Degradable agents targeting bcr-abl and uses thereof

Abstract

The application discloses a degradation agent targeting BCR-ABL and application thereof, and the degradation agent has the following general formula (I). The degradation agent can effectively reduce the BCR-ABL protein level in K562 cells, causes cell apoptosis, and can be used for treating various tumor diseases caused by wild type, mutant BCR-ABL abnormal expression, including chronic myelocytic leukemia, acute lymphoblastic leukemia and the like.

Description

Technical Field

[0001] This invention relates to a novel degradative agent targeting BCR-ABL and its uses. This invention belongs to the field of pharmaceutical technology. Background Technology

[0002] Chronic myelogenous leukemia (CML) is a myeloproliferative neoplasm originating from hematopoietic stem cells, accounting for approximately 40% of all adult leukemia cases. The initiating mutation in CML is the formation of the BCR-ABL fusion gene. The Abelson (ABL) tyrosine kinase gene on chromosome 9 and the BCR gene on chromosome 22 translocate and fuse to create a new chromosome segment called the Philadelphia chromosome (Ph). The protein encoded by this fusion gene has abnormal tyrosine kinase activity, leading to abnormal cell proliferation and the appearance of a large number of neutrophils at various stages of the body, thus inducing the development of CML. Patients with this disease often experience anemia and a significant bleeding tendency. The BCR-ABL fusion gene on the Philadelphia chromosome (Ph) is detectable in approximately 90% of patients with chronic myeloid leukemia (CML) and 30%–35% of patients with acute lymphoblastic leukemia (ALL), but is not expressed in normal cells, making it a suitable target for treatment. In recent years, several small-molecule inhibitors targeting BCR-ABL protein kinase activity have been designed and used clinically for targeted therapy of CML with good results. These include imatinib, dasatinib, nilotinib, and ponatinib, as well as the allosteric modulator asciminib. While these drugs have yielded clinical benefits, they have also led to acquired resistance. Approximately 60% of CML-BP patients and almost all ALL patients relapse within weeks or months after treatment. Acquired resistance is known to result from point mutations in the BCR-ABL kinase domain, dysregulation of resistance gene expression, and signaling pathway commutation. More than 70 point mutations have been observed in CML patients who develop resistance after initial treatment. The most common mutations in imatinib-treated relapsed patients are T315I, E225K, Y253F, and M351T; the most common mutations in dasatinib-treated patients include T315I, F317L / V / I / C, and V299L. Although bosutinib and ponatinib can inhibit various BCR-ABL1 mutants and have good efficacy in patients with PH+ CML in slow, accelerated, or fulminant phases, their clinical application is limited due to serious adverse reactions such as gastrointestinal toxicity, elevated transaminases, rash, thrombocytopenia, or arterial thrombosis. Therefore, there is an urgent clinical need for drugs with good safety profiles that can address abnormal BCR-ABL gene expression or resistance mutations.

[0003] Targeted protein degradation (PROTAC) technology, exemplified by protein hydrolysis-targeted chimeras, is a novel strategy that selectively knocks out target proteins using small molecules. It induces ubiquitination of the target protein by attaching an E3 ligase ligand to the structure of the target protein ligand, thereby enabling the protein to be recognized and degraded by the cellular protein quality control system. Unlike traditional protein function modulators, including inhibitors, targeted protein degraders can achieve a similar effect to "knockout" specific genes, overcoming the problems of incomplete inhibition, negative feedback regulation, and the tendency to induce drug resistance mutations associated with small molecule inhibitors. Furthermore, because the target protein's function is restored only after re-expression in vivo, its duration of action is longer, allowing for smaller dosages and improved drug safety. These properties are particularly suitable for targeted therapy of BCR-ABL, and especially advantageous for treating CML and AML associated with drug-resistant BCR-ABL mutations. Summary of the Invention

[0004] The purpose of this invention is to provide a BCR-ABL-targeting degradative agent and its uses.

[0005] To achieve the above objectives, the present invention employs the following technical means:

[0006] In one aspect, the present invention relates to compounds of general formula (I), or pharmaceutically acceptable salts thereof:

[0007]

[0008] in,

[0009] R1 and R2 are independently selected from H, D, and C. 1-6 Alkyl or C 1-6 Haloalkyl; or R1 and R2 together form =O or =S;

[0010] L does not exist or L is selected from -(CH2). 0-4 C(R5)(R6)-;

[0011] R5 is selected from H, D, and C. 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 Alkyne group, -(CH2) 0-4 C 3-8 Cycloalkyl, -(CH2) 0-4 4-6 membered heterocyclic groups, -(CH2) 0-4 C 6-10 Aryl, -(CH2) 0-4 5-10 aryl groups, -(CH2) 1-6 C(O)ORa -(CH2) 1-6 C(O)NR b R c -(CH2) 1-6 OC(O)R a -(CH2) 1-6 N(R b )C(O)R a -(CH2) 1-6 OC(O)OR a -(CH2) 1-6 N(R b )C(O)OR a -(CH2) 1-4 OR a -(CH2) 1-4 SR a Or natural amino acid side chains;

[0012] R6 is selected from H or D;

[0013] Alternatively, R5 and R6 together form =O or =S;

[0014] R3 is selected from H, D, and C. 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0015] R4 is selected from H, D, and -C(O)R a -C(O)OR a -C(O)CNR b R c -C(O)C(R) f )NR d R e -C 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0016] Alternatively, R3 and R5 can connect together to form -(CH2). 3-5 ;

[0017] Alternatively, R3 and R4, together with the N atoms attached to them, can form a 5-6 membered heterocyclic group;

[0018] Where R a R b and R c Independently selected from H and C 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 cycloalkyl, 4-8 membered heterocyclic, C 6-10Aryl or 5-10 heteroaryl groups, which may be optionally substituted with 1, 2, 3 or more halogens, OH, or NH2;

[0019] R d and R e Independently selected from H, -C(O)R a or -C(O)OR a ;

[0020] R f Selected from C 1-6 Alkyl, C 1-6 Haloalkyl, -(CH2) 1-4 C(O)R a Or -(CH2) 1-4 C(O)OR a .

[0021] Preferably, R1 is selected from H or D;

[0022] R2 is selected from H or D;

[0023] Alternatively, R1 and R2 together form =O or =S;

[0024] L does not exist or L is -(CH2). 0-4 C(R5)(R6)-;

[0025] R5 is selected from H, D, and C. 1-6 Alkyl, C 1-6 Haloalkyl, -(CH2) 1-4 Phenyl, -(CH2) 1-6 C(O)OR a -(CH2) 1- 6C(O)NR b R c -(CH2) 1-6 N(R b )C(O)R a -(CH2) 1-6 N(R b )C(O)OR a Or -(CH2) 1-4 SR a ;

[0026] R6 is selected from H or D;

[0027] Alternatively, R5 and R6 together form =O or =S;

[0028] R3 is selected from H, D, and C. 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0029] R4 is selected from H, D, and C(O)Ra C(O)OR a C(O)C(R) f )NR d R e C 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0030] Alternatively, R3 and R5 can connect together to form -(CH2). 3-5 ;

[0031] Alternatively, R3 and R4, together with the N atoms attached to them, can form a 5-6 membered heterocyclic group;

[0032] Where R a R b and R c Independently selected from H and C 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 6-10 Aryl or 5-6 heteroaryl, which may optionally be substituted by 1, 2, 3 or more halogens, OH, NH2;

[0033] R d and R e Independently selected from H, C(O)R a or C(O)OR a ;

[0034] R f Selected from C 1-6 Alkyl, C 1-6 Haloalkyl, -(CH2) 1-4 C(O)R a Or -(CH2) 1-4 C(O)OR a .

[0035] Preferably, R1 is H;

[0036] R2 is H;

[0037] Or R1 and R2 together form = 0;

[0038] L does not exist or L is -C(R5)(R6)-, -CH2C(R5)(R6)-, or -CH2CH2C(R5)(R6)-;

[0039] Where R5 is selected from H, CH2CH2CH3, CH2CH(CH3)2, CH2Ph, CH2C(O)OC(CH3)3, CH2C(O)NH2, CH2C(O)NHC(CH3)3, CH2CH2CH2CH2NHC(O)CH3, CH2CH2CH2CH2NHC(O)CH2CH3, CH2CH2CH2CH2NHC(O)C H(OH)CH3, CH2CH2CH2CH2NHC(O)C(OH)(CH3)2, CH2CH2CH2CH2NHC(O)CH=CHCH3, CH2CH2CH2CH2NHC(O)Ph, CH2CH2CH2CH2NHC(O)Py, CH2CH2CH2CH2NHC(O)OC(CH3)3 or CH2SC(CH3)3;

[0040] R6 is H;

[0041] Or R5 and R6 together form =0;

[0042] R3 is selected from H or CH3;

[0043] R4 is selected from H, CH3, C(O)CH3, C(O)CH2CH3, C(O)CH2C(CH3)3, C(O)CH(OH)CH3, C(O)C(OH)(CH3)2, C(O)CH=CHCH3, C(O)Ph, C(O)Py, C(O)OC(CH3)3,

[0044] Or R3 and R5 can connect together to form -CH2CH2CH2-;

[0045] Alternatively, R3 and R4, together with the N atoms they are attached to, can form a morpholino group.

[0046] Preferably, the compound has the structure of general formula (II):

[0047]

[0048] in,

[0049] L stands for -(CH2) 0-4 CH(R5)-;

[0050] R5 is selected from H, D, and C. 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 Alkyne group, -(CH2) 0-4 C 3-8 Cycloalkyl, -(CH2)0-4 4-6 membered heterocyclic groups, -(CH2) 0-4 C 6-10 Aryl, -(CH2) 0-4 5-10 aryl groups, -(CH2) 1-6 C(O)NR b R c Or natural amino acid side chains;

[0051] R3 is selected from H, D, and C. 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0052] R4 is selected from H, D, and C(O)R a C(O)OR a C(O)CNR b R c C(O)C(R) f )NR d R e C 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0053] Alternatively, R3 and R5 can connect together to form -(CH2). 3-5 ;

[0054] Alternatively, R3 and R4, together with the N atoms attached to them, can form a 5-6 membered heterocyclic group;

[0055] Where R a R b and R c Independently selected from H and C 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 cycloalkyl, 4-8 membered heterocyclic, C 6-10 Aryl or 5-10 heteroaryl groups, which may be optionally substituted with 1, 2, 3 or more halogens, OH, or NH2;

[0056] R d and R e Independently selected from H, C(O)R a or C(O)OR a ;

[0057] R f Selected from C 1-6 Alkyl, C 1-6 Haloalkyl, -(CH2) 1-4 C(O)R a Or -(CH2) 1-4 C(O)ORa .

