PROTAC Chimeric Compounds, Their Preparation Methods and Uses

Abstract

The present invention relates to PROTAC chimeric compounds, their preparation methods and uses, in particular to compounds represented by general formula (I), their pharmaceutically acceptable salts, or stereoisomers, and their applications in the treatment of diseases such as tumors, immunity or inflammation. The present invention also relates to pharmaceutical preparations, pharmaceutical compositions of the compounds and their applications.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and particularly relates to PROTAC chimeric compounds and their use in the preparation of drugs for treating or preventing diseases such as tumors. Background Art

[0002] The technology of proteolysis targeting chimeras (PROTAC) originated from the discovery by scientists of the ubiquitin (Ub)-regulated protein degradation process. In eukaryotic cells, efforts have been continuously made to maintain appropriate protein levels, and thousands of proteins are being generated and degraded at every moment. The key factor for maintaining protein balance is a small protein molecule called ubiquitin. When it is linked to a protein, it causes these proteins to be transported to the proteasome for degradation.

[0003] Targeted protein degradation is an emerging direction in the field of drug research and development. Protein-targeted degradation drugs attempt to design small molecules into a new type of drug. The role of traditional small molecules is to block the function of proteins, while the role of protein-targeted degraders is to degrade these proteins by transporting them into the proteasome.

[0004] Dr. Craig Crews and Raymond Deshaies designed a series of bifunctional chimeric molecules based on peptide-based compounds to induce the degradation of methionyl aminopeptidase 2 (MetAP-2), and formally proposed the PROTAC concept and applied for the related patent WO2002020740A3. However, since these compounds that play a connecting role based on large and cumbersome peptides are difficult to enter cells, the first-generation PROTACs failed.

[0005] In 2008, the Crews team designed the second-generation PROTACs based on the E3 ubiquitin ligase MDM2 that can be used to degrade the androgen receptor (AR).

[0006] In 2015, the Crews team designed a new generation of PROTACs based on the novel E3 ubiquitin ligase VHL and CRBN ligands.

[0007] Hematopoietic progenitor kinase 1 (HPK1), also known as MAP4K1 (mitogen-activated protein kinase kinase kinase kinase 1), is a serine / threonine kinase and is a member of the MAP4K family. In addition, there are 5 other members in its family, including MAP4K2, MAP4K3, MAP4K4, MAP4K5, and MAP4K6.

[0008] The main processes by which HPK1 participates in regulating TCR are as follows: (1) TCR binds to extracellular antigen through MHC, thereby activating the TCR pathway to transmit signals to downstream adaptor protein molecules; (2) The adaptor protein tyrosine kinases Lck and Zap70 activate SLP76, which then phosphorylates HPK1; (3) Activated HPK1 then phosphorylates the receptor protein SLP-76; (4) The phosphorylation reaction of SLP-76 provides multiple protein-binding sites for the 14-3-3 (TCR pathway inhibitory protein) receptor protein to form a complex; (5) The complex phosphorylated by SLP-76 participates in the downregulation of the Erk signaling pathway and leads to the ubiquitination and degradation process of SLP76, resulting in a decrease in the TCR signaling pathway and T cell proliferation. In summary, HPK1 can negatively regulate the TCR signaling pathway. Therefore, HPK1 can serve as a new regulatory mechanism for T cell-mediated immune responses and become a new immune anti-tumor target. HPK1 can bind to many adaptor proteins, such as the SLP-76 family, CARD11, HIS, HIP-55, the GRB2 family, LAT, the CRK family, etc., and interact with each other to activate the JNK / SAPK signaling pathway of hematopoietic stem cells, thereby negatively regulating the TCR pathway. The blockade of the Erk MAPK pathway is the inhibitory mechanism for negatively regulating TCR-induced IL-2 gene transcription.

[0009] HPK1 can bind to many adaptor proteins, such as the SLP-76 family, CARD11, HIS, HIP-55, the GRB2 family, LAT, the CRK family, etc., and interact with each other to activate the JNK / SAPK signaling pathway of hematopoietic stem cells, thereby negatively regulating the TCR pathway. MAP4K3 is also known as GLK kinase, and its biological function is exactly opposite to that of HPK1. GLK can promote the activation of the TCR pathway by binding to downstream adaptor proteins.

[0010] To better meet the huge clinical needs, the present invention aims to provide a PROTAC compound with a novel structure and HPK1 inhibitory / degrading activity, which has good physicochemical properties and drug-like characteristics. The compound of the present invention or its pharmaceutically acceptable salt has good safety, efficacy, and high bioavailability. Therefore, the compound of the present invention has good application potential in the treatment of diseases mediated by HPK1. Summary of the Invention

[0011] One object of the present invention is to provide a targeted PROTAC compound with good HPK1 inhibitory / degrading activity or its pharmaceutically acceptable salt or stereoisomer. The compound of the present invention also has good performance in terms of HPK1 selectivity, can induce the production of IL-2, and further has good HPK1 protein degradation ability. In addition, the compound of the present invention also has good physicochemical properties (such as solubility, physical and / or chemical stability), good pharmacokinetic properties (such as good bioavailability, good metabolic stability, appropriate half-life and duration of action), good safety (low toxicity (such as reduced cardiotoxicity) and / or fewer side effects), and is less likely to develop drug resistance.

[0012] Another object of the present invention is to provide the use of the compound or its pharmaceutically acceptable salt or stereoisomer in the treatment of diseases such as tumors, immunity, or inflammation.

[0013] The compound of the present invention has the structure shown in the general formula (I):

[0014] [B-L] n -HPK1 ligand (I),

[0015] or its pharmaceutically acceptable salt or stereoisomer,

[0016] wherein:

[0017] The HPK1 ligand is, for example, an HPK1 inhibitor; B is a degrading tag, such as an E3 ligase ligand; L is a linking group between B and the HPK1 ligand; n is the number of degrading tags linked to the HPK1 ligand, selected from 1, 2, or 3.

[0018] The choice of the linking site, the number of linkages of the group binding to the E3 ligase, and the linking site on the HPK1 inhibitor will all affect the activity of the compound.

[0019] S1. HPK1 ligand

[0020] In the first aspect, the HPK1 ligand according to the present invention is a compound of the following formula (H-I). The first aspect includes the following schemes:

[0021] Scheme 1: A compound of general formula (H-I):

[0022]

[0023] or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

[0024] wherein:

[0025] W is selected from CR 1 or N;

[0026] The CyB ring is selected from a 4- to 10-membered cycloalkyl, a 4- to 10-membered heterocyclic group, a 5- to 8-membered aryl, or a 5- to 8-membered heteroaryl;

[0027] R 1 is independently selected from hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, -CN, -NO2, or -OR 1a , and the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by halogen, hydroxy, -C 1-8 alkoxy, cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0028] R 2 is selected from hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, oxo, -CN, -NO2, -OR 2a , -SO2R 2a , -COR 2a , -CO2R 2a , -CONR 2a R 2b , -C(=NR 2a )NR 2b R 2c , -NR 2a R 2b , -NR 2a COR 2b , -NR 2a CONR 2b R 2c , -NR 2a CO2R 2b , -NR 2a SONR 2b R 2c , -NR2a SO2NR 2b R 2c 、 or -NR 2a SO2R 2b , wherein the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by halogen, hydroxy, -C 1-8 alkoxy, cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0029] R 2a 、 R 2b and R 2c are each independently hydrogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl, wherein the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by at least one substituent R 2d ; or

[0030] (R 2a and R 2b ), (R 2b and R 2c ), or (R 2c and R 2a ) together with one or more atoms to which they are attached form a 3 - to 9 - membered ring, the ring containing 0, 1, or 2 heteroatoms independently selected from nitrogen, oxygen, or optionally oxidized sulfur as one or more ring members, and the ring is optionally substituted by at least one substituent R 2e ;

[0031] wherein R 2d and R 2e are each independently hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, oxo group, -CN, -NO2, -OR 2f 、 -SO2R 2f 、 -COR 2f 、 -CO2R 2f 、 -CONR 2f R 2g 、 -C(=NR 2f )NR 2g R 2h 、 -NR 2f R2g , -NR 2f COR 2g , -NR 2f CONR 2g R 2h , -NR 2f CO2R 2g , -NR 2f SONR 2g R 2h , -NR 2f SO2NR 2g R 2h , or -NR 2f SO2R 2g , said -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by at least one substituent selected from halogen, -C 1-8 alkyl, -OR 2i , -NR 2i R 2j , cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0032] R 2f , R 2g , R 2h , R 2i and R 2j are each independently hydrogen, -C 1-8 alkyl, C 1-8 alkoxy -C 1-8 alkyl -, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0033] R 3 is selected from hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, -CN, or -NO2;

[0034] R 4 is independently hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, oxo group, -CN, -NO2, -OR 4a , -SO2R 4a , -SO2NR 4a R 4b , -COR 4a , -CO2R4a 、 -CONR 4a R 4b 、 -C(=NR 4a )NR 4b R 4c 、 -NR 4a R 4b 、 -NR 4a COR 4b 、 -NR 4a CONR 4b R 4c 、 -NR 4a CO2R 4b 、 -NR 4a SONR 4b R 4c 、 -NR 4a SO2NR 4b R 4c 、 or -NR 4a SO2R 4b , wherein the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted with at least one substituent R 4d ;

[0035] R 4a 、 R 4b and R 4c are each independently hydrogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl, wherein the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted with at least one substituent R 4e ;

[0036] R 4d and R 4e are each independently hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, oxo group, -CN, -NO2, -OR 4f 、 -SO2R 4f 、 -SO2NR 4f R 4g 、 -COR 4f 、 -CO2R 4f 、 -CONR 4f R4g , -C(=NR 4f )NR 4g R 4h , -NR 4f R 4g , -NR 4f COR 4g , -NR 4f CONR 4g R 4h , -NR 4f CO2R 4f , -NR 4f SONR 4f R 4g , -NR 4f SO2NR 4g R 4h , or -NR 4f SO2R 4g , wherein the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by at least one substituent selected from halogen, -C 1-8 alkyl, -OR 4i , -NR 4i R 4j , cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0037] R 4f , R 4g , R 4h , R 4i and R 4j are each independently hydrogen, -C 1-8 alkyl, C 1-8 alkoxy-C 1-8 alkyl-, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0038] s is 0, 1 or 2, provided that the valence theory is satisfied;

[0039] t is 0, 1, 2, 3 or 4, provided that the valence theory is satisfied;

[0040] m is 0, 1, 2, 3 or 4, provided that the valence theory is satisfied;

[0041] CyD is selected from cycloalkyl, heterocyclic group, aryl, or heteroaryl.

[0042] Scheme 2: The compound according to Scheme 1, wherein R 1 is selected from hydrogen, halogen, -C 1-8Alkyl, preferably hydrogen, halogen, -C 1-6 Alkyl, more preferably hydrogen.

[0043] Embodiment 3: A compound according to any one of Embodiments 1-2, wherein CyB is selected from a 4- to 10-membered heterocyclic group, a 5- to 8-membered aryl group, or a 5- to 8-membered heteroaryl group;

[0044] Embodiment 4: The compound according to Embodiment 3, wherein the CyB ring is selected from a 5- to 6-membered nitrogen-containing heterocyclic group, a 5- to 6-membered oxygen-containing heterocyclic group, or a 5- to 6-membered nitrogen-containing heteroaryl group.

[0045] Embodiment 5: The compound according to Embodiment 4, wherein the CyB ring is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, pyrrolidinyl, piperidinyl, or pyridyl.

[0046] Embodiment 6: The compound according to Embodiment 5, wherein the CyB ring is selected from

[0047] Embodiment 7: The compound according to Embodiment 4, wherein the CyB ring is selected from

[0048] Embodiment 8: The compound according to any one of Embodiments 1-7, wherein the R 2 is selected from hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, oxo group, -CN, -NO2, -OR 2a , -SO2R 2a , -COR 2a , -CO2R 2a , -CONR 2a R 2b , -C(=NR 2a )NR 2b R 2c , -NR 2a R 2b , -NR 2a COR 2b , -NR 2a CONR 2b R 2c , -NR 2a CO2R 2b , -NR 2a SONR 2b R 2c , -NR 2a SO2NR 2b R 2c , or -NR2a SO2R 2b wherein the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by halogen, hydroxy, -C 1-8 alkoxy, cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0049] R 2a 、R 2b 、and R 2c are each independently hydrogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl, and the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by at least one halogen, -C 1-8 alkyl, -CN, hydroxy, or -NO2;

[0050] t is selected from 0, 1, 2, 3 or 4;

[0051] Furthermore,

[0052] R 2 is selected from -C 1-8 alkyl, preferably -C 1-3 alkyl, more preferably methyl or ethyl; t is 0 or 1.

[0053] Scheme 9: A compound according to any one of Schemes 3 - 8, wherein the is selected from: (including )

[0054] Scheme 10: A compound according to any one of Schemes 3 - 8, wherein the is selected from

[0055]

[0056] Scheme 11: A compound according to any one of Schemes 1 - 10, wherein the R 3 is selected from hydrogen, halogen, -C 1-8 alkyl, preferably halogen, more preferably fluorine; s is selected from 0 or 1.

[0057] Scheme 12: A compound according to any one of Schemes 1 - 11, wherein R 4Independently is hydrogen, halogen, -CN, -C 1-8 alkyl, and said -C 1-8 alkyl is optionally substituted by at least one substituent R 4d ;

[0058] R 4d independently is hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, oxo group, -CN, -NO2, -OR 4f , -SO2R 4f , -SO2NR 4f R 4g , -COR 4f , -CO2R 4f , -CONR 4f R 4g , -C(=NR 4f )NR 4g R 4h , -NR 4f R 4g , -NR 4f COR 4g , -NR 4f CONR 4g R 4h , -NR 4f CO2R 4f , -NR 4f SONR 4f R 4g , -NR 4f SO2NR 4g R 4h , or -NR 4f SO2R 4g , and said -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by at least one substituent selected from halogen, -C 1-8 alkyl, -OR 4i , -NR 4i R 4j ;

[0059] R 4f , R 4g , R 4h , R 4i , and R 4j each independently is hydrogen, -C 1-8 alkyl, C 1-8 alkoxy-C1-8 alkyl-, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0060] m is selected from 0, 1, 2, or 3.

[0061] Scheme 13: The compound according to Scheme 12, wherein the R 4 is -CN, halogen, preferably fluorine, chlorine, bromine, more preferably fluorine, chlorine.

[0062] Scheme 14: The compound according to Scheme 12, wherein the R 4 is -C 1-8 alkyl, preferably -C 1-3 alkyl, more preferably methyl or ethyl.

[0063] Scheme 15: The compound according to Scheme 12, wherein the R 4 is -C 4f alkyl substituted by -NR 4g R 1-8 wherein R 4f and R 4g are independently -C 1-8 alkyl, preferably methyl.

[0064] Scheme 16: The compound according to Scheme 15, wherein the R 4 is

[0065] Scheme 17: The compound according to Scheme 12, wherein the R 4 is -OR 4f wherein R 4f is selected from hydrogen, -C 1-8 alkyl, preferably -C 1-3 alkyl, more preferably methyl or ethyl.

[0066] Scheme 18: The compound according to Scheme 17, wherein the R 4 is

[0067] Scheme 19: The compound according to any one of Schemes 12 - 17, wherein the R 4 is methyl, ethyl, fluorine, chlorine, -CN, m is selected from 0, 1, or 2.

[0068] Scheme 20: The compound according to Scheme 12, wherein the R 4 is -C 1-8 alkyl, preferably -C 1-3 alkyl, more preferably methyl, ethyl, which is substituted by at least one halogen.

[0069] Embodiment 21: The compound according to Embodiment 20, wherein the halogen is fluorine, chlorine, bromine, preferably fluorine, chlorine.

[0070] Embodiment 22: The compound according to Embodiment 20 or 21, wherein the R 4 is -CHF2, -CH2F, -CF3, -CH2CHF2, -CH2CH2F, or -CH2CF3, preferably -CF3.

[0071] Embodiment 23: The compound according to Embodiments 1-22, wherein the CyD is selected from a 5-membered heteroaryl ring, a benzene ring, a 6-membered heteroaryl ring, a C 8-12 heterocyclic group, and the heteroaryl ring and the heterocyclic group contain 0, 1, or 2 heteroatoms independently selected from nitrogen, oxygen, or optionally oxidized sulfur as one or more ring members.

[0072] Embodiment 24: The compound according to Embodiment 23, wherein the CyD is selected from X1, X2, and X3 are each independently selected from CH, C, or N.

[0073] Embodiment 25: The compound according to Embodiment 23, wherein the CyD is selected from X1, X2, and X3 are each independently selected from C or N.

[0074] Embodiment 26: The compound according to Embodiment 25, wherein the CyD is selected from a benzene ring.

[0075] Embodiment 27: The compound according to Embodiment 24, wherein the CyD is selected from X1, X2, and X3 are each independently selected from CH, C, or N, provided that at least one of X1, X2, and X3 is N.

[0076] Embodiment 28: The compound according to Embodiment 25, wherein the CyD is selected from X1, X2, and X3 are each independently selected from C or N, provided that X1, X2, and X3 cannot all be C at the same time.

[0077] Embodiment 29: The compound according to Embodiment 27, the CyD is selected from pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, or triazinyl, preferably

[0078] Embodiment 30: The compound according to Embodiment 28, wherein the CyD is selected from

[0079]

[0080] Embodiment 31: A compound according to Embodiment 23, wherein the CyD is selected from 5-membered heteroaryl rings, said heteroaryl ring containing 0, 1 or 2 heteroatoms independently selected from nitrogen, oxygen or optionally oxidized sulfur as one or more ring members.

[0081] Embodiment 32: A compound according to Embodiment 31, wherein the CyD is selected from

[0082] Embodiment 33: A compound according to any one of Embodiments 1-32, wherein W is selected from CH or N.

[0083] In a second aspect, the HPK1 ligand according to the present invention is a compound of the following formula (H-II). The second aspect includes the following embodiments:

[0084] Embodiment 1: A compound represented by the general formula (H-II):

[0085]

[0086] or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

[0087] wherein R 1 , CyB, R 2 , t, R 3 , s, R 4 , m and CyD are each as defined in the first aspect above.

[0088] Embodiment 2: A compound according to Embodiment 1, wherein the is preferably

[0089] Embodiment 3: A compound according to Embodiment 1 or 2, wherein the R 1 and R 3 are each hydrogen.

[0090] Embodiment 4: A compound according to any one of Embodiments 1-3, wherein the CyD is pyridyl, preferably

[0091] Embodiment 5: A compound according to any one of Embodiments 1-4, wherein the R 4 is hydrogen.

[0092] In a third aspect, the HPK1 ligand according to the present invention is a compound of the following formula (H-III). The third aspect includes the following embodiments:

[0093] Embodiment 1: A compound represented by the general formula (H-III):

[0094]

[0095] Wherein:

[0096] W, R 1 , R 3 , s, R 4 , m and CyD are each as defined in the first aspect above;

[0097] R X is selected from H and C 1-8 alkyl, wherein said C 1-8 alkyl is optionally substituted by one or more substituents independently selected from deuterium, tritium, halogen, -OH, -CN, -NR Xa R Xb , -OR Xa , -CO-NHR Xb , -CO-NR Xa R Xb and 3-10 membered cycloalkyl; and

[0098] R Xa , R Xb are each independently selected from C 1-8 alkyl.

[0099] Embodiment 2: A compound according to Embodiment 1, wherein R X is selected from H and C 1-4 alkyl, wherein said C 1-4 alkyl is optionally substituted by 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, -OH, -CN, -NR Xa R Xb , -OR Xa , -CO-NHR Xb , -CO-NR Xa R Xb and 3-7 membered cycloalkyl; and

[0100] R Xa , R Xb are each independently selected from C 1-4 alkyl.

[0101] Embodiment 3: A compound according to Embodiment 1 or 2, wherein R X is selected from H and methyl, ethyl, isopropyl, isobutyl, sec-butyl and tert-butyl, wherein said methyl, ethyl, isopropyl, isobutyl, sec-butyl and tert-butyl are optionally substituted by 1, 2 or 3 substituents independently selected from deuterium, F, Cl, -OH, -CN, -NR Xa R Xb , -OR Xa , -CO-NHR Xb , -CO-NR Xa R Xbsubstituted by substituents of 3- to 6-membered cycloalkyl; and

[0102] R Xa 、R Xb are each independently methyl.

[0103] Embodiment 4: A compound according to any one of Embodiments 1-3, wherein R X is selected from: H, methyl, -CD3, ethyl, isopropyl, -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3,

[0104] Embodiment 5: A compound represented by the general formula (H-III):

[0105]

[0106] wherein:

[0107] W, R 1 、R 3 、s, R 4 、m and CyD are each as defined in the first aspect above;

[0108] R X is selected from C 1-8 alkyl, or C Xa R Xb alkyl substituted by -NR 1-8 ;

[0109] R Xa and R Xb are each independently selected from C 1-8 alkyl, preferably methyl.

[0110] Embodiment 6: The compound according to Embodiment 5, wherein the said R X is selected from methyl,

[0111] In the fourth aspect, the HPK1 ligand according to the present invention is a compound of the following formula (H-IV). The fourth aspect includes the following embodiments:

[0112] Embodiment 1: A compound represented by the general formula (H-IV)

[0113]

[0114] wherein the said R 1 、R 3 、s, R 4 、m, CyD and R X are each as defined in the third aspect above.

[0115] In a fifth aspect, the HPK1 ligand according to the present invention is a compound of formula (H-V) below. The fifth aspect includes the following embodiments:

[0116] Embodiment 1: A compound represented by the general formula (H-V):

[0117]

[0118] or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

[0119] wherein:

[0120] R 4 is independently hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, oxo group, -CN, -NO2, -OR 4a , -SO2R 4a , -SO2NR 4a R 4b , -COR 4a , -CO2R 4a , -CONR 4a R 4b , -C(=NR 4a )NR 4b R 4c , -NR 4a R 4b , -NR 4a COR 4b , -NR 4a CONR 4b R 4c , -NR 4a CO2R 4b , -NR 4a SONR 4b R 4c , -NR 4a SO2NR 4b R 4c , or -NR 4a SO2R 4b , and the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted with at least one substituent R 4d ;

[0121] R 4a , R 4b and R 4c are each independently hydrogen, -C1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl, said -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by at least one substituent R 4e ;

[0122] R 4d and R 4e are each independently hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, oxo group, -CN, -NO2, -OR 4f , -SO2R 4f , -SO2NR 4f R 4g , -COR 4f , -CO2R 4f , -CONR 4f R 4g , -C(=NR 4f )NR 4g R 4h , -NR 4f R 4g , -NR 4f COR 4g , -NR 4f CONR 4g R 4h , -NR 4f CO2R 4f , -NR 4f SONR 4f R 4g , -NR 4f SO2NR 4g R 4h , or -NR 4f SO2R 4g , said -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by at least one substituent selected from halogen, -C 1-8 alkyl, -OR 4i , -NR 4i R 4j , cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0123] R 4f , R4g , R 4h , R 4i and R 4j are each independently hydrogen, -C 1-8 alkyl, C 1-8 alkoxy-C 1-8 alkyl-, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0124] m is 0, 1, 2, 3, or 4, provided that the valence theory is satisfied;

[0125] CyD is selected from cycloalkyl, heterocyclic group, aryl or heteroaryl.

[0126] Scheme 2: The compound according to Scheme 1, wherein the CyD is selected from a 5-membered heteroaromatic ring, a benzene ring, a 6-membered heteroaromatic ring, C 8-12 heterocyclic group, and the heteroaromatic ring and the heterocyclic group contain 0, 1, or 2 heteroatoms independently selected from nitrogen, oxygen, or optionally oxidized sulfur as one or more ring members.

[0127] Scheme 3: The compound according to Scheme 2, wherein X1, X2, and X3 are each independently selected from CH, C, or N.

[0128] Scheme 4: The compound according to Scheme 2, wherein X1, X2, and X3 are each independently selected from C or N.

[0129] Scheme 5: The compound according to Scheme 4, wherein the CyD is selected from a benzene ring.

[0130] Scheme 6: The compound according to Scheme 3, wherein X1, X2, and X3 are each independently selected from CH, C, or N, provided that at least one of X1, X2, and X3 is N.

[0131] Scheme 7: The compound according to Scheme 4, wherein X1, X2, and X3 are each independently selected from C or N, provided that X1, X2, and X3 cannot all be C at the same time.

[0132] Scheme 8: The compound according to Scheme 6, wherein the CyD is selected from pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, or triazinyl, preferably

[0133] Scheme 9: The compound according to Scheme 7, wherein

[0134]

[0135] Embodiment 10: A compound according to Embodiment 2, wherein the CyD is selected from 5-membered heteroaryl rings, and the heteroaryl ring contains 0, 1, or 2 heteroatoms independently selected from nitrogen, oxygen, or optionally oxidized sulfur as one or more ring members.

[0136] Embodiment 11: A compound according to Embodiment 10, wherein the CyD is selected from

[0137] Embodiment 12: A compound according to any one of Embodiments 1 - 11, wherein R 4 is independently hydrogen, halogen, -CN, or -C 1-8 alkyl, and the -C 1-8 alkyl is optionally substituted by at least one substituent R 4d ;

[0138] R 4d is independently hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, oxo, -CN, -NO2, -OR 4f -, -SO2R 4f -, -SO2NR 4f R 4g -, -COR 4f -, -CO2R 4f -, -CONR 4f R 4g -, -C(=NR 4f )NR 4g R 4h -, -NR 4f R 4g -, -NR 4f COR 4g -, -NR 4f CONR 4g R 4h -, -NR 4f CO2R 4f -, -NR 4f SONR 4f R 4g -, -NR 4f SO2NR 4g R 4h -, or -NR 4f SO2R 4g , and the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is each optionally substituted by at least one selected from halogen, -C 1-8alkyl, -OR 4i , -NR 4i R 4j is substituted by a substituent of cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0139] R 4f , R 4g , R 4h , R 4i and R 4j are each independently hydrogen, -C 1-8 alkyl, C 1-8 alkoxy-C 1-8 alkyl-, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0140] m is selected from 0, 1, 2 or 3.

[0141] Embodiment 13: The compound according to Embodiment 12, wherein the R 4 is -CN, halogen, preferably fluorine, chlorine, bromine, more preferably fluorine or chlorine.

[0142] Embodiment 14: The compound according to Embodiment 12, wherein the R 4 is -C 1-8 alkyl, preferably -C 1-3 alkyl, more preferably methyl or ethyl.

[0143] Embodiment 15: The compound according to any one of Embodiments 12 - 14, wherein the R 4 is methyl, ethyl or fluorine; m is selected from 0 or 1.

[0144] In a sixth aspect, the HPK1 ligand according to the present invention is a compound of the following formula (H-VI). The sixth aspect includes the following embodiments:

[0145] Embodiment 1: A compound represented by the general formula (H-VI):

[0146]

[0147] or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

[0148] wherein:

[0149] R 5 is selected from hydrogen, halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, -CN, -NO2, -OR 2a , -SO2R 2a , -COR 2a, -CO2R 2a , -CONR 2a R 2b , -C(=NR 2a )NR 2b R 2c , -NR 2a R 2b , -NR 2a COR 2b , -NR 2a CONR 2b R 2c , -NR 2a CO2R 2b , -NR 2a SONR 2b R 2c , -NR 2a SO2NR 2b R 2c , or -NR 2a SO2R 2b , wherein the -C 1-8 alkyl, -C 2-8 alkenyl or -C 2-8 alkynyl is each optionally substituted by halogen, hydroxy, -C 1-8 alkoxy, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl;

[0150] R 1 , R 2a , R 2b , R 2c , R 4 , m and CyD are each as defined in the first aspect above.

[0151] Aspect 2: A compound according to Aspect 1, wherein R 5 is -C 1-8 alkyl, preferably -C 1-3 alkyl, more preferably methyl or ethyl.

[0152] In a seventh aspect, an HPK1 ligand according to the present invention is a compound of the following formula (H-VII). The seventh aspect includes the following embodiments:

[0153] Aspect 1: A compound represented by the general formula (H-VII):

[0154]

[0155] or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof,

[0156] wherein:

[0157] X4 is selected from O or an optionally oxidized S heteroatom;

[0158] W’ is CR 6 or N;

[0159] R 6 is selected from halogen, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, heteroaryl, oxo group, -CN, -NO2, -OR 2a , -SO2R 2a , -COR 2a , -CO2R 2a , -CONR 2a R 2b , -C(=NR 2a )NR 2b R 2c , -NR 2a R 2b , -NR 2a COR 2b , -NR 2a CONR 2b R 2c , -NR 2a CO2R 2b , -NR 2a SONR 2b R 2c , -NR 2a SO2NR 2b R 2c , or -NR 2a SO2R 2b , wherein the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by halogen, hydroxy, -C 1-8 alkoxy, cycloalkyl, heterocyclic group, aryl, or heteroaryl;

[0160] R 2a , R 2b , R 2c , R 4 , m and CyD are each as defined in the first aspect above.

[0161] Embodiment 2: A compound according to Embodiment 1, wherein X4 is O or S, preferably S.

[0162] Embodiment 3: A compound according to Embodiment 1 or 2, wherein W’ is CR 6 .

[0163] Embodiment 4: A compound according to any one of Embodiments 1 - 3, wherein R 6Selected from halogen, -CN, -NO2 or -NR 2a R 2b .

[0164] Embodiment 5: A compound according to Embodiment 4, wherein R 6 is -NR 2a R 2b .

[0165] Embodiment 6: A compound according to any one of Embodiments 1-5, wherein R 2a and R 2b are each independently hydrogen or -C 1-8 alkyl, preferably -C 1-3 alkyl, more preferably methyl or ethyl.

[0166] Embodiment 7: A compound according to Embodiment 5, wherein R 6 is -NH2.

[0167] Embodiment 8: A compound according to any one of Embodiments 1-7, wherein the CyD is selected from C 3-10 cycloalkyl, benzene ring, 5- to 6-membered monocyclic heteroaryl ring, 9- to 10-membered heteroaryl ring or 5- to 10-membered heterocyclic group, and the heteroaryl ring and heterocyclic group contain 1 or 2 heteroatoms independently selected from nitrogen, oxygen or optionally oxidized sulfur as ring members.

