.15 mmol) . The mixture was stirred at 25℃ for 2 hrs. The reaction mixture was quenched by the addition of water (3 mL) at 25℃ and extracted with DCM (5 mL × 3) . The combined organic layers were washed with brine (15 mL) , dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give I-008-2. LCMS [M+H] ⁺: 222.3.

Step 3: To a solution of I-008-2 (50.0 mg, 0.226 mmol) in DMSO (1 mL) was added INT-1 (31.4 mg, 0.226 mmol) and K ₂CO ₃ (46.8 mg, 0.338 mmol) . The mixture was stirred at 100℃for 2 hrs. The reaction was concentrated and purified by prep-HPLC to afford I-008. ¹H NMR (400 MHz, DMSO-d ₆) δ 11.18 (s, 1H) , 9.81 (s, 1H) , 9.22 (s, 1H) , 8.72 (q, J = 5.6, 5.0 Hz, 1H) , 8.02 (s, 1H) , 7.98 (q, J = 4.5 Hz, 1H) , 7.89 (d, J = 8.3 Hz, 1H) , 7.65 (dd, J = 8.4, 1.6 Hz, 1H) , 7.54 (s, 1H) , 6.86 (d, J = 2.5 Hz, 1H) , 2.83 (d, J = 4.4 Hz, 3H) , 2.74 (d, J = 4.6 Hz, 3H) . LCMS [M+H] ⁺: 325.3.

Example 7: Preparation of N-methyl-4- ( (7- (methylsulfonyl) quinazolin-2-yl) amino) -1H-pyrrole-2-carboxamide (I-009)

Step 1: A mixture of 7-bromo-2-chloroquinazoline (244 mg, 1.01 mmol) and INT-1 (140 mg, 1.01 mmol) in DMSO (5.0 mL) was stirred at 25℃ for 1 hr, followed by the addition of KF (87.6 mg, 1.51 mmol) . The resulting mixture was stirred at 120℃ for 2 hrs. The mixture was cooled to 25℃ and pour it into water (10 mL) . The mixture was put in ultrasound bath for 10 min and filtered to afford I-009-1. LCMS [M+H] ⁺: 346.0.

Step 2: A mixture of I-009-1 (60.0 mg, 133 μmol) , copper (II) trifluoroacetate (38.6 mg, 133 μmol) , methylsulfinyloxysodium (85.7 mg, 839 μmol) , N, N'-dimethylethane-1, 2-diamine (8.22 mg, 93.3 μmol, 10.0 μL) in DMSO (1 mL) was stirred at 140℃ for 2 hrs. The reaction mixture was diluted with EtOAc (5 mL) and filtered. The filtrate was diluted with water (3.0 mL) and extracted with EtOAc (15 mL × 3) . The combined organic layers were washed with brine (3 mL) , dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-009. ¹H NMR (400 MHz, D ₂O+DMSO-d ₆) δ 9.30 (s, 1H) , 8.09 (d, J = 8.3 Hz, 2H) , 7.69 (dd, J = 8.3, 1.8 Hz, 1H) , 7.48 (s, 1H) , 7.03 -6.93 (m, 1H) , 3.29 (s, 3H) , 2.74 (s, 3H) . LCMS [M+H] ⁺: 346.20.

Example 8: Preparation of 4- ( (7- (1H-pyrazol-4-yl) quinazolin-2-yl) amino) -N-methyl-1H-pyrrole-2-carboxamide (I-011)

Step 1: A mixture of I-009-2 (130 mg, 288 μmol) , 4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (112 mg, 577 μmol) , XPhos (27.5 mg, 57.7 μmol) , Pd (dppf) Cl ₂ (21.1 mg, 28.8 μmol) and Cs ₂CO ₃ (282 mg, 866 μmol) in dioxane (5 mL) and H ₂O (1 mL) was degassed and purged with N ₂ for 3 times, and then the mixture was stirred at 100℃ for 12 hrs under N ₂. The reaction mixture was diluted with EtOAc (5 mL) , and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-010. ¹H NMR (400 MHz, DMSO-d ₆) δ 13.10 (s, 1H) , 11.13 (s, 1H) , 9.58 (s, 1H) , 9.07 (s, 1H) , 8.44 (s, 1H) , 8.15 (s, 1H) , 7.96 (q, J = 4.2 Hz, 1H) , 7.79 (d, J = 8.5 Hz, 1H) , 7.77 (s, 1H) , 7.58 (dd, J = 8.3, 1.7 Hz, 1H) , 7.50 (s, 1H) , 6.85 (s, 1H) , 2.74 (d, J = 4.5 Hz, 3H) . LCMS [M+H] ⁺: 334.2.

Example 9: Preparation of N- (2- (dimethylamino) ethyl) -4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxamide (I-014)

Step 1: To a solution of methyl 4-amino-1H-pyrrole-2-carboxylate (1.08 g, 6.99 mmol) in DMF (25 mL) was added K ₂CO ₃ (1.45 g, 10.48 mmol) , 2-chloroquinazoline (1.15 g, 6.99 mmol) . The mixture was stirred at 80℃ for 16 hrs. The reaction mixture was washed with water (30 mL) and extracted with EtOAc (15 mL × 3) . The combined organic layers were washed with brine (30 mL × 2) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel chromatography to afford I-014-1.

Step 2: To a solution of I-014-1 (1.3 g, 4.61 mmol) in THF (46 mL) was added aqueous LiOH (1 M, 46 mL) . The mixture was stirred at 85℃ for 12 hrs. The reaction mixture was neutralized by HCl (1 M) and then extracted with EtOAc (40 mL × 3) . The combined organic layers were washed with brine (50 mL) , dried over anhydrous sodium sulfate, and concentrated to give I-014-2. LCMS [M-H] ⁺: 253.0.

Step 3: To a solution of I-014-2 (100 mg, 393 μmol) in DMF (1 mL) was added N', N'-dimethylethane-1, 2-diamine (36.4 mg, 412.9 μmol, 45.1 uL) , HOBt (58.46 mg, 432.66 μmol) , EDCI (82.94 mg, 432.66 μmol) and DIPEA (101.6 mg, 786.65 μmol, 137.02 μL) . The resulting mixture was degassed and purged with N ₂ for 3 times and stirred at 25℃ for 16 hrs under N ₂ atmosphere. The reaction mixture was diluted with water (2 mL) and extracted with EtOAc (3 mL × 3) . The combined organic layers were washed with brine (5.0 mL × 2) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-014. ¹H NMR (400 MHz, CD ₃OD) δ 9.07 (s, 1H) , 7.79 (d, J = 8.0 Hz, 1H) , 7.73 (t, J = 8.2 Hz, 1H) , 7.64 (d, J = 8.5 Hz, 1H) , 7.52 (s, 1H) , 7.29 (t, J = 7.4 Hz, 1H) , 6.96 (d, J = 1.6 Hz, 1H) , 3.50 (t, J = 6.8 Hz, 2H) , 2.57 (t, J = 6.9 Hz, 2H) , 2.32 (s, 6H) . LCMS [M+H] ⁺: 325.0.

Example 10: Preparation of N- (2- (4-methylpiperazin-1-yl) ethyl) -4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxamide (I-015)

A mixture of I-014-2 (100 mg, 393.3 μmol) , 2- (4-methylpiperazin-1-yl) ethan-1-amine (59.1 mg, 412.9 μmol) , EDCI (82.94 mg, 432.66 μmol) , DIPEA (101.6 mg, 786.65 μmol, 137.02 μL) and HOBt (58.46 mg, 432.66 μmol) in DMF (1 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 25℃ for 3 hrs under N ₂ atmosphere. The reaction mixture was quenched by with water (5 mL) , concentrated under reduced pressure and purified by prep-HPLC to afford I-015. ¹H NMR (400 MHz, CD ₃OD) δ 9.08 (s, 1H) , 7.79 (d, J = 8.0 Hz, 1H) , 7.73 (ddd, J = 8.5, 6.8, 1.5 Hz, 1H) , 7.64 (d, J = 8.5 Hz, 1H) , 7.51 (s, 1H) , 7.30 (d, J = 7.4 Hz, 1H) , 6.95 (d, J = 1.6 Hz, 1H) , 3.50 (d, J = 6.8 Hz, 2H) , 2.69 -2.43 (m, 10H) , 2.29 (s, 3H) . LCMS [M+H] ⁺: 380.3.