[0058] Preferably, L is -(CH2). 0-4 CH(R5)-;

[0059] R5 is selected from H, D, and C. 1-6 Alkyl, C 1-6 Haloalkyl, -(CH2) 1-4 Phenyl or -(CH2) 1-6 C(O)NR b R c ;

[0060] R3 is selected from H, D, and C. 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0061] R4 is selected from H, D, and C(O)R a C(O)OR a C(O)C(R) f )NR d R e C 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0062] Alternatively, R3 and R5 can connect together to form -(CH2). 3-5 ;

[0063] Alternatively, R3 and R4, together with the N atoms attached to them, can form a 5-6 membered heterocyclic group;

[0064] Where R a R b and R c Independently selected from H and C 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 6-10 Aryl or 5-6 heteroaryl, which may optionally be substituted by 1, 2, 3 or more halogens, OH, NH2;

[0065] R d and R e Independently selected from H, -C(O)R a or -C(O)OR a ;

[0066] R f Selected from C 1-6 Alkyl, C 1-6 Haloalkyl, -(CH2) 1-4 C(O)R a Or -(CH2)1-4 C(O)OR a .

[0067] Preferably, L is selected from -CH(R5)-, -CH2CH(R5)-, or -CH2CH2CH(R5)-;

[0068] R5 is selected from H, -CH2CH2CH3, -CH2CH(CH3)2, -CH2Ph, -CH2C(O)NH2 or -CH2C(O)NHC(CH3)3;

[0069] R3 is selected from H or CH3;

[0070] R4 is selected from H, -CH3, -C(O)CH3, -C(O)CH2CH3, -C(O)CH2C(CH3)3, -C(O)CH(OH)CH3, -C(O)C(OH)(CH3)2, -C(O)CH=CHCH3, -C(O)Ph, -C(O)Py, -C(O)OC(CH3)3,

[0071] Or R3 and R5 can connect together to form -CH2CH2CH2-;

[0072] Alternatively, R3 and R4, together with the N atoms they are attached to, can form a morpholino group.

[0073] Preferably, the compound has the structure of general formula (III):

[0074]

[0075] in,

[0076] L stands for -(CH2) 0-4 CH(R5)-;

[0077] R5 is selected from H, D, and C. 1-6 Alkyl, C 1-6 Haloalkyl, -(CH2) 1-4 Phenyl or -(CH2) 1-6 C(O)NR b R c ;

[0078] R3 is selected from H, D, and C. 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0079] R4 is selected from H, D, and -C(O)R a -C(O)OR a C 1-6 Alkyl or C 1-6 Halogenated alkyl groups;

[0080] Alternatively, R3 and R5 can connect together to form -(CH2). 3-5 ;

[0081] Alternatively, R3 and R4, together with the N atoms attached to them, can form a 5-6 membered heterocyclic group;

[0082] Where R a and R b Independently selected from H and C 1-6 Alkyl, C 1-6 Haloalkyl, C 2-6 alkenyl, C 2-6 alkynyl or C 6-10 Aryl;

[0083] R c Selected from C 1-6 Alkyl or C 1-6 Halogenated alkyl groups.

[0084] Preferably, L is selected from -CH(R5)-, -CH2CH(R5)-, or -CH2CH2CH(R5)-;

[0085] R5 is selected from H, -CH2CH2CH3, -CH2CH(CH3)2, -CH2Ph ​​or -CH2C(O)NHC(CH3)3;

[0086] R3 is selected from H or -CH3;

[0087] R4 is selected from H, -CH3, -C(O)CH3, -C(O)CH2CH3, -C(O)CH2C(CH3)3, -C(O)CH=CHCH3, -C(O)Ph or -C(O)OC(CH3)3;

[0088] Or R3 and R5 can connect together to form -CH2CH2CH2-;

[0089] Alternatively, R3 and R4, together with the N atoms they are attached to, can form a morpholino group.

[0090] Preferably, the compound is selected from the following:

[0091]

[0092]

[0093]

[0094] Furthermore, the present invention also provides a pharmaceutical composition comprising any of the compounds described in any one of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0095] Furthermore, the present invention also proposes the use of the said compound, or a pharmaceutically acceptable salt thereof, or the said pharmaceutical composition in the preparation of a medicament for treating neoplastic diseases caused by abnormal expression of BCR-ABL or by drug-resistant mutations.

[0096] Preferably, the disease is selected from chronic myeloid leukemia, acute lymphoblastic leukemia, atypical chronic myeloid leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, B-cell lymphoblastic leukemia / lymphoma, lymphoblastic lymphoma, Tatton-Brown-Rachman syndrome, or acute promyelocytic leukemia.

[0097] Compared with the prior art, the beneficial effects of the present invention are:

[0098] This invention designs and synthesizes a series of BCR-ABL-targeting degraders by adding hydrophobic tags to known small-molecule inhibitors of BCR-ABL. These degraders can effectively reduce BCR-ABL protein levels in K562 cells, inducing apoptosis. They can be used to treat various neoplastic diseases caused by abnormal expression of wild-type and mutant BCR-ABL, including chronic myeloid leukemia and acute lymphoblastic leukemia. Attached Figure Description

[0099] Figure 1A-1C The effect of the compound of this invention on the content of BCR-ABL protein in K562 cells was detected by Western Blot experiment. Detailed Implementation

[0100] The present invention will be further described below with reference to specific embodiments. However, these embodiments are only for illustrating the present invention and do not constitute any limitation on the scope of protection of the present invention. Those skilled in the art should understand that modifications or substitutions can be made to the details and form of the technical solutions of the present invention without departing from the spirit and scope of the present invention, but all such modifications and substitutions fall within the scope of protection of the present invention.

[0101] definition

[0102] Chemical definition

[0103] The definitions of specific functional groups and chemical terms are described in more detail below.

[0104] When listing a range of values, it is assumed that each value and a subrange within that range will be included. For example, "C 1-6 Alkyl groups include C1, C2, C3, C4, C5, C6, and C6. 1-6 C 1-5 C 1-4 C 1-3 C1-2 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 alkyl.

[0105] “C 1-6 "Alkyl" refers to a straight-chain or branched saturated hydrocarbon group having 1 to 6 carbon atoms. In some embodiments, C 1-4 Alkyl groups are preferred. C 1-6 Specific examples of alkyl groups include: methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). Alkyl groups may be optionally substituted with one or more substituents, for example, by 1 to 5 substituents, 1 to 3 substituents, or 1 substituent. Common alkyl abbreviations include: Me (-CH3), Et (-CH2CH3), iPr (-CH(CH3)2), nPr (-CH2CH2CH3), n-Bu (-CH2CH2CH2CH3), or i-Bu (-CH2CH(CH3)2).

[0106] “C 2-6 "Alkenyl" refers to a straight-chain or branched hydrocarbon group having 2 to 6 carbon atoms and at least one carbon-carbon double bond. In some embodiments, C 2-4 Alkenyl groups are preferred. C 2-6 Specific examples of alkenyl groups include: vinyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), pentenyl (C5), pentadienyl (C5), hexenyl (C6), and so on. An alkenyl group may be optionally substituted with one or more substituents, for example, by 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0107] “C 2-6 "Alkyne" refers to a straight-chain or branched hydrocarbon group having 2 to 6 carbon atoms, at least one carbon-carbon triple bond, and optionally one or more carbon-carbon double bonds. In some embodiments, C 2-4 The alkynyl group is preferred. C 2-6Specific examples of alkynyl groups include, but are not limited to: ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), pentyynyl (C5), hexynyl (C6), and so on. The alkynyl group may be optionally substituted by one or more substituents, for example, by 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0108] "Halogen" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0109] Therefore, "C" 1-6 "Halogenated alkyl" and "C" 1-4 "Halogenated alkyl" refers to the above "C" 1-6 "alkyl" and "C" 1-4 "alkyl" is substituted with one or more halogen groups. In some embodiments, C 1-4 Haloalkyl groups are particularly preferred, and C4 groups are more preferred. 1-2 Halogenated alkyl groups. Exemplary alkyl halogenated groups include, but are not limited to: -CF3, -CH2F, -CHF2, -CHFCH2F, -CH2CHF2, -CF2CF3, -CCl3, -CH2Cl, -CHCl2, 2,2,2-trifluoro-1,1-dimethyl-ethyl, etc. The alkyl halogenated group can be substituted at any available connection point, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0110] “C 3-12 "Cycloalkyl" refers to a non-aromatic cyclic hydrocarbon group having 3 to 12 ring carbon atoms and zero heteroatoms, which can be monocyclic, bicyclic, or polycyclic, and optionally contains a double bond. "C 4-12 "Cycloalkyl" refers to a non-aromatic cyclic hydrocarbon group having 4 to 12 ring carbon atoms and zero heteroatoms, which is monocyclic, bicyclic, or polycyclic, and optionally contains a double bond. In some embodiments, C is preferred. 3-6 cycloalkyl, more preferably C 5-6 Cycloalkyl groups. Exemplary cycloalkyl groups include, but are not limited to: cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptanetrienyl (C7), etc. The cycloalkyl group can be substituted at any available connection point, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0111] "4-10 membered heterocyclic group" refers to a 4- to 10 membered non-aromatic ring system having a cyclic carbon atom and 1 to 4 cyclic heteroatoms. In some embodiments, a 5- to 10 membered heterocyclic group is preferred, which is a 5- to 10 membered non-aromatic ring system having a cyclic carbon atom and 1 to 4 cyclic heteroatoms. In some embodiments, a 4- to 6 membered heterocyclic group is preferred, which is a 4- to 6 membered non-aromatic ring system having a cyclic carbon atom and 1 to 3 cyclic heteroatoms. In some embodiments, a 5- to 6 membered heterocyclic group is preferred, which is a 5- to 6 membered non-aromatic ring system having a cyclic carbon atom and 1 to 3 cyclic heteroatoms. The heterocyclic group can be substituted at any available connection point, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0112] Exemplary 3-membered heterocyclic groups containing one heteroatom include, but are not limited to: azircyclopropane, oxacyclopropane, and thiorenyl. Exemplary 4-membered heterocyclic groups containing one heteroatom include, but are not limited to: azircyclobutane, oxacyclobutane, and thiorenyl. Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to: tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolidinyl, and pyrrolidin-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to: dioxasulfuranyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to: triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to: piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to: piperazinyl, morpholinyl, dithianyl, and dioxane. Exemplary 6-membered heterocyclic groups containing three heteroatoms include, but are not limited to: triazinanyl. Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to: azirheptanyl, oxaheptanyl, and thianyl.