[0168] Embodiment 9: A compound according to Embodiment 8, wherein the CyD is a 9- to 10-membered bicyclic heteroaryl ring containing 1 or 2 nitrogen heteroatoms, preferably

[0169] In some embodiments, the present invention provides a compound of formula (I) as described above according to the first to seventh aspects, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein the HPK1 ligand compound is selected from those shown in Table 1-1 below:

[0170] Table 1-1:

[0171]

[0172]

[0173]

[0174] The present invention provides the above-described HPK1 ligand compound, or a pharmaceutically acceptable salt thereof, stereoisomer, and the HPK1 ligand compound is preferably selected from the HPK1 ligand exemplified compounds disclosed herein, as shown in Table 1-2 below:

[0175] Table 1-2:

[0176]

[0177]

[0178] S2. Degrade tag B

[0179] In some embodiments, B is a group that binds to an E3 ligase selected from von Hippel-Lindau (VHL), Cereblon, XIAP, E3A, MDM2, anaphase promoting complex (APC), UBR5 (EDD1), SOCS / BC-box / eloBC / CUL5 / RING, LNXp80, CBX4, CBLL1, HACE1, HECTD1, HECTD2, HECTD3, HECW1, HECW2, HERC1, HERC2, HERC3, HERC4, HUWE1, ITCH, NEDD4, NEDD4L, PPIL2, PRPF19, PIAS1, PIAS2, PIAS3, PIAS4, RANBP2, RNF4, RBX1, SMURF1, SMURF2, STUB1, TOPORS, TRIP12, UBE3A, UBE3B, UBE3C, UBE4A, UBE4B, UBOX5, UBR5, WWP1, WWP2, Parkin, A20 / TNFAIP3, AMFR / gp78, ARA54, β-TrCP1 / BTRC, BRCA1, CBL, CHIP / STUB1, E6, E6AP / UBE3A, F-box protein 15 / FBXO15, FBXW7 / Cdc4, GRAIL / RNF128, HOIP / RNF31, cIAP-1 / HIAP-2, cIAP-2 / HIAP-1, cIAP(pan), ITCH / AIP4, KAP1, MARCH8, MindBomb1 / MIB1, MindBomb2 / MIB2, MuRF1 / TRIM63, NDFIP1, NEDD4, NleL, Parkin, RNF2, RNF4, RNF8, RNF168, RNF43, SART1, Skp2, SMURF2, TRAF-1, TRAF-2, TRAF-3, TRAF-4, TRAF-5, TRAF-6, TRIM5, TRIM21, TRIM32, UBR5 or ZNRF3.

[0180] Further, the B is a group that binds to an E3 ligase selected from VHL, Cereblon, MDM2 or cIAP.

[0181] In some embodiments, the B has the general formula shown below:

[0182]

[0183] Wherein:

[0184] Each G is independently selected from CR C2 R C3 、NR C2 、CO, or SO2;

[0185] Y is selected from a bond or NH;

[0186] p is selected from 0, 1, or 2;

[0187] Each of W1, W2, W3, W4, and W5 is independently optionally selected from N or CR C4 ;

[0188] Each X5 is independently selected from O or S;

[0189] Each V1 is independently absent or selected from NH, O, S, SO, SO2, SO2NR C2 、CO, CO2, C(O)NR C2 、C(S)NR C2 、NR C2 、NR C2 CO, NR C2 CONR C3 、-C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl, and the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by R C4 Substituted, preferably each V1 is independently absent, -O-, -CH2-, -CH=CH-, or -NH-;

[0190] Each V2 is independently selected from CR C2 R C3 、NR C2 、O or S;

[0191] Each Z is independently selected from hydrogen, halogen, hydroxyl, amino, -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl, and the -C 1-8 alkyl, -C 2-8 alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclic group, aryl, or heteroaryl is each optionally substituted by R C5 Substituted;

[0192] R C1 , R C2 , R C3 , R C4 and R C5 is selected from hydrogen, carboxyl, cyano, nitro, halogen atoms, -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 Alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR C6 、-SO2R C6 、-SO2NR C6 R C7 、-COR C6 、-CO2R C6 、-CONR C6 R C7 、-POR C6 R C7 、-NR C6 R C7 、-NR C6 COR C7 、-NR C6 CONR C7 R C8 、-NR C6 CO2R C7 、-NR C6 SO2NR C7 R C8 , or -NR C6 S02R C7 , the -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 The alkynyl, cycloalkyl, heterocyclyl, aryl or heteroaryl groups are each optionally substituted with at least one substituent R C9 replace;

[0193] R C6 , R C7 , R C8 and R C9 Selected from hydrogen, halogen, hydroxy, amino, -C 1-8 Alkyl, -C 2-8 Alkenyl, -C 2-8 alkynyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

[0194] Further,

[0195] G is selected from CH2, CO, SO2, NH or NC 1-6 alkyl;

[0196] X5 is selected from O or S;

[0197] V1 is absent, NH, O, -C 1-8 alkyl, or -C 2-8 alkenyl, preferably absent, -O-, -CH2-, -CH=CH-, or -NH-.

[0198] V2 is selected from NH, N-C 1-6 alkyl, N-C 6-10 aryl, N-3-10 membered heterocyclic group, N-5-10 membered heteroaryl, N-C 3-10 cycloalkyl, O or S;

[0199] Z is selected from C 1-6 alkyl, C 3-10 cycloalkyl, halogen or hydrogen;

[0200] R C1 is selected from hydrogen, C 1-3 alkyl, hydroxy or -CH2-3-10 membered heterocyclic group;

[0201] W1, W2, W3, W4 and W5 are each independently optionally selected from N or -CR C4 , each of said R C4 is independently selected from hydrogen, halogen, C 1-3 alkyl, halo C 1-3 alkyl, hydroxy, deuterated C 1-3 alkyl, or Preferably, W1, W2, W3, W4 and W5 are selected from -CR C4 , and R C4 is selected from hydrogen, halogen (fluorine, chlorine), methyl, or

[0202] In some embodiments, said B is selected from:

[0203]

[0204]

[0205] V1 is absent, -O-, -CH2-, -CH=CH-, or -NH-.

[0206] In some embodiments, said B is selected from:

[0207]

[0208] And V1 is absent, or selected from -O-, -CH2-, -CH=CH-, or -NH-.

[0209] In some embodiments, said B is selected from:

[0210]

[0211] and

[0212] V1 is absent, -O-, -CH2-, or -NH-, preferably absent.

[0213] Further, said B is selected from:

[0214]

[0215] and V1 is absent, or is selected from -O-, -CH2-, or -NH-, preferably absent.

[0216] In some embodiments, said B is as follows:

[0217]

[0218] preferably In some embodiments, said B is as follows:

[0219]

[0220] In other embodiments, said B has the general formula as follows:

[0221] wherein:

[0222] CyD, R 4 and m are as defined in the first aspect of the above "S1. HPK1 Ligand" section.

[0223] In some such embodiments, said CyD is phenyl; R 4 is -OR 4f wherein R 4f is selected from hydrogen, -C 1-8 alkyl, preferably -C 1-3 alkyl, more preferably methyl or ethyl; and m is 1.

[0224] In some such embodiments, said B is as follows:

[0225]

[0226] S3. Linker L

[0227] L is a linking group between group B and the HPK1 ligand.

[0228] In some embodiments, said L is (LNK) u wherein:

[0229] Each LNK is independently absent or selected from C 1-8 alkylene, C 2-8 alkynylene, or cycloalkyl, heterocyclic group, heteroaromatic ring or aryl optionally substituted with one or more halogens or C 1-8 alkyl, wherein the cycloalkyl is preferably selected from wherein the heterocyclic group is preferably selected from:

[0230] and wherein the aryl is preferably selected from a benzene ring;

[0231] m is an integer between 1 and 8; and

[0232] u is an integer between 1 and 20.

[0233] In some embodiments, the L is (LNK) u , wherein:

[0234] Each LNK is independently absent, or selected from C 1-8 alkylene, C 2-8 alkynylene, or cycloalkyl, heterocycloalkyl, heteroaromatic ring or aromatic ring optionally substituted with one or more halogens or C 1-8 alkyl, wherein the cycloalkyl is selected from wherein the heterocycloalkyl is selected from:

[0235] and wherein the aromatic ring is selected from a benzene ring;

[0236] m is an integer between 1 and 8;

[0237] u is an integer between 1 and 20.

[0238] In some embodiments of the present invention, the L is selected from:

[0239]

[0240] In some embodiments of the present invention, the L is selected from:

[0241]

[0242] In some embodiments of the present invention, the L is selected from:

[0243]

[0244] In some embodiments of the present invention, the L is:

[0245]

[0246] In some embodiments of the present invention, the L is selected from:

[0247]

[0248] Preferably

[0249] In some embodiments of the present invention, the L is selected from:

[0250]

[0251]

[0252] In some embodiments of the present invention, the L is selected from:

[0253]

[0254] In some embodiments of the present invention, the L is selected from:

[0255]

[0256] In some embodiments of the present invention, the L is selected from:

[0257]

[0258] In some embodiments of the present invention, the L is selected from:

[0259]

[0260] In some embodiments of the present invention, the compound of general formula (I) is a compound of formula (II) as follows:

[0261]

[0262] Wherein:

[0263] The W, R 1 , R 2 , t, R 3 , s, CyB, R 4 , m and CyD are each as defined in the first aspect of the "S1. HPK1 Ligand" section above; and

[0264] The L, B and n are as defined herein.

[0265] In particular, the B is as defined in the section "S2. Degradation tag B". In particular, the L is as defined in the section "S3. Linker L".

[0266] In some such embodiments, the CyB is selected from

[0267] In some embodiments, the CyB is selected from

[0268] In some embodiments, the CyB is Preferably

[0269] In some embodiments, the is selected from (including )

[0270] In some embodiments, the is selected from

[0271] In some embodiments, the is Preferably

[0272] In some embodiments, the W is N.

[0273] In some embodiments, the R 1 and R 3 are each hydrogen.

[0274] In some embodiments, the CyD is pyridyl, preferably

[0275] In some embodiments, the R 4 is hydrogen.

[0276] In some embodiments, the L is

[0277] In some embodiments, the B is:

[0278] Preferably And

[0279] V1 is absent, -O-, -CH2-, or -NH-, preferably absent.

[0280] Preferably, the B is More preferably

[0281] In some embodiments, n is 1.

[0282] In some embodiments of the present invention, the compound of formula (I) is a compound of formula (III) as follows:

[0283]

[0284] Wherein:

[0285] Said R 4 , m and CyD are each as defined in the second aspect of the "S1. HPK1 Ligand" section above; and

[0286] Said L, B and n are as defined herein.

[0287] In particular, said B is as defined in the "S2. Degradation Tag B" section. In particular, said L is as defined in the "S3. Linker L" section.

[0288] In some embodiments of the present invention, the compound of formula (I) is a compound of formula (IV) as follows:

[0289]

[0290] Wherein:

[0291] Said W, R 1 , R 3 , s, R 4 , m and CyD are each as defined in the first and second aspects of the "S1. HPK1 Ligand" section above; and

[0292] Said R X is as defined in the second aspect of the "S1. HPK1 Ligand" section above;

[0293] Said L, B and n are as defined herein.

[0294] In particular, said B is as defined in the "S2. Degradation Tag B" section. In particular, said L is as defined in the "S3. Linker L" section.

[0295] In some such embodiments, W is N.

[0296] In some embodiments, said R 1 is selected from hydrogen or -C 1-3 alkyl, preferably hydrogen.

[0297] In some embodiments, said R 3 is selected from hydrogen or -C 1-3 alkyl, preferably hydrogen.

[0298] In some embodiments, s is 0 or 1.

[0299] In some embodiments, R X is selected from C 1-4 alkyl, wherein the C 1-4 alkyl is optionally substituted with 1, 2, or 3 substituents independently selected from deuterium, F, Cl, Br, -OH, -CN, -NR Xa R Xb or -OR Xa . In some such embodiments, the C 1-4 alkyl is selected from methyl, ethyl, isopropyl, isobutyl, sec-butyl, and tert-butyl, preferably methyl or ethyl.

[0300] In some embodiments, R Xa and R Xb are each independently selected from C 1-4 alkyl, preferably methyl.

[0301] In some embodiments, R X is selected from methyl or ethyl, wherein the methyl or ethyl is optionally substituted with 1, 2, or 3 substituents independently selected from deuterium, F, Cl, -OH, -CN, and -OCH3. In some embodiments, R X is selected from methyl or ethyl, wherein the methyl or ethyl is optionally substituted with 1, 2, or 3 substituents independently selected from deuterium, F, and -OCH3. In some embodiments, R X is selected from methyl, -CD3, ethyl, -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, or preferably methyl, -CD3, ethyl, -CH2CH2F, -CH2CHF2, -CH2CF3, or

[0302] In some embodiments, the CyD is selected from wherein X1, X2, and X3 are each independently selected from CH, C, or N, provided that at least one of X1, X2, and X3 is N.

[0303] In some embodiments, the CyD is selected from X1, X2, and X3 are each independently selected from C or N, provided that X1, X2, and X3 cannot all be C simultaneously.

[0304] In some embodiments, the CyD is selected from pyridyl, preferably

[0305] In some embodiments, R 4Independently is hydrogen or -C 1-3 alkyl, preferably hydrogen.

[0306] In some embodiments, m is 0 or 1.

[0307] In some embodiments, said L is

[0308] In some embodiments, said B is:

[0309] Preferably And

[0310] V1 is absent, -O-, -CH2-, or -NH-, preferably absent.

[0311] Preferably, said B is More preferably

[0312] In some embodiments, n is 1.

[0313] The present invention provides the compounds described above, or pharmaceutically acceptable salts, stereoisomers thereof, said compounds being selected from the exemplary compounds disclosed herein, as shown in Table 2 below:

[0314] Table 2:

[0315]

[0316]

[0317]

[0318]

[0319]

[0320]

[0321]

[0322]

[0323]

[0324]

[0325]

[0326]

[0327]

[0328]

[0329] Pharmaceutical Composition and Use

[0330] The present invention also provides a pharmaceutical composition, which comprises any compound according to the present invention or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. The pharmaceutical composition may optionally contain one or more pharmaceutical carriers and be formulated into any pharmaceutically acceptable pharmaceutical preparation.

[0331] The present invention also provides a pharmaceutical preparation of any compound according to the present invention or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof. The pharmaceutical preparation may optionally contain one or more pharmaceutical carriers.

[0332] The compound of general formula (I) according to the present invention or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof has excellent high selectivity inhibitory activity against HPK1 and can treat and / or prevent diseases such as tumors, immunity, and inflammation.

[0333] The present invention provides a method for degrading / inhibiting HPK1 activity, which comprises administering to an individual a compound according to the present invention or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, including the compound of general formula (I) or the specific compounds exemplified herein.

[0334] The compounds provided by the present invention have good HPK1 inhibitory / degrading activity and have good physicochemical properties and drug-forming characteristics. The compounds of the present invention have good potential for treating diseases mediated by HPK1.

[0335] WO2021057872A1 discloses PROTAC compounds formed by MAP4Ks family inhibitors and Cereblon protein ligands. The inventors surprisingly found that the compounds of the present invention show improved HPK1 selectivity.

[0336] The present invention also provides a method for treating a patient suffering from a disease that can be regulated by HPK1, which comprises administering to the patient an effective amount of a compound according to the present invention (such as a compound conforming to general formula (I) or the specific compounds exemplified herein) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0337] The present invention also provides a method for inhibiting / degrading HPK1 activity in a patient in need of HPK1 activity inhibition, which comprises administering to the patient an effective amount of a compound according to the present invention (such as a compound conforming to general formula (I) or the specific compounds exemplified herein) or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0338] The present invention also provides the use of any compound according to the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of a medicament for treating or preventing HPK1-mediated related diseases. HPK1 has a negative feedback regulatory effect in the T cell-mediated signaling pathway. Therefore, the compound according to the present invention can be used as an anti-tumor drug in the treatment of cancer or non-cancerous proliferative diseases. Further, the HPK1-mediated diseases include, but are not limited to, lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, breast cancer, ductal carcinoma of the breast, head and neck cancer, endometrial cancer, corpus cancer, rectal cancer, liver cancer, kidney cancer, renal pelvic cancer, esophageal cancer, esophageal adenocarcinoma, glioma, prostate cancer, thyroid cancer, female reproductive system cancer, carcinoma in situ, lymphoma, neurofibromatosis, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, oral cancer, pharyngeal cancer, multiple myeloma, leukemia, non-Hodgkin lymphoma, villous adenoma of the large intestine, melanoma, cell tumor and sarcoma, and myelodysplastic syndrome.

[0339] The present invention also protects a method for preventing and / or treating HPK1-mediated diseases and related diseases, which includes administering to a subject a therapeutically effective amount of a compound according to the present invention, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a composition comprising the compound according to the present invention. Further, the HPK1-mediated diseases and related diseases are selected from lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, ovarian cancer, peritoneal cancer, breast cancer, ductal carcinoma of the breast, head and neck cancer, endometrial cancer, corpus cancer, rectal cancer, liver cancer, kidney cancer, renal pelvic cancer, esophageal cancer, esophageal adenocarcinoma, glioma, prostate cancer, thyroid cancer, female reproductive system cancer, carcinoma in situ, lymphoma, neurofibromatosis, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, oral cancer, pharyngeal cancer, multiple myeloma, leukemia, non-Hodgkin lymphoma, villous adenoma of the large intestine, melanoma, cell tumor and sarcoma, and myelodysplastic syndrome.

[0340] The present invention also provides a method for treating a cancer patient, which includes administering to the patient an effective amount of a compound according to the present invention (for example, a compound of formula (I)), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof.

[0341] Definition

[0342] The compounds described in the present invention are named according to the chemical structural formula. If the naming of the compound does not conform to the chemical structural formula when representing the same compound, the chemical structural formula shall prevail.

[0343] In the present invention, unless otherwise specified, scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. However, for a better understanding of the present invention, the definitions of some terms are provided below. When the definitions and explanations of the terms provided by the present invention are different from those commonly understood by those skilled in the art, the definitions and explanations provided by the present invention shall prevail.

[0344] As used herein (including the appended aspects), unless the context clearly dictates otherwise, singular terms such as "a", "an", and "the" include their corresponding plural referents.

[0345] Unless the context clearly dictates otherwise, the term "or" is used to mean "and / or" and may be used interchangeably therewith.

[0346] The term "optionally" or "optionally" or "optional" means that the subsequently described event or condition may but need not occur, and the description includes the case where the event or condition occurs and the case where the event or condition does not occur.

[0347] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include deuterium and variants of hydrogen, provided that the valence of the particular atom is normal and the resulting compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on an aromatic group. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the type and number of substituents may be arbitrary based on what is chemically achievable.

[0348] The term "alkyl" refers to a hydrocarbon group selected from straight-chain and branched-chain saturated hydrocarbon groups containing 1 to 18 (such as 1 to 12, further such as 1 to 10, still further such as 1 to 8, or 1 to 6, or 1 to 4, or 1 to 3, or 1 to 2) carbon atoms.

[0349] The term "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), and iodine (I).

[0350] The term "haloalkyl" refers to an alkyl group in which one or more hydrogens are replaced by one or more halogen atoms (such as fluorine, chlorine, bromine, and iodine). Examples of haloalkyls include halo-C 1-8 alkyl, halo-C 1-6 alkyl, or halo-C 1-4 alkyl, but are not limited to -CF3, -CH2Cl, -CH2CF3, -CHCl2, etc.

[0351] The term "alkenyl" refers to a hydrocarbyl group selected from straight-chain and branched-chain hydrocarbyl groups containing at least one C═C double bond and from 2 to 18 (such as from 2 to 8, further such as from 2 to 6) carbon atoms.

[0352] The term "alkynyl" refers to a hydrocarbyl group selected from straight-chain and branched-chain hydrocarbyl groups containing at least one C≡C triple bond and from 2 to 18 (such as from 2 to 8, further such as from 2 to 6) carbon atoms.

[0353] The term "alkyloxy" or "alkoxy" refers to an alkyl group as defined above attached to the parent molecular moiety through an oxygen atom. Examples of alkyloxy (e.g., C 1-6 alkyloxy or C 1-4 alkyloxy) include but are not limited to methoxy, ethoxy, isopropoxy, propoxy, n-butoxy, tert-butoxy, pentyloxy, hexyloxy, and the like.

[0354] The term "alkoxy-alkyl-" refers to an alkyl group as defined above substituted by an alkoxy group as defined above. Examples of alkoxy-alkyl- (e.g., C 1-8 alkoxy-C 1-8 alkyl-) include but are not limited to methoxymethyl, ethoxymethyl, isopropoxymethyl, or propoxymethyl, and the like.

[0355] The term "cycloalkyl" refers to a hydrocarbyl group selected from saturated cyclic hydrocarbyl groups, said saturated cyclic hydrocarbyl groups including monocyclic and polycyclic (e.g., bicyclic and tricyclic) groups, including fused cycloalkyl, bridged cycloalkyl, or spirocycloalkyl.

[0356] For example, cycloalkyl can contain from 3 to 12 (such as from 3 to 10, further such as from 3 to 8, further such as from 3 to 6, 3 to 5, or 3 to 4) carbon atoms. Even further for example, cycloalkyl can be selected from monocyclic groups containing from 3 to 12 (such as from 3 to 10, further such as from 3 to 8, 3 to 6) carbon atoms. Examples of monocyclic cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. Specifically, examples of saturated monocyclic cycloalkyl (e.g., C 3-8 cycloalkyl) include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl.

[0357] The term "spirocycloalkyl" refers to a cyclic structure containing carbon atoms and formed by at least two rings sharing one atom. For example, 7- to 12-membered spirocycloalkyl refers to a cyclic structure containing 7 to 12 carbon atoms and formed by at least two rings sharing one atom.

[0358] The term "fused cycloalkyl" refers to a fused ring containing carbon atoms and formed by two or more rings sharing two adjacent atoms. For example, a 4- to 10-membered fused cycloalkyl refers to a fused ring containing 4 to 10 ring carbon atoms and formed by two or more rings sharing two adjacent atoms.

[0359] The term "bridged cycloalkyl" refers to a cyclic structure containing carbon atoms and formed by two rings sharing two atoms that are not adjacent to each other. For example, a 7- to 10-membered bridged cycloalkyl refers to a cyclic structure containing 7 to 12 carbon atoms and formed by two rings sharing two atoms that are not adjacent to each other.

[0360] The term "cycloalkenyl" refers to a non-aromatic cyclic alkyl group having 3 to 10 carbon atoms, having a single ring or multiple rings and having at least one double bond and preferably 1 to 2 double bonds. In one embodiment, the cycloalkenyl is cyclopentenyl (1-cyclopent-1-enyl, 1-cyclopent-2-enyl, 1-cyclopent-3-enyl) or cyclohexenyl (1-cyclohex-1-enyl, 1-cyclohex-2-enyl, 1-cyclohex-3-enyl), preferably cyclohexenyl.

[0361] The term "cycloalkynyl" refers to a non-aromatic cycloalkyl group having 5 to 10 carbon atoms, having a single ring or multiple rings and having at least one triple bond.

[0362] The term "aryl", used alone or in combination with other terms, refers to a group selected from:

[0363] - 5- and 6-membered carbocyclic aromatic rings, such as phenyl;

[0364] - bicyclic ring systems, such as 7- to 12-membered bicyclic ring systems, wherein at least one ring is a carbocyclic and aromatic, such as naphthyl and indanyl; and

[0365] - tricyclic ring systems, such as 10- to 15-membered tricyclic ring systems, wherein at least one ring is a carbocyclic and aromatic, such as fluorenyl.

[0366] The terms "aromatic hydrocarbon ring" and "aryl" are used interchangeably herein. In some embodiments, the monocyclic or bicyclic aromatic hydrocarbon ring has 5 to 10 ring-forming carbon atoms (i.e., C 5-10 aryl). Examples of monocyclic or bicyclic aromatic hydrocarbon rings include, but are not limited to, phenyl, naphthalen-1-yl, naphthalen-2-yl, anthryl, phenanthryl, etc. In some embodiments, the aromatic hydrocarbon ring is a naphthalene ring (naphthalen-1-yl or naphthalen-2-yl) or a phenyl ring. In some embodiments, the aromatic hydrocarbon ring is a phenyl ring.

[0367] The term "heteroaryl" refers to a group selected from:

[0368] - A 5-, 6- or 7-membered aromatic monocyclic ring containing at least one heteroatom, such as 1 to 4 heteroatoms, or in some embodiments 1 to 3 heteroatoms, or in some embodiments 1 to 2 heteroatoms, said heteroatoms being selected from nitrogen (N), sulfur (S) and oxygen (O), and the remaining ring atoms being carbon;

[0369] - A 7- to 12-membered bicyclic ring containing at least one heteroatom, such as 1 to 4 heteroatoms, or in some embodiments 1 to 3 heteroatoms, or in other embodiments 1 or 2 heteroatoms, said heteroatoms being selected from N, O and S, the remaining ring atoms being carbon, and wherein at least one ring is aromatic and there is at least one heteroatom in the aromatic ring; and

[0370] - An 11- to 14-membered tricyclic ring containing at least one heteroatom, such as 1 to 4 heteroatoms, or in some embodiments 1 to 3 heteroatoms, or in other embodiments 1 or 2 heteroatoms, said heteroatoms being selected from N, O and S, the remaining ring atoms being carbon, and wherein at least one ring is aromatic and there is at least one heteroatom in the aromatic ring.

[0371] When the total number of S and O atoms in the heteroaryl exceeds 1, those heteroatoms are not adjacent to each other. In some embodiments, the total number of S and O atoms in the heteroaryl is not greater than 2. In some embodiments, the total number of S and O atoms in the aromatic heterocycle is not greater than 1. When the heteroaryl contains more than one heteroatom ring member, the heteroatoms may be the same or different. One or more nitrogen atoms in the ring of the heteroaryl may be oxidized to form N-oxides. As used herein, the term "C-linked heteroaryl" means that the heteroaryl is linked to the core molecule by a bond from a C-atom of the heteroaryl ring.

[0372] The term "heteroaryl" in some embodiments refers to a monocyclic or bicyclic aromatic heterocycle having 5, 6, 7, 8, 9 or 10 ring members, wherein 1, 2, 3 or 4 heteroatom ring members are independently selected from nitrogen (N), sulfur (S) and oxygen (O), and the remaining ring members are carbon. In some embodiments, the monocyclic or bicyclic aromatic heterocycle is a monocyclic or bicyclic containing 1 or 2 heteroatom ring members independently selected from nitrogen (N), sulfur (S) and oxygen (O). In some embodiments, the monocyclic or bicyclic aromatic heterocycle is a 5- to 6-membered heteroaryl ring that is monocyclic and has 1 or 2 heteroatom ring members independently selected from nitrogen (N), sulfur (S) and oxygen (O). In some embodiments, the ring of the monocyclic or bicyclic aromatic heterocycle is an 8- to 10-membered heteroaryl ring that is bicyclic and has 1 or 2 heteroatom ring members independently selected from nitrogen, sulfur and oxygen.

[0373] "Heterocyclic group", "heterocycle" or "heterocyclic" are interchangeable and refer to a non-aromatic heterocyclic group containing one or more heteroatoms selected from nitrogen, oxygen or optionally oxidized sulfur as ring members and the remaining ring members are carbon, including monocyclic rings, fused rings, bridged rings and spiro rings, i.e., containing monocyclic heterocyclic groups, bridged heterocyclic groups, spiro heterocyclic groups, and fused heterocyclic groups. The term "optionally oxidized sulfur" as used herein refers to S, SO or SO2.

[0374] The term "monocyclic heterocyclic group" refers to a monocyclic group in which at least one ring member is a heteroatom selected from nitrogen, oxygen or optionally oxidized sulfur. The heterocycle can be saturated or partially saturated.

[0375] Representative examples of exemplary monocyclic 4- to 10-membered heterocyclic groups include, but are not limited to, the following groups:

[0376] where the wavy line indicates the point of attachment.

[0377] The term "spiro heterocyclic group" refers to a 5- to 20-membered polycyclic heterocyclic group having rings connected by a common carbon atom (referred to as the spiro atom), containing one or more heteroatoms selected from nitrogen, oxygen or optionally oxidized sulfur as ring members and the remaining ring members are carbon. One or more of the rings of the spiro heterocyclic group may contain one or more double bonds, but none of the rings has a completely conjugated π electron system. Preferably, the spiro heterocyclic group is 6 to 14-membered, and more preferably 7 to 12-membered. Depending on the number of common spiro atoms, the spiro heterocyclic group is divided into monospiro heterocyclic group, dispiro heterocyclic group, or polyspiro heterocyclic group, and preferably refers to monospiro heterocyclic group or dispiro heterocyclic group, and more preferably 4-membered / 4-membered, 3-membered / 5-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospiro heterocyclic group.

[0378] The term "fused heterocyclic group" refers to a 5- to 20-membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms (carbon and carbon atoms or carbon and nitrogen atoms) with another ring, containing one or more heteroatoms selected from nitrogen, oxygen or optionally oxidized sulfur as ring members and the remaining ring members are carbon. One or more of the rings of the fused heterocyclic group may contain one or more double bonds, but none of the rings has a completely conjugated π electron system. Preferably, the fused heterocyclic group is 6 to 14-membered, preferably 7 to 12-membered and more preferably 7 to 10-membered. Depending on the number of member rings, the fused heterocyclic group is divided into bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclic groups, preferably refers to bicyclic or tricyclic fused heterocyclic groups, and more preferably 5-membered / 5-membered, or 5-membered / 6-membered bicyclic fused heterocyclic groups.

[0379] The term "bridged heterocyclic group" or "bridged heteroaryl group" refers to a 5- to 14-membered polycyclic heterocyclic group in which every two rings in the system share two non-adjacent atoms, containing one or more heteroatoms selected from nitrogen, oxygen, or optionally oxidized sulfur as ring members, and the remaining ring members being carbon. One or more rings of the bridged heterocyclic group may contain one or more double bonds, but no ring has a fully conjugated π-electron system. Preferably, the bridged heterocyclic group is 6- to 14-membered, and more preferably 7- to 10-membered. Depending on the number of member rings, the bridged heterocyclic group is divided into bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclic groups, and preferably refers to bicyclic, tricyclic, or tetracyclic bridged heterocyclic groups, and more preferably bicyclic or tricyclic bridged heterocyclic groups.