Example 11: Preparation of N- (1-methylpiperidin-4-yl) -4- (quinazolin-2-ylamino) -1H-pyrrole-2-carboxamide (I-017)

A mixture of I-014-2 (100 mg, 393 μmol) , 1-methylpiperidin-4-amine (59.15 mg, 412.99 μmol) , EDCI (82.9 mg, 432 μmol) , DIPEA (101 mg, 786 μmol) and HOBt (58.4 mg, 432 μmol) in DMF (1 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 25℃for 3 hrs under N ₂ atmosphere. The reaction mixture was diluted by water (5 mL) , concentrated under reduced pressure, and purified by prep-HPLC to afford I-017. ¹H NMR (400 MHz, CD ₃OD) δ 9.09 (s, 1H) , 7.81 (d, J = 8.0 Hz, 1H) , 7.75 (ddd, J = 8.4, 6.9, 1.5 Hz, 1H) , 7.66 (d, J = 8.5 Hz, 1H) , 7.54 (s, 1H) , 7.31 (ddd, J = 8.0, 6.9, 1.2 Hz, 1H) , 7.01 (d, J = 1.6 Hz, 1H) , 3.93 -3.81 (m, 1H) , 2.94 (d, J = 11.9 Hz, 2H) , 2.33 (s, 3H) , 2.19 (t, J = 11.9 Hz, 2H) , 1.97 (d, J = 12.5 Hz, 2H) , 1.69 (qd, J = 12.2, 3.8 Hz, 2H) . LCMS [M-H] ⁺: 351.2.

Example 12: Preparation of N-methyl-3- (quinazolin-2-ylamino) benzamide (I-018)

To a solution of 3-amino-N-methylbenzamide (100 mg, 665 μmol) in i-PrOH (2 mL) was added 2-chloroquinazoline (131 mg, 799 μmol) and TsOH (229mg, 1.33 mmol) at 20℃. The mixture was stirred at 60℃ for 4 hrs. The reaction mixture was concentrated and purified by Prep-HPLC to afford I-018. ¹H NMR (400 MHz, CD ₃OD) δ 9.19 (d, J = 0.7 Hz, 1H) , 8.35 (dt, J = 2.2, 1.0 Hz, 1H) , 8.11 -8.05 (m, 1H) , 7.86 (dd, J = 8.0, 1.5 Hz, 1H) , 7.80 (ddd, J = 8.3, 6.7, 1.4 Hz, 1H) , 7.74 (dd, J = 8.5, 1.1 Hz, 1H) , 7.44 -7.41 (m, 2H) , 7.39 (ddd, J = 8.0, 6.8, 1.3 Hz, 1H) , 2.95 (s, 3H) . LCMS [M+H] ⁺: 278.1.

Example 13: Preparation of 4- ( (7- (imidazo [1, 2-a] pyrimidin-3-yl) quinazolin-2-yl) amino) -N-methyl-1H-pyrrole-2-carboxamide (I-034)

Step 1: A mixture of 7-bromo-2-chloroquinazoline (300 mg, 1.23 mmol) , imidazo [1, 2-a]pyrimidine (176 mg, 1.48 mmol) , Pd (OAc) ₂ (27.7 mg, 0.123 mmol) , tetrabutylammonium chloride (68.5 mg, 0.246 mmol) and K ₂CO ₃ (511 mg, 3.70 mmol) in DMF (3 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 2 hrs under N ₂ atmosphere. The mixture was filtrated and washed with water (3 mL × 3) . The organic phase was concentrated under vacuum to give I-034-1. LCMS [M+H] ⁺: 282.2.

Step 2: To a solution of I-034-1 (50.0 mg, 0.359 mmol) in DMSO (1 mL) was added INT-1 (100 mg, 0.355 mmol) and K ₂CO ₃ (73.6 mg, 0.532 mmol) . The mixture was stirred at 100℃for 1 hr. The mixture was concentrated and purified by prep-HPLC to afford I-034. ¹H NMR (400 MHz, DMSO-d ₆) δ 11.14 (s, 1H) , 9.77 (s, 1H) , 9.26 -9.16 (m, 2H) , 8.65 (dd, J = 4.1, 1.9 Hz, 1H) , 8.20 (s, 1H) , 7.99 (d, J = 8.3 Hz, 1H) , 7.98 -7.92 (m, 1H) , 7.86 (s, 1H) , 7.62 (dd, J =8.2, 1.7 Hz, 1H) , 7.53 (s, 1H) , 7.20 (dd, J = 6.9, 4.1 Hz, 1H) , 6.88 (s, 1H) , 2.74 (d, J = 4.6 Hz, 3H) . LCMS [M+H] ⁺: 385.4.

Example 14: Preparation of N-methyl-4- ( (7- (5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyridin-3-yl) quinazolin-2-yl) amino) -1H-pyrrole-2-carboxamide (I-035)

Step 1: A mixture of I-009-1 (20.0 mg, 0.0580 mmol) , bis (pinacolato) diboron (29.3 mg, 0.116 mmol) , Pd (dppf) Cl ₂ (4.23 mg, 0.06 mmol) and KOAc (17.0 mg, 0.173 mmol) in dioxane (1 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 12 hrs under N ₂ atmosphere. The mixture was concentrated to give I-035-1. LCMS [M+H] ⁺: 312.1.

Step 2: A mixture of I-035-1 (22.8 mg, 0.073 mmol) , 3-bromo-5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyridine (22.1 mg, 0.110 mmol) , Pd (dppf) Cl ₂ (5.36 mg, 0.07 mmol) , and Cs ₂CO ₃ (71.7 mg, 0.220 mmol) in dioxane (1 mL) and H ₂O (0.4 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 12 hrs under N ₂ atmosphere. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (10 mL × 3) . The combined organic layers were washed with brine (20 mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-035. ¹H NMR (400 MHz, DMSO-d ₆) δ 11.13 (s, 1H) , 9.73 (s, 1H) , 9.18 (s, 1H) , 7.96 (q, J = 4.7, 4.2 Hz, 1H) , 7.90 (d, J = 8.3 Hz, 1H) , 7.65 (s, 1H) , 7.49 (s, 1H) , 7.42 (dd, J = 8.2, 1.7 Hz, 1H) , 6.85 (s, 1H) , 4.14 (t, J = 5.4 Hz, 2H) , 2.93 (t, J = 5.3 Hz, 2H) , 2.74 (d, J =4.5 Hz, 3H) , 1.99 -1.87 (m, 4H) . LCMS [M+H] ⁺: 388.2.

Example 15: Preparation of N-methyl-4- ( (7- (3-methyl-1H-pyrazol-4-yl) quinazolin-2-yl) amino) -1H-pyrrole-2-carboxamide (I-036)

A mixture of I-009-1 (100 mg, 0.14 mmol) , 3-methyl-4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (57.7 mg, 0.28 mmol) , Cs ₂CO ₃ (136 mg, 0.42 mmol) , Pd (dppf) Cl ₂ (10.2 mg, 0.02 mmol) in 1, 4-dioxane (5 mL) and H ₂O (1 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 12 hrs under N ₂ atmosphere. The reaction mixture was diluted with EtOAc (5 mL) and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-036. ¹H NMR (400 MHz, DMSO-d ₆) δ 12.84 (s, 1H) , 11.10 (s, 1H) , 9.61 (s, 1H) , 9.11 (s, 1H) , 7.97 (q, J = 4.5 Hz, 1H) , 7.83 (d, J = 8.3 Hz, 1H) , 7.60 (s, 1H) , 7.45 (dd, J = 8.3, 1.7 Hz, 2H) , 6.87 (s, 1H) , 2.74 (d, J = 4.5 Hz, 4H) . LCMS [M+H] ⁺: 348.3.

Example 16: Preparation of 4- ( (6- (1H-pyrazol-4-yl) quinazolin-2-yl) amino) -N-methyl-1H-pyrrole-2-carboxamide (I-037)

A mixture of I-005-1 (10.0 mg, 28.9 μmol) , (1H-pyrazol-4-yl) boronic acid (11.2 mg, 57.8 μmol) , Pd (dppf) Cl ₂ (2.11 mg, 2.89 μmol) , Cs ₂CO ₃ (28.2 mg, 86.7 μmol) in dioxane (2 mL) and H ₂O (0.4 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃for 12 hrs under N ₂ atmosphere. The reaction mixture was diluted with EtOAc (3 mL) and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-037. ¹H NMR (400 MHz, DMSO-d ₆) δ 12.98 (s, 1H) , 11.10 (s, 1H) , 9.62 (s, 1H) , 9.12 (s, 1H) , 8.26 (s, 1H) , 8.08 -7.94 (m, 4H) , 7.59 (d, J = 9.2 Hz, 1H) , 7.45 (s, 1H) , 6.88 (t, J = 2.3 Hz, 1H) , 2.74 (d, J = 4.5 Hz, 3H) . LCMS [M+H] ⁺: 334.2.

Example 17: Preparation of 5- ( (7- (1H-pyrazol-4-yl) quinazolin-2-yl) amino) -N-methylisothiazole-3-carboxamide (I-038)

Step 1: To a solution of 5-nitroisothiazole-3-carboxylic acid (300 mg, 1.72 mmol) , MeNH ₂ (107 mg, 3.44 mmol) and DIEA (1.13 mL, 6.89 mmol) in DMF (6.0 mL) was added HATU (1310 mg, 3.44 mmol) . The mixture was stirred at 20℃ for 12 hrs. The reaction mixture was added water (5.0 mL) , then extracted with EtOAc (10 mL × 3) . The combined organic layers were washed with brine (5.0 mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by pre-TLC to afford I-038-1. LCMS [M+H] ⁺: 188.1.