[0113] “C 6-10 "Aryl" refers to a group having a 4n+2 aromatic ring system (e.g., having 6 or 10 shared π electrons arranged in a ring) of monocyclic or bicyclic arrangement, providing 6-10 ring carbon atoms and zero heteroatoms in the aromatic ring system. In some embodiments, the aryl group has six ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, the aryl group has ten ring carbon atoms ("C... 10 "Aryl"; for example, naphthyl, such as 1-naphthyl and 2-naphthyl). The aryl group can be substituted at any available junction, for example, 1 to 5 substituents, 1 to 3 substituents or 1 substituent.

[0114] "5-10-membered heteroaryl" refers to a group comprising a 5-10-membered monocyclic or bicyclic 4n+2 aromatic ring system (e.g., having 6 or 10 shared π electrons arranged in a ring) having a cyclic carbon atom and 1-4 cyclic heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, a 5-6-membered heteroaryl is preferred, which is a group comprising a 5-6-membered monocyclic 4n+2 aromatic ring system (e.g., having 6 shared π electrons arranged in a ring) having a cyclic carbon atom and 1-4 cyclic heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur; in some embodiments, a 5-membered heteroaryl is preferred, which is a group comprising a 5-membered monocyclic 4n+2 aromatic ring system (e.g., having 6 shared π electrons arranged in a ring) having a cyclic carbon atom and 1-4 cyclic heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In some embodiments, a 6-membered heteroaryl group is preferred, which is a group of a 6-membered monocyclic 4n+2 aromatic ring system (e.g., having 6 shared π electrons arranged in a ring) having a cyclic carbon atom and 1-4 cyclic heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. The heteroaryl group can be substituted at any available connection point, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0115] Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to: pyrrole, furanyl, and thiophene. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to: imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to: triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to: tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to: pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to: pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to: triazinyl and tetraazinyl, respectively.

[0116] "Natural amino acids" are the basic structural units that make up proteins and are the foundation for later protein modifications in organisms. There are 20 essential natural amino acids for the human body. "Natural amino acid side chains" refer to the substituents on the α-carbon atom of natural amino acids. The 20 most common natural amino acids and their side chains are shown in Table 1 below:

[0117] Table 1

[0118] Gly G glycine -H Ala A alanine <![CDATA[-CH3]]> Val V Valine <![CDATA[-CH-(CH3)2]]> Leu L Leucine <![CDATA[-CH2-CH(CH3)2]]> Ile I Isoleucine <![CDATA[-CH(CH3)-CH2-CH3]]> Phe F Phenylalanine <![CDATA[-CH2-C6H5]]> Trp W Tryptophan <![CDATA[-C8NH6]]> Tyr Y Tyrosine <![CDATA[-CH2-C6H4-OH]]> Asp D Aspartic acid <![CDATA[-CH2-COOH]]> Asn N Asparagine <![CDATA[-CH2-CONH2]]> Glu E glutamic acid <![CDATA[-(CH2)2-COOH]]> Lys K Lysine <![CDATA[-(CH2)4-NH2]]> Gln Q glutamine <![CDATA[-(CH2)2-CONH2]]> Met M Methionine <![CDATA[-(CH2)2-S-CH3]]> Ser S Serine <![CDATA[-CH2-OH]]> Thr T threonine <![CDATA[-CH(CH3)-OH]]> Cys C Cysteine <![CDATA[-CH2-SH]]> Pro P proline <![CDATA[-C3H6]]> His H Histidine <![CDATA[-CH2-C3H3N2]]> Arg R Arginine <![CDATA[-(CH2)3-NHC(NH)NH2]]>

[0119] The present invention also includes D-amino acids with the opposite configuration to those of natural amino acids.

[0120] Other definitions

[0121] The term "pharmaceutically acceptable salt" refers to those salts that, within the bounds of reliable medical judgment, are suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., and in proportion to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of this invention include salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts formed by amino groups with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid; or salts formed with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid; or salts formed using methods used in the art, such as ion exchange methods. Other pharmaceutically acceptable salts include: adipic acid salts, alginate salts, ascorbate salts, aspartate salts, benzenesulfonate salts, benzoate salts, bisulfate salts, borate salts, butyrate salts, camphorate salts, camphor sulfonate salts, citrate salts, cyclopentylpropionate salts, diglucuronate salts, dodecyl sulfate salts, ethanesulfonate salts, formate salts, fumarate salts, gluconate salts, glyceryl phosphate salts, glucuronate salts, hemisulfate salts, heptarate salts, hexanoate salts, hydroiodate salts, 2-hydroxy-ethanesulfonate salts, lactobionate salts, lactate salts, laurate salts, lauryl sulfate salts, malate salts, maleate salts, malonate salts, methanesulfonate salts, 2-naphthalenesulfonate salts, nicotinate salts, nitrate salts, oleate salts, oxalate salts, palmitate salts, dihydroxynaphthalate salts, pectin ester salts, persulfate salts, 3-phenylpropionate salts, phosphate salts, picrate salts, p-pentanoate salts, propionate salts, stearate salts, succinate salts, sulfate salts, tartrate salts, thiocyanate salts, p-toluenesulfonate salts, undecanoate salts, valerate salts, etc. Pharmaceutically acceptable salts derived from suitable bases include alkali metals, alkaline earth metals, ammonium, and nitrogen. + (C 1-4 Alkyl)4 salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Further pharmaceutically acceptable salts, where appropriate, include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, lower alkyl sulfonate, and aryl sulfonate.

[0122] Chemical synthesis route of this invention: (1)

[0124] (2)

[0126] (3)

[0128] (4)

[0130] (5)

[0132] (6)

[0134]

[0135] Example 1: Preparation of N-(2-chloro-6-methylphenyl)-2-((2-methyl-6-(piperazin-1-yl)pyrimidin-4-yl)amino)thiazole-5-carboxamide (2)

[0136]

[0137] 395 mg (1 mmol) of compound 1 and 1.734 g (20 mmol) of piperazine, 20 ml of 1,4-dioxane, and 280 μl of TEA were added to a 100 ml two-necked flask. The mixture was heated to 130 °C and reacted for 6 h to give a white solid. The suspension was washed three times with ethyl acetate, filtered, and dried to give 326 mg of a white powdery solid, with a crude yield of 73%. 1 H NMR (400MHz, DMSO) δ9.96 (s, 1H), 8.28 (s, 1H), 7.39 (d, J = 7.4Hz, 1H), 7.33–7.21 (m, 2H), 6.09(s,2H),3.53(d,J=33.1Hz,4H),2.79(s,4H),2.51(s,1H),2.41(s,3H),2.25(s,3H). 13 C NMR(101MHz,DMSO)δ165.62,162.89,160.48,157.41,141.39,139.29,134.00,132.93,12 9.49,128.64,127.47,126.16,83.12,45.11,44.42,43.56,26.03,18.78.LRMS(ESI+):m / z calculated forC 20 H 22 ClN7OS(M+H + ):444.1368; found444.1362.

[0138] Example 2: Preparation of tert-butyl carbamate (3, DSC0BGly)

[0139]

[0140] 222 mg (0.5 mmol) of compound 2, 105 mg (0.6 mmol) of Boc-Gly-OH, 285 mg of HATU (0.75 mmol), 180 μl of NMM, and 3 ml of DMF were added to a 25 ml round-bottom flask and stirred at 25 °C for 1 h. 100 ml of saturated saline solution was added, and the mixture was extracted twice with 100 ml of ethyl acetate. The organic phases were combined, dried over anhydrous Na2SO4, and the solvent was removed under reduced pressure to obtain a yellow oily substance. Silica gel column chromatography was performed using EA as the eluent. After evaporation of the solvent, 70 mg of a white powdery solid was obtained, with a yield of 23% and a maximum temperature of 160.0–160.4 °C. 1 H NMR (400MHz, DMSO-d6) δ11.51(s,1H),9.89(s,1H),8.23(s,1H),7.40(d,J=6.8Hz,1H),7.32–7.16(m,2H),6. 78(t,J=5.8Hz,1H),6.08(s,1H),3.84(d,J=5.8Hz,2H),3.56(s,8H),2.43(s,3H),2.25(s,3H),1.40(s,9H). 13 C NMR (101MHz, DMSO) δ167.69,165.26,162.55,162.24,159.95,157.00,155.83,140.87,138.86,133.54,132.47, 129.08,128.23,127.05,125.80,82.81,77.98,43.38,43.29,43.20,43.15,41.77,28.24,25.62,18.35.MS(ESI + )m / z calculated for C 27 H 34 ClN8O4S(M+H) + :601.21; found 601.21

[0141] Example 3: Preparation of (S)-(4-amino-1-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-1,4-dioxobutane-2-yl)tert-butyl carbamate (4,DSCOBAsn)

[0142]