[0380] The term "alkylene" refers to a divalent alkyl group as defined above, which refers to a saturated straight-chain or branched divalent hydrocarbon group having a length of 1 to 18 carbon atoms (C 1-18 ), wherein the alkylene may be optionally and independently substituted with one or more of the following substituents. In another embodiment, the alkylene has 1 to 8 carbon atoms (C 1-8 ) or 1 to 6 carbon atoms (C 1-6 ). Examples of alkylene include but are not limited to methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), etc.

[0381] The term "alkenylene" refers to a divalent alkenyl group as defined above, which refers to a straight-chain or branched divalent hydrocarbon group having a length of two to eight carbon atoms (C 2-8 ) and having at least one unsaturated position, i.e., a carbon-carbon sp 2 double bond, wherein the alkenylene may be optionally and independently substituted with one or more of the substituents described in this application and includes groups having "cis" and "trans" orientations or "E" and "Z" orientations. Examples include but are not limited to vinylidene (-CH=CH-), allylidene (-CH2CH=CH-), etc.

[0382] The term "alkynylene" refers to a divalent alkynyl group as defined above, which refers to a straight-chain or branched divalent hydrocarbon group having a length of three to eight carbon atoms (C 3-8 ) and having at least one unsaturated position, i.e., a carbon-carbon sp triple bond, wherein the alkynylene may be optionally and independently substituted with one or more of the substituents described in this application. Examples include but are not limited to propargylene (propargylidene, -CH2C≡C-), etc.

[0383] The term "cycloalkylene" refers to a divalent cycloalkyl group as defined above. The term "heteroarylene" refers to a divalent heteroaryl group as defined above. The term "arylene" refers to a divalent aryl group as defined above. The term "heteroarylene" refers to a divalent heteroaryl group as defined above.

[0384] The compounds disclosed herein may contain asymmetric centers and can therefore exist as enantiomers. "Enantiomers" refer to two stereoisomers of a compound that are non-superimposable mirror images of each other. Where the compounds disclosed herein have two or more asymmetric centers, they can additionally exist as diastereomers. Enantiomers and diastereomers belong to the broader class of stereoisomers. All such possible stereoisomers, including substantially pure resolved enantiomers, their racemic mixtures, and mixtures of diastereomers, are intended to be included. All stereoisomers of the compounds disclosed herein and / or their pharmaceutically acceptable salts are intended to be included. Unless otherwise specifically mentioned, reference to an isomer applies to any possible isomer. Whenever the isomeric composition is not specified, all possible isomers are included.

[0385] As used herein, the term "substantially pure" means that the target stereoisomer contains no more than 35% by weight (such as no more than 30%, further such as no more than 25%, even further such as no more than 20%) of any other one or more stereoisomers. In some embodiments, the term "substantially pure" means that the target stereoisomer contains no more than 10% by weight (e.g., no more than 5%, such as no more than 1%) of any other one or more stereoisomers.

[0386] When the compounds disclosed herein contain an olefinic double bond, such double bonds are intended to include both E and Z geometric isomers unless otherwise specified.

[0387] When the compounds disclosed herein contain a disubstituted cyclohexyl or cyclobutyl group, the substituents found on the cyclohexyl or cyclobutyl ring can adopt cis and trans forms. The cis form means that both substituents are found on the upper side of the 2 substituent positions on the carbon, while the trans means that they are on opposite sides.

[0388] "Pharmaceutically acceptable salts" refer to those salts that are suitable for use in contact with the tissues of humans and lower animals within the scope of reasonable medical judgment, without undue toxicity, irritation, allergic response, etc., and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts can be prepared in situ during the final isolation and purification of the compounds disclosed herein, or separately by reacting the free base functional group with a suitable organic acid or by reacting an acidic group with a suitable base.

[0389] Alternatively, if the compounds disclosed herein are obtained as acid addition salts, the free base can be obtained by basifying a solution of the acid salt. Conversely, if the product is a free base, addition salts, such as pharmaceutically acceptable addition salts, can be produced by dissolving the free base in a suitable organic solvent and / or water and treating the solution with an acid, in accordance with conventional procedures for preparing acid addition salts from basic compounds. Those skilled in the art will recognize the various synthetic methods available for preparing non-toxic pharmaceutically acceptable addition salts without undue experimentation.

[0390] As defined herein, "its pharmaceutically acceptable salts" includes salts of at least one compound of formula (I) and salts of stereoisomers of the compound of formula (I), such as salts of enantiomers and / or diastereoisomers.

[0391] Unless otherwise specified, when a group has one or more connectable sites, any one or more of these sites of the group can be connected to other groups by chemical bonds. When the connection mode of the chemical bond is non-specific and there is an H atom at the connectable site, then when connecting the chemical bond, the number of H atoms at this site will correspondingly decrease with the number of connected chemical bonds to become a group with the corresponding valence. The chemical bond connecting the site to other groups can be represented by a straight solid line bond , or a wavy line For example, the straight solid line bond in -OCH3 indicates connection to other groups through the oxygen atom in this group; The wavy line in indicates connection to other groups through the #1 and ##1 carbon atoms in this phenyl group;

[0392] When applied to animals, humans, experimental subjects, cells, tissues, organs or biological fluids, the terms "administer", "administering", "treating" and "treatment" mean the contact of an exogenous agent, therapeutic agent, diagnostic agent or composition with the animal, human, subject, cell, tissue, organ or biological fluid. Treatment of cells encompasses the contact of a reagent with the cells, as well as the contact of the reagent with a fluid that contacts the cells. The terms "administer" and "treatment" also mean in vitro and ex vivo treatment of, for example, cells by a reagent, diagnostic agent, binding compound or by another cell. The term "subject" as used herein includes any living organism, preferably an animal, more preferably a mammal (such as a rat, mouse, dog, cat, and rabbit) and most preferably a human.

[0393] The term "effective amount" or "therapeutically effective amount" refers to an amount of an active ingredient, such as a compound, that is sufficient to effect such treatment of a disease, disorder or symptom when the compound is administered to a subject to treat the disease or at least one clinical symptom of the disease or disorder.

[0394] The term "disease" refers to any disease, discomfort, illness, symptom or indication, and can be used interchangeably with the term "condition" or "disorder".

[0395] Throughout the specification and the following aspects, unless the context requires otherwise, the terms "comprise" and variations such as "comprises" and "comprising" are intended to indicate the presence of the features hereinafter, but do not exclude the presence or addition of one or more other features. When used herein, the term "comprise" may be replaced by the terms "contain", "include" or sometimes by "have".

[0396] Throughout the specification and the following aspects, the term "C n-m " indicates a range including the endpoints, where n and m are integers and indicate the number of carbons. Examples include C 1-8 、C 1-6 etc.

[0397] Unless otherwise clearly defined elsewhere in this document, all other technical and scientific terms used herein have the meanings commonly understood by those of ordinary skill in the art to which this invention pertains.

[0398] Compound identification and characterization.

[0399] Examples give the preparation of representative compounds represented by formula (I) and related structure identification data. 1 The HNMR spectra were measured using a Bruker instrument (400 MHz), and the chemical shifts are expressed in ppm. Tetramethylsilane internal standard (0.00 ppm) was used. 1 Presentation method of H NMR: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublets, dt = doublet of triplets. When coupling constants are provided, their unit is Hz.

[0400] The mass spectra were obtained using an LC / MS instrument, and the ionization mode can be ESI or APCI. Unless otherwise specified, the test methods are as follows:

[0401] Time (min) Flow Rate (mL / min) <![CDATA[% A (0.1% FA H2O)]]> % B (Acetonitrile) 1 0.00 1 85 15 2 2.50 1 5 95 3 4.00 1 5 95 4 4.01 1 85 15 5 5.00 1 85 15

[0402] Thin layer chromatography silica gel plates use Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates, thin layer chromatography (TLC).

[0403] The specifications of the silica gel plates used are 0.15 mm - 0.2 mm, and the specifications for separating and purifying products by thin layer chromatography are 0.4 mm - 0.5 mm. Column chromatography generally uses Yantai Huanghai silica gel 200 - 300 mesh silica gel as the carrier.

[0404] In the following examples, unless otherwise specified, all temperatures are in degrees Celsius. Unless otherwise specified, various starting materials and reagents are commercially available or synthesized according to known methods. Commercially available raw materials and reagents are used directly without further purification, and commercially available manufacturers include, but are not limited to, Aldrich Chemical Company, ABCR GmbH&Co.KG, Acros Organics, Shanghai Bide Pharmaceutical Technology Co., Ltd., and Shanghai Shaoyuan Reagent Co., Ltd.

[0405] DMSO-d6: deuterated dimethyl sulfoxide.

[0406] DIEA: N,N-diisopropylethylamine.

[0407] Rt: retention time.

[0408] Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

[0409] For the purification of compounds, an eluent system of column chromatography and thin-layer chromatography is used, and the system is selected from: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: dichloromethane and ethyl acetate system, D: dichloromethane and ethanol system, where the volume ratio of the solvents varies according to the polarity of the compound, and a small amount of acidic or basic reagents can also be added for conditioning, such as acetic acid or triethylamine, etc.

[0410] Biological assays

[0411] The pharmacological properties of the compounds of the present invention can be confirmed by numerous biological assays. The following exemplary biological assays have been carried out with the compounds of the present invention.

[0412] Experimental Example 1. In vitro enzyme activity detection method for HPK1

[0413] The ADP-Glo method was used to detect kinase activity and measure the IC50 value to evaluate the inhibitory ability of the compound on human HPK1.

[0414] Enzyme buffer conditions: 50 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.01% Brij35, 2 mM DTT.

[0415] Configuration of kinase and substrate mixture: The working concentration of the kinase in the HPK1 reaction solution is 7 nM, and the working concentration of ATP is 20 μM.

[0416] Test procedure:

[0417] Dilute the compound with DMSO in a dilution plate. The maximum starting concentration of the compound is 1 μM, with 10 concentration gradient points (4-fold dilution).

[0418] Dilute the above 10 concentration gradient points of the compound 50-fold into the kinase reaction buffer and shake on an oscillator for 20 minutes. Prepare the kinase with the enzyme reaction buffer and add 2 μL of HPK1 kinase to each well of the reaction plate. The concentration of HPK1 kinase in the reaction solution is 7 nM.

[0419] Add 1 μL of the compound diluted in the buffer to each well, seal the plate with a sealing film, centrifuge at 1000 g for 30 seconds, and let it stand at room temperature for 10 minutes. Prepare the ATP solution with the enzyme reaction buffer and add 2 μL of the ATP solution (the working concentration of ATP is 20 μM) to the reaction plate. Seal the plate with a sealing film, centrifuge at 1000 g for 30 seconds, and react at room temperature for 60 minutes. Transfer 4 μL of ADP-Glo to a 384-well reaction plate, centrifuge at 1000 rpm / min for 1 minute, and incubate at 25 °C for 40 minutes. Transfer 8 μL of the Detection solution to a 384-well reaction plate, centrifuge at 1000 rpm / min for 1 minute, and incubate at 25 °C for 40 minutes. Use a BMG microplate reader to read the RLU (Relative luminescence unit) signal, and the signal intensity is used to characterize the activity level of the kinase.

[0420] The kinase activity data is expressed as the ratio of the kinase activity of the test compound to the kinase activity of the blank group (containing only DMSO), and the IC 50 value is obtained by curve fitting using Prism software (GraphPad 7.0).

[0421] The following table shows the HPK1 IC 50 value

[0422]

[0423] The HPK1 IC 50 of the compounds of the present invention is not greater than 60 nM, and the HPK1 IC 50 of most compounds of the present invention is not greater than 20 nM, indicating that the compounds of the present invention all have good inhibitory activity against HPK1 and have good application potential in the treatment of diseases mediated by HPK1.

[0424] Experimental Example 2. IL2 Activity Detection Method

[0425] The ability of the compound to activate T cells was evaluated by testing the EC50 value of human PBMC IL2 activation using the ELISA method.

[0426] (1) Pretreatment of 96-well plates in the IL2 kit: Coat the plate with 5 μg / ml CD3 antibody, 100 μl per well, overnight at 4°C.

[0427] (2) PBMC cells (10 5 / well). The compound was diluted with DMSO in a dilution plate, with the maximum starting concentration of the compound being 10 μM, 8 concentration gradient points (3-fold dilution), added to a 96-well plate, and 5 μg / ml CD28 antibody was added at the same time. Incubate for 24 h.

[0428] (3) Wash the ELISA plate twice with PBS, add 100 μl / well ELISA diluent, then add 100 μl of the standard and the incubated sample obtained in step 2, and incubate at room temperature for 2 h.

[0429] (4) Wash each well three times, adding 400 μl of washing buffer to each well and blotting with clean absorbent paper for the last time.

[0430] (5) Add 200 μl of IL2 conjugate to each well, cover with new tape, and incubate at room temperature for 2 h. Repeat step (4).

[0431] (6) Protect from light, add 200 μl of substrate to each well and incubate at room temperature for 20 min.

[0432] (7) Add 50 μL of stop solution to each well. The color in the well should change from blue to yellow. The ELISA reader reads the 450 nM signal, and the signal intensity is used to characterize the degree of IL2 activity. The EC50 value is obtained by fitting with Prism software.

[0433] The following table shows the effect of the compounds of the present invention on inducing IL2 production.

[0434] Compound <![CDATA[IL2 EC 50 (nM)]]> Compound <![CDATA[IL2 EC 50 (nM)]]> C003 110.9 C132 34.58 C006 178.8 C134 41.68 C017 323.5 C135 44.1 C018 102.2 C139 103.86 C021 37.9 C153 37.07 C022 95.6 C154 103.71 C024 196 C160 45.47 C110 55.59 C162 27.33 C122 59.85 C170 23.81 C131 41.68 C196 34.68

[0435] Experimental Example 3. HPK1 protein degradation detection

[0436] The WB method was used to test HPK1 protein degradation and DC50 / Dmax to evaluate the in vitro degradation ability of the compounds.

[0437] RAMOS cell line was selected as the degradation cell line. Culture conditions: RMPI 1640 medium, plus 10% FBS, 37°C, 5% CO2.

[0438] Experimental steps:

[0439] (1) Cell plating: In a 6 cm culture dish, 4 ml of culture medium, 2 × 10 cells were plated. 6 Pieces / dish.

[0440] (2) Drug addition: Dilute the compound (stock solution concentration 10 mM) with DMSO in the dilution plate. The maximum starting concentration of the compound is 10 μM, with 7 concentration gradient points (3-fold dilution). Take 4 μL of each and add it to the above culture dish, and incubate for 16 h.

[0441] (3) Collect the cells and add lysis buffer, centrifuge at 12000 rpm / min for 20 min, and collect the supernatant. After quantification with a BCA quantification kit, add 5×loading buffer and denature for 10 min.

[0442] (4) Add 10 μg of denatured protein to the SDS-PAGE gel, perform electrophoresis, transfer the membrane, block with 5% BSA for 1 h, incubate with the primary antibody overnight at 4 °C, wash the primary antibody, add the secondary antibody and incubate at room temperature for 1 h, and wash the secondary antibody.

[0443] Imaging: Scan the membrane with a dual-color fluorescence imaging system. Use Image J software to quantify the Western results and calculate DC 50 and Dmax by comparing with the control group.

[0444] The following table shows the HPK1 degradation activity of the compounds of the present invention

[0445] Compound <![CDATA[DC 50 (nM)]]> Dmax (%) Compound <![CDATA[DC 50 (nM)]]> Dmax (%) C024 2.1 77 C162 29.16 86.50 C022 55 86 C167 23.88 66.78 C084 1.21 93.00 C170 52.25 87.09 C085 385 56.4 C177 632.3 51.57 C092 21.36 -- C181 214 64.38 C110 9.349 93.00 C195 18.7 80.69 C115 23.54 66.4 C196 8.8 93.78 C131 385.6 52 C197 5.28 77.54 C132 29.34 73 C198 32.92 91.84 C134 7.219 78 C199 83.01 83.77 C135 175.8 63 C201 30.63 76.10 C137 229.6 58.44 C203 5.99 91.74 C139 52.36 80.90 C204 58.02 85.42 C140 197.6 50.68 C205 89.93 83.00 C153 616.2 52.47 C206 190.3 73.00 C160 12.65 88.85 C209 29.03 84.00

[0446] The compounds of the present invention have good HPK1 protein degradation ability.

[0447] Experimental Example 4. Determination of Pharmacokinetic Properties

[0448] Female Balb / c mice (weight 18 - 22 g) are fasted overnight before the experiment. Dissolve the test compound in the solvent DMSO:PEG400:30% captisol:HCl = 5:20:70:5 (v / v / v / v), and administer it by single-dose gavage at 10 mg / kg or by intravenous injection at 2 mg / kg. Blood is collected from the orbital venous plexus at 15 minutes, 30 minutes, and 1, 2, 4, 6, 8, and 24 hours after dosing. Approximately 0.08 mL of blood is collected at each time point and placed in a 1.5 mL centrifuge tube containing EDTA-2K anticoagulant. The blood sample is centrifuged within 2 hours (3200 g, 10 minutes, 4 °C) to obtain plasma samples. The plasma samples are stored frozen at -70 °C to -80 °C in an ultra-low temperature refrigerator before sample processing. Before sample processing, take the plasma samples out of the refrigerator and thaw them at room temperature. Then, add 20 μL of each plasma sample to a 96-well plate, and then add 120 μL of acetonitrile containing internal standard to precipitate proteins. After vortex mixing, centrifuge at 4 °C, 4950 g for 15 minutes, take the supernatant, mix it with an equal volume of 0.1% formic acid aqueous solution, and perform LC-MS / MS analysis.

[0449] Before the experiment, SD rats weighing about 220 g (half male and half female) were fasted overnight. The compound of the present invention was formulated in a solvent of DMSO:PEG400:30% captisol:HCL = 5:20:70:5 (v / v / v / v), and administered by single intravenous injection (2 mg / mL) at 2 mL / kg and by single intragastric administration at 10 mL / kg at doses of 5 mg / kg, 2 mg / kg, and 1 mg / kg of the compound of the present invention. Blood was collected from the orbital venous plexus before dosing and at 0.083 (only collected for intravenous dosing), 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after dosing, approximately 0.150 mL at each time point, and placed in a 1.5 mL centrifuge tube containing EDTA-2K anticoagulant. The blood samples were centrifuged within 1 hour (3200 g, 10 min, 4 °C) to obtain plasma samples. The plasma samples were stored frozen at -70 °C to -80 °C in an ultra-low temperature freezer before processing. Before sample processing, the plasma samples were taken out of the refrigerator and thawed at room temperature. Then, 20 μL of plasma samples were taken and added to a 96-well plate, and then 120 μL of acetonitrile containing an internal standard was added to precipitate proteins. After vortex mixing, it was centrifuged at 4 °C and 4700 g for 15 minutes, and the supernatant was mixed with 0.1% formic acid water in equal volume for LC-MS / MS analysis.

[0450] The results of the pharmacokinetic determination of single-dose administration in mice are shown in the following table.

[0451]

[0452] The results show that the compound of the present invention has good exposure (AUC) both by oral and intravenous administration, and has good oral half-life and oral bioavailability.

[0453] Preparation Examples

[0454] The present invention will be further described in detail below with reference to specific examples, but these examples do not limit the scope of the present invention. The following examples are used to understand the method and core idea of the present invention. For those skilled in the art, any possible changes or substitutions made without departing from the concept of the present invention fall within the protection scope of the present invention. The experimental methods without specific conditions noted in the examples of the present invention are usually conventional conditions or conditions recommended by the raw material or commodity manufacturer; the reagents without the source noted are usually conventional reagents that can be purchased through commercial channels.

[0455] Synthesis of Intermediates

[0456] Preparation of Intermediate warhead 3

[0457]

[0458] Step 1: Preparation of Intermediate 2

[0459] Compound 1 (10 g, 30.8 mmol) was dissolved in tetrahydrofuran (100 mL). NaH (1.8 g, 46.2 mmol) was added at 0 °C, and the mixture was stirred at 0 °C for 0.5 h under nitrogen protection. Then benzenesulfonyl chloride (8 g, 46.2 mmol) was added in an ice bath, and the mixture was stirred at room temperature for 3 h. After the reaction was completed, the reaction solution was poured into ice water, and compound 2 was obtained by filtration. LCMS (ESI) m / z: 464.2 [M+H] + 。

[0460] Step 2: Preparation of intermediate 3

[0461] Compound 2 (1.0 g, 2.15 mmol) and compound 1 (0.975 g, 2.585 mmol) were dissolved in dioxane (22 mL). Potassium carbonate (0.595 g, 4.305 mmol) dissolved in water (7.5 mL) was mixed with the above solution, and then Pd(dppf)Cl2·CH2Cl2 (315 mg, 0.43 mmol) was added. The mixture was stirred at 80 °C for 1 h under nitrogen protection. LCMS showed that there was no raw material remaining. The reaction solution was cooled to room temperature, extracted three times with dichloromethane, the organic phases were combined, washed twice with saturated brine, the organic phase was dried over sodium sulfate, filtered, and the filtrate was concentrated. The crude product was obtained as intermediate 3 by column chromatography. LCMS (ESI) m / z: 589.2 [M+H] + 。

[0462] Step 3: Preparation of intermediate 4

[0463] Compound 3 (7 g, 11.9 mmol) was dissolved in 4M hydrochloric acid dioxane solution (70 mL), and the reaction solution was stirred at room temperature for 2 h. After the reaction was completed, the solvent was directly evaporated to dryness to obtain the crude product, and the crude product was washed with ether to obtain intermediate 4. LCMS (ESI) m / z: 488.9 [M+H] + 。

[0464] Step 4: Preparation of intermediate warhead 3

[0465] Compound 4 (3.5 g, 6.68 mmol) was dissolved in a mixed solvent of acetonitrile (140 mL) and dioxane (36 mL). Then, acetaldehyde (2.64 mL, 13.36 mmol) and acetic acid (0.38 mL, 6.68 mmol) were added successively. The reaction mixture was stirred at room temperature for 1 hour, and then sodium cyanoborohydride (1470 mg, 23.38 mmol) was added at room temperature. The mixture was further stirred at 40 °C for 4 hours, and the previous operation was repeated, and then stirred at 40 °C for 12 hours. After the reaction was completed, the pH of the mixture was adjusted to 14 with ammonia water. After extraction with dichloromethane, the organic layer was washed with brine, dried over anhydrous sodium sulfate, and concentrated in vacuo to obtain the crude product. The crude product was purified by column chromatography to obtain intermediate warhead 3. DCM was added, dissolved, ammonia water was added, the organic phase was separated, and concentrated. LCMS (ESI) m / z: 515.0 [M+H] + 。

[0466] Example 1: Preparation of Compound C001

[0467]

[0468] Step 1:

[0469] 2,5-Dibromo-3-methylpyridine (5 g, 20.1 mmol), 1-tert-butoxycarbonylpiperazine (5.6 g, 30.1 mmol) and K2CO3 (8.3 g, 60.2 mmol) were dissolved in dimethyl sulfoxide (50 mL). The mixture was stirred at 120 °C for 36 hours. LCMS showed that the reactants were partially converted and the product was detected. After the reaction mixture was cooled to room temperature, water (100 mL) was added for quenching, and the mixture was extracted with ethyl acetate (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product. The crude product was purified by silica gel chromatography to obtain Compound 001a. LCMS: [M+H] + = 356.2.

[0470] Step 2:

[0471] 001a (2.8 g, 7.89 mmol), bis(pinacolato)diboron (3 g, 11.83 mmol), potassium acetate (2.32 g, 23.66 mmol) and Pd(dppf)Cl2 (576 mg, 0.789 mmol) were dissolved in dimethyl sulfoxide (30 mL). The mixture was stirred at 80 °C for 2 hours under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. After the reaction mixture was cooled to room temperature, water (100 mL) was added for quenching, and the mixture was extracted with ethyl acetate (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the residue. The residue was purified by silica gel chromatography to obtain Compound 001b. LCMS: [M+H]+ = 322.4。

[0472] Step 3:

[0473] Dissolve warhead 3 (200 mg, 0.623 mmol), 001b (287 mg, 0.623 mmol), potassium carbonate (258 mg, 1.87 mmol) and Brettphos-Pd-G3 (56 mg, 0.0623 mmol) in dimethyl sulfoxide (2 mL). Stir the mixture at 100 °C overnight under a nitrogen atmosphere. LCMS shows that the reactants are completely converted and the product is detected. After the reaction solution is cooled to room temperature, add water (50 mL) to quench it, and extract with ethyl acetate (30 mL × 2). Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain a residue. Purify the residue by silica gel chromatography to obtain compound 001c. LCMS: [M+H] + = 702.5。

[0474] Step 4:

[0475] Dissolve 001c (250 mg, 0.357 mmol) in hydrochloric acid dioxane solution (5 mL, 4 M), and stir overnight at room temperature. LCMS shows that the reactants are completely converted and the product is detected. Distill the reaction solution under reduced pressure to obtain the crude product 001d. LCMS: [M+H] + = 472.2。

[0476] Step 5:

[0477] Dissolve 001d (16 mg, 0.028 mmol), 001e (9 mg, 0.036 mmol), HATU (16 mg, 0.042 mmol) and N,N-diisopropylethylamine (15 mg, 0.112 mmol) in dimethyl sulfoxide (2 mL) at room temperature, and stir the reaction overnight at room temperature. LCMS shows that the reactants are completely converted and the product is detected. Add water (50 mL) to the reaction solution to quench it, and extract with dichloromethane (30 mL × 2). Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain a residue. Purify by Pre-TLC to obtain compound C001. LCMS: Rt = 2.190 min; LCMS: [M+H] + = 688.4。

[0478] Example 2: Preparation of compound C002

[0479]

[0480] Dissolve 002a (35 mg, 0.062 mmol), 002b (19 mg, 0.081 mmol), DIEA (32 mg, 0.248 mmol) and HATU (30.3 mg, 0.08 mmol) in dimethyl sulfoxide (2 mL), and stir the mixture at room temperature for 16 hours. LCMS shows that the reactants are completely converted and the product is detected. After the reaction solution is cooled to room temperature, add water (50 mL) to quench it, and extract with dichloromethane (30 mL * 2). Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product. Purify by Pre-HPLC to obtain compound C002. LCMS: Rt = 2.099 min; [M + H] + = 785.3.

[0481] Example 3: Preparation of compound C003

[0482]

[0483] Dissolve 003a (33 mg, 0.058 mmol), 003b (21 mg, 0.075 mmol), DIEA (30 mg, 0.232 mmol) and HATU (29 mg, 0.075 mmol) in dimethyl sulfoxide (2 mL), and stir the mixture at room temperature for 16 hours. LCMS shows that the reactants are completely converted and the product is detected. After the reaction solution is cooled to room temperature, add water (50 mL) to quench it, and extract with dichloromethane (30 mL * 2). Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product. Purify by Pre-HPLC to obtain compound C003. LCMS: [M + H] + = 819.3.

[0484] Example 4: Preparation of compound C004

[0485]

[0486] Step 1:

[0487] Dissolve 4a (500 mg, 2.63 mmol) and 4b (795 mg, 3.95 mmol) in DMF (10 mL), add NaH (210 mg, 5.26 mmol) under ice bath, and stir at room temperature for 12 h. TLC shows that the raw materials are completely converted and the product is detected. Add 30 mL of water to the reaction solution, extract with ethyl acetate (20 mL * 3), dry the organic phase with anhydrous sodium sulfate, filter, and concentrate the filtrate in vacuo to obtain the residue. Purify the residue by column chromatography to obtain compound 4c.

[0488] Step 2:

[0489] 4c (970 mg, 2.61 mmol), B2Pin2 (1.33 g, 5.23 mmol), KOAc (770 mg, 7.84 mmol) and Pd(dppf)Cl2 (212 mg, 0.26 mmol) were dissolved in dioxane (20 mL) at room temperature. The mixture was purged with nitrogen three times, heated to 80 °C and stirred at this temperature for 3 h. TLC showed that the starting materials were completely converted and the product was detected. After the reaction was completed, the reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography to obtain compound 4d.

[0490] Step 3:

[0491] 4d (136 mg, 0.33 mmol), 4e (100 mg, 0.22 μmol), BrettPhos Pd G3 (20 mg, 22 μmol) and K2CO3 (90 mg, 0.65 mmol) were dissolved in DMSO / H2O (4 mL / 0.8 mL) at room temperature. The mixture was purged with nitrogen three times, heated to 100 °C and stirred at this temperature for 12 h. LCMS showed that the reactants were completely converted and the product was detected. 30 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by Prep-TLC to obtain compound 4f.

[0492] Step 4:

[0493] 4f (123 mg, 75.8 μmol) was dissolved in DCM (3 mL), and TFA (1 mL) was added with stirring. The mixture was stirred at room temperature for 1 h. LCMS showed that the reactants were completely converted and the product was detected. The reaction mixture was distilled under reduced pressure, and the crude product was dissolved in MeOH (3 mL). Ammonia in methanol solution (1 mL) was added with stirring, and the mixture was stirred at room temperature for 1 h. LCMS showed that the reactants were completely converted and the product was detected. The reaction mixture was distilled under reduced pressure to obtain the crude product of compound 4g.

[0494] Step 5:

[0495] 4g (83 mg, 0.17 mmol), 4h (69 mg, 0.26 mmol), DIEA (67 mg, 0.51 mmol) and HATU (130 mg, 0.34 mmol) were dissolved in DMSO (2 mL), and the mixture was stirred at room temperature for 12 h. LCMS showed that the reactants were completely converted and the product was detected. 30 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by Prep-TLC to obtain the final product C004. LCMS: Rt = 1.918 min, [M+H] + = 737.4.