Step 2: To a solution of I-038-1 (340 mg, 1.54 mmol) in EtOH (4.0 mL) was added NH ₄Cl (247 mg, 4.62 mmol) in H ₂O (1 mL) , followed by the addtion of iron powder (430 mg, 7.71 mmol) . The mixture was stirred at 25℃ for 6 hrs. The reaction mixture was filtered and then extracted with EtOAc (3.0 mL × 3) . The combined organic layers were dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give I-038-2. LCMS [M+Na] ⁺: 180.1.

Step 3: To a solution of I-038-2 (200 mg, 1.27 mmol) and INT-2 (420 mg, 1.27 mmol) in dioxane (5.0 mL) was added Pd ₂ (dba) ₃ (116 mg, 0.127 mmol) , Xantphos (147 mg, 0.254 mmol) and Cs ₂CO ₃ (829 mg, 2.54 mmol) under N ₂ atomosphere. The reaction mixture was stirred at 120℃ for 2 hrs under microwave irratation. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-038. ¹H NMR (400 MHz, DMSO-d ₆) δ 13.21 (s, 1H) , 11.91 (s, 1H) , 9.36 (s, 1H) , 8.58 (s, 1H) , 8.43 (dd, J = 9.3, 4.8 Hz, 1H) , 8.25 (s, 1H) , 8.07 (s, 1H) , 8.01 (d, J = 8.4 Hz, 1H) , 7.84 (d, J = 8.4 Hz, 1H) , 7.33 (s, 1H) , 2.77 (d, J = 4.8 Hz, 3H) . LCMS [M+H] ⁺: 352.0.

Example 18: Preparation of N- (4- ( (methylamino) methyl) phenyl) -7- (1H-pyrazol-4-yl) quinazolin-2-amine (I-039)

Step 1: To a solution of 4- ( (methylamino) methyl) aniline (500 mg, 3.67 mmol) in THF (5.0mL) was added Boc ₂O (0.843 mL, 3.671 mmol) at 0℃. The reaction mixture was warmed to room temperature and stirred for 2 h. The reaction mixture was concentrated to afford I-039-1.

Step 2: To a mixture of I-039-1 (100 mg, 0.423 mmol) and 7-bromo-2-chloroquinazoline (124mg, 0.508 mmol) in DMSO (1 mL) was added potassium fluoride (36.9mg, 0.635 mmol) . The resulting mixture was stirred at 120℃ for 2 hrs. The reaction mixture was concentrated under reduced pressure and purified by prep-TLC to afford I-039-2. LCMS [M+H] ⁺: 443.2.

Step 3: A mixture of I-039-2 (200 mg, 0.056 mmol) , 2-methylpropan-2-yl 4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyrazole-1-carboxylate (151.3 mg, 0.514 mmol) , Pd (dppf) Cl ₂ (31.4 mg, 0.043 mmol) and K ₂CO ₃ (177.7 mg, 1.286 mmol) in THF (2.0 mL) and H ₂O (0.4 mL) was stirred at 80℃ for 2 h under N ₂ atmosphere. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (10 mL × 3) . The combined organic layers were washed with brine (20 mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-039-3.

Step 4: To a solution of I-039-3 (100 mg, 0.303 mmol) in MeOH (5.0 mL) was added HCl/MeOH (3 mL) . The mixture was stirred at 20℃ for 2 hrs. The reaction was concentrated under reduced pressure and purified by prep-HPLC to afford I-039. ¹H NMR (400 MHz, D2O+DMSO-d ₆) δ 9.19 (s, 1H) , 8.27 (s, 2H) , 7.98 (d, J = 8.6 Hz, 2H) , 7.91 (d, J = 8.4 Hz, 1H) , 7.85 (d, J = 1.4 Hz, 1H) , 7.69 (dd, J = 8.3, 1.6 Hz, 1H) , 7.44 -7.40 (m, 2H) , 4.05 (s, 2H) , 2.56 (s, 3H) . LCMS [M+H] ⁺: 331.1.

Example 19: Preparation of 4- ( (7- (1H-pyrazol-4-yl) quinazolin-2-yl) amino) -N-methylbenzamide (I-040)

Step 1: To a solution of 4-amino-N-methylbenzamide (300 mg, 1.20 mmol) in 2-methylpropan-2-ol (15 mL) was added INT-2 (661 mg, 1.72 mmol) and TsOH (516 mg, 2.0 mmol) . The mixture was stirred at 100℃ for 2 hrs. The reaction was filtered and washed with EtOAc (10 mL) to afford I-040-1. LCMS [M+H-Boc] ⁺: 345.2

Step 2: A solution of I-040-1 (100 mg, 0.19 mmol) in MeOH/HCl (10 mL) was stirred at 25℃ for 1 hr. The reaction was concentrated under vacuum and purified by prep-HPLC to give I-040. ¹H NMR (400 MHz, DMSO-d ₆) δ 10.21 (s, 1H) , 9.27 (s, 1H) , 8.38 (s, 2H) , 8.33 (s, 1H) , 8.05 (d, J = 8.4 Hz, 2H) , 7.94 (d, J = 8.3 Hz, 2H) , 7.84 (d, J = 7.0 Hz, 2H) , 7.75 (dd, J = 8.3, 1.6 Hz, 1H) , 2.79 (d, J = 2.2 Hz, 3H) , 2.54 (s, 1H) . LCMS [M+H] ⁺: 345.2.

Example 20: Preparation of 3- ( (6- (1H-pyrazol-4-yl) quinazolin-2-yl) amino) -N-methylbenzamide (I-041)

Step 1: A mixture of INT-4 (400 mg, 1.27 mmol) , 3-amino-N-methylbenzamide (209 mg, 1.39 mmol) and K ₂CO ₃ (526 mg, 3.81 mmol) in DMSO (2 mL) was stirred at 100℃ for 2 hrs. The mixture was filtered and the filtrate was concentrated under reduced pressure to give I-041-1.

Step 2: A solution of I-041-1 (400 mg, 0.933 mmol) in HCl/MeOH (5.0 mL) was stirred at 25℃ for 2 hrs. The reaction mixture was concentrated and purified by pre-HPLC to afford I-041. ¹H NMR (400 MHz, DMSO-d ₆) δ 13.02 (s, 1H) , 9.94 (s, 1H) , 9.27 (s, 1H) , 8.39 -8.28 (m, 3H) , 8.18 -8.15 (m, 1H) , 8.14 -8.09 (m, 2H) , 8.04 (s, 1H) , 7.69 -7.64 (m, 1H) , 7.40 (d, J = 5.2 Hz, 2H) , 2.80 (d, J = 4.5 Hz, 3H) . LCMS [M+H] ⁺: 345.2.

Example 21: Preparation of 5- ( (6- (1H-pyrazol-4-yl) quinazolin-2-yl) amino) -N-methylisothiazole-3-carboxamide (I-042)

Step 1: To a solution of 5-nitroisothiazole-3-carboxylic acid (200 mg, 1.15 mmol) in DMF (2 mL) was added HATU (524 mg, 1.38 mmol) and DIPEA (0.570 mL, 3.45 mmol) and MeNH ₂·HCl (93.1 mg, 1.38 mmol) . The mixture was stirred at 25℃ for 2 hrs. The reaction mixture was quenched with water (10 mL) and extracted with EtOAc (5.0 mL × 3) . The combined organic layers were washed with brine (10 mL) , dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give I-042-1. LCMS [M+H] ⁺: 188.3.

Step 2: To a solution of I-042-1 (200 mg, 1.07 mmol) in EtOH (2 mL) and H ₂O (0.40 mL) was added iron powder (298 mg, 5.34 mmol) and NH ₄Cl (171 mg, 3.21 mmol) . The mixture was stirred at 20℃ for 6 hrs. The mixture was filtrated, and the filtrate was concentrated under vacuum to give I-042-2. LCMS [M+H] ⁺: 158.2.

Step 3: A mixture of INT-4 (100 mg, 0.318 mmol) , I-042-2 (49.9 mg, 0.318 mmol) , Pd (OAc) ₂ (7.13 mg, 0.032 mmol) , Xantphos (36.8 mg, 0.064 mmol) and Cs ₂CO ₃ (207 mg, 0.635 mmol) in dioxane (1 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 110℃ for 1 hr under N ₂ atmosphere via microwave irritation. The mixture was filtrated to give I-042-3. LCMS [M+H] ⁺: 436.3.