[0143] 222 mg (0.5 mmol) of compound 2 was reacted with 139 mg (0.6 mmol) of Boc-L-Asn-OH, and the procedure was the same as that for compound 3, to give 50 mg of white powdery solid, yield 15%, MP 179.9-180.1 °C. 1 H NMR (400MHz, DMSO-d6) δ11.52(s,1H),9.88(s,1H),8.23(s,1H),7.40(d,J=7.5Hz,1H),7.26(ddd,J=28.2,14.3,8.3Hz,4H),6.85(s,1H),6.08(s ,1H),4.75(q,J=8.1,7.6Hz,1H),3.74–3.41(m,8H),2.58(dd,J=15.0,7 .5Hz,1H),2.43(s,3H),2.28(d,J=6.5Hz,1H),2.24(s,3H),1.38(s,9H). 13 CNMR(101MHz,DMSO-d6)δ171.39,169.74,165.23,162.53,162.31,159.93,157.00,154.93,140.86,138.84,133.53,132. 45,129.04,128.19,127.02,125.78,82.87,78.29,47.06,44.18,43.58,43.18,41.30,37.50,28.19,25.59,18.31.MS(ESI + )m / z calculated for C 29 H 37 ClN9O5S(M+H) + :658.23; found 658.23

[0144] Example 4: Preparation of (R)-(1-(4-(6-(5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-1-oxohexyl-3-yl)tert-butyl carbamate (5,DSC0BNva)

[0145]

[0146] 222 mg (0.5 mmol) of compound 2 was reacted with 130 mg (0.6 mmol) of Boc-L-Nva-OH, following the same procedure as compound 3, to give 185 mg of white powdery solid, yield 58%, MP 186.1-186.3 °C. 1 H NMR (400MHz, DMSO-d6) δ11.52(s,1H),9.88(s,1H),8.23(s,1H),7.40(d,J=6. 5Hz,1H),7.30–7.24(m,2H),6.96(d,J=8.2Hz,1H),6.07(s,1H),4.40(q,J=8.5 Hz,1H),3.65–3.46(m,8H),2.69(s,2H),2.43(s,3H),2.24(s,3H),1.50(dd,J =17.7,8.2Hz,2H),1.38(s,9H),1.26(d,J=27.0Hz,2H),0.86(d,J=7.3Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ170.73,165.23,162.54,162.27,159.94,157.01,155.35,140.86,138.85,133.53,132.46,129.06,128.20 ,127.03,125.80,82.86,78.03,49.91,44.15,43.71,43.25,41.13,38.26,33.62,28.22,25.58,18.59,18.32,13.73.MS(ESI+)m / z calculated for C 31 H 42 ClN8O4S(M+H) + :643.26; found 643.26

[0147] Example 5 Preparation of tert-butyl(R)-(3-(tert-butylthio)-1-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-1-oxopropyl-2-yl)carbamate (6, DSC0BCys)

[0148]

[0149] 222 mg (0.5 mmol) of compound 2 was reacted with 166 mg (0.6 mmol) of Boc-L-Cys-OH, following the same procedure as compound 3, to give 195 mg of white powdery solid, yield 55%, MP 128.8-129.0 °C.

[0150] 1 H NMR (400MHz, DMSO) δ11.52(s,1H),9.88(s,1H),8.23(s,1H),7.40(d,J=7.6Hz,1H),7.27(d,J=11.2Hz,3H),6.08(s, 1H),4.48(dd,J=16.3,8.0Hz,1H),3.69(s,8H),2.86–2.80(m,2H),2.43(s,3H),2.24(s,3H),1.38(d,J=5.2Hz,18H).

[0151] 13 C NMR(101MHz,DMSO-d6)δ168.80,165.18,162.51,162.51,156.99,154.94,154.92,138.82,133.51,132.43,131.62,129.02, 128.68,128.16,126.99,82.88,78.43,52.57,50.72,44.26,43.12,42.03,38.23,30.69,29.83,28.14,25.52,18.28.MS(ESI + )m / z calculated for C 32 H 44 ClN8O4S2(M+H) + 703.26; found 703.29

[0152] Example 61: Preparation of tert-butyl carbamate (7, DSC0BPhe)

[0153]

[0154] 222 mg (0.5 mmol) of compound 2 was reacted with 159 mg (0.6 mmol) of Boc-L-Phe-OH, following the same procedure as compound 3, to give 195 mg of white powdery solid, yield 55%, MP 118.3-118.5 °C. 1H NMR (400MHz, DMSO-d6) δ11.53(s,1H),9.90(s,1H),8.22(s,1H),7.40(d,J=7.7Hz,1H),7.32–7.16(m,8H),6.01(s ,1H),4.62(q,J=7.7Hz,1H),3.68–3.38(m,8H),2.89(s,1H),2.73(s,1H),2.41(s,3H),2.23(s,3H),1.32(s,9H). 13 C NMR(101MHz,DMSO-d6)δ170.17,165.20,162.15,159.94,156.99,155.11,140.85,138.85,137.62,133.54,132.46,129.48,1 29.05,128.11,127.03,126.38,125.82,82.79,78.14,51.43,44.22,43.35,43.08,41.10,38.26,28.17,25.56,18.32.MS(ESI + )m / z calculated for C 34 H 40 ClN8O4S(M+H) + :691.26; found 691.27

[0155] Example 7: Preparation of (S)-2-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazine-1-carbonyl)pyrrolidine-1-carboxylic acid tert-butyl ester (8, DSC0BPro)

[0156]

[0157] 222 mg (0.5 mmol) of compound 2 was reacted with 129 mg (0.6 mmol) of Boc-L-Pro-OH, and the procedure was the same as that for compound 3, to give 105 mg of white powdery solid, yield 33%, MP 159.3-159.5 °C. 1H NMR (400MHz, DMSO-d6) δ11.52(s,1H),9.88(s,1H),8.23(s,1H),7.40(d,J=7.3Hz,1H),7.30–7.23(m,2H),6.0 8(s,1H),4.65–4.61(m,1H),3.57(d,J=17.9Hz,8H),2.43(s,3H),2.24(s,3H),1.83–1.71(m,6H),1.38(s,9H). 13 C NMR(101MHz,DMSO-d6)δ170.67,170.33,165.20,162.27,159.87,156.98,153.11,140.79,138.80,133.50,132.42,128.99, 128.14,126.97,125.77,78.37,78.19,56.40,46.49,46.32,43.50,43.36,40.92,29.91,28.05,25.54,23.02,18.27.MS(ESI + )m / z calculated for C 30 H 38 ClN8O4S(M+H) + :641.24; found 641.26

[0158] Example 8 Preparation of tert-butyl (4-(4-(6-(5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-4-oxobutyl)carbamate (9, DSC0BGABA)

[0159]

[0160] 222 mg (0.5 mmol) of compound 2 was reacted with 122 mg (0.6 mmol) of Boc-GABA-OH, and the procedure was the same as that for compound 3, to give 50 mg of white powdery solid, yield 16%, MP 110.1-110.3℃. 1 H NMR (400MHz, DMSO-d6) δ11.51(s,1H),9.88(s,1H),8.22(s,1H),7.40(d,J=7.3Hz,1H),7.34–7.15(m,2H ),6.81(s,1H),6.06(s,1H),3.55(s,8H),2.93(q,J=6.9Hz,6H),2.42(s,3H),2.24(s,3H),1.37(s,9H). 13CNMR(101MHz,DMSO-d6)δ170.58,165.21,162.53,162.26,156.97,155.62,138.83,133.52,13 2.43,129.03,127.01,125.77,77.41,41.14,29.64,28.26,25.58,25.08,18.30.MS(ESI+)m / z calculated for C 29 H 38 ClN8O4S(M+H) + :629.24; found 629.24

[0161] Example 9: Preparation of tert-butyl carbamate (10, DSC0Bβ-Ala)

[0162]

[0163] 222 mg (0.5 mmol) of compound 2 was reacted with 114 mg (0.6 mmol) of Boc-β-Ala-OH, following the same procedure as compound 3, to give 208 mg of a yellow powdery solid, yield 68%, MP 89.0-89.2 °C. 1 H NMR (400MHz, DMSO-d6) δ11.52(s,1H),9.88(s,1H),8.23(s,1H),7.40(d,1H),7.32–7.16(m,2H),6.71( s,1H),6.07(s,1H),3.57(d,J=12.7Hz,8H),3.22–3.11(m,4H),2.42(s,3H),2.24(s,3H),1.37(s,9H). 13 C NMR(101MHz,DMSO-d6)δ169.43,165.25,164.65,162.36,159.97,156.03,155.51,140.88,138.87,133.56,132.48,1 29.07,128.22,127.05,125.81,82.83,77.69,44.57,44.23,43.54,43.19,38.26,30.79,28.26,25.60,18.33.MS(ESI + )m / z calculated for C 28 H 36 ClN8O4S(M+H)+ :615.23; found 615.24

[0164] Example 10: Preparation of tert-butyl (2-(4-(6-(5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-2-oxoethyl)(methyl)carbamate (11, DSC0BSar)

[0165]

[0166] 222 mg (0.5 mmol) of compound 2 and 114 mg (0.6 mmol) of Boc-Sar-OH were reacted, and 285 mg of HATU (0.75 mmol), 180 μl of NMM, and 3 ml of DMF were added to a 25 ml round-bottom flask. The mixture was stirred at 25 °C for 1 h, and 100 ml of saturated brine was added. The mixture was extracted twice with 100 ml of ethyl acetate. The organic phases were combined and dried over anhydrous Na2SO4. After removing the solvent under reduced pressure, a yellow oily substance was obtained. The substance was then subjected to silica gel column chromatography with EA / EtOH = 5:1 as the eluent. After evaporating the solvent, 10 mg of a white powdery solid was obtained, with a yield of 3% and a maximum temperature of 138.5-138.7 °C. 1 H NMR (400MHz, DMSO-d6) δ11.53(s,1H),9.89(s,1H),8.23(s,1H),7.40(d,J=7.3Hz,1H),7.28(s,2H),6 .07(s,1H),4.07(s,2H),3.56(s,8H),2.79(d,J=12.4Hz,3H),2.42(s,3H),2.24(s,3H),1.40(s,9H). 13 C NMR (101MHz, DMSO) δ167.03,165.33,162.61,160.04,157.05,138.90,133.57,132.51,131.61,129.73,129.12,128.73,12 8.28,127.09,125.84,78.88,78.65,65.13,50.24,49.71,43.39,43.35,35.46,28.14,28.05,26.61,25.62,18.35.MS(ESI + )m / z calculated for C 28 H 36 ClN8O4S(M+H) + :615.23; found 615.23

[0167] Example 11: Preparation of N-(2-chloro-6-methylphenyl)-2-((6-(4-(2-(dimethylamino)-2-oxoacetyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide (12, DSC0NN)

[0168]

[0169] 444 mg (1.0 mmol) of compound 2 and 140 mg (1.2 mmol) of N,N-dimethyloxalic acid were reacted, with the same procedure as for compound 3, to give 520 mg of white powdery solid, yield 96%, MP 125.1-125.3 °C. 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),9.90(s,1H),8.22(s,1H),7.39(dd,J=7.4,2.1Hz,1H) ,7.29–7.24(m,2H),6.09(s,1H),3.64–3.56(m,8H),2.92(s,6H),2.42(s,3H),2.23(s,3H). 13 C NMR (101MHz, DMSO) δ165.51,164.89,164.39,163.64,162.42,160.18,157.20,141.01,139.01,133.6 3,132.60,129.24,128.41,127.20,125.96,83.19,45.98,44.79,43.94,43.06,38.42,25.72,18.45.