[0496] 1 1H NMR (500 MHz, DMSO-d6) δ 12.05 (d, J = 2.8 Hz, 1H), 10.52 (s, 1H), 8.85 (d, J = 2.1 Hz, 2H), 8.38 (d, J = 2.2 Hz, 2H), 8.12 (d, J = 2.7 Hz, 1H), 8.10 (s, 1H), 7.69–7.63 (m, 2H), 7.47 (dd, J = 8.2, 2.1 Hz, 1H), 5.43 (dt, J = 7.5, 3.7 Hz, 1H), 3.95 (d, J = 19.7 Hz, 1H), 3.78 (q, J = 9.4, 6.3 Hz, 1H), 3.64 (dt, J = 12.7, 6.4 Hz, 3H), 3.06–2.96 (m, 3H), 2.75 (q, J = 7.5, 6.7 Hz, 2H), 2.41 (d, J = 10.0 Hz, 2H), 2.30 (s, 3H), 2.04 (q, J = 14.0, 10.0 Hz, 8H), 1.80 (s, 2H), 1.23 (d, J = 3.6 Hz, 1H), 1.04 (t, J = 7.2 Hz, 3H).

[0497] Example 5: Preparation of Compound C005

[0498]

[0499] Step 1:

[0500] Dissolve 5a (1 g, 11.48 mmol) and TEA (1.74 g, 17.22 mmol) in DCM (20 mL), add Boc2O (3.26 g, 14.92 mmol) under ice bath, and stir at room temperature for 2 h. TLC shows complete conversion of the starting material and the product is detected. Distill the reaction mixture under reduced pressure to obtain a residue. Purify the residue by column chromatography to obtain Compound 5b.

[0501] Steps 2 - 6:

[0502] Steps 2 - 6 are carried out according to the methods described in Steps 1 - 5 of Example 4, except that Compound 5b and 5c are used to replace 4b and 4a in Step 1 of Example 4 respectively, and Compound 5i is used to replace Compound 4h in Step 5 of Example 4, to obtain Compound C005. LCMS: Rt = 1.773 min, [M + H] + = 689.4, purity = 98.77%.

[0503] 11H NMR (500 MHz, DMSO-d6) δ 12.16–11.95 (m, 1H), 10.44 (d, J = 12.9 Hz, 1H), 8.98–8.72 (m, 2H), 8.48–8.33 (m, 2H), 8.17–8.04 (m, 2H), 7.59 (dd, J = 23.1, 8.2 Hz, 2H), 7.41 (dd, J = 20.2, 8.2 Hz, 2H), 5.65 (d, J = 49.4 Hz, 1H), 4.26 (s, 1H), 3.97 (ddd, J = 31.1, 12.8, 4.7 Hz, 1H), 3.83 (dt, J = 19.8, 6.6 Hz, 2H), 3.73 (dd, J = 9.7, 6.5 Hz, 2H), 3.65–3.53 (m, 1H), 3.01 (s, 2H), 2.71 (dt, J = 24.4, 6.6 Hz, 2H), 2.42 (s, 1H), 2.27 (d, J = 35.2 Hz, 3H), 2.05 (s, 5H), 1.05 (t, J = 7.2 Hz, 3H).

[0504] Example 6: Preparation of Compound C006

[0505]

[0506] Step 1:

[0507] Dissolve 6a (500 mg, 24.00 mmol), 6b (922 mg, 48.00 mmol) and TEA (486 mg, 48.00 mmol) in DMF (20 mL), raise the temperature to 100 °C and stir at this temperature for 12 h. TLC shows that the raw materials are completely converted and the product is detected. Add 30 mL of water to the reaction solution, extract with ethyl acetate (100 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate in vacuo to obtain a residue. Distill the reaction solution under reduced pressure to obtain a residue. Purify the residue by column chromatography to obtain compound 6c.

[0508] Step 2:

[0509] Dissolve 6d (500 mg, 2.63 mmol) and 6e (380 mg, 4.21 mmol) in DMF (40 mL), add NaH (632 mg, 15.80 mmol) under ice bath, and stir at room temperature for 12 h. TLC shows that the raw materials are completely converted and the product is detected. Add 100 mL of water to the reaction solution, extract with ethyl acetate (100 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate in vacuo to obtain a residue. Distill the reaction solution under reduced pressure to obtain a residue. Purify the residue by column chromatography to obtain compound 6f.

[0510] Step 3:

[0511] Dissolve 6f (430 mg, 1.65 mmol), B2Pin2 (840 mg, 3.31 mmol), KOAc (485 mg, 4.96 mmol) and Pd(dppf)Cl2 (134 mg, 0.16 mmol) in dioxane (15 mL) at room temperature. Replace nitrogen three times, raise the temperature to 80 °C and stir at this temperature for 3 h. TLC shows that the raw materials are completely converted and the product is detected. After the reaction is completed, the reaction solution is cooled to room temperature, filtered, and the filtrate is concentrated in vacuo to obtain a residue. Purify the residue by column chromatography to obtain compound 6g.

[0512] Step 4:

[0513] Dissolve 6g (300 mg, 0.98 mmol), 6h (300 mg, 0.65 mmol), BrettPhos Pd G3 (59 mg, 65 μmol) and K2CO3 (270 mg, 1.95 mmol) in DMSO / H2O (12 mL / 3 mL) at room temperature. Replace nitrogen three times, raise the temperature to 100 °C and stir at this temperature for 12 h. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water and 3 mL of formic acid to the reaction solution, extract with dichloromethane (50 mL × 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate in vacuo to obtain a residue. Purify the residue by column chromatography to obtain compound 6i.

[0514] Step 5:

[0515] Dissolve 6i (80 mg, 0.14 mmol) and 6j (65 mg, 0.20 mmol) in TFA (5 mL) and stir at room temperature for 1 h. LCMS shows that the reactants are completely converted and the product is detected. Distill the reaction solution under reduced pressure to obtain a crude product, dissolve it in DMSO (2 mL), add DIEA (53 mg, 0.41 mmol) and HATU (103 mg, 0.27 mmol), and stir at room temperature for 2 h. LCMS shows that the reactants are completely converted and the product is detected. Purify by preparative chromatography to obtain the final product C006. LCMS: Rt = 1.795 min, [M+H] + = 663.4, purity = 94.00%.

[0516] 11H NMR (500 MHz, DMSO-d6) δ 12.06 (d, J = 2.7 Hz, 1H), 10.77 (s, 1H), 9.72 (s, 1H), 8.86 (s, 1H), 8.81 (d, J = 2.3 Hz, 1H), 8.41 (dd, J = 2.5, 1.0 Hz, 1H), 8.39 (s, 1H), 8.16 (s, 1H), 8.13 (d, J = 2.7 Hz, 1H), 8.11 (d, J = 0.7 Hz, 1H), 7.36–7.28 (m, 2H), 6.64 (d, J = 9.0 Hz, 2H), 5.70 (d, J = 7.6 Hz, 1H), 4.99 (s, 2H), 4.44–4.22 (m, 2H), 3.16 (d, J = 11.6 Hz, 2H), 2.79–2.68 (m, 1H), 2.64–2.58 (m, 2H), 2.38 (s, 3H), 2.17–2.04 (m, 6H), 1.86 (qd, J = 12.2, 4.6 Hz, 1H), 1.10 (t, J = 7.2 Hz, 3H).

[0517] Example 7: Preparation of Compound C008

[0518]

[0519] According to the method described in Steps 2-5 of Example 6, except that in Step 2 of Example 6, Compound 8a and K2CO3 were used to replace 6d and NaH respectively and the reaction temperature was set at 70 °C, Compound C008 was obtained. LCMS: Rt = 1.741 minutes, [M+H] + = 667.4.

[0520] Example 8: Preparation of Compound C009

[0521]

[0522] Step 1:

[0523] Dissolve Compound 9a (1.0 g, 5.3 mmol), N-Boc-1,2-ethylenediamine (1.02 g, 6.38 mmol), and K2CO3 (1.47 g, 10.6 mmol) in DMSO (20 mL) and react at 120 °C for 6 hours. TLC showed that the raw materials were completely converted and the product was detected. Add 100 mL of water to the reaction solution, extract with ethyl acetate (100 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate under vacuum to obtain a residue. Distill the reaction solution under reduced pressure to obtain a residue. Purify the residue by column chromatography (PE:EA = 5:1) to obtain Compound 9b. [M+H] + = 230.0.

[0524] Step 2:

[0525] Dissolve compound 9b (0.5 g, 1.52 mmol), B2Pin2 (578.9 mg, 2.28 mmol), KOAc (447 mg, 4.56 mmol) and Pd(dppf)Cl2 (111 mg, 0.152 mmol) in dioxane (5 mL) at room temperature. Replace nitrogen three times, raise the temperature to 80 °C and stir at this temperature for 3 h. TLC shows that the raw materials are completely converted and the product is detected. After the reaction is completed, the reaction solution is cooled to room temperature, filtered, and the filtrate is concentrated under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 9c. [M+H] + = 378.0.

[0526] Step 3:

[0527] Dissolve compound 9c (0.4 g, 1.06 mmol), Warhead3 (300 mg, 0.58 mmol), BrettPhos Pd G3 (60 mg, 66 μmol) and K2CO3 (270 mg, 1.98 mmol) in DMSO / H2O (4 mL / 1 mL) at room temperature. Replace nitrogen three times, raise the temperature to 100 °C and stir at this temperature for 12 h. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water and 3 mL of formic acid to the reaction solution, extract with dichloromethane (50 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 9d.

[0528] Step 4:

[0529] Dissolve compound 9d in 10 ml of HCl / dioxane and stir at room temperature for 30 minutes to obtain the crude product of compound 9e. [M+H] + = 446.

[0530] Step 5:

[0531] Dissolve compound 9e (130 mg, 0.29 mmol), 3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (68 mg, 0.29 mmol), HATU (165 mg, 0.435 mmol) and DIEA (75 mg, 0.58 mmol) in 10 ml of DMSO and react at room temperature for 1 hour. LCMS shows that the reactants are completely converted and the product is detected. Purify by preparative chromatography to obtain the final product C009. [M+H] + = 662.4, purity: 95.3%.

[0532] 11H NMR (400 MHz, MeOD) δ 8.75 (s, 1H), 8.59 (d, J = 12.9 Hz, 2H), 8.36 (s, 1H), 8.21 (s, 1H), 8.04 (s, 1H), 7.86 (s, 1H), 7.80–7.74 (m, 1H), 7.57–7.50 (m, 2H), 4.71–4.60 (m, 1H), 3.89 (t, J = 6.7 Hz, 2H), 3.77 (s, 6H), 3.30–3.14 (m, 4H), 2.80 (t, J = 6.7 Hz, 2H), 2.41 (d, J = 13.5 Hz, 7H), 1.40 (t, J = 7.3 Hz, 3H).

[0533] Example 9: Preparation of Compound C010

[0534]

[0535] Step 1:

[0536] Dissolve compound 010a (10 g, 53 mmol) in anhydrous THF (200 ml). At 0 °C, add LiHMDS (80.6 mL, 80.65 mmol) and react for 0.5 h. Then add 2-fluoro-3-methyl-5-bromopyridine (10.07 g, 53 mmol) to the above system and react at room temperature for 3 h. LCMS shows that the reactants are completely converted and the product is detected. Add 100 mL of water to the reaction solution, extract with ethyl acetate (100 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate under vacuum to obtain a residue. Distill the reaction solution under reduced pressure to obtain a residue. Purify the residue by column chromatography to obtain compound 010b. [M + H] + = 356.1.

[0537] Step 2:

[0538] Dissolve compound 010b (3.0 g, 8.45 mmol), B2Pin2 (3.2 g, 12.67 mmol), KOAc (2.485 g, 25.3 mmol) and Pd(dppf)Cl2 (618 mg, 0.845 mmol) in dioxane (30 mL) at room temperature, displace nitrogen three times, raise the temperature to 110 °C and stir at this temperature for 3 h. TLC shows that the raw materials are completely converted and the product is detected. After the reaction is completed, cool the reaction solution to room temperature, filter, and concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 010c. [M + H] + = 404.0.

[0539] Step 3:

[0540] Dissolve compound 010c (2.14 g, 5.3 mmol), Warhead 3 (2.72 g, 5.3 mmol), Pd(dppf)Cl2 (387 mg, 5.3 mmol) and K2CO3 (1.46 g, 10.6 mmol) in DMSO / H2O (20 mL / 5 mL) at room temperature. Replace nitrogen three times, raise the temperature to 100 °C and stir at this temperature for 12 h. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water and 3 mL of formic acid to the reaction solution, extract with dichloromethane (50 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 010d. [M+H] + = 712.

[0541] Step 4:

[0542] Dissolve compound 010d in 20 ml of ethanol, then add sodium hydroxide (60 mg, 1.4 mmol) and stir at 50 °C for 15 minutes. LCMS shows that the reactants are completely converted and the product is detected. Concentrate the reaction solution under vacuum to obtain the crude product of compound 010e. [M+H] + = 572.

[0543] Step 5:

[0544] Dissolve compound 010e in 10 ml of HCl / dioxane and stir at room temperature for 30 minutes to obtain the crude product of compound 010f. [M+H] + = 472.

[0545] Step 6:

[0546] Dissolve compound 010f (400 mg, 0.85 mmol), methyl 6-bromohexanoate (195 mg, 0.93 mmol), K2CO3 (586.5 mg, 4.25 mmol) and KI (705.5 mg, 4.25 mmol) in ACN and react at 90 °C for 6 hours. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water to the reaction solution and extract with dichloromethane (50 mL * 3). Dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Obtain compound 010g by column chromatography.

[0547] Step 7:

[0548] Dissolve compound 010g (200 mg, 0.33 mmol) and LiOH (79 mg, 3.3 mmol) in a 10 ml system of THF / H2O = 1:1 and stir at room temperature for 20 hours. LCMS shows that the reactants are completely converted and the product is detected. Directly dry by evaporation to obtain compound 010h. [M+H] += 586。

[0549] Step 8:

[0550] Dissolve compound 010h (100 mg, 0.17 mmol), (2S,4R)-1-((S)-2-amino-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide (90 mg, 0.34 mmol), HATU (130 mg, 0.75 mmol), and DIEA (122 mg, 0.044 mmol) in DMF (2 ml), react at room temperature for 2 hours. LCMS shows that the reactants are completely converted and the product is detected. Purify by preparative chromatography to obtain the final product C010. LCMS: Rt = 2.180 min; [M+H] + = 998.7。

[0551] Example 10: Preparation of compound C011

[0552]

[0553] Step 1:

[0554] Dissolve compound 010f (400 mg, 0.85 mmol, prepared according to Steps 1-5 of Example 9) in CAN (10 ml), add tert-butyl 3-(methanesulfonyloxymethyl)azetidine-1-carboxylate (226 mg, 0.85 mmol), DIEA (548 mg, 4.25 mmol), and KI (706 mg, 4.25 mmol), and react at 100 °C for 16 hours. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water and dichloromethane (50 mL * 3) to the reaction solution for extraction. Dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate in vacuo to obtain a residue. Compound 011g is obtained by column chromatography.

[0555] Step 2:

[0556] Dissolve compound 011g in 10 ml of HCl / dioxane and stir at room temperature for 30 minutes to obtain the crude product of compound 011h.

[0557] Step 3:

[0558] Compound 011h (100 mg, 0.185 mmol) was dissolved in DMF (2 ml), 3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (48 mg, 0.185 mmol), HATU (105 mg, 0.27 mmol), and DIEA (95 mg, 0.74 mmol) were added, and the reaction was carried out at room temperature for 1 hour. LCMS showed that the reactants were completely converted and the product was detected. It was purified by preparative chromatography to obtain C011.

[0559] Example 11: Preparation of Compound C017

[0560]

[0561] Step 1:

[0562] Warhead 3 (167 mg, 0.142 mmol), 4-N-Boc-aminocyclohexanone (113 mg, 0.532 mmol), and acetic acid (21 mg, 0.142 mmol) were dissolved in dichloromethane / dimethyl sulfoxide (4 mL / 4 mL), and the mixture was stirred and reacted at 35 °C for 16 hours. Then sodium triacetoxyborohydride (376 mg, 1.77 mmol) was added to the mixture and the reaction was continued by stirring at 35 °C for 2 hours. LCMS showed that the reactants were completely converted and the product was detected. After the reaction solution was cooled to room temperature, saturated ammonium chloride solution (50 mL) was added for quenching, and it was extracted with dichloromethane (30 mL * 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a residue. The residue was purified by silica gel chromatography to obtain compound 017a. LCMS: [M + H] + = 669.4.

[0563] Step 2:

[0564] 017a (105 mg, 0.157 mmol) was dissolved in dichloromethane / dimethyl sulfoxide (1 mL / 5 mL), and the reaction was carried out by stirring at room temperature for 16 hours. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was distilled under reduced pressure to obtain compound 017b. LCMS: [M + H] + = 569.4.

[0565] Step 3:

[0566] Compound 017b (45 mg, 0.079 mmol), 017c (24 mg, 0.103 mmol), HATU (45 mg, 0.118 mmol) and N,N - diisopropylethylamine (41 mg, 0.316 mmol) were dissolved in dimethyl sulfoxide (2 mL). The mixture was stirred at room temperature overnight. LCMS showed that the reactants were completely converted and the product was detected. After the reaction solution was cooled to room temperature, water (50 mL) was added to quench it, and it was extracted with dichloromethane (30 mL * 2). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated in vacuo to obtain a residue. It was purified by Pre - TLC to obtain compound C017. LCMS: [M + H] + = 785.6. 1 H NMR (500 MHz, DMSO - d6) δ 12.06 (s, 1H), 10.42 (s, 1H), 8.96 (s, 1H), 8.85 (s, 1H), 8.39 (s, 1H), 8.30 (s, 1H), 8.24 (d, J = 6.7 Hz, 1H), 8.17–8.06 (m, 2H), 7.80 (d, J = 10.4 Hz, 1H), 7.74 (d, J = 6.8 Hz, 1H), 7.47 (q, J = 8.0 Hz, 2H), 4.47–4.39 (m, 1H), 4.07–3.90 (m, 2H), 3.87–3.79 (m, 2H), 3.27–3.10 (m, 5H), 2.82–2.56 (m, 6H), 2.38 (s, 3H), 2.33–2.06 (m, 6H), 2.03–1.73 (m, 7H), 1.66–1.48 (m, 4H), 1.18–1.04 (m, 3H).

[0567] Example 12: Preparation of compound C023

[0568]

[0569] Step 1:

[0570] 23a (1 g, 5.20 mmol) and 23b (645 mg, 10.39 mmol) were dissolved in NMP (20 mL), and NaH (416 mg, 10.39 mmol) was added. The temperature was raised to 110 °C and stirred at this temperature for 12 h. TLC showed that the starting materials were completely converted and the product was detected. The reaction solution was distilled under reduced pressure to obtain a residue. The residue was purified by column chromatography to obtain compound 23c.

[0571] Step 2:

[0572] Dissolve 23c (1.25 g, 5.73 mmol), B2Pin2 (2.91 g, 10.47 mmol), KOAc (1.69 g, 17.20 mmol) and Pd(dppf)Cl2 (465 mg, 0.57 mmol) in dioxane (25 mL) at room temperature. Replace nitrogen three times, then raise the temperature to 80 °C and stir at this temperature for 3 h. TLC shows that the raw materials are completely converted and the product is detected. After the reaction is completed, cool the reaction solution to room temperature, filter, concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 8d.

[0573] Step 3:

[0574] Dissolve 23d (345 mg, 1.30 mmol), 23e (400 mg, 0.87 mmol), BrettPhos Pd G3 (79 mg, 87 μmol) and K2CO3 (360 mg, 2.60 mmol) in DMSO / H2O (10 mL / 2 mL) at room temperature. Replace nitrogen three times, then raise the temperature to 100 °C and stir at this temperature for 12 h. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water to the reaction solution, extract with dichloromethane (50 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 23f.

[0575] Step 4:

[0576] Dissolve 23f (382 mg, 0.68 mmol) in ACN (10 mL), add IBX (379 mg, 1.36 mmol), raise the temperature to 80 °C and stir at this temperature for 2 h. LCMS shows that most of the raw materials have been converted and the product is detected. After the reaction is completed, cool the reaction solution to room temperature, filter, concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 23g.

[0577] Step 5:

[0578] Dissolve 23g (50 mg, 89 μmol) and 23h (36 mg, 0.18 mmol) in DMSO / DCM (2 mL / 2 mL) at room temperature, add 0.1 mL of acetic acid, and stir at room temperature for 1 hour. Then add sodium borohydride acetate (95 mg, 0.45 mmol) and stir at room temperature for 1 h. LCMS shows that most of the raw materials have been converted and the product is detected. Add 30 mL of water to the reaction solution, extract with dichloromethane (20 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Purify the residue by Prep-TLC to obtain compound 23i.

[0579] Step 6:

[0580] Dissolve 23i (15 mg, 20.1 μmol) in TFA (3 mL) and stir at room temperature for 1 hour. LCMS shows that the reactants are completely converted and the product is detected. Distill the reaction solution under reduced pressure to obtain the crude compound 23j.

[0581] Step 7:

[0582] Dissolve 23j (10 mg, 20.2 μmol), 23k (8 mg, 30.3 μmol), DIEA (8 mg, 60.5 μmol) and HATU (15 mg, 40.3 mmol) in DMSO (2 mL) and stir at room temperature for 12 hours. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water to the reaction solution, extract with dichloromethane (20 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Purify by preparative chromatography to obtain the final product C023. LCMS: Rt = 1.489 minutes, [M + H] + = 766.4.

[0583] Example 13: Preparation of compound C025

[0584]

[0585] Step 1:

[0586] Dissolve 25a (445 mg, 1.30 mmol), 8b (300 mg, 0.65 mmol), KOAc (320 mg, 3.25 mmol) and Pd(dppf)Cl2 (53 mg, 65.1 μmol) in DMSO (10 mL), displace nitrogen three times, raise the temperature to 100 °C and stir at this temperature for 3 h. LCMS shows that the starting materials are completely converted. Cool the reaction solution to room temperature, add BrettPhos G3 (59 mg, 65.1 μmol) and K2CO3 (270 mg, 1.95 mmol). Displace nitrogen three times, raise the temperature to 100 °C and stir at this temperature for 12 h. LCMS shows that the reactants are completely converted and the product is detected. Add 100 mL of water to the reaction solution, extract with ethyl acetate (100 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Distill the reaction solution under reduced pressure to obtain a residue. Purify the residue by column chromatography to obtain compound 25c.

[0587] Step 2:

[0588] Dissolve 25c (85 mg, 0.12 mmol) in TFA (3 mL) and stir at room temperature for 1 hour. LCMS shows that the reactants are completely converted and the product is detected. Distill the reaction solution under reduced pressure to obtain the crude compound 23j.

[0589] Step 3:

[0590] Dissolve 25e (1 g, 6.20 mmol) and 25f (1.59 g, 12.41 mmol) in dioxane / 60% KOH aq (3 mL / 0.2 mL), and stir at room temperature for 12 h. TLC shows that the reactants are completely converted and the product is detected. Add 30 mL of water to the reaction solution, extract with ethyl acetate (50 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 25g.

[0591] Step 4:

[0592] Dissolve 25g (1.09 g, 3.77 mmol) in MeOH (5 mL), add hydrochloric acid in methanol solution (10 mL), and stir at room temperature for 12 h. Distill the reaction solution under reduced pressure to obtain the crude compound 25h.

[0593] Step 5:

[0594] Dissolve 25h (425 mg, 3.62 mmol) and 25i (500 mg, 1.81 mmol) in MeOH / DIEA (5 mL / 1 mL), heat to 90 °C and stir at this temperature for 12 h. LCMS shows that the raw materials are completely converted. Cool the reaction solution to room temperature, add 30 mL of water to the reaction solution, adjust the pH to 7 with 1N HCl, extract with ethyl acetate (50 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Distill the reaction solution under reduced pressure to obtain a residue. Purify the residue by column chromatography to obtain compound 25j.

[0595] Step 6:

[0596] Dissolve 25j (32 mg, 82.0 μmol), 25d (25 mg, 54.6 μmol), DIEA (22 mg, 163.9 μmol) and HATU (42 mg, 109.3 μmol) in DMSO (2 mL), and stir at room temperature for 12 h. LCMS shows that the reactants are completely converted and the product is detected. Purify by preparative chromatography to obtain the final product C025. LCMS: Rt = 1.737 min, [M + H] + = 829.6.

[0597] Example 14: Preparation of compound C026

[0598]

[0599] Step 1:

[0600] Dissolve 26a (837 mg, 3.62 mmol) in DCM (10 mL), add TFA (1 mL), and stir at room temperature for 1 hour. Distill the reaction mixture under reduced pressure to obtain the crude compound 26b.

[0601] Step 2:

[0602] Dissolve 26b (474 mg, 3.62 mmol) and 26c (500 mg, 1.81 mmol) in MeOH / DIEA (5 mL / 1 mL), heat to 90 °C and stir at this temperature for 12 h. LCMS shows that the raw materials are completely converted. Cool the reaction mixture to room temperature, add 30 mL of water to the reaction mixture, adjust the pH to 7 with 1 N HCl, extract with ethyl acetate (50 mL × 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Distill the reaction mixture under reduced pressure to obtain a residue. Purify the residue by column chromatography to obtain compound 26d.

[0603] Step 3:

[0604] Dissolve 26d (32 mg, 82.0 μmol), 25d (25 mg, 54.6 μmol), DIEA (22 mg, 163.9 μmol), and HATU (42 mg, 109.3 μmol) in DMSO (2 mL), and stir at room temperature for 12 h. LCMS shows that the reactants are completely converted and the product is detected. Purify by preparative chromatography to obtain the final product C026. LCMS: Rt = 1.902 minutes, [M+H] + = 827.1.

[0605] Example 15: Preparation of compound C027

[0606]

[0607] Step 1:

[0608] Dissolve compound 027a (900 mg, 2.64 mmol) in HCl / dioxane (10 mL, 4 M) at room temperature and stir for 1 hour. LCMS shows that the reactants are completely converted and the product is detected. Distill the reaction mixture under reduced pressure to obtain compound 027b. LCMS: [M+H] + = 242.1.

[0609] Step 2:

[0610] Compound 027b (731 mg, 2.62 mmol) and DIEA (339 mg, 2.62 mmol) were dissolved in dichloromethane / dimethyl sulfoxide (5 mL / 5 mL) and stirred for 10 min. Subsequently, compound 027c (672 mg, 3.15 mmol) and acetic acid (157 mg, 2.62 mmol) were added to the reaction mixture, and the reaction was stirred at 35 °C for 16 h. Then, NaBH(OAc)3 (1.67 g, 7.87 mmol) was added to the reaction mixture and stirred at 35 °C for 1 h. LCMS showed that the reactants were completely converted and the product was detected. The reaction mixture was poured into saturated sodium bicarbonate solution (60 mL), and extracted with dichloromethane (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated in vacuo to obtain the crude product. The crude product was purified by flash silica gel column chromatography to obtain compound 027d. LCMS: [M+H] + = 439.2.

[0611] Step 3:

[0612] Compound 027d (510 mg, 1.16 mmol), potassium acetate (228 mg, 2.33 mmol), bis(pinacolato)diboron (384 mg, 1.51 mmol) and Pd(dppf)Cl2 (85 mg, 0.116 mmol) were dissolved in dimethyl sulfoxide (5 mL). The reaction mixture was stirred at 100 °C for 2 h under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. After the reaction mixture was cooled to room temperature, water (60 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (20 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated in vacuo to obtain the crude product. The crude product was purified by flash silica gel column chromatography to obtain compound 027e. LCMS: [M+H] + = 405.2.

[0613] Step 4:

[0614] Compound 027e (54 mg, 0.159 mmol), warhead 3 (50 mg, 0.097 mmol), potassium carbonate (44 mg, 0.318 mmol) and BrettPhos-Pd-G3 (49 mg, 0.005 mmol) were dissolved in dimethyl sulfoxide (2 mL). The reaction mixture was stirred at 100 °C for 3 h under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. After the reaction mixture was cooled to room temperature, water (50 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (20 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated in vacuo to obtain the crude product. The crude product was purified by flash silica gel column chromatography to obtain compound 027f. LCMS: [M+H] + = 655.4.

[0615] Step 5:

[0616] Compound 027f (25 mg, 0.0382 mmol) was dissolved in trifluoroformic acid / dichloromethane (1 mL / 5 mL) at room temperature and stirred for 2 hours. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was distilled under reduced pressure to obtain compound 027g. LCMS: [M+H] + = 555.2

[0617] Step 6:

[0618] Compound 027g (21 mg, 0.038 mmol), 027h (12 mg, 0.045 mmol), DIEA (24 mg, 0.189 mmol) and HATU (19 mg, 0.049 mmol) were dissolved in dimethyl sulfoxide (2 mL). The mixture was stirred at room temperature for 16 hours. LCMS showed that the reactants were completely converted and the product was detected. After the reaction solution was cooled to room temperature, water (50 mL) was added for quenching, and the mixture was extracted with dichloromethane (30 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain a crude product. The crude product was purified by Pre-TLC to obtain compound C027. LCMS: [M+H] + = 805.5

[0619] Example 16: Preparation of compound C028

[0620]

[0621] Step 1:

[0622] 28a (50 mg, 89 μmol) and 28b (38 mg, 0.18 mmol) were dissolved in DMSO / DCM (2 mL / 2 mL) at room temperature. 0.1 mL of acetic acid was added, and the mixture was stirred at room temperature for 1 hour. Sodium triacetoxyborohydride (95 mg, 0.45 mmol) was added, and the mixture was stirred at room temperature for 1 h. LCMS showed that most of the starting materials had been converted and the product was detected. 30 mL of water was added to the reaction solution, and the mixture was extracted with dichloromethane (20 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by Prep-TLC to obtain compound 28c.

[0623] Step 2:

[0624] 28c (15 mg, 19.7 μmol) was dissolved in TFA (3 mL) and stirred at room temperature for 1 hour. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was distilled under reduced pressure to obtain crude compound 28d.

[0625] Step 3:

[0626] Dissolve 28d (10 mg, 18.9 μmol), 28e (8 mg, 28.3 μmol), DIEA (8 mg, 56.6 μmol) and HATU (15 mg, 37.8 μmol) in DMSO (2 mL), and stir at room temperature for 12 hours. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water to the reaction solution, extract with dichloromethane (20 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Purify by preparative chromatography to obtain the final product C028. LCMS: Rt = 1.522 minutes, [M + H] + = 780.5.