Step 4: To a solution of I-042-3 (50.0 mg, 0.115 mmol) in MeOH (1 mL) was added HCl (6 M, 0.096 mL, 0.574 mmol) . The mixture was stirred at 25℃ for 16 hrs. The mixture was filtrated, and the obtained solid was purified by prep-HPLC to afford I-042. ¹HNMR (400 MHz, DMSO-d ₆) δ 12.2-11.8 (m, 1H) , 9.42 (s, 1H) , 8.50 -8.38 (m, 1H) , 8.24 (d, J = 7.2 Hz, 4H) , 7.87 (d, J = 8.0 Hz, 1H) , 7.34 (s, 1H) , 2.77, (d, J = 4.8 Hz, 3H) . LCMS [M+H] ⁺: 352.3.

Example 22: Preparation of N- (4- ( (methylamino) methyl) phenyl) -6- (1H-pyrazol-4-yl) quinazolin-2-amine (I-043)

Step 1: To a solution of I-039-1 (100 mg, 0.423 mmol) in DMSO (1.0 mL) was added INT-4 (133 mg, 0.423 mmol) and K ₂CO ₃ (146 mg, 1.06 mmol) . The mixture was stirred at 100℃ for 2 hrs under N ₂ atmosphere. The reaction was diluted with water (10 mL) and extracted with EtOAc (8 mL × 3) . The combined organic phases were washed with brine and dried over anhydrous sodium sulfate, concentrated, and purified by prep-HPLC to give I-043-1. LCMS [M+H] ⁺: 515.3.

Step 2: A solution of I-043-1 (180 mg, 0.340 mmol) in HCl /MeOH (5.0 mL) was stirred at 25℃ for 2 hrs. The reaction was concentrated and purified by prep-HPLC to give I-043. ¹H NMR (400 MHz, CD ₃OD) δ 9.08 (s, 1H) , 8.02 -7.93 (m, 4H) , 7.85 (d, J = 8.1 Hz, 2H) , 7.59 (d, J = 8.6 Hz, 1H) , 7.25 (d, J = 8.1 Hz, 2H) , 3.72 (s, 2H) , 2.37 (s, 3H) . LCMS [M+H] ⁺: 331.4.

Example 23: Preparation of N- (1- (2-methyl-2-azaspiro [3.3] heptan-6-yl) -1H-pyrazol-4-yl) -6- (1H-pyrazol-4-yl) quinazolin-2-amine (I-044)

Step 1: To a solution of tert-butyl 6-hydroxy-2-azaspiro [3.3] heptane-2-carboxylate (2.00 g, 9.37 mmol) and PPh ₃ (3.69 g, 14.0 mmol) in THF (25mL) was added DIAD (2.84 g, 14.0 mmol) at 0℃ under N ₂ atmosphere. The mixture was stirred at 25℃ for 18 hrs. The mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL × 3) . The combined organic layers were washed with brine (100 mL) , dried over Na ₂SO ₄, and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel chromatography to afford I-044-1. LCMS [M+H] ⁺: 309.1.

Step 2: To a solution of I-044-1 (2.50 g, 8.10 mmol) in THF (20 mL) was added LiAlH ₄ (1.08 g, 28.3 mmol) under N ₂ atmosphere. The mixture was stirred at 25℃ for 8 hrs. The reaction mixture was diluted with water (50 mL) and extracted with EtOAc (50 mL × 3) . The combined organic layers were washed with brine (100 mL) , dried over Na ₂SO ₄, and filtered. The filtrate was concentrated and purified by silica gel chromatography to give I-044-2.

Step 3: To a mixture of I-044-2 (200 mg, 1.47 mmol) ) and INT-4 (260 mg, 1.47 mmol) in DMSO (15 mL) was added conc. HCl (12 M, 0.12 mL, 1.47 mmol) . The mixture was stirred at 80℃ for 3 hrs. The reaction mixture was concentrated under reduced pressure and purified by prep-HPLC to afford I-044. ¹H NMR (400 MHz, DMSO-d ₆) δ 11.10 (s, 1H) , 9.62 (s, 1H) , 9.06 (s, 1H) , 7.97 (q, J = 4.7, 3.9 Hz, 1H) , 7.78 (s, 2H) , 7.66 (s, 1H) , 7.48 (s, 1H) , 6.82 (d, J = 4.1 Hz, 1H) , 3.98 (t, J = 7.0 Hz, 2H) , 2.73 (d, J = 4.5 Hz, 3H) , 2.57 (t, J = 8.1 Hz, 2H) , 2.11 (p, J = 7.5 Hz, 2H) . LCMS [M+H] ⁺: 387.4.

Example 24: Preparation of N- (4- ( (methylamino) methyl) phenyl) -7- (5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyridin-3-yl) quinazolin-2-amine (I-045)

Step 1: To a solution of (4-iodophenyl) methanamine (10.0 g, 42.9 mmol) and TEA (8.95 mL, 64.4 mmol) in THF (100 mL) was added Boc ₂O (14.8 mL, 64.4 mmol) at 0℃. The mixture was stirred at 25℃ for 18 hrs. The reaction mixture was diluted with water (300 mL) and extracted with EtOAc (100 mL × 3) . The combined organic layers were washed with brine (100 mL) , dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give I-045-1. LCMS [M+H- ^tBu] ⁺: 278.0.

Step 2: To a stirred suspension of NaH (0.60 g, 15.07 mmol) in THF (30 mL) was added I-045-1 (2.0 g, 6.0 mmol) at 0℃, followed by the addition of CH ₃I (2.13 g, 15.0 mmol) . The mixture was stirred at 25℃ for 18 hrs. The reaction mixture was quenched with aqueous NaHCO ₃ (50 mL) , extracted with EtOAc (50 mL × 3) . The combined organic layers were washed with brine (50 mL) , dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give I-045-2. LCMS [M+H- ^tBu] ⁺: 291.9.

Step 3: A mixture of 7-bromoquinazolin-2-amine (2.50 g, 11.2 mmol) , 4, 4, 5, 5-tetramethyl-2- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1, 3, 2-dioxaborolane (3.12 g, 12.2 mmol) , KOAc (3.29 g, 33.5 mmol) , Pd (dppf) Cl ₂ (0.41 g, 0.558 mmol) in dioxane (25 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 80℃ for 2 hrs under N ₂ atmosphere. The mixture was concentrated under vacuum, diluted in DCM (20 mL) , and filtered. The filtrate was concentrated, and the obtained residue was washed with PE (10 mL × 3) to afford I-045-3. LCMS [M+H] ⁺: 190.0.

Step 4: A mixture of I-045-3 (500 mg, 1.84 mmol) , 3-bromo-5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyridine (532 mg, 2.65 mmol) , Pd (dppf) Cl ₂ (96.8 mg, 0.132 mmol) , K ₃PO ₄ (1.68 g, 7.94 mmol) in dioxane (5 mL) and H ₂O (1 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 2 hrs under N ₂ atmosphere. The mixture was poured into water (10 mL) and filtrated. The filtrate was extracted with EtOAc (10 mL × 3) . The combined organic layers were washed with brine (10 mL) , dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give I-045-4. LCMS [M+H] ⁺: 266.3.

Step 5: A mixture of I-045-4 (200 mg, 0.482 mmol) , I-045-2 (184 mg, 0.531 mmol) , CuI (9.19 mg, 0.048 mmol) , K ₂CO ₃ (200 mg, 1.45 mmol) and methyl [ (1R, 2R) -2- (methylamino) cyclohexyl] amine (13.7 mg, 0.096 mmol) in DMF (1 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 2 hrs under N ₂ atmosphere. The reaction mixture was diluted with NH ₄Cl (5 mL) and extracted with EtOAc (5 mL × 3) . The combined organic layers were washed with brine (5 mL) , dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give I-045-5. LCMS [M+H] ⁺: 485.4.

Step 6: A mixture of I-045-5 (5.0 mg, 10.3 μmol) in HCl/MeOH (2 mL) was stirred at 25℃for 1 hr under N ₂ atmosphere. The reaction was filtered, concentrated under reduced pressure and purified by prep-HPLC to afford I-045. ¹H NMR (400 MHz, CD ₃OD) δ 8.39 (brs, 2H) , 8.05 -7.98 (m, 2H) , 7.92 -7.86 2 (m, 1H) , 7.75 (d, J = 8.50 Hz, 1H) , 7.39 -7.45 (m, 2H) , 7.21 -7.29 (m, 1H) , 4.06 -4.19 (m, 4H) , 2.96 (d, J = 5.50 Hz, 2 H) , 2.68 (d, J = 1.50 Hz, 3 H) , 1.97 -2.06 (m, 4H) . LCMS [M+H] ⁺: 385.3.