[0170] MS(ESI + )m / z calculated for C 24 H 28 ClN8O3S(M+H) + 543.17; found 573.16

[0171] Example 122: Preparation of 2-((6-(4-(L-phenylalanyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)-N-(2-chloro-6-methylphenyl)thiazol-5-amide (13, DSC0Phe)

[0172]

[0173] 2 ml of 1,4-dioxane was added to the solid compound 7 (0.12 mmol) in the reaction flask and stirred to dissolve. Then, 2 ml of 1,4-dioxane solution containing 4 M HCl was added, and the mixture was stirred at room temperature for 1.5 h. The solvent was removed by vacuum evaporation to obtain 70 mg of white powder solid, with a yield of 100%. 1 H NMR(400MHz,DMSO-d6)δ9.94(s,1H),8.36(s,2H),8.25(s,1H),7.37–7.23(m,9H),6.01( s,1H),4.62(d,J=33.6Hz,2H),4.22(s,1H),3.30–3.08(m,8H),2.40(s,3H),2.23(s,3H). 13 C NMR (101MHz, DMSO) δ167.01,165.25,162.02,159.93,159.25,157.03,138.84,134.46,133.97,133.54,133.17,129.78, 129.63,129.06,128.68,127.46,125.52,123.64,82.83,51.90,49.89,44.21,42.87,42.74,37.04,25.57,18.33.MS(ESI + )m / z calculated for C 29 H 31 ClN8O2S(M+H) + :590.20; found 620.20

[0174] Example 13 Preparation of tert-butyl(1-((((R)-1-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-4-methyl-1-oxopentane-2-yl)amine)-4-methyl-1-oxopentane-2-yl)carbamate (14, DSC0Bleu-leu)

[0175]

[0176] 222 mg (0.5 mmol) of compound 2 was reacted with 124 mg (0.6 mmol) of Boc-leu-leu-OH, and the procedure was the same as that for compound 3, to give 133 mg of white powdery solid, yield 58%, MP 162.3-162.5 °C. 1H NMR (400MHz, DMSO-d6) δ11.53(s,1H),9.88(s,1H),8.22(s,1H),7.40(d,J=8.2Hz,1H),7.31–7.20(m,2H),6.89(s,1H),6.06(s,1H ),4.79(s,1H),3.82(d,J=4.7Hz,2H),3.65–3.41(m,8H),2.42(s,3H),2.24(s,3H),1.59(s,4H),1.34(m,11H),0.91–0.81(m,12H). 13 C NMR (101MHz, DMSO) δ172.16,170.26,165.22,164.63,162.54,159.94,157.01,155.24,140.85,138.85,133.54,132.46,129.06,128.21,127 .04,125.81,82.85,78.11,46.46,44.18,43.60,43.15,40.85,38.26, 28.16,25.58,24.30,23.99,23.23,22.98,21.72,21.54,18.32.MS(ESI + )m / z calculated for C 37 H 53 ClN9O5S(M+H) + 770.36; found 770.35

[0177] Example 14: Preparation of N-(2-chloro-6-methylphenyl)-2-((6-(4-(leucyl-L-leucine)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)thiazole-5-carboxamide (15, DSC0leu-leu).

[0178]

[0179] Compound 14 was dissolved in 2 ml of 1,4-dioxane in a reaction flask. 2 ml of 1,4-dioxane solution in HCl (4 M) was added to the reaction flask. The mixture was stirred at room temperature for 1-1.5 h. The solvent was removed by vacuum distillation, and the solid was concentrated to obtain 210 mg of white powder. The product was unpurified, with a crude yield of 100% and a maximum temperature of 176.2-178.0 °C. 1H NMR (400MHz, DMSO-d6) δ9.95(s,1H),8.73(d,J=7.9Hz,1H),8.24(s,1H),8.15(d,J=5.3Hz,2H),7.90(d,J=8.3Hz,1H),7.39(d,J=7.1Hz,1H),7. 32–7.18(m,2H),6.12(s,1H),4.78(m,1H),3.94(t,J=8.4Hz,8H),3.39( s,1H),2.44(s,3H),2.23(s,3H),1.59–1.48(m,6H),0.88–0.80(m,12H). 13 C NMR (101MHz, DMSO) δ172.17,170.32,170.18,169.99,168.68,156.74,155.24,138.82,137.63,133.48,132.45,131.67,129.05,128.72, 127.03,66.38,58.97,50.66,47.07,46.48,44.01,43.63,28.15,24.06,23.49,23.22,23.11,22.97,22.74,21.94,21.63,18.33.MS(ESI + )m / z calculated for C 32 H 44 ClN9O3S(M+H) + :670.27; found 670.30

[0180] Example 15 Preparation of bis-tert-butyl(6-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-aminopyrimidin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)(R)-dicarbamate (16, DSC0DB2Lys)

[0181]

[0182] 444 mg (1 mmol) of compound 2 was reacted with 380 μL (1.2 mmol) of Boc-D-Lys(Boc)-OH, with 570 mg (1.5 mmol) of HATU added as a condensing agent, 320 μL of NMM as a base, and 4 mL of DMF as a solvent. The reaction process was the same as that of compound 3. 333 mg of solid was obtained, with a yield of 43.1%. 1H NMR (400MHz, DMSO-d6) δ11.53(s,1H),9.89(s,1H),8.23(s,1H),7.40(d,J=7.6 Hz,1H),7.31~7.24(m,2H),6.94(d,J=8.1Hz,1H),6.76(t,J=5.6Hz,1H),6.08(s 1H),4.36(s,1H),3.63~3.48(m,8H),2.89(s,2H),2.43(s,3H),2.24(s,3H),1.53~1.47(m,2H),1.37(s,18H),1.28~1.23(m,4H). 13 C NMR(101MHz,DMSO)δ171.15,165.68,163.00,162.72,160.38,157.46,156.08,15 5.80,141.31,139.29,133.99,132.91,129.50,128.64,127.48,126.24,83.28,7 8.48,77.82,50.65,44.62,44.12,43.70,41.58,40.61,40.40,40.19,39.98,39. 77,39.56,39.36,38.71,31.46,29.70,28.74,28.67,26.03,23.01,18.76.MS(ESI + )m / z calculated for C 36 H 50 ClN9O6S(M+H) + :772.3367; found:772.3389

[0183] Example 16: Preparation of N-(2-chloro-6-methylphenyl)-2-((2-methyl-6-(4-(2-morpholinoacetyl)piperazin-1-yl)pyrimidin-4-yl)amino)thiazole-5-carboxamide (17, DSC0Mor)

[0184]

[0185] 222 mg (0.5 mmol) of compound 2 and 87 mg (0.6 mmol) of morpholine-4-ylacetic acid were reacted, with the same procedure as for compound 3, to give 136 mg of white powdery solid, yield 48%, MP 192.7-193.0 °C. 1H NMR (400MHz, DMSO-d6) δ11.52(s,1H),9.91(s,1H),8.21(s,1H),7.39(dd,J=7.4,2.0Hz,1H),7.30 –7.24(m,2H),6.06(s,1H),3.61–3.56(m,8H),3.54(s,4H),3.19(s,2H),2.42(s,7H),2.23(s,3H). 13 C NMR(101MHz,DMSO-d6)δ167.68,165.37,162.65,162.54,160.10,157.07,140.96,138.94,133.60,132.55,129.18,1 28.35,127.14,125.88,82.94,66.30,60.88,56.19,53.06,45.92,44.60,44.02,43.34,40.94,25.68,18.41.MS(ESI + )m / z calculated for C 26 H 32 ClN8O3S(M+H) + :571.20; found 571.20

[0186] Preparation of Example 17 (18, DSC0XGly)

[0187]

[0188] 530 mg (0.882 mmol) of compound 3, after deBoc removal, was reacted with 149 μL (1.2 mmol) of 3,3-dimethylbutyric acid. Then, 570 mg (1.5 mmol) of HATU, 700 μL of NMM, and 6 mL of DMF were added, following the same procedure as for compound 3. A total of 274 mg of product 18 was obtained, with a yield of 51.9%. 1 H NMR (400MHz, DMSO) δ11.52(s,1H),9.89(s,1H),8.23(s,1H),7.41(d,J=7.5Hz,1H),7.31~7.24(m,2H) ,6.08(s,1H),3.99(d,J=3.9Hz,2H),3.57(s,8H),2.43(s,3H),2.24(s,3H),2.06(s,2H),0.98(s,9H). 13C NMR (101MHz, DMSO) δ171.46,167.87,165.70,162.98,162.68,160.39,157.44,139.29,133.97,132.90,129.50,128.68,127.48,126.25 ,83.26,56.51,49.09,43.84,43.59,40.82,40.59,40.38,40.17,39.96,39.75,39.55,39.34,30.96,30.16,26.03,19.01,18.76.MS(ESI + )m / z calculated for C 28 H 36 ClN8O3S(M+H) + :599.23; found 599.23