[0627] Example 17: Preparation of Compound C030

[0628]

[0629] Step 1:

[0630] Dissolve 30a (2 g, 5.86 mmol), B2Pin2 (2.98 mg, 11.72 mmol), KOAc (1.73 g, 17.58 mmol) and Pd(dppf)Cl2 (475 mg, 0.59 mmol) in dioxane (40 mL) at room temperature, displace nitrogen three times, raise the temperature to 90 °C and stir at this temperature for 3 h. TLC shows that the starting materials are completely converted and the product is detected. After the reaction is completed, the reaction solution is cooled to room temperature, filtered, and the filtrate is concentrated under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 30b.

[0631] Step 2:

[0632] Dissolve 30b (631 mg, 1.63 mmol), 30c (500 mg, 1.08 mmol), BrettPhos Pd G3 (98 mg, 0.11 mmol) and K2CO3 (450 mg, 3.25 mmol) in DMSO / H2O (10 mL / 2 mL) at room temperature, displace nitrogen three times, raise the temperature to 90 °C and stir at this temperature for 12 h. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water to the reaction solution, extract with dichloromethane (50 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 30d.

[0633] Step 3:

[0634] Dissolve 28c (354 mg, 0.52 mmol) in TFA (3 mL) and stir at room temperature for 1 hour. LCMS shows that the reactants are completely converted and the product is detected. Distill the reaction solution under reduced pressure to obtain the crude compound 30e.

[0635] Step 4:

[0636] Dissolve 30e (50 mg, 0.11 mmol) and 30f (53 mg, 0.22 mmol) in DMSO / DCM (2 mL / 2 mL) at room temperature, add 0.1 mL of acetic acid, and stir at room temperature for 1 hour. Add sodium triacetoxyborohydride (70 mg, 0.33 mmol), and stir at room temperature for 1 h. LCMS shows that most of the starting materials have been converted and the product is detected. Add 30 mL of water to the reaction solution, extract with dichloromethane (20 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate in vacuo to obtain a residue. Purify the residue by Prep-TLC to obtain compound 30g.

[0637] Step 5:

[0638] Dissolve 30g (57 mg, 83.8 μmol) in DCM (3 mL), add TFA (1 mL), and stir at room temperature for 1 hour. LCMS shows that the reactants are completely converted and the product is detected. Distill the reaction solution under reduced pressure to obtain the crude compound 30h.

[0639] Step 6:

[0640] Dissolve 30h (49 mg, 82.8 μmol), 30i (33 mg, 124.2 μmol), DIEA (32 mg, 248.4 μmol) and HATU (63 mg, 165.6 μmol) in DMSO (2 mL), and stir at room temperature for 1 hour. LCMS shows that the reactants are completely converted and the product is detected. Add 30 mL of water to the reaction solution, extract with dichloromethane (20 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate in vacuo to obtain a residue. Purify the residue by Prep-TLC to obtain the final product C030. LCMS: Rt = 1.570 min, [M + H] + = 830.6.

[0641] Example 18: Preparation of compound C032

[0642]

[0643] Step 1:

[0644] 032a (100 mg, 0.212 mmol) and 032b (66 mg, 0.275 mmol) were dissolved in dichloromethane / dimethyl sulfoxide (4 mL / 4 mL), and the mixture was stirred at 35 °C for 16 h. Then sodium triacetoxyborohydride (225 mg, 1.06 mmol) was added to the mixture and stirring was continued at 35 °C for 2 h. LCMS showed complete conversion of the reactants and the product was detected. After the reaction solution was cooled to room temperature, saturated ammonium chloride solution (50 mL) was added for quenching, and extraction was carried out with dichloromethane (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated in vacuo to obtain a residue. The residue was purified by silica gel chromatography to obtain compound 032c. LCMS: [M+H] + = 695.5.

[0645] Step 2:

[0646] 032c (60 mg, 0.086 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added at room temperature, and the mixture was stirred for 2 h. LCMS showed complete conversion of the reactants and the product was detected. The reaction solution was distilled under reduced pressure to obtain compound 032d. LCMS: [M+H] + = 595.4.

[0647] Step 3:

[0648] 32d (51 mg, 0.086 mmol), 032e (24 mg, 0.087 mmol), and DIEA (55 mg, 0.426 mmol) were dissolved in dimethyl sulfoxide (2 mL) at room temperature, and the reaction solution was stirred at 80 °C for 2 h. LCMS showed complete conversion of the reactants and the product was detected. After the reaction solution was cooled to room temperature, water (50 mL) was added for quenching, and extraction was carried out with methanol / dichloromethane (20 mL / 20 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated in vacuo to obtain a residue. The residue was purified by Pre-TLC to obtain compound C032. LCMS: [M+H] + = 851.6. 11H NMR (500 MHz, DMSO-d6) δ 12.04 (s, 1H), 11.09 (s, 1H), 8.96 (d, J = 2.2 Hz, 1H), 8.84 (s, 1H), 8.38 (s, 1H), 8.30–8.28 (m, 1H), 8.11 (d, J = 2.5 Hz, 1H), 8.09 (s, 1H), 7.70–7.66 (m, 1H), 7.34 (dd, J = 11.7, 7.9 Hz, 2H), 5.09 (dd, J = 12.6, 5.5 Hz, 2H), 4.24 (s, 2H), 3.26 (d, J = 3.7 Hz, 2H), 3.23–3.10 (m, 6H), 3.01–2.96 (m, 2H), 2.92–2.74 (m, 4H), 2.66–2.56 (m, 2H), 2.46 (s, 2H), 2.39–2.33 (m, 4H), 2.11–1.96 (m, 8H), 1.80–1.72 (m, 2H), 1.71–1.60 (m, 4H), 1.04 (t, J = 7.2 Hz, 3H).

[0649] Example 19: Preparation of Compound C033

[0650]

[0651] Step 1:

[0652] Dissolve 33a (1 g, 5.68 mmol) and 33b (1.37 g, 8.52 mmol) in THF (20 mL), add NaH (570 mg, 14.21 mmol), and stir at room temperature for 12 h. TLC shows that the raw materials are completely converted and the product is detected. Add 100 mL of water to the reaction solution, extract with ethyl acetate (100 mL * 3), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate under vacuum to obtain a residue. Distill the reaction solution under reduced pressure to obtain a residue. Purify the residue by column chromatography to obtain compound 33c.

[0653] Step 2:

[0654] Dissolve 33c (1.37 g, 4.32 mmol), B2Pin2 (2.19 g, 8.64 mmol), KOAc (1.27 g, 12.96 mmol), and Pd(dppf)Cl2 (350 mg, 0.43 mmol) in dioxane (30 mL) at room temperature, displace nitrogen three times, raise the temperature to 80 °C, and stir at this temperature for 3 h. TLC shows that the raw materials are completely converted and the product is detected. After the reaction is completed, cool the reaction solution to room temperature, filter, and concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography to obtain compound 33d.

[0655] Step 3:

[0656] 33d (80 mg, 0.22 mmol), 33e (75 mg, 0.15 mmol), BrettPhos Pd G3 (14 mg, 15 μmol), and K2CO3 (60 mg, 0.44 mmol) were dissolved in DMSO / H2O (2 mL / 0.5 mL) at room temperature. The mixture was purged with nitrogen three times, then heated to 90 °C and stirred at this temperature for 12 h. LCMS showed that the reactants were completely converted and the product was detected. 30 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (50 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography to obtain compound 33f.

[0657] Step 4:

[0658] 33g (69 mg, 0.13 mmol) was dissolved in DCM (3 mL), and TFA (1 mL) was added. The mixture was stirred at room temperature for 1 h. LCMS showed that the reactants were completely converted and the product was detected. The reaction mixture was distilled under reduced pressure to obtain crude compound 33h.

[0659] Step 5:

[0660] 33g (56 mg, 0.13 mmol), 33h (53 mg, 0.19 mmol), NMI (37 mg, 0.45 mmol), and TCFH (44 mg, 0.16 mmol) were dissolved in DMSO (2 mL). The mixture was stirred at room temperature for 1 h. LCMS showed that the reactants were completely converted and the product was detected. 30 mL of water was added to the reaction mixture, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by Prep-TLC to obtain the final product C033. LCMS: Rt = 1.719 min, [M+H] + = 683.4.

[0661] Example 20: Preparation of compound C034

[0662]

[0663] Step 1:

[0664] 034a (1 g, 5.35 mmol), 034b (1.2 g, 5.35 mmol), and cesium carbonate (1.74 g, 5.35 mmol) were dissolved in acetonitrile (10 mL). The reaction mixture was stirred at room temperature for 16 h. TLC showed that the reactants were completely converted and a new spot was detected. The reaction mixture was distilled under reduced pressure to obtain a crude product. The crude product was purified by silica gel chromatography to obtain compound 034c.

[0665] Step 2:

[0666] Dissolve 034c (1.212 g, 3.67 mmol) in dimethyl sulfoxide (5 mL), add potassium acetate (1.08 g, 11.01 mmol), bis(pinacolato)diboron (1.21 g, 4.77 mmol) and Pd(dppf)Cl2 (0.27 g, 0.367 mmol) at room temperature. The reaction mixture was stirred at 100 °C for 2 hours under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. The reaction mixture was distilled under reduced pressure to obtain the crude product. It was purified by silica gel chromatography to obtain compound 034d. LCMS: [M - Bu + H] + = 278.1

[0667] Step 3:

[0668] Mix 034d (64 mg, 0.176 mmol), warhead 3 (70 mg, 0.136 mmol), potassium carbonate (56 mg, 0.407 mmol) and BrettPhos-Pd-G3 (12 mg, 0.0136 mmol). The reaction mixture was stirred at 90 °C for 16 hours under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. After the reaction mixture was cooled to room temperature, water (50 mL) was added for quenching, and it was extracted with dichloromethane (20 mL × 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product. It was purified by silica gel chromatography to obtain compound C034e. LCMS: [M + H] + = 546.4

[0669] Step 4:

[0670] Dissolve compound 034e (42 mg, 0.077 mmol) in trifluoroacetic acid / dichloromethane (1 mL / 5 mL) at room temperature and stir for 2 hours. LCMS showed that the reactants were completely converted and the product was detected. The reaction mixture was distilled under reduced pressure to obtain compound 034f. LCMS: [M + H] + = 446.3

[0671] Step 5:

[0672] Compound 034f (34 mg, 0.076 mmol), 034g (25 mg, 0.092 mmol), DIEA (50 mg, 0.381 mmol) and HATU (38 mg, 0.099 mmol) were dissolved in dimethyl sulfoxide (2 mL), and the mixture was stirred at room temperature for 16 h. LCMS showed that the reactants were completely converted and the product was detected. After the reaction solution was cooled to room temperature, water (50 mL) was added for quenching, and the mixture was extracted with dichloromethane (20 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product. The crude product was purified by Pre-TLC to obtain compound C034. LCMS: Rt = 2.200 min; [M+H] + = 696.4.

[0673] Example 21: Preparation of compound C038

[0674]

[0675] According to the method described in Example 19, except that 38b was used instead of 33b in step 1, the final product C038 was obtained. LCMS: Rt = 1.796 min, [M+H] + = 711.4.

[0676] Example 22: Preparation of compound C039

[0677]

[0678] According to the method described in Example 20, except that N-Boc-3-aminopropyl bromide was used instead of 034b and DMF was used instead of acetonitrile in step 1, compound C039 was obtained. LCMS: [M+H] + = 710.4. 11H NMR (500 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.53 (s, 1H), 8.81 (s, 1H), 8.76 (t, J = 5.6 Hz, 1H), 8.36 (s, 1H), 8.12 (s, 1H), 8.08 (d, J = 2.7 Hz, 1H), 8.03 (d, J = 8.2 Hz, 2H), 7.99 (d, J = 2.1 Hz, 1H), 7.87 (dd, J = 8.4, 2.1 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.09 (d, J = 8.2 Hz, 1H), 4.27 (s, 2H), 4.15 (d, J = 6.0 Hz, 2H), 3.76 (ddd, J = 13.6, 8.2, 5.8 Hz, 1H), 3.63 (dt, J = 11.8, 6.0 Hz, 1H), 3.54–3.47 (m, 2H), 3.25 (s, 1H), 3.05 (s, 2H), 2.75 (q, J = 7.2, 6.6 Hz, 2H), 2.30 (s, 3H), 2.21–1.96 (m, 8H), 1.12–1.03 (m, 3H).

[0679] Compound 039b: LCMS: [M - Bu + H] + = 292.2. Compound 039c: LCMS: [M + H] + = 560.3. Compound 039d: LCMS: [M + H] + = 460.3.

[0680] Example 23: Preparation of Compound C040

[0681]

[0682] Except that in step 1, 4-(N-Boc-amino)-1-butanol was used instead of 034b, THF was used to replace acetonitrile, and the reaction was carried out under an ice bath, according to the method described in Example 20, Compound C040 was obtained. LCMS: Rt = 1.998 min; [M + H] + = 724.4.

[0683] Compound 039b: LCMS: [M - Bu + H] + = 292.2. Compound 040c: LCMS: [M + H] + = 560.3. Compound 040d: LCMS: [M + H] + = 460.4.

[0684] Example 24: Preparation of Compound C041

[0685]

[0686] Compound C041 was obtained according to the method described in Example 19, except that 41b was used instead of 33b in Step 1. LCMS: Rt = 1.911 min, [M+H] + = 725.4.

[0687] Example 25: Preparation of Compound C042

[0688]

[0689] Step 1:

[0690] Compound 042a (21 mg, 0.046 mmol), 042b (14 mg, 0.059 mmol), DIEA (30 mg, 0.23 mmol) and HATU (23 mg, 0.059 mmol) were dissolved in dimethyl sulfoxide (2 mL). The mixture was stirred at room temperature for 16 h. LCMS showed complete conversion of the reactants and the product was detected. After the reaction solution was cooled to room temperature, water (50 mL) was added for quenching, and the mixture was extracted with dichloromethane (20 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product. The crude product was purified by Pre-TLC to obtain Compound C042. LCMS: [M+H] + = 676.4.

[0691] 1 H NMR (500 MHz, DMSO-d6) δ 12.02 (s, 1H), 10.43 (s, 1H), 8.82 (s, 1H), 8.68 (d, J = 5.6 Hz, 1H), 8.38 (s, 1H), 8.15 (s, 1H), 8.12 (d, J = 2.4 Hz, 1H), 8.04 (d, J = 7.5 Hz, 2H), 7.82 (d, J = 1.8 Hz, 1H), 7.76–7.72 (m, 1H), 7.53–7.44 (m, 2H), 7.09 (d, J = 8.7 Hz, 1H), 4.14 (t, J = 6.0 Hz, 2H), 3.83 (t, J = 6.7 Hz, 2H), 3.61–3.47 (m, 4H), 3.26 (s, 1H), 3.15–3.07 (m, 2H), 2.72 (t, J = 6.6 Hz, 2H), 2.31 (s, 3H), 2.07 (dd, J = 12.5, 6.1 Hz, 2H), 1.28 (dd, J = 16.9, 6.6 Hz, 6H), 1.25–1.21 (m, 3H).

[0692] Example 26: Preparation of Compound C044

[0693]

[0694] Compound C044 was obtained according to the method described in Example 19, except that 44b was used instead of 33b in Step 1. LCMS: Rt = 1.767 min, [M+H] + = 697.4.

[0695] Example 27: Preparation of Compound C091

[0696]

[0697] Step 1:

[0698] Dissolve N-Boc-4-methylenepiperidine (2 g, 10.14 mmol) in 9-BBN (20 mL) (0.5 M, dissolved in THF) at room temperature. Stir the mixture at 60 °C for 3 h under a nitrogen atmosphere. Then, distill off THF under reduced pressure, add DMF (N,N-dimethylformamide) (S0, 20 mL) and water (S1, 2 mL) as solvents, and then add 1-bromo-4-iodobenzene (2.6 g, 9.19 mmol), potassium carbonate (2.8 g, 20.28 mmol), and Pd(dppf)Cl2 (0.74 g, 1.01 mmol). Stir the mixture at 60 °C for 2 h under nitrogen protection. TLC shows that the reactants are completely converted and a new spot is formed. Pour the reaction solution into water (100 mL), and then extract with ethyl acetate (30 mL × 2). Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product. Purify by silica gel chromatography to obtain Compound 91a.

[0699] Step 2:

[0700] Dissolve Compound 91a (1.32 g, 3.73 mmol) in dioxane (20 mL) at room temperature. Add bis(pinacolato)diboron (1.42 g, 5.59 mmol), potassium acetate (1.10 g, 11.19 mmol), and Pd(dppf)Cl2 (0.27 g, 0.37 mmol). Stir the mixture at 100 °C for 2 h under a nitrogen atmosphere. TLC shows that the reactants are completely converted and a new spot is detected. Pour the reaction solution into water (50 mL), and then extract with ethyl acetate (30 mL × 2). Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product. Purify by silica gel chromatography to obtain Compound 91b.

[0701] Step 3:

[0702] Compound 91b (1.5 g, 3.73 mmol) was dissolved in toluene (10 mL) and water (5 mL) at room temperature. 5-Bromo-2-iodopyridine (1.32 g, 4.65 mmol), sodium carbonate (1.35 g, 12.69 mmol) and tetrakis(triphenylphosphine)palladium (0.24 g, 0.21 mmol) were added. The mixture was stirred at 90 °C for 16 h under a nitrogen atmosphere. TLC showed that the reactants were completely converted and a new spot was detected. The reaction solution was poured into water (50 mL), and then extracted with ethyl acetate (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography to obtain compound 91c.

[0703] Step 4:

[0704] Compound 91c (100 mg, 0.23 mmol) was dissolved in dioxane (5 mL) at room temperature. Potassium acetate (68 mg, 0.69 mmol), bis(pinacolato)diboron (88 mg, 0.35 mmol) and Pd(dppf)Cl2 (0.017 g, 0.023 mmol) were added. The mixture was stirred at 100 °C for 2 h under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was filtered, and the filtrate was concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography to obtain compound 91d. LCMS: [M+H] + = 397.2.

[0705] Step 5:

[0706] Compound warhead3 (95 mg, 0.18 mmol) was dissolved in DMSO (2 mL) and water (0.2 mL) at room temperature. Compound 91d (90 mg, 0.23 mmol), potassium carbonate (75 mg, 0.54 mmol) and BrettPhos-G3-Pd (16 mg, 0.018 mmol) were added. The mixture was stirred at 90 °C for 16 h under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was poured into water (50 mL), and then extracted with DCM (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography to obtain compound 91e. LCMS: [M+H] + = 647.5.

[0707] Step 6:

[0708] Compound 91e (118 mg, 0.18 mmol) was dissolved in dichloromethane (10 mL) at room temperature. Trifluoroacetic acid (1 mL) was added. The mixture was stirred at room temperature for 2 h. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was rotary evaporated to obtain crude compound 91f. LCMS: [M+H]+ = 547.3。

[0709] Step 7:

[0710] Dissolve compound 91f (33 mg, 0.060 mmol) in DMSO (2 mL) at room temperature, add 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (21 mg, 0.078 mmol), HATU (30 mg, 0.079 mmol) and DIPEA (R3, 39 mg, 0.30 mmol). Stir the mixture at room temperature under a nitrogen atmosphere for 16 hours. LCMS shows that the reactants are completely converted and the product is detected. Pour the reaction solution into water (50 mL), then extract with DCM (30 mL * 2). Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product. Purify by Pre-TLC to obtain compound C091 (9.69 mg, yield: 18.72%).

[0711] LCMS: [M + H] + = 797.4; 1 H NMR (500 MHz, DMSO-d6) δ 12.15 (s, 1H), 10.51 (s, 1H), 9.52 (d, J =

[0712] 1.7 Hz, 1H), 9.02 (s, 1H), 8.68 (dd, J = 8.4, 2.2 Hz, 1H), 8.42 (s, 1H), 8.18 (d, J = 2.6 Hz, 1H), 8.15–8.07 (m, 4H), 7.64 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 1.9 Hz, 1H), 7.39 (dd, J = 8.2, 1.9 Hz, 1H), 7.35 (d, J = 8.2 Hz, 2H), 4.50–4.41 (m, 1H), 4.29–4.23 (m, 1H), 3.79–3.73 (m, 1H), 3.66–3.52 (m, 2H), 3.01 (d, J = 10.6 Hz, 3H), 2.75 (q, J = 7.5, 6.8 Hz, 3H), 2.62 (dt, J = 13.7, 6.3 Hz, 3H), 2.39 (q, J = 7.3 Hz, 3H), 2.11–1.99 (m, 6H), 1.88 (s, 1H), 1.72 (s, 1H), 1.64–1.57 (m, 1H), 1.04 (t, J = 7.2 Hz, 3H).

[0713] Example 28: Preparation of compound C096

[0714]

[0715] Compound 91f (33 mg, 0.060 mmol) was dissolved in DMSO (2 mL) at room temperature. 2-(2,6-Dioxopiperidin-3-yl)-4-fluoroisoindoline-1,3-dione (22 mg, 0.078 mmol) and DIPEA (39 mg, 0.30 mmol) were added. The mixture was stirred at 80 °C for 4 h under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was poured into water (50 mL), and then extracted with DCM (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product. C096 (14.37 mg, yield: 28.06%) was obtained by Pre-TLC (MeOH / DCM = 13%). LCMS: [M+H] + = 803.5; 1 H NMR (500 MHz, DMSO-d6) δ 12.16 (s, 1H), 11.09 (s, 1H), 9.53 (s, 1H), 9.03 (s, 1H), 8.68 (d, J = 8.1 Hz, 1H), 8.43 (s, 1H), 8.14 (dd, J = 19.3, 11.7 Hz, 5H), 7.67 (t, J = 7.8 Hz, 1H), 7.38 (d, J = 7.9 Hz, 2H), 7.35–7.25 (m, 2H), 5.09 (dd, J = 12.7, 5.4 Hz, 1H), 4.31–4.23 (m, 1H), 3.70 (d, J = 9.5 Hz, 2H), 3.02 (d, J = 9.9 Hz, 2H), 2.91–2.80 (m, 3H), 2.67 (d, J = 6.0 Hz, 2H), 2.59 (d, J = 18.4 Hz, 1H), 2.45–2.39 (m, 2H), 2.13–1.95 (m, 7H), 1.80–1.70 (m, 3H), 1.56–1.39 (m, 3H), 1.05 (t, J = 7.0 Hz, 3H).

[0716] Example 29: Preparation of Compound C084

[0717]

[0718] The first step

[0719] Compound C084a (2 g, 10.31 mmol) was dissolved in DMF (10 mL) at room temperature. Cesium carbonate (4.03 g, 12.37 mmol) and 1-methylpiperidin-4-yl 4-methylbenzenesulfonate (1.61 g, 11.34 mmol) were added, and the mixture was stirred at room temperature for 16 h. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was filtered, and the filtrate was concentrated in vacuo to obtain crude compound C084b (2.14 g, yield 99.79%).

[0720] The second step

[0721] Compound C084c (530 mg, 1.21 mmol) was dissolved in dioxane (20 mL) at room temperature. Bis(pinacolato)diboron (368.72 mg, 1.45 mmol), potassium acetate (356 mg, 3.63 mmol) and Pd(dppf)Cl2·DCM (99 mg, 0.12 mmol) were added, and the mixture was stirred at 90 °C for 6 h under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, and the filter cake was washed with dioxane (20 mL). The filtrate was taken to obtain crude compound C084d (480 mg, 1.188 mmol), which was directly used for the next step. LCMS: [M+H] + = 404.3.

[0722] The third step

[0723] Compound C084d (52 mg, 0.12 mmol) was dissolved in DMSO (2 mL) and water (0.2 mL) at room temperature. C084e (62 mg, 0.12 mmol), potassium carbonate (50 mg, 0.36 mmol) and BrettPhos-G3-Pd (10 mg, 0.011 mmol) were added, and the mixture was stirred at 90 °C for 16 h under a nitrogen atmosphere. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was poured into water (30 mL), then extracted with dichloromethane (20 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography (methanol:dichloromethane = 0-14%) to obtain compound C084f (37 mg, 0.055 mmol).

[0724] The fourth step

[0725] Compound C084f (30 mg, 0.054 mmol) was dissolved in dichloromethane (5 mL) at room temperature, and TFA (1 mL) was added. The mixture was stirred at room temperature for 2 h. LCMS showed that the reactant was completely converted and the product was detected. The reaction solution was poured into water (30 mL), and then extracted with dichloromethane (20 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain crude compound C084g (24 mg, 0.053 mmol).

[0726] The fifth step

[0727] Compound C084g (24 mg, 0.053 mmol) was dissolved in DMSO (2 mL) at room temperature, and 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (19 mg, 0.071 mmol), HATU (27 mg, 0.071 mmol) and DIPEA (35 mg, 0.27 mmol) were added. The mixture was stirred at room temperature for 16 h. LCMS showed that the reactant was completely converted and the product was detected. The reaction solution was poured into water (50 mL), and then extracted with DCM (30 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product. It was purified by Pre-HPLC to obtain compound C084 (7.06 mg, yield: 18.07%). LCMS: [M+H] + = 792.4.

[0728] Example 30: Preparation of compound C085

[0729]

[0730] The first step

[0731] Compound C085a (1.8 g, 6.07 mmol), C085b (1.6 g, 9.11 mmol), and K2CO3 (2.1 g, 15.18 mmol) were dissolved in DMSO (40 mL). Nitrogen was displaced three times, and the temperature was raised to 100 °C and stirred at this temperature for 4 h. TLC showed that the raw materials were completely converted and the product was detected. 50 mL of water was added to the reaction solution, and it was extracted with EA (50 mL×3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (PE:EA = 10:1) to obtain compound C085c (1.09 g, 39.7% yield).

[0732] The second step

[0733] Compound C085c (1.09 g, 2.41 mmol), KOAc (0.71 g, 7.23 mmol), B2pin2 (1.22 g, 4.82 mmol), and Pd(dppf)Cl2 (176 mg, 0.24 mmol) were dissolved in dioxane (20 mL). The mixture was purged with nitrogen three times and then heated to 90 °C and stirred for 2 h. LCMS showed complete conversion of the reactants and the detection of the product. After the reaction, the mixture was filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (DCM:MeOH = 20:1) to give C085d (180 mg, 15.0% yield).

[0734] The third step

[0735] C085d (119 mg, 0.29 mmol), warhead 3 (100 mg, 0.19 mmol), Pd(PPh3)4 (22 mg, 0.02 mmol), and K2CO3 (79 mg, 0.57 mmol) were dissolved in DMSO and H2O (3 mL / 0.5 mL). The mixture was purged with nitrogen three times and then heated to 90 °C and stirred for 12 h. LCMS showed complete conversion of the reactants and the detection of the product. After the reaction, the mixture was filtered, 50 mL of water was added, and the mixture was extracted with DCM (30 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by column chromatography (DCM:MeOH = 4:1) to give C085e (72 mg, 56.3% yield).

[0736] The fourth step

[0737] C085e (73 mg, 0.11 mmol) was dissolved in DCM and TFA (3 mL / 0.5 mL), and the mixture was stirred at room temperature for 1 h. LCMS showed complete conversion of the reactants and the detection of the product. The reaction mixture was distilled under reduced pressure to obtain the crude compound C085f.

[0738] The fifth step

[0739] C085f, C085g (44.3 mg, 0.17 mmol), and NMI (31.6 mg, 0.39 mmol) were dissolved in DMSO (2 mL). Under stirring, TCFH (337 mg, 0.13 mmol) was added, and the mixture was stirred at room temperature for 2 h. LCMS showed complete conversion of the reactants and the detection of the product. 50 mL of water was added to the reaction mixture, and the mixture was extracted with DCM (30 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The residue was purified by preparative plate (DCM:NH3 / MeOH = 12:1) to give the final product C085 (31.54 mg, 35.06% yield). LCMS: Rt = 1.817 min, [M+H] + = 818.6

[0740] Example 31: Preparation of Compound C092

[0741]

[0742] Except that C092e was used instead of C084e in the third step, according to the method described in Example 29, Compound C092 (7.06 mg, yield: 18.07%) was obtained. LCMS: [M+H] + = 791.4.

[0743] Example 32: Preparation of Compound C098

[0744]

[0745] Except that 98a was used instead of C085a in the first step, Compound C098 was obtained by a method similar to that described in Example 30. LCMS: Rt = 2.120 min; [M+H] + = 820.5.

[0746] Example 33: Preparation of Compound C108

[0747]

[0748] First Step

[0749] Compound 108-1 (2 g, 10.31 mmol), Compound 108-2 (3.72 g, 20.62 mmol), Cs2CO3 (10.08 g, 30.93 mmol), and KI (0.17 g, 1.03 mmol) were dissolved in DMF (40 mL). Nitrogen was displaced three times, and the mixture was heated to 60 °C and stirred at this temperature for 12 h. TLC showed that the raw materials were completely converted and the product was detected. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (PE:EA = 1:3) to obtain Compound 108-3 (3.03 g).

[0750] Second Step

[0751] Compound 108-3 (600 mg, 2.16 mmol), compound 108-4 (500 mg, 1.08 mmol), Pd(dppf)Cl2 (176 mg, 0.22 mmol), and K2CO3 (299 mg, 2.16 mmol) were dissolved in dioxane and H2O (10 mL / 3 mL). Nitrogen was displaced three times, and the mixture was heated to 80 °C and stirred for 3 h. LCMS showed that the reactants were completely converted and the product was detected. After the reaction, the mixture was filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (PE:EA = 1:1) to obtain compound 108-5 (280 mg, 53.2% yield).

[0752] The third step

[0753] Compound 108-5 (100 mg, 0.2 mmol), compound 108-6 (120 mg, 0.3 mmol), BrettPhos PdG3 (18 mg, 0.02 mmol), and K2CO3 (83 mg, 0.6 mmol) were dissolved in dioxane and H2O (3 mL / 0.5 mL). Nitrogen was displaced three times, and the mixture was heated to 90 °C and stirred for 12 h. LCMS showed that the reactants were completely converted and the product was detected. After the reaction, the mixture was filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (pure EA) to obtain compound 108-7 (87 mg, 67.8% yield).