Example 25: Preparation of N- (1- (2-methyl-2-azaspiro [3.3] heptan-6-yl) -1H-pyrazol-4-yl) -7- (1H-pyrazol-4-yl) quinazolin-2-amine (I-046)

A mixture of INT-3 (180 mg, 0.465 mmol) , 1- (2-methyl-2-azaspiro [3.3] heptan-6-yl) -1H-pyrazol-4-amine (126.4 mg, 0.658) and 4-methylbenzenesulfonic acid (100 mg, 0.572 mmol) in dioxane (5 mL) was stirred at 100℃ for 2 hrs. The mixture is concentrated and purified by prep-HPLC to afford I-046. ¹H NMR (400 MHz, CD ₃OD) δ 9.04 (s, 1H) , 8.27 (s, 1H) , 8.19 (s, 2H) , 7.88 (s, 1H) , 7.81 (d, J = 8.4 Hz, 1H) , 7.73 (s, 1H) , 7.61 (dd, J = 8.3, 1.6 Hz, 1H) , 4.76 (p, J =8.3 Hz, 1H) , 3.44 (s, 2H) , 3.35 (s, 2H) , 2.73 (s, 2H) , 2.71 (s, 2H) , 2.35 (s, 3H) . LCMS [M+H] ⁺: 387.2.

Example 26: Preparation of N- (1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-yl) -7- (1H-pyrazol-4-yl) quinazolin-2-amine (I-047)

To a solution of INT-2 (100 mg, 0.24 mmol) , 1- (1-methylpiperidin-4-yl) -1H-pyrazol-4-amine (43.6 mg, 0.24 mmol) in i-PrOH (5 mL) was added TsOH (41.7 mg, 0.24 mmol) . The mixture was stirred at 100℃ for 2 hrs. The reaction mixture was filtered, and the filtrate was concentrated and purified by prep-HPLC to afford I-047. ¹H NMR (400 MHz, DMSO-d ₆) δ13.14 (s, 1H) , 9.63 (s, 1H) , 9.10 (s, 1H) , 8.56 -8.07 (m, 3H) , 7.88 (s, 1H) , 7.82 (d, J = 8.3 Hz, 1H) , 7.67 (s, 1H) , 7.62 (dd, J = 8.4, 1.6 Hz, 1H) , 4.12 (p, J = 7.2 Hz, 1H) , 2.93 -2.84 (m, 2H) , 2.22 (s, 3H) , 2.10 -1.96 (m, 6H) . LCMS [M+H] : 375.5.

Example 27: Preparation of N4- (7- (1H-pyrazol-4-yl) quinazolin-2-yl) -N2- (2-methyl-2-azaspiro [3.3] heptan-6-yl) pyridine-2, 4-diamine (I-048)

Step 1: To a solution of tert-butyl 6-amino-2-azaspiro [3.3] heptane-2-carboxylate (896 mg, 4.22 mmol ) , 2-fluoro-4-nitropyridine (600 mg, 4.22 mmol) in DMF (12 mL) was added K ₂CO ₃ (1.45 g, 10.5 mmol) . The mixture was stirred at 90℃ for 12 hrs. The reaction mixture was diluted with water (20 mL) and extracted with DCM (25 mL × 3) . The combined organic layers were washed with brine (20 mL) , dried over Na ₂SO ₄, and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel chromatography to afford I-048-1.

Step 2: To a solution of I-048-1 (1.00 g, 2.99 mmol) in MeOH (15 mL) was added Pd/C 10%(0.06 g, 0.598 mmol) under N ₂ atmosphere. The suspension was degassed and purged with H ₂ for 3 times. The mixture was stirred under H ₂ (15 psi) at 25℃ for 2 hr. The reaction mixture was filtered and concentrated under reduced pressure to give I-048-2.

Step 3: A mixture of I-048-2 (450 mg, 1.47 mmol) ) and INT-3 (465 mg, 1.47 mmol) in dioxane (15 mL) and added Pd (dba) ₃ (676 mg, 0.73 mmol) , Xantphos (342 mg, 0.591 mmol) and Cs ₂CO ₃ (1.44 g, 4.43 mmol) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 3 hrs under N ₂. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (30 mL × 3) . The combined organic layers were washed with brine (100 mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by silica gel chromatography to afford I-048-3.

Step 4: To a solution of I-048-3 (100 mg, 9.27 mmol) in H ₂O (5.0 mL) was added HCl (12 M, 4.0 mL) . The mixture was stirred at 25℃ for 5 hrs. The reaction mixture was concentrated under reduced pressure to give I-048-4. LCMS [M+H] ⁺: 399.3.

Step 5: A mixture of I-048-4 (50.0 mg, 0.125 mmol) , formaldehyde (16.9 μL, 0.62 mmol) , AcOH (35.9 μL, 0.627 mmol) and NaBH ₃CN (7.89 mg, 0.125 mmol) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 25℃ for 5 hrs under N ₂ atmosphere. The combined organic layers were washed with brine (20 mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by HPLC to give I-048. ¹H NMR (400 MHz, CD ₃OD) δ 9.20 (d, J = 2.6 Hz, 1H) , 8.55 (brs, 2H) , 8.04 -7.90 (m, 3H) , 7.81 -7.67 (m, 2H) , 7.02 (d, J = 6.1 Hz, 1H) , 4.31 -4.22 (m, 2H) , 4.19 -4.10 (m, 3H) , 3.04 -2.80 (m, 5H) , 2.43 -2.31 (m, 2H) . LCMS [M+H] ⁺: 413.4.

Example 28: Preparation of N- (2- (1-methylazetidin-3-yl) -1, 2, 3, 4-tetrahydroisoquinolin-6-yl) -7- (1H-pyrazol-4-yl) quinazolin-2-amine (I-049)

Step 1: A mixture of 6-bromo-1, 2, 3, 4-tetrahydroisoquinoline (887 mg, 5.19 mmol) , 6-bromo-1, 2, 3, 4-tetrahydroisoquinoline (1.0 g, 4.72 mmol) , NaBH (OAc) ₃ (1.10 g, 5.19 mmol) , acetic acid (28.3 mg, 471 μmol, 26.9 μL) in DCM (10 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 20℃ for 8 hrs under N ₂ atmosphere. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (30mL × 3) . The combined organic layers were washed with brine (5 mL) , dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give I-049-1. LCMS [M+H] ⁺: 367.3.

Step 2: A mixture of I-049-1 (350 mg, 1.19 mmol) , INT-5 (522 mg, 1.42 mmol) , Cs ₂CO ₃ (1.16 g, 3.56 mmol) , Xantphos (274 mg, 474.03 μmol, 0.4 eq) and Pd ₂ (dba) ₃ (217 mg, 237 μmol, 0.2 eq) in dioxane (1 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 140℃ for 5 hrs under N ₂ atmosphere. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc mL (30mL × 3) . The combined organic layers were washed with brine (5mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-049-2. LCMS [M+H] ⁺: 582.3.

Step 3: A solution of I-049-2 (200 mg, 2.18 mmol) in HCl/dioxane (1 M, 21.78 mL) was stirred at 25℃ for 5 hrs under N ₂ atmosphere. The reaction was concentrated under reduced pressure to give I-049-3. LCMS [M+H] ⁺: 398.4.

Step 4: A mixture of I-049-3 (60.0 mg, 2.84 mmol) , paraformaldehyde (60.0 mg, 3.41 mmol) , NaBH (OAc) ₃ (63.20 mg, 3.13 mmol) , AcOH (7.08 mg, 284 μmol, 16.2 μL) in DCM (10 mL) was degassed and purged with N ₂ for 3 times. The mixture was filtered, concentrated, and purified by prep-HPLC to afford I-049. ¹H NMR (400 MHz, CD ₃OD) δ 9.09 (s, 1H) , 8.49 (s, 1H) , 8.20 (s, 2H) , 7.87 -7.82 (m, 2H) , 7.74 -7.63 (m, 3H) , 7.08 (d, J = 8.9 Hz, 1H) , 4.28 (dd, J = 10.5, 7.2 Hz, 2H) , 4.04 (dd, J = 9.5, 6.5 Hz, 2H) , 3.60 (s, 2H) , 3.49 (p, J = 6.8 Hz, 1H) , 3.01 (t, J = 6.1 Hz, 2H) , 2.94 (s, 3H) , 2.73 (t, J = 6.0 Hz, 2H) . LCMS [M+H] ⁺: 412.3.

Example 29: Preparation of 4- ( (7- (1H-pyrazol-4-yl) quinazolin-2-yl) amino) -N- (1-methylpiperidin-4-yl) benzamide (I-050)

Step 1: To a solution of 4-iodobenzoyl chloride (3.0 g, 11.2 mmol) in DCM (20.0 mL) was added 1-methylpiperidin-4-amine (1.29 g, 11.2 mmol) and TEA (4.69 mL, 33.7 mmol) . The resulting reaction was stirred at 25℃ for 3 hrs. The mixture was filtered, and the filtrate was concentrated under reduced pressure to afford I-050-1.