[0189] Example 18: Preparation of 1-(4-(6-(5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-1-oxopentane-2-yl)carbamate tert-butyl ester (19, DSC0BPA)

[0190]

[0191] 222 mg (0.5 mmol) of compound 2 was reacted with 130 mg (0.6 mmol) of 2-((tert-butoxycarbonyl)amino)valerate, following the same procedure as compound 3, to give 243 mg of white powdery solid, yield 75%, MP 82.4-82.6 °C. 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),9.92(s,1H),8.22(s,1H),7.40(dd,J=7.4,2.0Hz,1H),7.30–7.24(m,2H),6.97(d,J=8.2Hz,1H),6.07(s,1H) ,3.62–3.57(m,5H),3.43(s,4H),3.42(s,2H),2.43(s,3H),2.24(s,3H), 1.55–1.46(m,2H),1.37(s,9H),1.24–1.15(m,2H),0.87(t,J=7.3Hz,3H). 13C NMR(101MHz,DMSO-d6)δ170.72,165.19,162.54,162.23,159.92,157.00,155.33,140.84,138.83,133.52,132.45,129.02,128 .18,127.00,125.79,82.86,78.01,49.88,44.15,43.69,43.23,41.10,38.23,33.62,28.20,25.52,18.55,18.28,13.69.MS(ESI + )m / z calculated for C 31 H 42 ClN8O4S(M+H) + :643.26; found 643.26

[0192] Example 19 Preparation of (S)-(1-(4-(6-(5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-4-methyl-1-oxopentane-2-yl)tert-butyl carbamate (20, DSC0Bleu)

[0193]

[0194] 222 mg (0.5 mmol) of compound 2 and 139 mg (0.6 mmol) of Boc-L-Leu-OH were reacted, with the same procedure as for compound 3, to give 101 mg of white powdery solid, yield 31%, MP 147.3-147.5 °C. 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),9.89(s,1H),8.23(s,1H),7.40(dd,J=7.5,2.0Hz,1H),7.32–7.21(m,2H),7.02(d,J=8.3Hz,1H),6.07(s,1 H),4.45–4.39(m,1H),3.65–3.48((m,8H),2.43(s,3H),2.24(s,3H),1. 62(s,1H),1.54–1.42(m,2H),1.37(s,9H),0.88(dt,J=12.3,6.3Hz,6H). 13CNMR(101MHz,DMSO)δ170.95,165.24,162.56,162.29,159.95,157.01,155.38,140.84,138.85,133.54,132.46,129.06,1 28.21,127.04,125.80,82.84,78.05,48.54,44.19,44.15,43.59,43.28,28.23,25.59,24.32,23.12,21.57,18.33.MS(ESI + )m / z calculated for C 31 H 42 ClN8O4S(M+H) + :657.27; found 657.27

[0195] Example 20 Preparation of di-tert-butyl(6-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)(S)-dicarbamate (21,DSC0B2Lys)

[0196]

[0197] 222 mg (0.5 mmol) of compound 2 was reacted with 208 mg (0.6 mmol) of Boc-L-Lys(Boc)-OH, following the same procedure as compound 3, to give 69 mg of white powdery solid with a yield of 15% and a maximum temperature of 158.7-160.0 °C. 1 H NMR (400MHz, DMSO) δ11.52(s,1H),9.88(s,1H),8.23(s,1H),7.40(dd,J=7.5,1. 9Hz,1H),7.30~7.23(m,2H),6.93(d,J=8.0Hz,1H),6.76(t,J=5.6Hz,1H),6.07(s 1H),4.36(m,1H),3.63~3.51(m,8H),2.92~2.86(m,2H),2.43(s,3H),2.2 4(s,3H),1.56~1.46(m,2H),1.38(s,9H),1.36(s,9H),1.28~1.23(m,4H); 13C NMR (101MHz, DMSO) δ170.69,165.23,162.54,162.26,159.92,157.00,155.62,155.35,140.85,138.83,133.52,132.45,129.04,128. 18,127.02,125.78,82.82,78.02,77.36,50.19,44.18,43.69,43.25,41.13,38.26,31.00,29.25,28.29,28.22,25.58,22.56,18.31. MS(ESI+)m / z calculated for C 36 H 50 ClN9O6S(M+H) + :772.3367; found:772.3370.

[0198] Example 21: Preparation of (S)-N,N'-(6-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)diacetamide (22,DSC0A2Lys)

[0199]

[0200] 222 mg (0.5 mmol) of compound 2 was reacted with 138 mg (0.6 mmol) of Ac-L-Lys(Ac)-OH, following the same procedure as compound 3, to give 117 mg of a white powdery solid with a yield of 36%. MPa: 145.1-146.3 °C. 1 H NMR (400MHz, DMSO) δ11.52 (s, 1H), 9.90 (s, 1H), 8.24 (s, 1H), 8.14 (d, J = 8.1Hz, 1H), 7. 79(t,J=5.6Hz,1H),7.39(d,J=7.5Hz,1H),7.29~7.23(m,2H),6.09(s,1H),4.71(dt,J= 7.0,7.3Hz,1H),3.68~3.52(m,8H),3.00(dt,J=6.4,6.7Hz,2H),2.43(s,3H),2.24(s,3 H),1.85(s,3H),1.78(s,3H),1.65~1.44(m,2H),1.40~1.34(m,2H),1.30~1.22(m,2H); 13C NMR (101MHz, DMSO) δ170.35,168.93,165.20,162.52,162.25,159.93,157.00,140.86,138.82,133.55,132.46,129.00,128.14,1 26.99,125.77,82.85,59.74,48.15,44.26,43.68,43.15,41.05,38.26,31.33,28.92,25.56,22.63,22.61,22.32,20.74,18.30. MS(ESI + )m / z calculated for C 30 H 38 ClN9O4S(M+H) + :656.2529; found:656.2568.

[0201] Example 22 Preparation of (S)-N,N'-(6-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)dipropionamide (23,DS-CO-LYS-PROP2)

[0202]

[0203] 21386 mg (0.5 mmol) of the compound was added to 3 mL of 4 M HCl solution of 1,4-Dioxane, followed by stirring with 3 mL of 1,4-Dioxane. After reacting at room temperature for 2 hours, the solvent was removed by distillation under reduced pressure, yielding a yellow powdery solid partially adhering to the flask wall. The solid was dissolved in 3 mL of DMF, and 320 μL of NMM, 90 μL of propanoic acid (1.2 mmol), and 570 mg (1.5 mmol) of HATU were added as a condensing agent. The reaction was carried out overnight at room temperature. The reaction solution was then extracted with NaCl aqueous solution using EA (20 mL x 3), and the organic phase was collected. Column chromatography was performed using EA and EtOH as eluents (EA ~ EA:EtOH = 3:1), yielding 22 mg of a white powdery solid with a yield of 6.4% and MPa of 136.3–137.8 °C. 1H NMR (400MHz, DMSO) δ11.51(s,1H),9.89(s,1H),8.24(s,1H),8.04(d,J=8.1Hz,1H),7.69(t,J=4. 2Hz,1H),7.39(d,J=7.4Hz,1H),7.29~7.23(m,2H),6.08(s,1H),4.71(dt,J=5.8,7.6Hz,1H),3.68 ~3.51(m,8H),3.01(dt,J=6.8,7.2Hz,2H),2.43(s,3H),2.24(s,3H),2.16~2.10(m,2H),2.04(q, J=7.6Hz,2H),1.46~1.49(m,2H),1.39~1.34(m,2H),0.99(t,J=7.3Hz,3H),0.97(t,J=7.3Hz,3H); 13 C NMR(101MHz,DMSO)δ172.68,172.66,170.38,166.96,165.21,162.54,162.26,159.93, 157.00,140.84,138.82,133.54,132.46,131.73,131.54,129.00,128.64,128.13,126 .99,125.78,82.85,67.41,48.07,44.23,43.69,43.18,41.06,38.17,31.34,29.81,28 .96,28.53,28.36,28.17,25.55,23.26,22.60,22.39,18.29,13.86,10.78,9.98,9.93. MS(ESI+)m / zcalculated for C 32 H 42 ClN9O4S(M+H) + :684.2842; found:684.2861.

[0204] Example 23 Preparation of (S)-N,N'-(6-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)diisonicotinamide (24,DS-C0-LYS-ISONIC2)

[0205]

[0206] The reactant was replaced with isoisotinic acid 295 mg (2.4 mmol), and the reaction procedure was the same as in step 23, yielding 33 mg of a white powdery solid with a yield of 4.2% and MPa of 129.2–132.4 °C. ¹H NMR (400 MHz, DMSO) δ 11.56 (s, 1H), 9.93 (s, 1H), 8.98 (d, J = 7.6 Hz, 1H), 8.79 (t, J = 5.7 Hz, 1H), 8.71 (dd, J = 4.4, 1.7 Hz, 2H), 8.68 (dd, J = 4.4, 1.7 Hz). 2H),8.27(s,1H),7.80(dd,J=4.4,1.7Hz,1H),7.30(dd,J=4.4,1.7Hz,1H),7.39(dd,J=7.5,1.9Hz,1H),7.29~7.22(m,2H),6.09(s,1H),4.95(d t,J=7.5,7.2Hz,1H),3.74~3.55(m,8H),3.33~3.27(m,1H),2.43(s,3H) ,2.25(s,3H),1.81~1.76(m,2H),1.63~1.54(m,2H),1.48~1.38(m,2H); 13 C NMR (101MHz, DMSO) δ170.04,165.24,164.78,164.59,162.58,162.27,159.99,157.02,150.17,150.13,141.55,140.87,138.85,133.57,132.49 ,129.02,128.15,127.01,125.81,121.49,121.20,82.90,59.77,49.54 ,44.36,43.68,43.21,41.22,39.01,30.71,28.67,25.58,22.89,18.33. MS(ESI+)m / z calculated for C 38 H 40 ClN 11 O4S(M+H) + :782.2747; found:782.2756.