[0754] The fourth step

[0755] Compound 108-7 (87 mg, 0.14 mmol) was dissolved in DCM and TFA (3 mL / 0.5 mL), and the mixture was stirred at room temperature for 1 h. LCMS showed that the reactants were completely converted and the product was detected. The reaction mixture was distilled under reduced pressure to obtain crude compound 108-8.

[0756] The fifth step

[0757] Compound 108-8, compound 108-9 (56.4 mg, 0.21 mmol), and NMI (40 mg, 0.49 mmol) were dissolved in DMSO (2 mL). TCFH (47 mg, 0.17 mmol) was added with stirring, and the mixture was stirred at room temperature for 2 h. LCMS showed that the reactants were completely converted and the product was detected. The final product C108 (37.73 mg, 34.85% yield) was obtained by preparative chromatography. LCMS: Rt = 2.000 min, [M+H] + = 777.5, purity = 99.51%.

[0758] Example 34: Preparation of compound C109

[0759]

[0760] Compound C109 (19.01 mg, 34.85% yield, purity = 99.14%) was prepared according to the method described in Example 33, except that compound 109-2 was used instead of 108-1 in the first step. LCMS: Rt = 2.145 min; [M+H] + = 765.3.

[0761] Example 35: Preparation of Compound C110

[0762]

[0763] Compound C110 (7.06 mg, yield: 18.07%) was prepared according to the method described in Example 29, except that methyl iodide was used instead of 1-methylpiperidin-4-yl 4-methylbenzenesulfonate in the first step. LCMS: [M+H] + = 707.4.

[0764] C110a: LCMS: M+H] + = 208.1. C110c: LCMS: [M+H] + = 404.3. C110f: LCMS: [M+H] + = 457.1.

[0765] Example 36: Preparation of Compound C122

[0766]

[0767] First Step

[0768] Ph3PMeBr (12.56 g, 35.16 mmol) was dissolved in THF (40 mL), and NaH (1.88 g, 46.88 mmol) was added with stirring in an ice bath. The nitrogen was displaced three times, and the temperature was raised to 90 °C and stirred at this temperature for 3 h. Compound C122-1 (5 g, 23.44 mmol) was dissolved in THF (40 mL), and after cooling to room temperature, it was added dropwise with stirring, and stirred overnight at room temperature. TLC showed that the raw materials were completely converted and the product was detected. 150 mL of water was added to the reaction solution in an ice bath, and extracted with EA (100 mL * 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (PE:EA = 3:1) to obtain compound C122-2 (3.83 g, 77.3% yield).

[0769] Second Step

[0770] Dissolve compound C122-2 (3.81 g, 18.04 mmol) in THF (50 mL). Add 9-BBN (30 mL, 0.5 M in THF) with stirring. Replace nitrogen three times, raise the temperature to 90 °C and stir at this temperature for 2 h. Cool to room temperature, rotary evaporate to remove the solvent, add compound 4 (2.7 g, 13.88 mmol), Pd(dppf)Cl2 (1.01 g, 1.39 mmol), K2CO3 (5.76 g, 41.64 mmol), DMF and H2O (50 mL / 15 mL). Replace nitrogen three times, raise the temperature to 90 °C and stir at this temperature for 2 h. LCMS shows that the reactants are completely converted and the product is detected. After the reaction is completed, filter. Add 150 mL of water to the filtrate, extract with EA (100 mL * 3). Dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography (PE:EA = 1:1) to obtain compound C122-5 (4.58 g).

[0771] The third step

[0772] Dissolve C122-5 (4.58 g, 15.77 mmol) in DCM (80 mL). Dropwise add benzyl bromide (3.78 g, 22.08 mmol) with stirring at room temperature and stir for 24 h. LCMS shows that the reactants are completely converted and the product is detected. Concentrate the reaction under vacuum to obtain the crude product of compound C122-6.

[0773] The fourth step

[0774] Dissolve compound C122-6 in MeOH (20 mL). Add NaBH4 (2.98 g, 78.9 mmol) with stirring in an ice bath and slowly raise the temperature to room temperature and stir for 24 h. LCMS shows that the reactants are completely converted and the product is detected. After the reaction is completed, add 150 mL of water in an ice bath, extract with EA (100 mL * 3). Dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain the crude product of compound C122-7 (4.58 g).

[0775] The fifth step

[0776] Dissolve compound C122-7 (6 g, 15.52 mmol) in MeOH (50 mL). Add Pd / C (500 mg) with stirring. Replace hydrogen three times and stir at room temperature for 12 h. LCMS shows that the reactants are completely converted and the product is detected. After the reaction is completed, filter. Distill the filtrate under reduced pressure to obtain the crude product of compound C122-8.

[0777] The synthesis method for the subsequent steps was the same as that described for the synthesis of compound C135 in Example 40. The final product C122 (10.57 mg, 33.27% yield, purity = 99.96%) was obtained. LCMS: Rt = 1.951 min, [M+H] + = 817.5.

[0778] Example 37: Preparation of compound C131

[0779]

[0780] The first step

[0781] Compound C131-1 (5 g, 25.71 mmol) and compound C131-2 (7.76 g, 38.56 mmol) were dissolved in DMF (70 mL). While stirring, KOtBu (5.77 g, 51.42 mmol) was added. The nitrogen was displaced three times, and the temperature was raised to 80 °C and stirred at this temperature for 4 h. TLC showed that the raw materials were completely converted and the product was detected. After the reaction was completed, it was filtered. 150 mL of water was added to the filtrate, and it was extracted with EA (100 mL * 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo to obtain a residue. The residue was purified by column chromatography (pure EA) to obtain compound C131-3 (6.92 g, 96.7% yield).

[0782] The synthesis method for the subsequent steps was the same as that of compound C122. The final product C131 (3.32 mg, 30.47% yield, purity = 93.26%) was obtained. LCMS: Rt = 1.831 min, [M+H] + = 805.5.

[0783] Example 38: Preparation of compound C132

[0784]

[0785] The first step

[0786] Compound C132a (1.4 g, 4.92 mmol) was dissolved in DMF (15 mL) at room temperature. 2-Fluoro-5-nitrotoluene (0.84 g, 5.41 mmol) and cesium carbonate (4.8 g, 14.73 mmol) were added, and the mixture was stirred at 120 °C for 5 h. TLC showed that the reactants were completely converted and a new spot was detected. The reaction solution was poured into water (100 mL), and then extracted with ethyl acetate (30 mL * 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 4%) to obtain compound C132b (689 mg, yield: 33.36%).

[0787] The second step

[0788] Compound C132b (393 mg, 0.94 mmol) was dissolved in methanol (5 mL) and dichloromethane (1 mL) at room temperature. Pd / C (10.00 mg, 0.094 mmol) (5%) was added, and the reaction mixture was stirred at room temperature for 16 h. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was filtered and concentrated in vacuo to give the crude compound C132c (364 mg, yield: 99.75%). LCMS: [M+H] + = 390.3

[0789] The third step

[0790] tert-Butyl nitrite (288 mg, 2.79 mmol, purity 100%) and cuprous bromide (334 mg, 2.33 mmol) were dissolved in acetonitrile (5 mL) at room temperature. The temperature was raised to 65 °C, and then compound C132c (364 mg, 0.93 mmol) dissolved in acetonitrile (10 mL) was added. The reaction mixture was stirred at 65 °C for 2 h under a nitrogen atmosphere. TLC showed that the reactant was completely converted and a new spot was detected. The reaction mixture was filtered and concentrated in vacuo to give the crude product. It was purified by silica gel chromatography (petroleum ether:ethyl acetate = 0 - 9%) to give compound C132d (73 mg, yield: 17.23%).

[0791] The fourth step

[0792] Compound C132d (73 mg, 0.16 mmol) was dissolved in dioxane (5 mL) at room temperature. Bis(pinacolato)diboron (82 mg, 0.32 mmol, purity 100%), Pd(dppf)Cl2·DCM (13 mg, 0.016 mmol) and potassium acetate (48 mg, 0.49 mmol) were added. The reaction mixture was stirred at 100 °C for [time not specified] h under a nitrogen atmosphere. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was filtered and concentrated in vacuo to give the crude product. It was purified by silica gel chromatography (petroleum ether:ethyl acetate = 0 - 6%) to give compound C132e (56 mg, yield: 69.50%). LCMS: [M+H] + = 501.4

[0793] The fifth step

[0794] Dissolve compound warhead 3 (56 mg, 0.11 mmol) in dioxane (5 mL) and water (1 mL) at room temperature. Add compound C132e (55 mg, 0.11 mmol), potassium carbonate (46 mg, 0.33 mmol) and Pd(PPh3)4 (13 mg, 0.011 mmol). Stir the reaction mixture at 80 °C for 16 h under a nitrogen atmosphere. LCMS shows that the reactants are completely converted and the product is detected. Filter the reaction mixture and concentrate it in vacuo to obtain the crude product. Purify it by silica gel chromatography (methanol:dichloromethane = 0 - 6%) to obtain compound C132f (86 mg, yield: 97.84%). LCMS: [M+H] + = 809.6.

[0795] Step 6

[0796] Dissolve compound C132f (86 mg, 0.11 mmol) in methanol (5 mL) and water (1 mL) at room temperature. Add NaOH (44 mg, 1.1 mmol). Stir the reaction mixture at room temperature for 2 h. LCMS shows that the reactants are completely converted and the product is detected. Pour the reaction mixture into water (30 mL), then extract with dichloromethane (30 mL×2). Dry the organic phase over anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product compound C132g (71 mg, yield 99.86%). LCMS: [M+H] + = 669.5.

[0797] Step 7

[0798] Dissolve compound C132g (71 mg, 0.11 mmol) in DCM (5 mL) at room temperature. Add trifluoroacetic acid (1 mL). Stir the reaction mixture at room temperature for 2 h. LCMS shows that the reactants are completely converted and the product is detected. Concentrate the reaction mixture in vacuo to obtain the crude product compound C132h (60 mg, yield: 99.38%). LCMS: [M+H] + = 569.5.

[0799] Step 8

[0800] Compound C132h (60 mg, 0.11 mmol) was dissolved in DMSO (3 mL) at room temperature. 4-Chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (39 mg, 0.15 mmol), HATU (55 mg, 0.14 mmol) and DIPEA (71 mg, 0.55 mmol) were added. The reaction mixture was stirred at room temperature for 16 h. LCMS showed that the reactants were completely converted and the product was detected. The reaction mixture was poured into water (50 mL), then extracted with dichloromethane (20 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to obtain the crude product. Purification by Pre-HPLC gave compound C132 (12.7 mg, yield: 14.60%). LCMS: [M+H] + = 819.5.

[0801] 1H NMR (500 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.52 (s, 1H), 8.80 (s, 1H), 8.35 (s, 1H), 8.12 (s, 1H), 8.08 (d, J = 2.3 Hz, 1H), 7.99 (d, J = 8.1 Hz, 2H), 7.65 (d, J = 8.2 Hz, 1H), 7.59 (s, 1H), 7.42 (d, J = 9.3 Hz, 1H), 7.15 (d, J = 8.0 Hz, 1H), 4.22 (m, 1H), 4.03–3.96 (m, 1H), 3.81–3.75 (m, 2H), 3.64 (m, 2H), 3.12–3.05 (m, 3H), 3.03–2.97 (m, 2H), 2.79–2.69 (m, 5H), 2.44–2.38 (m, 2H), 2.37 (s, 3H), 2.17–1.93 (m, 10H), 1.70–1.57 (m, 3H), 1.55–1.41 (m, 3H), 1.04 (t, J = 7.1 Hz, 3H).

[0802] Example 39: Preparation of compound C134

[0803]

[0804] Except that in the first step was used instead of 2-fluoro-5-nitrotoluene, according to the method described in Example 38, compound C134 (12.7 mg, yield: 14.60%) was obtained. LCMS: [M+H] + = 824.4.

[0805] C134c: LCMS: [M+H] + = 390.3. C134e: LCMS: [M+H] += 501.4. C134f: LCMS: [M+H] + = 809.6. C134g: LCMS: [M+H] + = 669.5. C134h: LCMS: [M+H] + = 569.5.

[0806] Example 40: Preparation of Compound C135

[0807]

[0808] The first step

[0809] Dissolve Compound C135-1 (2 g, 7.08 mmol), Compound C135-2 (1.69 g, 10.62 mmol), and DIEA (1.83 g, 14.16 mmol) in ACN (30 mL), raise the temperature to 90 °C and stir at this temperature for 3 h. TLC shows that the raw materials are completely converted and the product is detected. Add 50 mL of water to the reaction solution, extract with EA (50 mL * 3), dry the organic phase with anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography (PE:EA = 4:1) to obtain Compound C135-3 (2.34 g, 78.4% yield).

[0810] The second step

[0811] Dissolve Compound C135-3 (2.34 g, 5.55 mmol) in MeOH (30 mL), add Pd / C (230 mg) with stirring, displace hydrogen three times, and stir at room temperature for 1 h. LCMS shows that the reactants are completely converted and the product is detected. After the reaction is completed, filter, and distill the filtrate under reduced pressure to obtain the crude product of Compound C135-4.

[0812] The third step

[0813] Dissolve CuBr (0.95 g, 6.65 mmol) and tBuONO (0.86 g, 8.31 mmol) in ACN (15 mL), displace nitrogen three times, raise the temperature to 65 °C and stir for 10 min. Take the crude product of C135-4 and dissolve it in ACN (15 mL), add it dropwise with stirring, and stir at the maintained temperature for 2 h. LCMS shows that the reactants are completely converted and the product is detected. After the reaction is completed, filter, and concentrate the filtrate under vacuum to obtain a residue. Purify the residue by column chromatography (PE:EA = 9:1) to obtain Compound C135-5 (335 mg, 13.3% yield).

[0814] The fourth step

[0815] Compound C135-5 (0.34 g, 0.74 mmol), KOAc (0.22 g, 2.22 mmol), B2pin2 (0.38 g, 1.48 mmol), and Pd(dppf)Cl2 (55 mg, 0.07 mmol) were dissolved in dioxane (5 mL). The mixture was purged with nitrogen three times and then heated to 90 °C and stirred for 2 h. LCMS showed that the reactants were completely converted and the product was detected. After the reaction was completed, the mixture was filtered, and the filtrate was concentrated under vacuum to obtain a residue. The residue was purified by column chromatography (PE:EA = 4:1) to give compound C135-6 (270 mg, 73.1% yield).

[0816] Step 5

[0817] Compound C135-6 (248 mg, 0.49 mmol), warhead 3 (170 mg, 0.33 mmol), Pd(PPh3)4 (38 mg, 0.07 mmol), and K2CO3 (127 mg, 0.99 mmol) were dissolved in DMSO and H2O (3 mL / 0.5 mL). The mixture was purged with nitrogen three times and then heated to 90 °C and stirred for 12 h. LCMS showed that the reactants were completely converted and the product was detected. After the reaction was completed, the mixture was filtered, 50 mL of water was added, and the mixture was extracted with DCM (30 mL * 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain a residue. The residue was purified by preparative plate (DCM:MeOH = 4:1) to give compound C135-7 (96 mg, 43.3% yield).

[0818] Step 6

[0819] Compound C135-7 (116 mg, 0.17 mmol) was dissolved in DCM and TFA (3 mL / 0.5 mL), and the mixture was stirred at room temperature for 1 h. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was distilled under reduced pressure to obtain crude compound C135-8.

[0820] Step 7

[0821] Compound C135-8, compound C135-9 (68.5 mg, 0.26 mmol), and NMI (49 mg, 0.6 mmol) were dissolved in DMSO (2 mL). TCFH (57 mg, 0.2 mmol) was added with stirring, and the mixture was stirred at room temperature for 2 h. LCMS showed that the reactants were completely converted and the product was detected. The final product was obtained by preparative chromatography (45.22 mg, 32.00% yield, purity = 99.81%). LCMS: Rt = 2.153 min, [M+H] + = 821.6.

[0822] Example 41: Preparation of Compound C136

[0823]

[0824] The first step

[0825] Dissolve Compound C136a (211 mg, 0.75 mmol) in DMF (5 mL) at room temperature, add p-fluoronitrobenzene (127 mg, 0.90 mmol) and potassium carbonate (311 mg, 2.25 mmol), and stir the reaction mixture at 100 °C for 3 hours. TLC shows that the reactants are completely converted and a new spot is detected. Pour the reaction mixture into water (50 mL), then extract with ethyl acetate (30 mL × 2). Dry the organic layer over anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product. Purify by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 14%) to obtain Compound C136b (193 mg, yield: 64.02%).

[0826] The second step

[0827] Dissolve Compound C136b (193 mg, 0.48 mmol) in methanol (5 mL) at room temperature, add Pd / C (20 mg, 0.19 mmol) (5%), and stir the reaction mixture at room temperature under a hydrogen atmosphere for 16 hours. LCMS shows that the reactants are completely converted and the product is detected. Filter the reaction mixture, and concentrate the filtrate in vacuo to obtain Compound C136c (178 mg, yield: 99.63%). LCMS: [M + H] + = 374.3

[0828] The third step

[0829] Dissolve tert-butyl nitrite (149 mg, 1.44 mmol) and CuBr (172 mg, 1.20 mmol) in acetonitrile (5 mL) at room temperature, heat to 65 °C, then add Compound C136c (178 mg, 0.48 mmol) dissolved in acetonitrile (5 mL). Stir the reaction mixture at 65 °C under a nitrogen atmosphere for 2 hours. TLC shows that the reactants are completely converted and a new spot is detected. Filter the reaction mixture, and concentrate in vacuo to obtain the crude product. Purify by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 7%) to obtain Compound C136d (43 mg, yield: 20.63%).

[0830] The fourth step

[0831] Compound C136d (43 mg, 0.098 mmol) was dissolved in 1,4-dioxane (5 mL) at room temperature. Bis(pinacolato)diboron (50 mg, 0.20 mmol), potassium acetate (29 mg, 0.30 mmol) and Pd(dppf)Cl2·DCM (8 mg, 0.0098 mmol) were added. The reaction mixture was stirred at 80 °C for 16 h under a nitrogen atmosphere. TLC showed complete conversion of the reactants and the detection of a new spot. The reaction mixture was filtered and concentrated in vacuo to give the crude product. It was purified by silica gel chromatography (petroleum ether:ethyl acetate = 0 - 10%) to give compound C136e (50 mg, yield: 100%).

[0832] The fifth step

[0833] Compound C136e (50 mg, 0.10 mmol) was dissolved in 1,4-dioxane (5 mL) and water (1 mL) at room temperature. C136f (50 mg, 0.10 mmol), tetrakis(triphenylphosphine)palladium(0) (23.11 mg, 0.020 mmol) and potassium carbonate (55.28 mg, 0.40 mmol) were added. The mixture was stirred at 80 °C under a nitrogen atmosphere. LCMS showed complete conversion of the reactants and the detection of the product. The reaction mixture was filtered and concentrated in vacuo to give the crude product. It was purified by silica gel chromatography (methanol:dichloromethane = 0 - 7%) to give compound C136g (70 mg, yield: 44.33%). LCMS: [M+H] + = 765.4.

[0834] The sixth step

[0835] Compound C136g (70 mg, 0.092 mmol) was dissolved in dichloromethane (2 mL) and DMSO (2 mL) at room temperature. Paraformaldehyde (8.3 mg, 0.28 mmol) and acetic acid (10 mg, 0.17 mmol) were added. The mixture was stirred at 35 °C for 2 h under a nitrogen atmosphere. Then sodium triacetoxyborohydride (98 mg, 0.46 mmol) was added and the mixture was stirred at 35 °C for 16 h. LCMS showed complete conversion of the reactants and the detection of the product. The reaction mixture was poured into water (50 mL), then extracted with dichloromethane (30 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. It was purified by silica gel chromatography (methanol:dichloromethane = 0 - 6%) to give compound C136h (22 mg, yield: 30.82%). LCMS: [M+H] + = 779.5.

[0836] The seventh step

[0837] Compound C136h (22 mg, 0.028 mmol) was dissolved in methanol (5 mL) and water (1 mL) at room temperature. NaOH (11 mg, 0.28 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was poured into water (20 mL), then extracted with dichloromethane (20 mL * 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give crude compound C136i (18 mg, yield 99.74%). LCMS: [M+H] + = 639.5.

[0838] The eighth step

[0839] Compound C136i (18 mg, 0.028 mmol) was dissolved in DCM (5 mL) at room temperature. TFA (5 mL) was added, and the reaction mixture was stirred at room temperature for 1 hour. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was concentrated in vacuo to give crude compound C136j (15 mg, yield 98.79%), which was used directly in the next step without purification. LCMS: [M+H] + = 539.4.

[0840] The ninth step

[0841] Compound C136j (15 mg, 0.028 mmol) was dissolved in DMSO (2 mL) at room temperature. 4-Chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (10 mg, 0.037 mmol), HATU (14 mg, 0.037 mmol) and DIEA (20 mg, 0.15 mmol) were added, and the reaction mixture was stirred at room temperature for 16 hours. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was poured into water (50 mL), then extracted with dichloromethane (20 mL * 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give a crude product. Purification by Pre-HPLC gave compound C136 (4.82 mg, yield: 21.95%). LCMS: Rt = 1.999 min; [M+H] + = 789.5.

[0842] Example 42: Preparation of compound C137

[0843]

[0844] The first step

[0845] The compound 1-Boc-4-fluoro-4-(hydroxymethyl)piperidine (2.1 g, 9.00 mmol) was dissolved in hydrochloric acid dioxane solution (10 mL) (4 M) at room temperature and stirred for 1 hour. The reaction solution was concentrated under vacuum to obtain the crude compound C137a (P0, 1.53 g, yield: 100%), which was directly used in the next step.

[0846] The second step

[0847] Compound C137a was dissolved in DMF (10 mL) at room temperature. p-Fluoronitrobenzene (1.53 g, 10.84 mmol) and potassium carbonate (3.74 g, 27.06 mmol) were added. The mixture was stirred at 80 °C for 3 hours under a nitrogen atmosphere. TLC showed that the reactants were completely converted and a new spot was detected. The reaction solution was poured into water (50 mL), and then extracted with ethyl acetate (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. It was purified by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 47%) to obtain compound C137b (1.3 g, yield: 56.74%).

[0848] The third step

[0849] Compound C137b (1.2 g, 4.72 mmol) was dissolved in acetonitrile (20 mL) at room temperature. IBX (3.95 g, 14.11 mmol) was added. The reaction solution was stirred at 55 °C for 6 hours. TLC showed that the reactants were completely converted and a new spot was detected. The reaction solution was filtered to obtain the filtrate, and the filtrate was rotary evaporated to obtain the crude compound C137c (1.19 g, yield: 99.96%).

[0850] The fourth step

[0851] Compound C137c (1.19 g, 4.72 mmol) was dissolved in dichloromethane (20 mL) and dimethyl sulfoxide (10 mL) at room temperature. tert-Butyl piperazine-1-carboxylate (1.3 g, 6.98 mmol) was added. The reaction solution was stirred at room temperature for 1 hour. Then NaBH(OAc)3 (3 g, 14.15 mmol) was added, and the mixture was stirred at room temperature for 16 hours. TLC showed that the reactants were completely converted and a new spot was detected. The reaction solution was poured into water (30 mL), and then extracted with dichloromethane (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. It was purified by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 23%) to obtain compound C137d (870 mg, yield: 43.65%)

[0852] The fifth step

[0853] Compound C137d (870 mg, 2.06 mmol) was dissolved in methanol (10 mL) and dichloromethane (2 mL) at room temperature. Wet palladium on carbon (219.23 mg, 2.06 mmol) (5%) was added, and the reaction mixture was stirred at room temperature for 16 hours under a hydrogen atmosphere. LCMS showed complete conversion of the reactant and detection of the product. The reaction mixture was filtered and concentrated in vacuo to obtain the crude product. It was purified by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 56%) to give compound C137e (360 mg, yield: 44.54%). LCMS: [M+H] + = 393.3.

[0854] Step 6

[0855] Copper(I) bromide (330 mg, 2.30 mmol) and tert-butyl nitrite (285 mg, 2.76 mmol) were dissolved in acetonitrile (5 mL) at room temperature. The temperature was raised to 65 °C, and then compound C137e (360 mg, 0.92 mmol) dissolved in acetonitrile (5 mL) was added. The reaction mixture was stirred at 65 °C for 2 hours under a nitrogen atmosphere. LCMS showed complete conversion of the reactant and detection of the product. The reaction mixture was filtered and the filtrate was concentrated in vacuo to obtain the crude product. It was purified by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 25%) to give compound C137f (113 mg, yield: 27.00%). LCMS: [M+H] + = 456.2.

[0856] Step 7

[0857] Compound C137f (113 mg, 0.25 mmol) was dissolved in dioxane (5 mL) at room temperature. Pd(dppf)Cl2·DCM (20 mg, 0.024 mmol), bis(pinacolato)diboron (127 mg, 0.50 mmol) and potassium acetate (74 mg, 0.75 mmol) were added, and the mixture was stirred at 80 °C for 16 hours under a nitrogen atmosphere. LCMS showed complete conversion of the reactant and detection of the product. The reaction mixture was filtered and concentrated in vacuo to obtain the crude product. It was purified by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 9%) to give compound C137g (99 mg, yield: 79.42%). LCMS: [M+H] + = 504.4.

[0858] Step 8

[0859] Dissolve compound warhead 3 (80 mg, 0.16 mmol) in dioxane (5 mL) and water (1 mL) at room temperature. Add C137g (99 mg, 0.20 mmol), potassium carbonate (67 mg, 0.48 mmol) and tetrakis(triphenylphosphine)palladium(0) (18 mg, 0.016 mmol). Stir the mixture at 80 °C for 3 hours under a nitrogen atmosphere. LCMS shows that the reactants are completely converted and the product is detected. Pour the reaction solution into water (50 mL), then extract with dichloromethane (30 mL × 2). Dry the organic layer over anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product. Purify by silica gel chromatography (methanol:dichloromethane = 0 - 6%) to obtain compound C137h (63 mg, yield: 49.99%). LCMS: [M+H] + = 812.5.

[0860] Step 9

[0861] Dissolve compound C137h (63 mg, 0.078 mmol) in methanol (5 mL) and water (1 mL) at room temperature. Add sodium hydroxide (32 mg, 0.80 mmol). Stir the reaction solution at room temperature for 2 hours. LCMS shows that the reactants are completely converted and the product is detected. Pour the reaction solution into water (50 mL), then extract with dichloromethane (30 mL × 2). Dry the organic layer over anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product compound C137i (52 mg, yield: 99.76%). LCMS: [M+H] + = 672.5.

[0862] Step 10

[0863] Dissolve compound C137i (52 mg, 0.077 mmol) in DCM (5 mL) at room temperature. Add TFA (1 mL). Stir the reaction solution at room temperature for 2 hours. LCMS shows that the reactants are completely converted and the product is detected. Concentrate the reaction solution in vacuo to obtain the crude product compound C137j (44 mg, yield: 99.43%), which is directly used in the next step. LCMS: [M+H] + = 572.5.

[0864] Step 11

[0865] Compound C137j (44 mg, 0.077 mmol) was dissolved in DMSO (2 mL) at room temperature. 4-Chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (23 mg, 0.086 mmol), HATU (38 mg, 0.10 mmol) and DIPEA (50 mg, 0.39 mmol) were added, and the reaction mixture was stirred at room temperature for 16 h. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was poured into water (50 mL), then extracted with dichloromethane (30 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated in vacuo to obtain the crude product. Purification by Pre-HPLC gave compound C137 (10.26 mg, yield: 16.18%). LCMS: [M+H] + = 822.5.

[0866] Example 43: Preparation of compound C140

[0867]

[0868] The first step

[0869] Compound C140e (50 mg, 0.10 mmol, which can be prepared according to the methods described in the first to fourth steps of Example 41) was dissolved in dioxane (5 mL) and water (1 mL) at room temperature. Compound C140f (42 mg, 0.10 mmol), Pd(dppf)Cl2·DCM (8 mg, 0.010 mmol) and potassium carbonate (42 mg, 0.30 mmol) were added, and the reaction mixture was stirred at 90 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered and concentrated in vacuo to obtain the crude product, compound C140g (71 mg, yield: 99.95%).

[0870] The second step

[0871] Compound C140g (71 mg, 0.099 mmol) was dissolved in methanol (5 mL) and water (1 mL) at room temperature. NaOH (40 mg, 0.99 mmol) was added, and the reaction mixture was stirred at room temperature for 2 h. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was filtered and concentrated in vacuo to obtain the crude product. Purification by silica gel chromatography (methanol:dichloromethane = 0 - 4%) gave compound C140h (44 mg, yield: 77.11%). LCMS: [M+H] + = 574.4.

[0872] The third step

[0873] Compound C140h (44 mg, 0.077 mmol) was dissolved in dioxane hydrochloride (5 mL, 4 M) at room temperature and stirred for 1 hour. LCMS showed that the reactant was completely converted and the product was detected. The reaction solution was concentrated in vacuo to give crude compound C140i (39 mg, yield: 99.70%). LCMS: [M+H] + = 474.3.

[0874] The fourth step

[0875] Compound C140i (39 mg, 0.076 mmol) was dissolved in DMSO (3 mL) at room temperature. 4-Chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (20 mg, 0.076 mmol), TCFH (28 mg, 0.099 mmol) and NMI (31 mg, 0.38 mmol) were added. The reaction solution was stirred at room temperature for 16 hours. LCMS showed that the reactant was completely converted and the product was detected. The reaction solution was poured into water (30 mL), then extracted with dichloromethane (20 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. Purification by Pre-HPLC gave compound C140 (5.71 mg, yield: 10.23%). LCMS: [M+H] + = 707.3.