Step 2: A mixture of I-050-1 (489 mg, 1.42 mmol) , INT-3 (420 mg, 1.42 mmol) , CuI (270 mg, 1.42 mmol) , Cs ₂CO ₃ (138 mg, 4.266 mmol) and methyl [ (1R, 2R) -2- (methylamino) cyclohexyl] amine (101 mg, 0.711 mmol) were dissolved in DMF (10.0 mL) and heated at 100℃for 12 hrs. The reaction mixture was filtered, added with water (10 mL) , and extracted with EtOAc (15 mL × 3) . The combined organic layers were washed with brine (10 mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by pre-HPLC to afford I-050-2. LCMS [M+H] ⁺: 512.3.

Step 3: A solution of I-050-2 (300 mg, 0.586 mmol) in HCl/MeOH (10.0 mL) was stirred at 25℃ for 1 hr. The reaction mixture was concentrated under reduced pressure to give I-050. ¹H NMR (400 MHz, DMSO-d ₆) δ 9.26 (s, 1H) , 8.31 (s, 2H) , 8.02 -7.93 (m, 3H) , 7.91 -7.83 (m, 3H) , 7.74 (dd, J = 8.4, 1.6 Hz, 1H) , 3.45 (d, J = 12.0 Hz, 2H) , 3.35 -3.21 (m, 1H) , 3.07 (t, J =12.7 Hz, 2H) , 2.75 (s, 3H) , 2.12 -1.96 (m, 2H) , 1.83 (q, J = 11.6 Hz, 2H) . LCMS [M+H] ⁺: 428.2.

Example 30: Preparation of N- (7-methoxy-2-methyl-1, 2, 3, 4-tetrahydroisoquinolin-6-yl) -7- (1H-pyrazol-4-yl) quinazolin-2-amine (I-051)

Step 1: To a solution of 4-bromo-3-methoxybenzaldehyde (10.0 g, 46.5 mmol) in toluene (100 mL) was added 2-amino-1, 1-dimethoxyethane (5.38 g, 51.1 mmol) and MgSO ₄ (8.40 g, 69.8 mmol) . The mixture was stirred at 100℃ for18 hrs. The reaction mixture was filtered and concentrated under reduced pressure to give I-051-1. ¹H NMR (400 MHz, CDCl ₃) δ 8.24 (s, 1H) , 7.59 (d, J = 8.0, Hz, 1H) , 7.41 (d, J = 1.6, Hz, 1H) , 7.11 (dd, J = 2.0, 8.0, Hz, 1H) , 4.70 (t, d, J = 5.2, Hz, 1H) , 3.97 (s, 3H) , 3.79 (d, J = 4.4, Hz, 2H) , 3.44 (s, 6H) .

Step 2: To a solution of I-051-1 (9.0 g, 29.8 mmol) in PPA (50 mL) and the mixture was stirred at 140℃ for 3 hrs. The reaction mixture was diluted with water (500 mL) and extracted with DCM (200 mL × 3) . The combined organic layers were washed with brine (500 mL) , dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give I-051-2.

Step 3: To a solution of I-051-2 (2.70 g, 11.3 mmol) in MeOH (1 mL) was added CH ₃I (2.74 g, 19.3 mmol) . The mixture was stirred at 25℃ for 16 hrs. The reaction mixture was concentrated under reduced pressure to give I-051-3. LCMS [M+H] ⁺: 252.0.

Step 4: To a solution of I-051-3 (4.0 g, 10.5 mmol) in MeOH (40 mL) was added NaBH ₄ (1.20 g, 31.7 mmol) at 0℃. The mixture was stirred at 25℃ for 16 hrs. The reaction mixture was quenched by saturated NH ₄Cl (40 mL) and extracted with EtOAc (50 mL × 3) . The combined organic layers were washed with brine (100 mL) , dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give I-051-4.

Step 5: A mixture of I-051-4 (50.0 mg, 0.20 mmol) , INT-3 (69.2 mg, 0.23 mmol) , Cs2CO3 (190 mg, 0.59 mmol) , Pd ₂ (dba) ₃ (17.9 mg, 0.02 mmol) and Xantphos (22.5 mg, 0.039 mmol) in dioxane (2 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100 ℃ for 3 hrs under N ₂ atmosphere. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (10 mL × 3) . The combined organic layers were washed with brine (30 mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-051-5. LCMS [M+H] ⁺: 471.5.

Step 6: A mixture of I-051-5 (30.0 mg, 0.064 mmol) in HCl/MeOH (5 mL) was stirred at 25℃ for 2 hrs. The reaction mixture was freeze-dried under reduced pressure to give I-051. ¹H NMR (400 MHz, DMSO-d ₆) δ 11.29 (s, 1H) , 9.34 (s, 1H) , 8.34 (s, 2H) , 8.09 -7.98 (m, 2H) , 7.93 (d, J = 1.5 Hz, 1H) , 7.82 (dd, J = 8.4, 1.6 Hz, 1H) , 6.99 (s, 1H) , 4.49 -4.41 (m, 1H) , 4.27 (dd, J = 15.5, 8.2 Hz, 1H) , 3.86 (s, 3H) , 3.66 (d, J = 9.8 Hz, 1H) , 3.39 -3.22 (m, 2H) , 3.07 -2.97 (m, 1H) , 2.92 (d, J = 4.6 Hz, 3H) . LCMS [M+H] ⁺: 387.2.

Example 31: Preparation of N- (2-methyl-4- (4-methylpiperazin-1-yl) phenyl) -7- (1H-pyrazol-4-yl) quinazolin-2-amine (I-052)

A mixture of 2-methoxy-4- (4-methylpiperazin-1-yl) aniline (80.2 mg, 362 μmol) , INT-2 (100 mg, 302 μmol) , TsOH (78.0 mg, 453 μmol) in i-PrOH (4 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 5 hrs under N ₂ atmosphere. The reaction mixture was quenched with water (10 mL) and extracted with EtOAc (30mL × 3) . The combined organic layers were washed with brine (5mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-052. ¹H NMR (400 MHz, CD ₃OD) δ 9.05 (s, 1H) , 8.48 (s, 1H) , 8.46 (s, 1H) , 8.18 (s, 2H) , 7.85 -7.78 (m, 2H) , 7.64 (dt, J = 8.3, 1.8 Hz, 1H) , 6.75 (t, J = 2.5 Hz, 1H) , 6.67 (dt, J = 8.9, 2.1 Hz, 1H) , 3.95 (s, 3H) , 3.38 -3.34 (m, 4H) , 3.11 (t, J = 5.0 Hz, 4H) , 2.72 (s, 3H) . LCMS [M+H] ⁺: 416.4.

Example 32: Preparation of N- (2-methoxy-4- (4-methylpiperazin-1-yl) phenyl) -7- (1H-pyrazol-4-yl) quinazolin-2-amine (I-053)

A mixture of INT-2 (100 mg, 302 μmol) , 2-methyl-4- (4-methylpiperazin-1-yl) aniline (80.2 mg, 362 μmol) , TsOH (78.0 mg, 453 μmol) in i-PrOH (4 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 5 hrs under N ₂ atmosphere. The reaction mixture was quenched with water (10 mL) and extracted with EtOAc (30mL × 3) . The combined organic layers were washed with brine (5mL) , dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and purified by prep-HPLC to afford I-052. ¹H NMR (400 MHz, CD ₃OD) δ 9.04 (s, 1H) , 8.54 (s, 1H) , 8.14 (s, 2H) , 7.82 (d, J = 8.4 Hz, 1H) , 7.74 (s, 1H) , 7.60 (dd, J = 8.3, 1.6 Hz, 1H) , 7.50 (d, J = 8.5 Hz, 1H) , 6.96 (d, J = 2.7 Hz, 1H) , 6.92 (dd, J = 8.8, 2.5 Hz, 1H) , 3.28 (t, J = 5.1 Hz, 6H) , 2.78 (t, J = 5.0 Hz, 4H) , 2.47 (s, 3H) , 2.30 (s, 3H) . LCMS [M+H] ⁺: 416.4.

Example 33: Preparation of 3- ( (7- (1H-pyrazol-4-yl) quinazolin-2-yl) amino) -N-methylbenzamide (I-054)

Step 1: To a solution of 7-bromo-2-chloroquinazoline (140 mg, 575 μmol) in i-PrOH (5.0 mL) was added 3-amino-N-methylbenzamide (104 mg, 690 μmol) and TsOH (198 mg, 1.15 mmol) at 20℃. The mixture was stirred at 100℃ for 2 hrs. The mixture was concentrated under vacuum. The crude product was triturated with EtOAc (10 mL) at 25℃ for 30 mins, and then was filtered to give I-054-1. LCMS [M+H] ⁺: 357.0.