[0207] Example 24 Preparation of (2E, 2'E)-N, N'-((S)-6-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)bis(but-2-enamide) (25,DS-C0-LYS-CROT2)

[0208]

[0209] The reactant was replaced with 207 mg (2.4 mmol) of crotonic acid. The reaction process was the same as in step 23, yielding 14 mg of white powdery solid with a yield of 2.0% and a maximum temperature of 137.5-138.9 °C. 1 H NMR (400MHz, DMSO) δ11.54(s,1H),9.90(s,1H),8.24(s,1H),8.19(d,J=8.2Hz,1H),7.85(t,J=5 .6Hz,1H),7.40(dd,J=7.4,1.9Hz,1H),7.30~7.23(m,2H),6.68~6.52(m,2H),6.07(s,1H),6.02( dd,J=15.8,2.2Hz,1H),5.87(dd,J=25.3,2.0Hz,1H),4.78(dt,J=5.4,8.2Hz,1H),3.67~3.52(m, 8H),3.08(dt,J=6.56.5Hz,2H),2.43(s,3H),2.24(s,3H),1.79~1.74(m,6H),1.69~1.35(m,6H); 13 C NMR(101MHz,DMSO)δ170.29,165.20,164.77,164.50,162.53,162.25,15 9.93,156.99,140.84,138.82,138.40,137.30,133.53,132.45,129.01,1 28.15,126.99,126.00,125.77,125.44,82.84,59.74,48.10,44.29,43.6 5,43.15,41.08,38.12,31.34,28.94,25.55,22.65,18.29,17.35,17.24. MS(ESI+)m / z calculated for C 34 H 42 ClN9O4S(M+H) + :708.2842; found:708.2864.

[0210] Example 25: Preparation of (S)-N,N'-(6-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)dibenzoamide (26,DS-C0-LYS-BEN2)

[0211]

[0212] The reactant was replaced with 293 mg (2.4 mmol) of benzoic acid. The reaction process was the same as in step 23, yielding 178 mg of a white powdery solid with a yield of 22.8% and a maximum temperature of 135.6-137.4 °C. 1 H NMR (400MHz, DMSO) δ11.53(s,1H),9.89(s,1H),8.61(d,J=7.6Hz,1H),8.45(t,J=5. 7Hz,1H),8.24(s,1H),7.91~7.88(m,2H),7.83~7.81(m,2H),7.55~7.39(m,6H),7.30 ~7.23(m,2H),6.08(s,1H),4.96~4.88(m,1H),3.75~3.54(m,8H),3.30~3.25(m,2H) ,2.43(s,3H),3.25(s,3H),1.81~1.73(m,2H),1.62~1.52(m,2H),1.47~1.39(m,2H); 13 C NMR (101MHz, DMSO) δ170.39,170.36,166.22,166.15,165.20,162.55,162.26,159.93,156.99 ,140.84,138.82,134.70,133.96,133.53,132.45,131.29,130.92,129.00,128.19,128.16,1 27.48,127.08,126.99,125.77,82.86,59.73,49.33,44.30,43.67,43.19,41.16,40.16,39.9 5,39.74,39.53,39.32,39.11,38.90,38.86,30.81,28.88,25.55,22.90,20.73,18.28,14.06. MS(ESI+)m / z calculated for C 40 H 42 ClN9O4S(M+H) + :780.2842; found:780.2855.

[0213] Example 26 Preparation of (2R, 2'R)-N, N'-((S)-6-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)bis(2-hydroxypropionamide) (27,DS-C0-LYS-LAC2)

[0214]

[0215] The reactant was replaced with 169 mg (2.4 mmol) of D-Lactic acid, and the reaction process was the same as in step 23, yielding 108 mg of white powdery solid with a yield of 15.1% and a maximum temperature of 135.6-137.1 °C. 1 H NMR (400MHz, DMSO) δ11.51(s,1H),9.90(s,1H),8.24(s,1H),7.74(d,J=4.0Hz,1H),7.65(t,J=5. 8Hz,1H),7.40(dd,J=7.4,1.9Hz,1H),7.30~7.23(m,2H),6.09(s,1H),5.59(d,J=5.0Hz,1H),5.4 3~5.42(d,J=2.4Hz,1H),4.77(dt,J=5.9,4.4Hz,1H),4.05~3.90(m,2H),3.65~3.54(m,8H),3.07 ~3.02(m,2H),2.43(s,3H),2.24(s,3H),1.70~1.57(m,2h),1.46~1.29(m,2H),1.21~1.17(m,6H); 13 C NMR (101MHz, DMSO) δ174.80,174.78,174.29,170.79,170.30,165.69,162.99,162.70,160. 40,157.48,141.33,139.30,134.01,132.93,129.48,128.62,127.46,126.24,83.32,67.72, 67.50,60.21,48.35,44.64,44.13,43.56,41.57,40.62,40.41,40.20,40.00,39.79,39.58, 39.37,38.23,32.17,29.39,26.03,22.47,21.60,21.35,21.21,20.77,20.67,18.77,14.54. MS(ESI+)m / z calculated for C 32 H 42ClN9O6S(M+H) + :716.2741; found:716.2744.

[0216] Example 27 Preparation of (S)-N,N'-(6-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)bis(2-hydroxy-2-methylpropionamide) (28,DS-C0-LYS-HY2)

[0217]

[0218] The reactant was changed to 250 mg (2.4 mmol) of 2-Hydroxyisobutyric acid. The reaction process was the same as in 23, yielding 238 mg of white powdery solid with a yield of 32.0% and a maximum temperature of 125.2-126.7 °C. 1 H NMR (400MHz, DMSO) δ11.51(s,1H),9.89(s,1H),8.25(s,1H),7.72(d,J=4.Hz,1H ),7.61(t,J=6.0Hz,1H),7.41~7.38(m,1H),7.29~7.23(m,2H),6.09(s,1H),5.54 (s,1H),5.30(s,1H),4.77(dt,J=5.8,4.3Hz,1H),3.69~3.55(m,8H),3.06(dt,J =6.9,7.3,2H),2.44(s,3H),2.25(s,3H),1.75~1.37(m,6H),1.29~1.23(m,12H); 13 C NMR(101MHz,DMSO)δ176.78,176.27,170.75,170.31,165.69,163.03,162.70,160.43,1 57.47,141.32,139.29,134.01,132.93,129.45,128.59,127.45,126.25,83.32,72.51,7 2.35,60.20,48.45,44.64,44.15,43.55,41.58,40.58,40.37,40.17,39.96,39.75,39. 54,39.33,38.47,32.30,29.41,28.47,28.28,27.80,26.01,22.33,21.18,18.76,14.52. MS(ESI+)m / z calculated for C 34H 46 ClN9O6S(M+H) + :744.3054; found:744.3074.

[0219] Example 28 Preparation of (S)-3-((tert-butoxycarbonyl)amino)-4-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-4-oxobutyrate tert-butyl ester (29)

[0220]

[0221] Using compound 2 (0.45 mmol, 200 mg) and tert-butyloxycarbonyl-L-aspartic acid-4-tert-butyl ester (0.54 mmol, 156 mg) as starting materials, and HATU (280 mg) as catalyst, NMM (140 μL) and DMF (3 mL) were added to a round-bottom flask and incubated at room temperature for 1 h. 50 mL of saturated brine was added, and the mixture was extracted twice with 50 mL of ethyl acetate. The combined organic phases were dried over anhydrous Na₂SO₄, and after removing the solvent under reduced pressure, the mixture was chromatographically analyzed by silica gel column chromatography (ethyl acetate: petroleum ether = 1:1) to give 81 mg of a white solid, yield 25.2%, MP 187.8–188.9 °C. 1 H NMR (400MHz, DMSO) δ11.52(s,1H),9.88(s,1H),8.23(s,1H),7.39(t,J=8.8Hz,2H),7.27(q,J=7.3Hz,2H),6.08(s,1H),4.73(dd,J=14. 8,7.3Hz,1H),3.73(s,4H),3.55(d,J=9.3Hz,1H),3.41(d,J=9.8Hz,3H),2.43(s,4H),2.24(s,3H),1.39(d,J=6.9Hz,18H),1.24(s,1H). 13 C NMR (101MHz, DMSO) δ170.02,169.28,165.69,162.99,162.77,160.37,157.49,155.24,141.30,139.30,133.99,132.92,12 9.50,127.48,83.37,80.46,78.99,47.54,44.55,44.04,43.59,41.85,38.20,28.62,28.17,26.04,18.76.LRMS(ESI+):m / z calculated for C 33 H 43 ClN8O6S(M+H +):715.2788; found 715.2815.

[0222] Example 29 Preparation of (S)-3-amino-4-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)-4-oxobutyrate tert-butyl ester (30)

[0223]

[0224] Compound 29 was dissolved in a dioxane solution containing 4M hydrochloric acid, stirred at room temperature for 1 hour, and the solvent was removed under reduced pressure to obtain a white solid, which can be directly used in the next reaction.

[0225] Example 30 Preparation of (S)-4-(4-(6-((5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)pyrimidin-4-yl)piperazin-1-yl)-3-(3,3-dimethylbutamido)-4-oxobutyrate tert-butyl ester (31)

[0226]

[0227] Using compound 30 (0.6 mmol, 369 mg) and tert-butylacetyl chloride (0.6 mmol, 85 μL) as starting materials, triethylamine (350 μL) as catalyst, and dichloromethane (3 mL) as solvent, the mixture was added to a round-bottom flask and incubated on ice for 3 h. 50 mL of saturated brine was added, and the mixture was extracted twice with 50 mL of dichloromethane. The combined organic phases were dried over anhydrous Na₂SO₄, and after removing the solvent under reduced pressure, the mixture was chromatographically analyzed on a silica gel column (ethyl acetate: petroleum ether = 3:1). After removing the solvent under reduced pressure, 18 mg of a white powdery solid was obtained, with a yield of 4.2%. MPa: 163.3–164.8 °C. 1 H NMR (400MHz, DMSO) δ11.52(s,1H),9.89(s,1H),8.23(s,1H),7.40(d,J=7.1Hz,1H),7.29(d,J=6.8Hz,2H),6.08(s,1H),3.83(s,1H),3.69(s,3H),3 .52(d,J=46.9Hz,4H),2.89(s,1H),2.70(d,J=8.8Hz,1H),2.51(s,1H),2 .43(s,4H),2.25(s,3H),2.00(d,J=2.8Hz,2H),1.37(s,9H),0.95(s,9H). 13C NMR (101MHz, DMSO) δ179.91,169.15,162.68,159.97,140.77,139.57,133.83,129.62,127.28,126 .55,83.85,83.46,80.78,80.52,48.70,44.64,31.01,30.20,28.16,26.03,18.77.LRMS(ESI+):m / z calculated forC 33 H 43 ClN8O5S(M+H + ):713.2995; found713.3033.