[0876] 1 H NMR (500 MHz, DMSO-d6) δ 9.01 (d, J = 207.9 Hz, 1H), 8.44–8.06 (m, 2H), 7.87–7.17 (m, 3H), 6.96 (d, J = 7.5 Hz, 1H), 4.22–3.79 (m, 4H), 3.52 (s, 1H), 3.00 (d, J = 11.9 Hz, 2H), 2.75 (s, 1H), 1.87–1.69 (m, 3H), 1.64–1.23 (m, 5H)

[0877] Example 44: Preparation of compound C153

[0878]

[0879] Compound C153a (30 mg, 0.054 mmol) was dissolved in DMSO (3 mL) at room temperature. C153b (14 mg, 0.056 mmol), TCFH (16 mg, 0.057 mmol) and NMI (23 mg, 0.28 mmol) were added. The reaction mixture was stirred at room temperature for 16 h. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was poured into water (50 mL), then extracted with dichloromethane (30 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. Purification by Pre-HPLC gave compound C153 (5.13 mg, yield: 12.06%). LCMS: [M+H] + = 783.6.

[0880] 1 H NMR (500 MHz, DMSO-d6) δ 11.92 (s, 1H), 10.38 (s, 1H), 8.77 (s, 1H), 8.34 (s, 1H), 8.12–8.00 (m, 4H), 7.34 (d, J = 7.9 Hz, 1H), 7.31 (s, 1H), 7.25 (d, J = 7.5 Hz, 1H), 7.07 (d, J = 8.8 Hz, 2H), 4.54–4.42 (m, 1H), 4.27–4.19 (m, 1H), 3.86–3.77 (m, 3H), 3.55–3.51 (m, 2H), 3.00 (d, J = 11.1 Hz, 2H), 2.82–2.65 (m, 5H), 2.39 (q, J = 7.2 Hz, 2H), 2.22 (s, 3H), 2.13–1.93 (m, 6H), 1.87–1.50 (m, 7H), 1.29–1.16 (m, 4H), 1.14–1.06 (m, 2H), 1.04 (t, J = 7.1 Hz, 3H).

[0881] Example 45: Preparation of compound C154

[0882]

[0883] Compound C154a (30 mg, 0.054 mmol) was dissolved in DMSO (3 mL) at room temperature. C154b (14 mg, 0.056 mmol), TCFH (16 mg, 0.057 mmol) and NMI (23 mg, 0.28 mmol) were added, and the reaction mixture was stirred at room temperature for 16 hours. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was poured into water (50 mL), then extracted with dichloromethane (30 mL * 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain the crude product. Purification by Pre-HPLC gave compound C154 (5.16 mg, yield: 12.06%). LCMS: [M+H] + = 787.6.

[0884] 1 H NMR (500 MHz, DMSO-d6) δ 11.92 (s, 1H), 10.53 (s, 1H), 8.77 (s, 1H), 8.34 (s, 1H), 8.14–7.99 (m, 4H), 7.50 (d, J = 6.8 Hz, 1H), 7.38 (d, J = 8.6 Hz, 2H), 7.07 (d, J = 8.5 Hz, 2H), 4.52–4.38 (m, 1H), 4.27–4.16 (m, 1H), 3.82 (d, J = 12.1 Hz, 2H), 3.76 (t, J = 6.5 Hz, 2H), 3.00 (d, J = 11.0 Hz, 2H), 2.78–2.35 (m, 4H), 2.39 (q, J = 7.1 Hz, 2H), 2.10–1.95 (m, 6H), 1.85–1.43 (m, 7H), 1.29–1.16 (m, 4H), 1.15–1.07 (m, 2H), 1.04 (t, J = 7.2 Hz, 3H).

[0885] Example 46: Preparation of compound C160

[0886]

[0887] The first step

[0888] Compound C160a (6.2 g, 31.95 mmol) was dissolved in DMF (50 mL) at room temperature. Cesium carbonate (12.49 g, 38.34 mmol) and iodomethane-d (5.09 g, 35.15 mmol) were added, and the mixture was stirred at room temperature for 16 hours. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to obtain the crude compound C160b (6.72 g, yield: 99.64%). LCMS: [M+H] + = 211.1.

[0889] The second step

[0890] Dissolve compound C160b (1 g, 2.15 mmol) in dioxane (10 mL) and water (3 mL) at room temperature. Add compound C160c (0.54 g, 2.58 mmol), potassium carbonate (0.89 g, 6.45 mmol) and Pd(dppf)Cl2·DCM (88 mg, 0.11 mmol). Stir the reaction mixture at 80 °C for 4 hours under a nitrogen atmosphere. LCMS shows that the reactants are completely converted and the product is detected. Filter the reaction mixture and concentrate it under vacuum to obtain the crude product. Purify it by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 30%) to obtain compound C160d (322 mg, yield: 35.47%). LCMS: [M+H] + = 422.4

[0891] The third step

[0892] Dissolve compound C160d (60 mg, 0.14 mmol) in dioxane (5 mL) and water (1 mL) at room temperature. Add compound C160e (59 mg, 0.15 mmol), tetrakis(triphenylphosphine)palladium (8 mg, 0.0071 mmol) and potassium carbonate (60 mg, 0.43 mmol). Stir the reaction mixture at room temperature for 16 hours. LCMS shows that the reactants are completely converted and the product is detected. Concentrate the reaction mixture under vacuum to obtain the crude product compound C160f (99 mg, yield: 99.32%). LCMS: [M+H] + = 699.4

[0893] The fourth step

[0894] Dissolve compound C160f (99 mg, 0.14 mmol) in methanol (S0, 5 mL) and water (1 mL) at room temperature. Add sodium hydroxide (56.00 mg, 1.40 mmol). Stir the reaction mixture at room temperature for 1 hour. LCMS shows that the reactants are completely converted and the product is detected. Concentrate the reaction mixture under vacuum to obtain the crude product. Purify it by silica gel chromatography (methanol: dichloromethane = 0 - 3%) to obtain compound C160g (70 mg, yield: 88.41%). LCMS: [M+H] + = 560.4

[0895] The fifth step

[0896] Compound C160 (70 mg, 0.13 mmol) was dissolved in methanol (5 mL) at room temperature, and dioxane hydrochloride (5 mL) was added. The reaction mixture was stirred at room temperature for 2 hours. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was concentrated under vacuum to obtain crude compound C160h (62 mg, yield: 99.94%). LCMS: [M+H] + = 460.5.

[0897] The sixth step

[0898] Compound C160h (62 mg, 0.12 mmol) was dissolved in DMSO (3 mL) at room temperature, and 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (39 mg, 0.15 mmol), TCFH (44 mg, 0.16 mmol) and NMI (50 mg, 0.61 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was poured into water (30 mL), and then extracted with dichloromethane (30 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a crude product. It was purified by Pre-HPLC to obtain compound C160 (22.68 mg, yield: 25.07%). LCMS: [M+H] + = 710.4. 1 H NMR (500 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.54 (s, 1H), 8.97 (s, 1H), 8.80 (s, 1H), 8.34 (d, J = 7.4 Hz, 2H), 8.09 (s, 1H), 8.03 (s, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.56 (s, 1H), 7.39 (d, J = 8.2 Hz, 1H), 6.97 (d, J = 8.9 Hz, 1H), 4.43 (dd, J = 25.0, 12.6 Hz, 3H), 3.81–3.72 (m, 1H), 3.66–3.51 (m, 2H), 2.86 (d, J = 12.2 Hz, 2H), 2.81–2.67 (m, 3H), 1.70 (dd, J = 49.1, 15.4 Hz, 6H), 1.31–0.96 (m, 7H).

[0899] Example 47: Preparation of compound C162

[0900]

[0901] The first step

[0902] C162 a (1.0 g, 4.5 mmol), C162 b (3.7 g, 4.1 mmol), a catalyst (150 mg, 0.2 mmol), potassium carbonate (1.7 g, 12.3 mmol), etc. were successively added into a 250 mL single-necked flask. Subsequently, 30 mL of dioxane and 10 mL of water were added as solvents. Then, after adding a condenser to the single-necked flask, nitrogen was replaced three times with a three-way valve and then heated to 80 °C for reaction for 3 h. After the reaction was completed as detected by LCMS, the reaction solution was filtered, and then extracted three times with water and dichloromethane (10 mL * 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (EA / PE = 30%). The product C162 c (1.05 g, 2.4 mmol) was obtained with a yield of 53.3%.

[0903] The second step

[0904] C162 c (95 mg, 0.22 mmol), C162 d (80 mg, 0.20 mmol), tetrakis(triphenylphosphine)palladium (23 mg, 0.02 mmol) catalyst, and potassium carbonate (82 mg, 0.60 mmol) were successively added into a 100 mL single-necked flask. Subsequently, 10 mL of dioxane and 1 mL of water were added as solvents. After the reactants were fully dissolved, a condenser was added to the single-necked flask and nitrogen was replaced three times. Then, the temperature was raised to 90 °C and the reaction was carried out overnight for 16 h. After the reaction was completed as detected by LCMS, the reaction solution was filtered and directly concentrated for use in the next step.

[0905] The third step

[0906] The crude product C162 e (crude) from the third step was put into a 100 mL reaction flask. After adding 5 mL of methanol solvent, an aqueous solution of sodium hydroxide (100 mg) was added. After the reaction continued at room temperature for about 2 h, a small amount of the reaction solution was taken. LCMS showed that the reaction was complete. After the reaction solution was filtered, it was extracted three times with dichloromethane. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel chromatography (MeOH / CH2Cl2 = 4%) to obtain the product C162 f (80 mg).

[0907] The fourth step

[0908] C162 f was put into a 100 mL single-necked flask. After adding HCl / dioxane (2 mol / L, 1 mL), some solids precipitated. 2 mL of methanol was added to assist dissolution. After the reaction was carried out at room temperature for 2 h, the reaction was completed as detected by LCMS. The solvent was directly rotary evaporated and used as the raw material for the next step.

[0909] The fifth step

[0910] C162 g (50 mg, 0.10 mmol), C162 h (35 mg, 0.13 mmol), TCFH (38 mg, 0.14 mmol), and 1-methylimidazole (34 mg, 0.41 mmol) were successively added into a 100 mL single-neck flask. After reacting overnight for 16 h at room temperature, LCMS showed that the reaction was complete. The final product C162 (20.78 mg, 0.02 mmol) was obtained by preparative chromatography separation, with a yield of 30%, [M+H] + = 721.3.

[0911] 1 H NMR (500 MHz, DMSO-d6) δ 11.96 (s, 1H), 10.51 (s, 1H), 8.96 (d, J = 2.2 Hz, 1H), 8.79 (s, 1H), 8.41–8.29 (m, 2H), 8.08 (d, J = 2.6 Hz, 1H), 8.05 (s, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 1.7 Hz, 1H), 7.39 (dd, J = 8.2, 1.7 Hz, 1H), 6.96 (d, J = 9.1 Hz, 1H), 4.51–4.37 (m, 3H), 4.23 (q, J = 7.3 Hz, 2H), 3.82–3.72 (m, 1H), 3.66–3.53 (m, 2H), 3.13–3.02 (m, 1H), 2.87 (t, J = 11.9 Hz, 2H), 2.75 (q, J = 7.7, 6.7 Hz, 2H), 1.84–1.58 (m, 6H), 1.43 (t, J = 7.3 Hz, 3H), 1.29–1.02 (m, 7H).

[0912] Example 48: Preparation of compound C163

[0913]

[0914] The first step

[0915] Compound C163a (1.78 g, 6.29 mmol) was dissolved in DMF (20 mL) at room temperature. 3,4-Difluoronitrobenzene (1 g, 6.29 mmol) and potassium carbonate (2.61 g, 18.87 mmol) were added. The reaction mixture was stirred at 85 °C for 3 hours under a nitrogen atmosphere. TLC showed that the reactants were completely converted and a new spot was detected. The reaction mixture was filtered and concentrated in vacuo to obtain the crude product. It was purified by silica gel chromatography (petroleum ether:ethyl acetate = 0 - 19%) to obtain compound C163b (1.5 g, yield: 56.61%).

[0916] The second step

[0917] Compound C163b (1.5 g, 3.56 mmol) was dissolved in methanol (30 mL) at room temperature. Pd / C (0.038 g, 0.36 mmol) (5%) was added, and the reaction mixture was stirred at room temperature under a hydrogen atmosphere for 4 hours. TLC showed complete conversion of the reactants and the detection of a new spot. The reaction mixture was filtered and concentrated in vacuo to obtain crude compound C163c (1.39 g, yield: 99.76%).

[0918] The third step

[0919] tert-Butyl nitrite (1.10 g, 10.65 mmol) and copper(I) bromide (R2, 1.53 g, 10.65 mmol) were dissolved in acetonitrile (10 mL) at room temperature. The temperature was raised to 65 °C, and then C163c (1.39 g, 3.55 mmol) dissolved in acetonitrile (10 mL) was added. The reaction mixture was stirred at 65 °C under a nitrogen atmosphere for 2 hours. TLC showed complete conversion of the reactants and the detection of a new spot. The reaction mixture was filtered and concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography (petroleum ether:ethyl acetate = 0 - 13%) to obtain compound C163d (296 mg, yield: 18.31%).

[0920] The fourth step

[0921] Compound C163d (296 mg, 0.65 mmol) was dissolved in dioxane (3 mL) at room temperature. Bis(pinacolato)diboron (0.33 g, 1.3 mmol), Pd(dppf)Cl2·DCM (53 mg, 0.065 mmol) and potassium acetate (0.19 g, 1.95 mmol) were added. The reaction mixture was stirred at 100 °C under a nitrogen atmosphere for 16 hours. LCMS showed complete conversion of the reactants and the detection of the product. The reaction mixture was filtered and concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography (petroleum ether:ethyl acetate = 0 - 9%) to obtain compound C163e (320 mg, yield: 97.98%). LCMS: [M+H] + = 503.3.

[0922] The fifth step

[0923] Compound C163e (50 mg, 0.10 mmol) was dissolved in dioxane (5 mL) and water (1 mL) at room temperature. Compound C163f (42 mg, 0.10 mmol), Pd(dppf)Cl2·DCM (8 mg, 0.010 mmol) and potassium carbonate (42 mg, 0.30 mmol) were added. The reaction mixture was stirred at 90 °C under a nitrogen atmosphere for 16 hours. The reaction mixture was filtered and concentrated in vacuo to obtain a crude product to obtain compound C163g (71 mg, yield: 99.95%).

[0924] The sixth step

[0925] Compound C163g (71 mg, 0.099 mmol) was dissolved in methanol (5 mL) and water (1 mL) at room temperature. NaOH (40 mg, 0.99 mmol) was added, and the reaction mixture was stirred at room temperature for 2 hours. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was filtered and concentrated in vacuo to obtain the crude product. It was purified by silica gel chromatography (methanol:dichloromethane = 0 - 4%) to obtain compound C163h (44 mg, yield: 77.11%). LCMS: [M+H] + = 574.4

[0926] The seventh step

[0927] Compound C163h (44 mg, 0.077 mmol) was dissolved in dioxane hydrochloride (5 mL, 4 M) at room temperature and stirred for 1 hour. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was concentrated in vacuo to obtain the crude compound C163i (39 mg, yield: 99.70%). LCMS: [M+H] + = 474.3

[0928] The eighth step

[0929] Compound C163i (39 mg, 0.076 mmol) was dissolved in DMSO (3 mL) at room temperature. 4-chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (20 mg, 0.076 mmol), TCFH (28 mg, 0.099 mmol) and NMI (31 mg, 0.38 mmol) were added, and the reaction mixture was stirred at room temperature for 16 hours. LCMS showed that the reactant was completely converted and the product was detected. The reaction mixture was poured into water (30 mL), then extracted with dichloromethane (20 mL * 2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to obtain the crude product. It was purified by Pre-HPLC to obtain compound C163 (5.71 mg, yield: 10.23%). LCMS: Rt = 2.019 min; [M+H] + = 724.4

[0930] Example 49: Preparation of compound C164

[0931]

[0932] Compound C164a (79 mg, 0.17 mmol) was dissolved in DMSO (3 mL) at room temperature. 3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-4-fluorobenzoic acid (56.16 mg, 0.22 mmol), HATU (97 mg, 0.26 mmol) and DIPEA (110 mg, 0.85 mmol) were added. The reaction mixture was stirred at room temperature for 16 h. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was poured into water (30 mL), then extracted with dichloromethane (20 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to obtain the crude product. The crude product was purified by Pre-HPLC to obtain compound C164 (4.33 mg, yield: 3.59%). LCMS: Rt = 2.196 min; [M+H] + = 691.4.

[0933] Example 50: Preparation of Compound C165

[0934]

[0935] Compound C165a (79 mg, 0.17 mmol) was dissolved in DMSO (3 mL) at room temperature. 3-(2,4-Dioxotetrahydropyrimidin-1(2H)-yl)-4-methylbenzoic acid (60 mg, 0.22 mmol), HATU (97 mg, 0.26 mmol) and DIPEA (110 mg, 0.85 mmol) were added. The reaction mixture was stirred at room temperature for 16 h. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was poured into water (30 mL), then extracted with dichloromethane (20 mL×2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to obtain the crude product. The crude product was purified by Pre-HPLC to obtain compound C165 (4.42 mg, yield: 3.61%). LCMS: Rt = 2.200 min; [M+H] + = 687.5.

[0936] Example 51: Preparation of Compound C167

[0937]

[0938] The first step

[0939] Compound C167a (6.2 g, 31.95 mmol) was dissolved in DMF (50 mL) at room temperature. Cesium carbonate (12.49 g, 38.34 mmol) and difluoroiodomethane (5.09 g, 35.15 mmol) were added, and the mixture was stirred at room temperature for 16 hours. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was filtered, and the filtrate was concentrated in vacuo to give crude compound C167b (6.72 g, yield: 99.64%). LCMS: [M+H] + = 211.1.

[0940] The second step

[0941] Compound C167b (1 g, 2.15 mmol) was dissolved in dioxane (10 mL) and water (3 mL) at room temperature. Compound C167c (0.54 g, 2.58 mmol), potassium carbonate (0.89 g, 6.45 mmol) and Pd(dppf)Cl2·DCM (88 mg, 0.11 mmol) were added, and the reaction mixture was stirred at 80 °C for 4 hours under a nitrogen atmosphere. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was filtered and concentrated in vacuo to give a crude product. It was purified by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 30%) to give compound C167d (322 mg, yield: 35.47%). LCMS: [M+H] + = 422.4.

[0942] The third step

[0943] Compound C167d (60 mg, 0.14 mmol) was dissolved in dioxane (5 mL) and water (1 mL) at room temperature. Compound C167e (59 mg, 0.15 mmol), tetrakis(triphenylphosphine)palladium (8 mg, 0.0071 mmol) and potassium carbonate (60 mg, 0.43 mmol) were added, and the reaction mixture was stirred at room temperature for 16 hours. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was concentrated in vacuo to give crude compound C167f (99 mg, yield: 99.32%). LCMS: [M+H] + = 699.4.

[0944] The fourth step

[0945] Compound C167f (99 mg, 0.14 mmol) was dissolved in methanol (S0, 5 mL) and water (1 mL) at room temperature. Sodium hydroxide (56.00 mg, 1.40 mmol) was added, and the reaction mixture was stirred at room temperature for 1 hour. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was concentrated in vacuo to give the crude product. Purification by silica gel chromatography (methanol:dichloromethane = 0 - 3%) gave compound C167g (70 mg, yield: 88.41%). LCMS: [M+H] + = 560.4.

[0946] The fifth step

[0947] Compound C167g (70 mg, 0.13 mmol) was dissolved in methanol (5 mL) at room temperature. Dioxane hydrochloride (5 mL) was added, and the reaction mixture was stirred at room temperature for 2 hours. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was concentrated in vacuo to give the crude compound C167h (62 mg, yield: 99.94%). LCMS: [M+H] + = 460.5.

[0948] The sixth step

[0949] Compound C167h (62 mg, 0.12 mmol) was dissolved in DMSO (3 mL) at room temperature. 4-Chloro-3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)benzoic acid (39 mg, 0.15 mmol), TCFH (44 mg, 0.16 mmol) and NMI (50 mg, 0.61 mmol) were added, and the reaction mixture was stirred at room temperature for 2 hours. LCMS showed complete conversion of the reactants and the product was detected. The reaction mixture was poured into water (30 mL), then extracted with dichloromethane (30 mL * 2). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give the crude product. Purification by Pre-HPLC gave compound C167 (22.68 mg, yield: 25.07%). LCMS: [M+H] + = 743.2.

[0950] 1 H NMR (500 MHz, DMSO-d6) δ 9.21 (s, 1H), 9.01–8.35 (m, 4H), 8.41–7.99 (m, 2H), 7.73–6.76 (m, 6H), 4.30–3.80 (m, 6H), 3.21–2.62 (m, 6H), 1.99–1.53 (m, 9H), 1.48–1.18 (m, 6H).

[0951] Example 52: Preparation of compound C169

[0952]

[0953] The first step

[0954] Dissolve C169a (2.0 g, 10.3 mmol) in 20 mL of DMF solvent. Add sodium hydride (1.2 g, 30.9 mmol) under an ice bath and react for 30 min. Then add C169b (3.5 g, 20.6 mmol). After reacting for about 2 h, LCMS shows that the reactants are completely converted and the reaction product is detected. Then end the reaction. Add saturated ammonium chloride aqueous solution to quench the reaction. Extract with ethyl acetate three times (20 mL * 3). Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate in vacuo. Purify by silica gel chromatography (EA / PE = 1:2). Obtain product C169c (2.2 g, 9.3 mmol, yield (90.3%)).

[0955] The second step

[0956] Add C169c (2.1 g, 8.9 mmol), C169d (3.7 g, 8.0 mmol), catalyst (295 mg, 0.4 mmol), potassium carbonate (3.35 g, 24.2 mmol), etc. into a 250 mL single-necked flask in sequence. Then add 60 mL of dioxane and 20 mL of water as solvents. After installing a condenser on the single-necked flask, displace nitrogen three times with a three-way joint and then heat to 80 °C and react for 3 h. After LCMS detects that the reaction is completed, filter the reaction solution, add water and dichloromethane to extract three times (10 mL * 3), collect the organic phase, dry with anhydrous sodium sulfate, filter, and concentrate. Then purify by silica gel chromatography (EA / PE = 30%). Obtain product C169e (1.13 g, 2.5 mmol), with a yield of 31.3%.

[0957] The third step

[0958] Add C169e (97 mg, 0.22 mmol), C169f (80 mg, 0.20 mmol), tetrakis(triphenylphosphine)palladium (23 mg, 0.02 mmol) catalyst and potassium carbonate (82 mg, 0.60 mmol) into a 100 mL single-necked flask in sequence. Then add 10 mL of dioxane and 1 mL of water as solvents. After fully dissolving the reactants, install a condenser on the single-necked flask and displace nitrogen three times. Then heat to 90 °C and react overnight for 16 h. After LCMS detects that the reaction is completed, filter the reaction solution and directly concentrate it for the next step.

[0959] The fourth step

[0960] Add the crude product C169 g (crude) from the third step into a 100 mL reaction flask. After adding 5 mL of methanol solvent, add an aqueous solution of sodium hydroxide (100 mg). After the reaction continues at room temperature for about 2 h, take a small amount of the reaction solution. LCMS shows that the reaction is complete. After filtering the reaction solution, extract it three times with dichloromethane, collect the organic phase, dry it over anhydrous sodium sulfate, filter, and concentrate it. Purify it by silica gel chromatography (MeOH / CH2Cl2 = 4%) to obtain the product C169 h (80 mg).

[0961] The fifth step

[0962] Add C169 h into a 100 mL single-necked flask. After adding HCl / dioxane (2 mol / L, 1 mL), some solids precipitate. Add 2 mL of methanol to assist dissolution. After reacting at room temperature for 2 h, detect by LCMS that the reaction is complete. Directly evaporate the solvent to dryness and use it as the raw material for the next step.

[0963] The sixth step

[0964] Add C169 i (50 mg, 0.10 mmol), C169 j (35 mg, 0.13 mmol), TCFH (38 mg, 0.14 mmol), and 1-methylimidazole (34 mg, 0.41 mmol) into a 100 mL single-necked flask in sequence. After reacting at room temperature overnight for 16 h, LCMS shows that the reaction is complete. The final product (31.2 mg, 0.02 mmol) C169 is separated by preparative chromatography, with a yield of 42%, [M+H] + = 735.3.

[0965] 1 H NMR (500 MHz, DMSO-d6) δ 11.96 (s, 1H), 10.53 (s, 1H), 8.95 (d, J = 2.4 Hz, 1H), 8.79 (s, 1H), 8.35 (s, 1H), 8.33 (dd, J = 9.0, 2.5 Hz, 1H), 8.08 (d, J = 2.6 Hz, 1H), 8.07 (s, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 1.9 Hz, 1H), 7.39 (dd, J = 8.2, 1.9 Hz, 1H), 6.97 (d, J = 9.1 Hz, 1H), 4.64 - 4.58 (m, 1H), 4.47 - 4.39 (m, 3H), 3.79–3.74 (m, 1H), 3.64–3.60 (m, 2H), 2.87 (t, J = 11.7 Hz, 2H), 2.79–2.73 (m, 3H), 1.80–1.59 (m, 6H), 1.48 (d, J = 6.7 Hz, 6H), 1.26–1.02 (m, 7H).

[0966] Example 53: Preparation of Compound C170

[0967]

[0968] According to the method described in the second to sixth steps of Example 51, except that Compound C170a was used instead of Compound C167b in the second step, Compound C170 (10.52 mg, yield: 7.72%) was obtained. LCMS: [M+H] + = 733.4 1 H NMR (500 MHz, DMSO-d6) δ 12.01 (s, 1H), 10.54 (s, 1H), 8.95 (d, J = 2.4 Hz, 1H), 8.79 (s, 1H), 8.36 (s, 1H), 8.32 (dd, J = 9.0, 2.5 Hz, 1H), 8.09 (s, 1H), 8.05 (s, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.39 (dd, J = 8.1, 2.0 Hz, 1H), 6.98 (d, J = 9.1 Hz, 1H), 4.41 (d, J = 13.1 Hz, 3H), 3.86–3.73 (m, 2H), 3.62 (dd, J = 17.1, 10.9 Hz, 2H), 2.87 (t, J = 12.4 Hz, 2H), 2.75 (q, J = 7.6, 6.8 Hz, 3H), 1.81–1.58 (m, 6H), 1.27–1.16 (m, 4H), 1.10 (tt, J = 7.9, 4.8 Hz, 5H), 1.01 (td, J = 7.3, 4.9 Hz, 2H).

[0969] C170c: LCMS: [M+H] + = 444.0. C170e: LCMS: [M+H] + = 723.3. C170f: LCMS: [M+H] + = 583.4. C170g: LCMS: [M+H] + = 483.3

[0970] Example 54: Preparation of Compound C172

[0971]

[0972] First step:

[0973] C172 a (95 mg, 0.22 mmol), C172 b (80 mg, 0.20 mmol), tetrakis(triphenylphosphine)palladium(0) (23 mg, 0.02 mmol) catalyst and potassium carbonate (82 mg, 0.60 mmol) were successively added into a 100 mL single-necked flask. Subsequently, 10 mL of dioxane and 1 mL of water were added as solvents. After the reactants were fully dissolved, the single-necked flask was equipped with a condenser and purged with nitrogen three times. Then the temperature was raised to 90 °C and the reaction was carried out overnight for 16 h. The reaction was monitored by LCMS. After the reaction solution was filtered, it was directly concentrated for the next step.

[0974] Step 2:

[0975] The crude product C172 c (crude) from the first step was charged into a 100 mL reaction flask. After adding 5 mL of methanol as a solvent, an aqueous solution of sodium hydroxide (100 mg) was added. After the reaction continued at room temperature for about 2 h, a small amount of the reaction solution was taken. LCMS showed that the reaction was complete. After the reaction solution was filtered, it was extracted three times with dichloromethane. The organic phases were collected, dried over anhydrous sodium sulfate, filtered, and concentrated. Purification was carried out by silica gel chromatography (MeOH / CH2Cl2 = 4%) to obtain the product C172 d (60 mg).

[0976] Step 3:

[0977] C172 d was charged into a 100 mL single-necked flask. After adding HCl / dioxane (2 mol / L, 1 mL), some solids precipitated. 2 mL of methanol was added to assist dissolution. After the reaction was carried out at room temperature for 2 h, the reaction was monitored by LCMS. After the solvent was directly rotary evaporated, it was used as the raw material for the next step.

[0978] Step 4:

[0979] C172 e (50 mg, 0.10 mmol), C172 f (35 mg, 0.13 mmol), TCFH (38 mg, 0.14 mmol), 1-methylimidazole (34 mg, 0.41 mmol) were successively charged into a 100 mL single-necked flask. After the reaction was carried out at room temperature overnight for 16 h, LCMS showed that the reaction was complete. The final product (33.2 mg, 0.02 mmol) C172 was separated by preparative chromatography, with a yield of 45%, [M+H] + = 738.3.

[0980] 11H NMR (500 MHz, DMSO-d6) δ 12.03 (s, 1H), 10.53 (s, 1H), 8.85 (s, 1H), 8.34 (s, 1H), 8.11 (d, J = 2.7 Hz, 1H), 8.07 (s, 1H), 8.02–7.95 (m, 2H), 7.65 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.39 (dd, J = 8.2, 1.9 Hz, 1H), 7.16 (t, J = 9.0 Hz, 1H), 4.53–4.40 (m, 1H), 4.23 (q, J = 7.3 Hz, 2H), 3.79–3.74 (m, 1H), 3.67–3.52 (m, 2H), 3.46 (d, J = 11.5 Hz, 2H), 2.84–2.66 (m, 5H), 1.85–1.77 (m, 3H), 1.73–1.47 (m, 4H), 1.43 (t, J = 7.3 Hz, 3H), 1.38–1.15 (m, 5H), 1.14–1.06 (m, 2H).

[0981] Example 55: Preparation of Compound C175

[0982]

[0983] The first step

[0984] Dissolve Compound C175a (45 mg, 0.090 mmol) in dioxane (50 mL) and water (5 mL) at room temperature. Add C175b (45 mg, 0.11 mmol), potassium carbonate (38 mg, 0.27 mmol), and tetrakis(triphenylphosphine)palladium(0) (10 mg, 0.0087 mmol, purity 100%). Stir the reaction mixture at 90 °C for 3 hours under a nitrogen atmosphere. TLC shows that the starting materials have completely reacted. Concentrate the reaction mixture in vacuo to obtain a crude product. Then dissolve it in dichloromethane (100 ml). Dry the organic layer over anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain crude Compound C175c (62 mg, yield: 96.57%), which is directly used in the next step.