Step 2: A mixture of I-054-1 (100 mg, 280 μmol) , 4- (4, 4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrazole (54.3 mg, 280 μmol) , K ₃PO ₄ (178 mg, 840 μmol) , Pd (dppf) Cl ₂ (20.5 mg, 28.0 μmol) in dioxane (1 mL) and H ₂O (0.25 mL) was degassed and purged with N ₂ for 3 times. The mixture was stirred at 100℃ for 2 hrs under N ₂ atmosphere. The mixture was filtrated, and the cake was diluted with DMSO (2 mL) . The mixture was filtrated and purified by prep-HPLC to afford I-054. ¹H NMR (400 MHz, DMSO-d ₆) δ 13.16 (s, 1H) , 9.91 (s, 1H) , 9.22 (s, 1H) , 8.50 (s, 1H) , 8.36 (dd, J = 8.2, 3.6 Hz, 1H) , 8.31 -8.23 (m, 2H) , 8.17 (s, 1H) , 7.90 (d, J = 8.4 Hz, 1H) , 7.85 (d, J = 1.6 Hz, 1H) , 7.71 (dd, J = 8.3, 1.7 Hz, 1H) , 7.43 -7.38 (m, 2H) , 2.81 (d, J = 4.5 Hz, 3H) . LCMS [M+H] ⁺: 345.1.

CDK20 Human CMGC kinase binding assay (KdELECT) :

For CDK20 assay, kinase-tagged T7 phage strains were prepared in an E. coli host derived from the BL21 strain. E. coli were grown to log-phase and infected with T7 phage and incubated with shaking at 32℃ until lysis. The lysates were centrifuged and filtered to remove cell debris. The remaining kinases were produced in HEK-293 cells and subsequently tagged with DNA for qPCR detection. Streptavidin-coated magnetic beads were treated with biotinylated small molecule ligands for 30 minutes at room temperature to generate affinity resins for kinase assays. The liganded beads were blocked with excess biotin and washed with blocking buffer (SeaBlock (Pierce) , 1%BSA, 0.05%Tween 20, 1 mM DTT) to remove unbound ligand and to reduce non-specific binding. Binding reactions were assembled by combining kinases, liganded affinity beads, and test compounds in 1x binding buffer (20%SeaBlock, 0.17x PBS, 0.05%Tween 20, 6 mM DTT) . Test compounds were prepared as 111X stocks in 100%DMSO. Kds were determined using an 11-point 3-fold compound dilution series with three DMSO control points. All compounds for Kd measurements are distributed by acoustic transfer (non-contact dispensing) in 100%DMSO. The compounds were then diluted directly into the assays such that the final concentration of DMSO was 0.9%. All reactions performed in polypropylene 384-well plate. Each was a final volume of 0.02 ml. The assay plates were incubated at room temperature with shaking for 1 hour and the affinity beads were washed with wash buffer (1x PBS, 0.05%Tween 20) . The beads were then re-suspended in elution buffer (1x PBS, 0.05%Tween 20, 0.5 μM nonbiotinylated affinity ligand) and incubated at room temperature with shaking for 30 minutes. The kinase concentration in the eluates was measured by qPCR. Kds were calculated with a standard dose-response curve using the Hill equation: Response = Background + (Signal –Background) / (1 + (Kd ^Hill Slope /Dose ^Hill Slope) ) . The Hill Slope was set to -1. Curves were fitted using a non-linear least square fit with the Levenberg-Marquardt algorithm.

An 11-point 3-fold serial dilution of each test compound was prepared in 100%DMSO at 100x final test concentration and subsequently diluted to 1x in the assay (final DMSO concentration = 1%) . Most Kds were determined using a compound top concentration = 30,000 nM.If the initial Kd determined was < 0.5 nM (the lowest concentration tested) , the measurement was repeated with a serial dilution starting at a lower top concentration. A Kd value reported as 40,000 nM indicates that the Kd was determined to be >30,000 nM.

The assay results are exhibited in Table 4, as well as Fig. 2 wherein x-axis represents the concentration of the compound (nM) , and y-axis represents assay signal.

Table 4

Compound No.	Kd (nM)
I-001	8700

In summary, structure-based drug discovery (SBDD) has been a mainstay method to identify hit molecules and perform lead optimization and predicted protein structures by AlphaFold has been considered as a powerful tool to identify hits for novel targets with no or limited structure information. Herein, we present an example of rapid identification of a CDK20 hit molecule by combining AlphaFold with our automated drug discovery AI engines PandaOmics Chemistry42 for the treatment of HCC within 30 days covering target selection, molecule generation, compound synthesis and biological testing.

CDK20 kinase activity assay:

A radiometric protein kinase assay (

ActivityAssay, available as a service product from Reaction Biology Corp. ) was used for measuring the kinase activity of the CDK20 kinases. This assay was performed in 96-well FlashPlates ^TM from PerkinElmer (Boston, MA, USA) in a 50 μl reaction volume. The reaction cocktail was pipetted in four steps in the following order: a. 25 μl of assay buffer (standard buffer/ [γ- ³³P] -ATP) ; b. 10 μl of ATP solution (in H ₂O) ; c.5 μl of test compound (in 10 %DMSO) ; d. 10 μl of enzyme/substrate mixture. The assay for CDK20 kinase contained 70 mM HEPES-NaOH pH 7.5, 3 mM MgCl ₂, 3 mM MnCl ₂, 3 μM Na-orthovanadate, 1.2 mM DTT, 50 μg/ml PEG ₂₀₀₀₀, 1.0 μM ATP, [γ- ³³P] -ATP (approx. 7×10 ⁵ cpm per well) , 200 ng/50 μl kinase protein, and substrate was 4.0 ug/50μl. The compounds were dissolved to 1×10 ^-3 M in volumes of 100%DMSO. The 1×10 ^-3 M stock solutions were subjected to a serial, semi-logarithmic dilution using 100 %DMSO as a solvent. The final volume of the assay was 50 μL All compounds were tested at 10 final assay concentrations in the range from 1×10 ^-5 M to 3×10 ^-10 M. The final DMSO concentration in the reaction cocktails was 1 %in all cases. The "low control" was defined as the value reflects unspecific binding of radioactivity to the plate in the absence of a protein kinase but in the presence of the substrate. The "high control" was defined as the full activity in the absence of any inhibitor. The difference between high and low control was taken as 100 %activity. As part of the data evaluation the low control value from a particular plate was subtracted from the high control value as well as from all 80 "compound values" of the corresponding plate.

The residual activities for each concentration and the compound IC ₅₀ values were calculated using Quattro Workflow V3.1.1 (Quattro Research GmbH, Munich, Germany; www. quattro-research. com) . The fitting model for the IC ₅₀ determinations was "Sigmoidal response (variable slope) " with parameters "top" fixed at 100 %and "bottom" at 0 %. The fitting method used was a least-squares fit.

The assay results shows that the compounds listed in Table 3 exhibited IC ₅₀ values (CDK20/CycT1) in the range from 10 nM to 10 μM, even in the range from 10 nM to 1 μM for compounds I-008, I-034, I-037, I-038, I-042, I-044, I-045, I-051, I-052, and I-053, and even more in the range from 10 nM to 100 nM for compounds I-11, I-035, I-036, I-039, I-040, I-046, I-047, I-048, I-049, and I-050.

More compounds listed in Table 1 have been or may be synthesized and can be or may be useful as CDK20 inhibitor.

The foregoing description is considered as illustrative only of the principles of the present disclosure. Further, since numerous modifications and changes will be readily apparent to those skilled in the art, it is not desired to limit the present disclosure to the exact construction and process shown as described above. Accordingly, all suitable modifications and equivalents may be considered to fall within the scope of the present disclosure as defined by the claims that follow.

All publications, patents and patent applications cited herein are incorporated by reference in their entirety into the disclosure.

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Claims

A compound of Formula (I)

or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof, wherein

each of X ¹, X ², X ³, and X ⁴ is independently N or CR ^A, in which R ^A is H or R ¹, with the proviso that no more than two of X ¹, X ², X ³, and X ⁴ are N;

ring A is a 5-or 6-membered aromatic or heteroaromatic ring which contains 0, 1, or 2 heteroatoms selected from the group consisting of N, O, and S, optionally further fused to one or two cyclic rings independently selected from -cycloalkyl, -heterocyclyl, -aryl, -heteroaryl ring;

R ¹ is H, halo, -CN, -NO ₂, -alkyl, -haloalkyl, -alkenyl, -alkynyl, - (CH ₂) _p-aryl, - (CH ₂) _p-heteroaryl, -cycloalkyl, -heterocyclyl, -aryl, -heteroaryl, -OH, -OR ^C, -SR ^C, -COR ^C, -CO ₂R ^C, -CONR ^CR ^D, -S (O) R ^C, -SO ₂R ^C, -SO ₃R ^C, -SO ₂NR ^CR ^D, -P (O) (OR ^C) ₂, or -NR ^CR ^D, in which each of R ^C and R ^D is independently H, -alkyl, -haloalkyl, -alkenyl, -alkynyl, - (CH ₂) _p-aryl, - (CH ₂) _p-heteroaryl, -cycloalkyl, -heterocyclyl, -aryl, or -heteroaryl, or R ^C and R ^D together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said heteroaryl, said heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic, or spiro bicyclic, contains 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by alkyl or oxo, and p is 1, 2, or 3;