[0228] Example 31 Preparation of benzyl N2-(tert-butoxycarbonyl)-N4-(tert-butyl)-L-asparagine (33)

[0229]

[0230] Using Boc-L-aspartic acid 1-benzyl ester (32, 1.0 mmol, 323 mg) and tert-butylamine (1.5 mmol, 157 μL) as starting materials, and HATU (1.5 mmol, 576 mg) as a catalyst, NMM (2.7 mmol, 280 μL) and DMF (2 ml) were added to a round-bottom flask. The mixture was stirred at room temperature for 1 h, and the reaction solution turned orange-red. 100 ml of saturated saline solution was added, and the mixture was extracted twice with 100 ml of ethyl acetate. The solution was then washed once with 100 ml of saturated sodium bicarbonate solution and once with 100 ml of 1 M hydrochloric acid aqueous solution. The solution was dried over anhydrous Na2SO4, and the solvent was removed under reduced pressure. After further solvent removal under reduced pressure, 384 mg of a white powdery solid was obtained, with a yield of 95.9%. LRMS(ESI+): m / z calculated for C 20 H 30 N2O5(M+H + ):379.2228; found 379.2247.

[0231] Example 32 Preparation of benzyl N4-(tert-butyl)-L-asparagine (34)

[0232]

[0233] Compound 33 was dissolved in a dioxane solution containing 4M hydrochloric acid, stirred at room temperature for 1 h, and the solvent was evaporated to obtain a white solid 34, which can be directly added to the next reaction. LRMS(ESI+): m / z calculated for C 15 H 22 N2O3(M+H +):279.1704; found 279.1721.

[0234] Example 33 Preparation of benzyl N4-(tert-butyl)-N2-(3,3-dimethylbutyryl)-L-asparagine (35)

[0235]

[0236] Using compound 34 (1.0 mmol, 278 mg) and 3,3-dimethylbutyric acid (1.0 mmol, 122 μL) as starting materials, and HATU (1.2 mmol, 456 mg) as catalyst, DIPEA (697 μL) and DMF (2 ml) were added to a round-bottom flask. The mixture was stirred at room temperature for 1 h, then 100 ml of saturated brine was added, and the mixture was extracted twice with 100 ml of ethyl acetate. The organic layer was then washed once with 100 ml of saturated sodium bicarbonate solution and once with 100 ml of 1 M hydrochloric acid aqueous solution. The mixture was dried over anhydrous Na₂SO₄, and the solvent was removed under reduced pressure. After further solvent removal under reduced pressure, 199 mg of a white powdery solid was obtained, with a yield of 52.9%. LRMS(ESI+): m / z calculated for C 21 H 32 N2O4(M+H + ):377.2435; found 377.2457.

[0237] Example 34N 4 Preparation of -(tert-butyl)-N2-(3,3-dimethylbutyryl)-L-asparagine (36)

[0238]

[0239] Using compound 35 (120 mg) as the starting material, 10% Pd / C (10 mg) and MeOH (5 mL) were added, and the reaction was catalyzed by H2 for 4 h. After filtration and removal of the solvent under reduced pressure, 69 mg of a white solid was obtained, with a yield of 75.6%. LRMS (ES-): m / z calculated for C 14 H 26 N2O4(MH + ):285.1819; found 285.1831.

[0240] Example 352 Preparation of ((6-(4-(N4-(tert-butyl)-N2-(3,3-dimethylbutyryl)-L-asparaginyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)-N-(2-chloro-6-methylphenyl)thiazol-5-carboxamide (37)

[0241]

[0242] Using compound 2 (0.2 mmol, 89 mg) and compound 36 (0.2 mmol, 57 mg) as raw materials, the same method as 3 was used to synthesize 4 mg of white solid, yield 2.8%, MP 167.4–168.0 °C. 1 H NMR(400MHz,DMSO)δ11.53(s,1H),9.89(s,1H),8.23(s,1H),7.74(m,3H),7.28(s,1H),6.09(s,1H),5.06(s,1H),3.64(s,4H) ,2.59(d,J=7.9Hz,2H),2.44(s,2H),2.24(s,4H),1.99(d,J=9.0Hz,2H),1.38(s,1H),1.25(s,2H),1.22(s,9H),0.94(s,9H). 13 C NMR (101MHz, DMSO) δ170.68,170.60,170.15,169.15,169.07,165.62,162.66,160.39,157.40,141.30,139.30,133.99,132.95, 130.11,129.49,129.12,127.46,83.33,50.44,50.33,48.75,45.71,44.79,31.00,30.11,28.97,25.97,18.76.LRMS(ESI+):m / z calculated for C 34 H 46 ClN9O4S(M+H + ):712.3155; found712.3141.

[0243] Example 36 Preparation of N-(6-(4-(2-((tert-butoxycarbonyl)amino)ethyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)-5-((2-chloro-6-methylphenyl)carbamoyl)thiazole-2-amine (38)

[0244]

[0245] Using compound 1 (0.45 mmol, 200 mg) and 1-(N-Boc-aminoethyl)piperazine (0.54 mmol, 124 mg) as starting materials, the same method as 2 was used to synthesize a white solid, yielding 110 mg, with a yield of 41.2%. LRMS(ESI+): m / z calculated for C 27 H 35 ClN8O3S(M+H +):587.2315; found 587.2322.

[0246] Example 372 Preparation of ((6-(4-(2-aminoethyl)piperazin-1-yl)-2-methylpyrimidin-4-yl)amino)-N-(2-chloro-6-methylphenyl)thiazol-5-carboxamide (39)

[0247]

[0248] Compound 38 was dissolved in a dioxane solution containing 4M hydrochloric acid, stirred at room temperature for 1 hour, and the solvent was evaporated to obtain a white solid, which can be directly added to the next reaction.

[0249] Example 38 Preparation of tert-butyl (S)-3-((tert-butoxycarbonyl)amino)-4-((2-(4-(6-(5-(((2-chloro-6-methylphenyl)carbamoyl)thiazolyl-2-yl)amino)-2-methylpyrimidin-4-yl)piperazin-1-yl)ethyl)amino)-4-oxobutyrate (40)

[0250]

[0251] Using compound 39 (0.11 mmol, 55 mg) and tert-butyloxycarbonyl-L-aspartic acid-4-tert-butyl ester (0.13 mmol, 37 mg) as starting materials, the same method as for compound 29 was used to synthesize 24 mg of a white solid, with a yield of 28.9%. MPa: 179.3–180.1 °C 1 H NMR(400MHz,DMSO)δ11.47(s,1H),9.88(s,1H),8.23(s,1H),7.69(s,3H),7.40(s,1H),7.29(s,2H),7.08(s,1H),6.06(s,1H) ,4.39–4.18(m,2H),4.14(s,1H),3.83(s,1H),3.51(s,4H),2.24(s,3H),1.65(s,1H),1.39(s,18H),1.29(s,4H),0.88(s,3H). 13C NMR (101MHz, DMSO) δ169.99,165.65,163.03,162.88,160.39,157.43,155.53,141.31,139.30,133.99,132.91,132.13,129.50,1 29.17,83.12,80.45,78.73,53.08,52.59,51.69,44.04,38.30,36.65,30.48,28.65,28.15,26.05,19.12,18.77.LRMS(ESI+):m / z calculated for C 35 H 48 ClN9O6S(M+H + ):758.3210; found 758.3226.

[0252] Example 39: Compound induces BCR-ABL degradation activity in K562 cells

[0253] Using the human myeloid leukemia cell line K562 as the research subject, multiple concentrations of the compound were administered over a 20-hour period. GNF-2 or dasatinib, ligands of the BCR-ABL protein, were used as controls. Western blotting was employed to detect the intracellular BCR-ABL protein content in K562 cells after administration, serving as an initial screening method for compound activity. Results are as follows: Figure 1A-1C As shown.

[0254] Example 40: Compound inhibits the proliferation activity of K562 cells.

[0255] Prepare 50 μL of K562 cell suspension (approximately 5000 cells / well) in a 96-well plate and pre-culture the plate in an incubator for 24 hours (37℃, 5% CO2). Add 50 μL of different concentrations of the test drug to the culture plate. Incubate for 48 hours. Add 10 μL of CCK-8 solution to each well and incubate for another 1–4 hours. Measure the absorbance at 450 nm using a microplate reader. The blank group contained culture medium and CCK-8 solution, with no cells; the negative control group contained cell suspension and CCK-8 solution, with no drug.

[0256]

[0257] The calculation results are shown in Table 2 below:

[0258] Table 2

[0259]

[0260]

Claims

1. Use of a BCR-ABL-targeting degrader in the preparation of a medicament for treating neoplastic diseases caused by abnormal expression or drug resistance mutations of BCR-ABL, wherein the BCR-ABL-targeting degrader is selected from the following compounds: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , or .

2. The use as described in claim 1, characterized in that, The BCR-ABL-targeting degradative agent is selected from the following compounds: or .

3. The use as described in claim 1, characterized in that, The neoplastic diseases caused by abnormal expression or drug resistance mutation of BCR-ABL are selected from chronic myeloid leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, B lymphocytic leukemia / lymphoma, lymphoblastic lymphoma, or Tatton-Brown-Rachman syndrome caused by abnormal expression or drug resistance mutation of BCR-ABL.

4. The use as described in claim 3, characterized in that, The chronic myeloid leukemia mentioned above is atypical chronic myeloid leukemia, the acute lymphoblastic leukemia mentioned above is acute lymphoblastic leukemia, and the acute myeloid leukemia mentioned above is acute promyelocytic leukemia.

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