[0985] The second step

[0986] Dissolve Compound C175c (62 mg, 0.086 mmol) in methanol (5 mL) and water (1 mL) at room temperature. Add sodium hydroxide (34 mg, 0.86 mmol). Stir the reaction mixture at room temperature for 2 hours. LCMS shows that the reactants have been completely converted and the product is detected. Filter the reaction mixture and concentrate in vacuo to obtain a crude product. Purify it by silica gel chromatography (methanol:dichloromethane = 0 - 4%) to obtain Compound C175d (42 mg, yield: 84.21%). LCMS: [M+H]+ = 577.5。

[0987] The third step

[0988] Dissolve compound C175d (42 mg, 0.073 mmol) in methanol (3 mL) at room temperature, add dioxane hydrochloride (3 mL, 4 M), and stir the reaction solution at room temperature for 2 hours. LCMS shows that the reactant is completely converted and the product is detected. Concentrate the reaction solution under vacuum to obtain crude compound C175e (37 mg, yield: 99.03%), which is directly used for the next step. LCMS: [M+H] + = 482.3。

[0989] The fourth step

[0990] Dissolve compound C175e (37 mg, 0.078 mmol) in DMSO (3 mL) at room temperature, add 4-chloro-3-(2,4-dioxotrihydropyrimidin-1(2H)-yl)benzoic acid (23 mg, 0.086 mmol), TCFH (R2, 33 mg, 0.12 mmol), and NMI (32 mg, 0.39 mmol), and stir the reaction solution at room temperature for 16 hours. LCMS shows that the reactant is completely converted and the product is detected. Pour the reaction solution into water (30 mL), then extract with dichloromethane (20 mL × 2). Dry the organic layer with anhydrous sodium sulfate, filter, and concentrate under vacuum to obtain a crude product. Purify by Pre-HPLC to obtain compound C175 (14.01 mg, yield: 23.46%). LCMS: Rt = 2.100 min; [M+H] + = 727.4。

[0991] Example 56: Preparation of compound C177

[0992]

[0993] The first step:

[0994] Add C177 a (60 mg, 0.13 mmol), C177 b (50 mg, 0.12 mmol), tetrakis(triphenylphosphine)palladium (14 mg, 0.01 mmol) catalyst, and potassium carbonate (51 mg, 0.36 mmol) to a 100 mL single-necked flask in sequence. Then add 10 mL of dioxane and 1 mL of water as solvents. After fully dissolving the reactants, install a condenser on the single-necked flask and displace with nitrogen three times. Then heat to 90 °C and react overnight for 16 h. Detect the completion of the reaction by LCMS. Filter the reaction solution and directly concentrate it for the next step.

[0995] The second step:

[0996] Put the crude product C177 c (crude product) from the first step into a 100 mL reaction flask, add 5 mL of methanol solvent, and then add an aqueous solution of sodium hydroxide (100 mg). After reacting at room temperature for about 2 h, take a small amount of the reaction solution. LCMS shows that the reaction is complete. After filtering the reaction solution, extract it three times with dichloromethane, collect the organic phase, dry it over anhydrous sodium sulfate, filter, and concentrate it. Purify it by silica gel chromatography (MeOH / CH2Cl2 = 4%) to obtain the product C177 d (60 mg).

[0997] The third step:

[0998] Put C177 d into a 100 mL single-necked flask. After adding HCl / dioxane (2 mol / L, 1 mL), some solids precipitate. Add 2 mL of methanol to assist dissolution. After reacting at room temperature for 2 h, detect that the reaction is complete by LCMS. Directly evaporate the solvent to dryness and use it as the raw material for the next step.

[0999] The fourth step:

[1000] Put C177 e (50 mg, 0.10 mmol), C177 f (35 mg, 0.13 mmol), TCFH (38 mg, 0.14 mmol), and 1-methylimidazole (34 mg, 0.41 mmol) into a 100 mL single-necked flask in sequence. After reacting at room temperature overnight for 16 h, LCMS shows that the reaction is complete. The final product (25.03 mg, 0.033 mmol) C177 is separated by preparative chromatography, with a yield of 33%, [M + H] + = 747.3.

[1001] 1 H NMR (500 MHz, DMSO-d6) δ 11.98 (s, 1H), 10.53 (s, 1H), 8.95 (s, 1H), 8.81 (s, 1H), 8.41 (s, 1H), 8.36 (d, J = 8.0 Hz, 1H), 8.10 (d, J = 2.7 Hz, 1H), 8.05 (s, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.39 (dd, J = 8.2, 2.0 Hz, 1H), 7.00 (d, J = 6.3 Hz, 1H), 4.51–4.34 (m, 3H), 4.06 (d, J = 7.1 Hz, 2H), 3.81–3.71 (m, 1H), 3.66–3.51 (m, 2H), 2.89 (t, J = 12.0 Hz, 2H), 2.81–2.68 (m, 3H), 1.84–1.58 (m, 6H), 1.36–1.02 (m, 8H), 0.60–0.52 (m, 2H), 0.45–0.37 (m, 2H).

[1002] Example 57: Preparation of Compound C179

[1003]

[1004] Except for using C179 a instead of C177 a in the first step, according to the method described in Example 56, C179 (25.03 mg, 0.033 mmol, yield 33%) was prepared, [M+H] + = 761.3.

[1005] 1 H NMR (500 MHz, DMSO-d6) δ 11.96 (s, 1H), 10.52 (s, 1H), 8.95 (s, 1H), 8.79 (s, 1H), 8.42–8.27 (m, 2H), 8.08 (s, 2H), 7.64 (d, J = 7.9 Hz, 1H), 7.56 (s, 1H), 7.39 (d, J = 7.6 Hz, 1H), 6.96 (d, J = 8.5 Hz, 1H), 4.87–4.74 (m, 1H), 4.54–4.28 (m, 3H), 3.83–3.52 (m, 4H), 2.92–2.71 (m, 5H), 2.21-2.06 (m, 2H), 2.04–1.91 (m, 2H), 1.88–1.62 (m, 9H), 1.26–1.01 (m, 7H).

[1006] Example 58: Preparation of Compound C181

[1007]

[1008] Except for using C181 a instead of C177 a in the first step, according to the method described in Example 56, C181 (25.03 mg, 0.033 mmol, yield 33%) was prepared. [M+H] + = 761.3.

[1009] Example 59: Preparation of Compound C195

[1010]

[1011] Except for using C195b instead of C169b and cesium carbonate instead of NaH in the first step, according to a method similar to that described in Example 52, C195 (29 mg, 0.04 mmol) was prepared, yield 67%, [M+H] + = 758.2.

[1012] 11H NMR (500 MHz, DMSO-d6) δ 12.01 (s, 1H), 10.52 (s, 1H), 8.99 (d, J = 2.5 Hz, 1H), 8.81 (s, 1H), 8.48 (s, 1H), 8.33 (dd, J = 9.0, 2.5 Hz, 1H), 8.19–8.08 (m, 2H), 7.64 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.39 (dd, J = 8.2, 2.0 Hz, 1H), 6.96 (d, J = 9.1 Hz, 1H), 6.42 (tt, J = 55.1, 3.9 Hz, 1H), 4.74 (td, J = 15.0, 3.7 Hz, 2H), 4.43 (dd, J = 24.0, 9.9 Hz, 3H), 3.82–3.71 (m, 1H), 3.68–3.50 (m, 2H), 2.87 (t, J = 11.8 Hz, 2H), 2.75 (q, J = 7.5, 6.7 Hz, 3H), 1.85–1.54 (m, 6H), 1.29–1.03 (m, 7H).

[1013] Example 60: Preparation of Compound C196

[1014]

[1015] Except using C196 b instead of C195 b in the first step, according to the method described in Example 59, Compound C196 was obtained. The final product C196 was obtained by preparative chromatography separation with a yield of 67%, [M+H] + = 776.2

[1016] 1 1H NMR (500 MHz, DMSO-d6) δ 12.05 (s, 1H), 10.52 (s, 1H), 9.00 (d, J = 2.4 Hz, 1H), 8.81 (s, 1H), 8.55 (s, 1H), 8.33 (dd, J = 9.0, 2.5 Hz, 1H), 8.18 (d, J = 2.5 Hz, 2H), 7.64 (d, J = 8.2 Hz, 1H), 7.56 (d, J = 2.0 Hz, 1H), 7.39 (dd, J = 8.2, 1.9 Hz, 1H), 6.96 (d, J = 9.1 Hz, 1H), 5.26 (q, J = 9.1 Hz, 2H), 4.43 (dd, J = 24.4, 11.2 Hz, 3H), 3.80–3.73 (m, 1H), 3.66–3.54 (m, 2H), 3.12–3.01 (m, 1H), 2.89 (s, 2H), 2.75 (q, J = 7.4, 6.6 Hz, 2H), 1.80–1.60 (m, 6H), 1.26–1.04 (m, 7H).

[1017] Example 61: Preparation of Compound C197

[1018]

[1019] Compound C197 can be obtained according to the method described in Example 59, except that fluoroethane is used instead of C195 b in the first step. The final product C197 is obtained by preparative chromatography separation. [M+H] + = 740.2.

[1020] Example 62: Preparation of Compound C198

[1021]

[1022] Compound C198 (10.52 mg, yield: 7.72%) was obtained according to the method described in Example 53, except that compound C198a was used instead of compound C170a in the first step.

[1023] Example 63: Preparation of Compound C199

[1024]

[1025] First step:

[1026] Place C199 a (1.5 g, 7.8 mmol), C199 b (1.3 g, 14.7 mmol), and TPP (2.4 g, 9.2 mmol) in a 100 mL three-necked flask. Subsequently, add 30 mL of freshly distilled tetrahydrofuran. Then, displace the system with nitrogen three times and stir at -20 °C for about 30 min. Gradually add 1.8 mL of DEAD (9.2 mmol) reagent. After reacting for 2 h, transfer the reaction to room temperature and react overnight. It was found that a new product was formed by TLC, and then the reaction was terminated. Add saturated ammonium chloride aqueous solution to quench the reaction, extract with ethyl acetate three times (20 mL * 3). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. Purification by column chromatography (EA / PE = 30%) gave the new product C199 c (850 mg).

[1027] Second to sixth steps:

[1028] The second to sixth steps were carried out in a similar manner to the second to sixth steps of Example 52 to obtain Compound C199 (29 mg, 0.04 mmol). [M+H] + = 764.3.

[1029] 11H NMR (500 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.51 (s, 1H), 8.93 (s, 1H), 8.80 (s, 1H), 8.40 (s, 1H), 8.35 (d, J = 6.3 Hz, 1H), 8.11 (s, 2H), 7.64 (d, J = 8.2 Hz, 1H), 7.58–7.53 (m, 1H), 7.42–7.35 (m, 1H), 7.01 (d, J = 6.5 Hz, 1H), 5.17–5.11 (m, 1H), 4.56–4.33 (m, 3H), 4.07–4.00 (m, 2H), 3.97–3.92 (m, 1H), 3.90–3.84 (m, 1H), 3.80–3.71 (m, 1H), 3.67–3.52 (m, 2H), 2.90 (t, J = 12.2 Hz, 2H), 2.81–2.67 (m, 3H), 2.47

[1030] –2.35 (m, 2H), 2.34–2.28 (m, 1H), 1.86–1.57 (m, 6H), 1.25–1.02 (m, 6H).

[1031] Example 64: Preparation of Compound C200

[1032]

[1033] According to the method of Example 63, Compound C200 can be obtained. The final product C200 was obtained by preparative chromatography separation, [M+H] + = 764.3.

[1034] Example 65: Preparation of Compound C201

[1035]

[1036] According to the method described in the second to sixth steps of Example 52, except that C201a was used instead of C169c in the second step, Compound C201 (31 mg, 0.04 mmol) was obtained, [M+H] + = 752.3.

[1037] 11H NMR (500 MHz, DMSO-d6) δ 11.96 (s, 1H), 10.51 (s, 1H), 8.96 (d, J = 2.0 Hz, 1H), 8.79 (s, 1H), 8.36 (s, 1H), 8.33 (dd, J = 9.0, 2.3 Hz, 1H), 8.09 (d, J = 2.6 Hz, 1H), 8.05 (s, 1H), 7.64 (d, J = 8.2 Hz, 1H), 7.55 (d, J = 1.7 Hz, 1H), 7.39 (dd, J = 8.2, 1.7 Hz, 1H), 6.96 (d, J = 9.0 Hz, 1H), 4.55–4.30 (m, 3H), 4.35 (t, J = 5.3 Hz, 2H), 3.83–3.69 (m, 3H), 3.66–3.53 (m, 2H), 3.27 (s, 3H), 2.87 (t, J = 12.1 Hz, 2H), 2.82–2.69 (m, 3H), 1.82–1.58 (m, 6H), 1.29–0.99 (m, 7H).

[1038] Example 66: Preparation of Compound C203

[1039]

[1040] Except that compound C203a was used instead of compound C170a in the first step, according to the method described in Example 53, compound C203 (10.52 mg, yield: 7.72%) was obtained. LCMS: [M+H] + = 709.2.

[1041] 1 1H NMR (500 MHz, DMSO-d6) δ 9.32–8.83 (m, 3H), 8.42–8.10 (m, 3H), 7.56 (d, J = 7.5 Hz, 1H), 7.41–7.08 (m, 2H), 4.08 (d, J = 34.1 Hz, 2H), 3.96–3.81 (m, 7H), 3.19–2.69 (m, 5H), 1.90–1.69 (m, 5H), 1.64–1.37 (m, 5H), 1.23 (d, J = 12.5 Hz, 1H).

[1042] Example 67: Preparation of Compound C204

[1043]

[1044] According to the method described in Example 66, compound C204 was obtained, and the final product C204 was obtained by preparative chromatography. [M+H] + = 688.8.

[1045] Example 68: Preparation of Compound C205

[1046]

[1047] The first step:

[1048] C205 a (0.2 g, 1.3 mmol), C205 b (0.5 g, 1.1 mmol), catalyst (40 mg, 0.05 mmol), potassium carbonate (0.5 g, 3.6 mmol), etc. were successively added into a 250 mL single-necked flask. Subsequently, 30 mL of dioxane and 10 mL of water were added as solvents. Then, after adding a condenser to the single-necked flask, nitrogen was replaced three times with a three-way valve and then heated to 80 °C for reaction for 3 h. After the reaction was completed by LCMS detection, the reaction solution was filtered, and then water and dichloromethane were added for extraction three times (10 mL * 3). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and then purified by silica gel chromatography (EA / PE = 30%). Product C205 c (0.1 g, 0.3 mmol) was obtained with a yield of 23%.

[1049] The second step:

[1050] C205 c (100 mg, 0.3 mmol), C205 d (70 mg, 0.2 mmol), tetrakis(triphenylphosphine)palladium (20 mg, 0.02 mmol) catalyst, and potassium carbonate (100 mg, 0.54 mmol) were successively added into a 100 mL single-necked flask. Subsequently, 10 mL of dioxane and 1 mL of water were added as solvents. After the reactants were fully dissolved, a condenser was added to the single-necked flask and nitrogen was replaced three times. Then, the temperature was raised to 90 °C and the reaction was carried out overnight for 16 h. After the reaction was completed by LCMS detection, it was purified by silica gel chromatography (MeOH / CH2Cl2 = 4%) to obtain product C205 e (39 mg).

[1051] The third step:

[1052] The product of the second step was placed in a 50 mL single-necked flask, and 10% Pd / C (30 mg) was added. After the system was replaced with hydrogen, the reaction was started at room temperature for 16 h. After the reaction was completed by LCMS detection, it was directly filtered through diatomaceous earth and then the methanol was evaporated to dryness for the next step.

[1053] The fourth step:

[1054] The crude product C205 f (crude product) from the third step was put into a 100 mL reaction flask. After adding 5 mL of methanol solvent, an aqueous solution of sodium hydroxide (100 mg) was added. After the reaction continued at room temperature for about 2 h, a small amount of the reaction solution was taken. LCMS showed that the reaction was complete. After filtering the reaction solution, it was extracted three times with dichloromethane. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and concentrated. It was purified by silica gel chromatography (MeOH / CH2Cl2 = 4%) to obtain the product C205 g (29 mg).

[1055] Step 5:

[1056] C205 g (29 mg, 0.06 mmol) was put into a 100 mL single-necked flask. After adding HCl / dioxane (2 mol / L, 1 mL), some solids precipitated. 2 mL of methanol was added to assist dissolution. After reacting at room temperature for 2 h, LCMS detected that the reaction was complete. The solvent was directly evaporated to dryness and used as the raw material for the next step.

[1057] Step 6:

[1058] C205 h (29 mg, 0.07 mmol), C205 I (19 mg, 0.07 mmol), TCFH (20 mg, 0.08 mmol), and 1-methylimidazole (20 mg, 0.24 mmol) were successively put into a 100 mL single-necked flask. After reacting at room temperature overnight for 16 h, LCMS showed that the reaction was complete. The final product (3.6 mg, 0.005 mmol) C205 was separated by preparative chromatography, and the yield was 7.1%. [M+H] + = 655.2.

[1059] Example 69: Preparation of compound C206

[1060]

[1061] Step 1

[1062] Compound C206a (4.5 g, 21.47 mmol) was dissolved in THF (10 mL), methanol (10 mL), and water (5 mL) at room temperature. Lithium hydroxide monohydrate (1.35 g, 32.20 mmol) was added, and the reaction solution was stirred at room temperature for 16 hours. LCMS showed that the reactants were completely converted and the product was detected. The reaction solution was concentrated in vacuo to obtain a crude product. Then it was dissolved in water (2 mL), 1N HCl solution was added to adjust to about pH 3, stirred and filtered. The filter cake was washed with water (30 mL) and then dried to obtain compound C206b (4.2 g, yield 100.03%). LCMS: [M+H] + = 196.1.

[1063] Step 2

[1064] Compound C206b (4.2 g, 21.47 mmol) was dissolved in THF (20 mL) under an ice bath. Compound C206c (2.54 g, 21.47 mmol), triphenylphosphine (5.63 g, 21.47 mmol) and DIAD (4.34 g, 21.47 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours under a nitrogen atmosphere. TLC showed that most of the reactants were converted and new spots were detected. The reaction mixture was poured into water (50 mL), then extracted with ethyl acetate (30 mL × 2). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 4%) to obtain compound C206c (2.14 g, yield: 33.69%).

[1065] The third step

[1066] Compound C206c (2.14 g, 7.23 mmol) was dissolved in DMF (20 mL) at room temperature. NaH (350 mg, 8.75 mmol) (60%) was added under an ice bath and stirred for 10 minutes, then methyl 3 - bromopropionate (1.81 g, 10.85 mmol) was added. The reaction mixture was stirred at room temperature for 16 hours. TLC showed that part of the reactants were converted and new spots were detected. The reaction mixture was concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography (petroleum ether: ethyl acetate = 0 - 9%) to obtain compound C206d (1.745 g, yield: 63.16%).

[1067] The fourth step

[1068] Compound C206d (1.7 g, 4.45 mmol) was dissolved in acetic acid (20 mL) at room temperature. Concentrated sulfuric acid (44 mg, 0.45 mmol) was added. The reaction mixture was stirred at 120 °C for 16 hours. LCMS showed that the reactants were completely converted and the product was detected. The reaction mixture was concentrated in vacuo to obtain a crude product. It was purified by silica gel chromatography (methanol: dichloromethane = 0 - 2%) to obtain compound C206e (0.9 g, yield: 75.5%). LCMS: [M + H] + = 268.4

[1069] The fifth step

[1070] Dissolve compound C206f (100 mg, 0.20 mmol) in DMSO (3 mL) at room temperature, add C206e (64 mg, 0.24 mmol), TCFH (84.17 mg, 0.30 mmol) and NMI (82.1 mg, 1 mmol), and stir the reaction mixture at room temperature for 16 hours. LCMS shows that the reactants are completely converted and the product is detected. Pour the reaction mixture into water (30 mL), then extract with dichloromethane (30 mL * 2). Dry the organic layer over anhydrous sodium sulfate, filter, and concentrate in vacuo to obtain the crude product. Purify by pre-HPLC to obtain compound C206 (4.4 mg, yield: 2.76%). LCMS: [M + H] + = 706.6.

[1071] 1 H NMR (500 MHz, DMSO-d6) δ 11.97 (s, 1H), 10.93 (s, 1H), 8.97 (d, J = 2.5 Hz, 1H), 8.79 (s, 1H), 8.36–8.30 (m, 2H), 8.08 (d, J = 2.3 Hz, 1H), 8.02 (s, 1H), 7.56–7.52 (m, 1H), 7.37 (d, J = 2.0 Hz, 1H), 7.32 (dd, J = 8.2, 2.1 Hz, 1H), 6.96 (d, J = 9.0 Hz, 1H), 4.48–4.39 (m, 3H), 4.31–4.24 (m, 1H), 3.93 (s, 3H), 2.87 (t, J = 12.4 Hz, 2H), 2.80–2.71 (m, 2H), 2.38–2.30 (m, 1H), 2.03–1.96 (m, 1H), 1.82–1.59 (m, 7H), 1.22–1.03 (m, 7H).

[1072] Example 70: Preparation of compound C209

[1073]

[1074] According to the method described in the second to sixth steps of Example 52, except that C209a is used instead of C169c in the second step, compound C209 (17.8 mg, 0.04 mmol) is obtained with a yield of 39%. [M + H] + = 737.3.

[1075] The compounds provided by the present invention and their applications have been introduced in detail above.

[1076] In this text, specific embodiments are used to illustrate the principles and implementation manners of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. It should be noted that for those of ordinary skill in the art, without departing from the principles of the present invention, several improvements and modifications can still be made to the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.

Claims

1. A compound represented by the general formula (I): [B-L] n -HPK1 ligand (I), or a pharmaceutically acceptable salt thereof, wherein: n is 1; the HPK1 ligand is a compound of general formula (H-I) or a compound of general formula (H-III): wherein: W is N; the CyB ring is selected from 4- to 6-membered cycloalkyl, 5- to 6-membered nitrogen-containing heterocyclic group, or 5- to 6-membered oxygen-containing heterocyclic group; R 1 and R 3 each is hydrogen; R 2 selected from -C 1-8 alkyl; R 4 Independently is hydrogen, halogen, -C 1-8 alkyl, -CN, -OR 4a , or -NR 4a R 4b , wherein said -C 1-8 alkyl is optionally substituted by at least one substituent R 4d substituted; R 4a and R 4b each independently is hydrogen; R 4d independently is a halogen; s is 0 or 1, provided that the valence theory is satisfied; t is 0 or 1, provided that the valence theory is satisfied; m is 0 or 1, provided that the valence theory is satisfied; CyD is selected from a benzene ring, a 5-membered heteroaromatic ring or a 6-membered heteroaromatic ring; R X Selected from H and C 1-8 alkyl, wherein said C 1-8 alkyl is optionally substituted by one or more substituents independently selected from deuterium, halogen, -OH, -CN, -NR Xa R Xb , -OR Xa , -CO-NHR Xb , -CO-NR Xa R Xb and 3- to 6-membered cycloalkyl; R Xa 、R Xb each independently selected from C 1-8 alkyl; B is selected from: and L is selected from:

2. The compound according to claim 1, wherein in the general formula (H-I), the CyB ring is selected from cyclobutyl, cyclopentyl, cyclohexyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, azetidinyl, pyrrolidinyl, or piperidinyl.

3. The compound according to claim 2, wherein the CyB ring is selected from 4. The compound according to claim 1, wherein in the general formula (H-I), R 2 is selected from -C 1-3 alkyl.

5. The compound according to claim 1, wherein in the general formula (H-I), is 6. The compound according to claim 1, wherein the compound of general formula (H-I) is a compound of general formula (H-V):

7. The compound according to claim 1, wherein in the general formula (H-III), R X is selected from C 1-4 alkyl, wherein the C 1-4 alkyl is optionally substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, Br, -OH, -CN, -NR Xa R Xb or -OR Xa .

8. The compound according to claim 7, wherein the C 1-4 alkyl group is selected from methyl, ethyl, isopropyl, isobutyl, sec-butyl and tert-butyl.

9. The compound according to claim 8, wherein said C 1-4 alkyl group is methyl or ethyl.

10. The compound according to claim 9, wherein R Xa and R Xb are each independently selected from C 1-4 alkyl.

11. The compound according to claim 10, wherein R Xa and R Xb are each methyl.

12. The compound according to claim 7, wherein R X is selected from methyl or ethyl, wherein the methyl or ethyl is optionally substituted with 1, 2 or 3 substituents independently selected from deuterium, F, Cl, -OH, -CN and -OCH3.

13. The compound according to claim 12, wherein R X is selected from methyl or ethyl, wherein the methyl or ethyl is optionally substituted with 1, 2 or 3 substituents independently selected from deuterium, F and -OCH3.

14. The compound according to claim 13, wherein R X is selected from methyl, -CD3, ethyl, -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, or 15. The compound according to claim 14, wherein R X is selected from methyl, -CD3, ethyl, -CH2CH2F, -CH2CHF2, -CH2CF3, or 16. The compound according to any one of claims 1-15, wherein CyD is selected from: a benzene ring; wherein X1, X2 and X3 are each independently selected from CH, C or N, provided that at least one of X1, X2 and X3 is N; and a 5-membered heteroaromatic ring, the heteroaromatic ring containing 1 or 2 heteroatoms independently selected from nitrogen, oxygen or sulfur as one or more ring members.

17. The compound according to claim 16, wherein the CyD is selected from a benzene ring, a pyridyl group, a pyrimidinyl group, a pyridazinyl group, a pyrazinyl group, a triazinyl group, 18. The compound according to claim 17, wherein the CyD is selected from a benzene ring, 19. The compound according to claim 18, wherein the CyD is a benzene ring, 20. A compound according to any one of claims 1 - 15, wherein R 4 is independently hydrogen, halogen, -C 1-3 alkyl, -CN, -OR 4a , or -NR 4a R 4b , wherein the -C 1-3 alkyl is optionally substituted with at least one halogen.

21. The compound according to claim 20, wherein R 4 is independently hydrogen, halogen or -C 1-3 alkyl, wherein the -C 1-3 alkyl is optionally substituted with at least one substituent selected from fluorine, chlorine or bromine.

22. The compound according to claim 21, wherein R 4 is independently hydrogen, methyl, ethyl, fluorine, chlorine, -CHF2, -CH2F, -CF3, -CH2CHF2, -CH2CH2F, or -CH2CF3.

23. The compound according to claim 22, wherein R 4 is independently hydrogen, methyl, ethyl, fluorine, or -CF3.

24. The compound according to claim 1, wherein the HPK1 ligand is selected from:

25. The compound according to claim 1, wherein the HPK1 ligand is selected from:

26. The compound according to any one of claims 1-15, wherein B is 27. The compound according to any one of claims 1-15, wherein said L is 28. A compound represented by the general formula (I): [B-L] n - ligand (I) of HPK1, or a pharmaceutically acceptable salt thereof, wherein the HPK1 ligand is a compound of general formula (H-I): wherein: W is N; For R 1 、R 3 and R 4 each represents hydrogen; CyD is pyridyl; L is B is n is 1; m is 0 or 1, provided that the valence theory is satisfied; and s is 0 or 1, provided that the valence theory is satisfied.

29. The compound according to claim 28, wherein is 30. A compound according to claim 28 or 29, wherein the CyD is 31. The compound according to claim 28, wherein B is 32. The compound according to claim 1, wherein the HPK1 ligand is a compound of general formula (H-III), R X is selected from methyl or ethyl, wherein the methyl or ethyl is optionally substituted with 1, 2 or 3 substituents independently selected from deuterium, F and -OCH3; CyD is selected from pyridyl; R 4 is hydrogen or -C 1-3 alkyl; L is and B is 33. The compound according to claim 32, wherein R X is selected from methyl, -CD3, ethyl, -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, or 34. The compound according to claim 33, wherein R X is selected from methyl, -CD3, ethyl, -CH2CH2F, -CH2CHF2, -CH2CF3, or 35. The compound according to claim 34, wherein R X is -CH2CHF2 or -CH2CF3.

36. A compound according to any one of claims 32 - 35, wherein the CyD is 37. The compound according to claim 36, wherein the CyD is 38. A compound according to any one of claims 32 - 35, wherein R 4 is hydrogen.

39. A compound according to any one of claims 32 - 35, wherein B is 40. A compound represented by the general formula (I): [B-L] n - The ligand (I) of HPK1, wherein: n, B and L are as defined in claim 1, 26 or 27, The HPK1 ligand is selected from: and the HPK1 ligand described in claim 24 or 25.

41. A compound, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

42. A pharmaceutical composition comprising the compound according to any one of claims 1-41, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

43. A pharmaceutical composition comprising the compound according to any one of claims 1-41, or a pharmaceutically acceptable salt thereof, and one or more therapeutically active ingredients.

44. Use of the pharmaceutical compound according to any one of claims 1-41 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to any one of claims 42-43 in the preparation of a drug for the treatment and / or prevention of HPK1-mediated diseases and related diseases.

45. The application according to claim 44, wherein the HPK1-mediated diseases and related diseases are selected from the group consisting of lung cancer, squamous cell carcinoma, bladder cancer, gastric cancer, peritoneal cancer, breast cancer, breast ductal carcinoma, head and neck cancer, rectal cancer, liver cancer, kidney cancer, renal pelvis cancer, esophageal cancer, esophageal adenocarcinoma, glioma, prostate cancer, thyroid cancer, female reproductive system cancer, lymphoma, neurofibromatosis, bone cancer, skin cancer, brain cancer, colon cancer, testicular cancer, gastrointestinal stromal tumor, oral cancer, pharyngeal cancer, multiple myeloma, leukemia, non-Hodgkin lymphoma, large intestinal villous adenoma, melanoma, cell tumor, sarcoma, and myelodysplastic syndrome.

46. The application according to claim 44, wherein the HPK1-mediated diseases and related diseases are selected from the group consisting of ovarian cancer, endometrial cancer, uterine body cancer, and carcinoma in situ.

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