R ² is H, halo, -C _1-6alkyl, -C _3-6cycloalkyl, or -C _1-6haloalkyl;

R ³ is R ^E, -NR ^ER ^B, or -CONR ^ER ^B, wherein each of R ^E and R ^B is independently H, -C _1-6alkyl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, -C _1-6alkylene-NR ^FR ^G, -C _1-6alkylene-C _3-6cycloalkyl, or -C _1- ₆alkylene-C _3-6heterocyclyl, or R ^E and R ^B together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said C _3-6heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic, or spiro bicyclic, contains 1 or 2 heteroatoms selected from the group consisting of N, O and S, and is unsubstituted or substituted by -C _1- ₆alkyl or oxo, and each of R ^F and R ^G is independently H or -C _1-6alkyl;

R ⁴ is independently H, -C _1-6alkyl, -OC _1-6alkyl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, or - (CH ₂) _q-C _3-6heterocyclyl, in which said C _3-6heterocyclyl is monocyclic, fused bicyclic, or spiro bicyclic, contains 1 or 2 heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by -C _1-6alkyl or oxo, and q is 1, 2, or 3; and

n is 0, 1, 2, 3, or 4.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in claim 1, wherein each of X ¹, X ², X ³, and X ⁴ is independently CR ^A, in which R ^A has the same meaning as defined above.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in claim 1, wherein one of X ¹, X ², X ³, and X ⁴ is N, and the remainder of X ¹, X ², X ³, and X ⁴ are independently CR ^A, in which R ^A has the same meaning as defined above.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein ring A is a 5-or 6-membered aromatic or heteroaromatic ring which contains 0, 1, or 2 heteroatoms independently selected from N and S, optionally fused to a 5-or 6-membered -cycloalkyl or heterocyclyl ring.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein ring A is a pyrrole, imidazole, benzene, pyridine, pyrazole, thiazole, isothiazole, or 1, 2, 3, 4-tetrahydroisoquinoline ring.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein ring A is a pyrrole ring.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein

is a group represented by the following formula:
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein

is a group represented by the following formula:
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein n is 0, 1, or 2.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein n is 1, and

is a group represented by the following formula:
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ¹ is independently H, halo, -CN, -NO ₂, -C _1-6alkyl, -C _1-6haloalkyl, -C _2-6alkenyl, -C _2-6alkynyl, - (CH ₂) _p-aryl, - (CH ₂) _p-heteroaryl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, aryl, heteroaryl, -OH, -OR ^C, -SR ^C, -COR ^C, -CO ₂R ^C, -CONR ^CR ^D, -S (O) R ^C, -SO ₂R ^C, -SO ₃R ^C, -SO ₂NR ^CR ^D, -P (O) (OR ^C) ₂, or -NR ^CR ^D, in which each of R ^C and R ^D is independently H, -C _1-6alkyl, -C _2-6alkenyl, -C _2-6alkynyl, - (CH ₂) _p-aryl, - (CH ₂) _p-heteroaryl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, aryl, or heteroaryl, or R ^C and R ^D together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said heteroaryl, said C _3-6heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic or spiro bicyclic, contains 1, 2, or 3 heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by C _1-6alkyl or oxo, and p is 1, 2, or 3.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ¹ is independently H, halo, -CN, -C _1-6alkyl, -C _1-6haloalkyl, - (CH ₂) _p- (5-or 6-membered aryl) , - (CH ₂) _p- (5-or 6-membered heteroaryl) , -C _3-6cycloalkyl, -C _3-6heterocyclyl, 5-or 6-membered aryl, 5-or 6-membered heteroaryl, -OR ^C, -SR ^C, -COR ^C, -CO ₂R ^C, -CONR ^CR ^D, -S (O) R ^C, -SO ₂R ^C, -SO ₂NR ^CR ^D, or -NR ^CR ^D, in which each of R ^C and R ^D is independently H or -C _1-6alkyl, or R ^C and R ^D together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said 5-to 10-membered heteroaryl, said C _3-6heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic or spiro bicyclic, contains 1, 2, or heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by C _1-6alkyl or oxo, and p is 1 or 2.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ¹ is independently H, halo, -CN, -C _1-3alkyl, -C _1-3haloalkyl, -CH ₂-phenyl, -CH ₂-pyridinyl, -O-phenyl, -O-pyridinyl, -OC _1-3alkyl, 2-oxopyrrolidin-1-yl, -CONHC _1-3alkyl, -SO ₂C _1-3alkyl, imidazolopyrimidinyl, tetrahydroimidazolopyridinyl, pyrazolyl, -C _3-6cycloalkyl, or -C _3- ₆heterocycyl, in which said C _3-6heterocycyl is monocyclic, fused bicyclic or spiro bicyclic, contains 1, 2, or 3 heteroatoms selected from N, O, and S, and is unsubstituted or further substituted by C _1-6alkyl or oxo.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ¹ is independently H, -Br, -CN, -CH ₃, -CF ₃, pyridin-2-ylmethyl, pyridin-2-yloxy, -OCH ₃, 2-oxopyrrolidin-1-yl, -CONHCH ₃, -SO ₂CH ₃, imidazo [1, 2-a] pyrimidin-3-yl, 5, 6, 7, 8-tetrahydroimidazo [1, 2-a] pyridin-3-yl, pyrazol-4-yl, 3-methyl-pyrazol-4yl, cyclopentyl, or oxetan-3-yl.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ² is H, -C _1-6alkyl, or -C _3-6cycloalkyl.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ³ is R ^E, -NHR ^E, -NR ^E’R ^B’, -CONHR ^E, or -CONR ^E’R ^B’; wherein R ^E is H, -C _1-6alkyl, -C _3- ₆cycloalkyl, -C _3-6heterocyclyl, -C _1-6alkylene-NHC _1-3alkyl, -C _1-6alkylene-N (C _1-3alkyl) ₂, -C _1- ₆alkylene-C _3-6cycloalkyl, or -C _1-6alkylene-C _3-6heterocyclyl, and R ^E’ and R ^B’ together with the nitrogen to which they are attached form a heterocyclic ring having 3 to 6 carbon atoms, in which each of said C _3-6heterocyclyl and said heterocyclic ring is monocyclic, fused bicyclic, or spiro bicyclic, contains 1 or 2 N heteroatoms, and is unsubstituted or substituted by -C _1-6alkyl.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ³ is a group represented by the following formula:
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ³ is a group represented by the following formula:
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ⁴ is H, -C _1-6alkyl, -C _3-6cycloalkyl, -C _3-6heterocyclyl, or - (CH ₂) _q-C _3-6heterocyclyl, in which said C _3-6heterocyclyl is monocyclic, fused bicyclic, or spiro bicyclic, contains 1 or 2 heteroatoms selected from the group consisting of N, O, and S, and is unsubstituted or further substituted by -C _1-6alkyl, and q is 1, 2, or 3.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein R ⁴ is H, (1-methylpiperidin-4-yl) methyl, or (2, 6-diazaspiro [3.3] heptan-2-yl) ethyl.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of the preceding claims, wherein the isotopic variant is a deuterated variant.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in claim 1, wherein the compound of Formula (I) is selected from a compound in table 1 or table 3.
A pharmaceutical composition for treating a CDK20-associated disease or condition, which comprises the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of claims 1 to 22, and a pharmaceutically acceptable carrier or excipient.
The pharmaceutical composition as claimed in claim 23, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.
The pharmaceutical composition as claimed in claim 24, which further comprises a second therapeutic agent useful for treating said disease or condition.
A method of treating a CDK20-associated disease or condition in a subject in need thereof, which comprises administering to the subject a therapeutically effective amount of the compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of claims 1 to 22.
The method as claimed in claim 26, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.
The method as claimed in claim 27, wherein the CDK20-associated disease or condition is hepatocellular carcinoma (HCC) .
A compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of claims 1 to 22 for use in the treatment of a CDK20-associated disease or condition.
The compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in claim 29, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.
Use of a compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of claims 1 to 22 in the manufacture of a medicament for treating a CDK20-associated disease or condition.
The use as claimed in claim 31, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.
A kit for treating a CDK20-associated disease or condition, which comprises a compound of Formula (I) or a pharmaceutically acceptable salt, solvate, stereoisomer, or isotopic variant thereof as claimed in any one of claims 1 to 22, a container, and optionally a package insert or label indicating a treatment.
The kit as claimed in claim 33, wherein the CDK20-associated disease or condition is a disease or condition selected from the group consisting of hepatocellular carcinoma (HCC) , colorectal cancer, glioblastoma, lung cancer, medulloblastoma, ovarian carcinoma, prostate cancer, and renal cell carcinoma.
The kit as claimed in claim 34, which further comprises a second therapeutical agent useful for treating said disease or disorder.