Synthesis of SHP2 and CDK4 / 6 dual-target inhibitory compounds and their preparation methods and applications

Abstract

The present invention belongs to the field of medicinal chemistry, and specifically relates to a synthesis of a dual-target inhibitor containing SHP2 and CDK4 / 6, and its preparation method and application. Contains three compounds of general formula I-III and pharmaceutically acceptable salts, enantiomers, diastereomers, tautomers, solvates, polymorphs or prodrugs thereof; the present invention prepares a new class of dual-target inhibitors of SHP2 and CDK4 / 6 through the method of pharmacophore fusion and rational drug design to solve the current problems of drug resistance and poor efficacy of SHP2 inhibitors and CDK4 / 6 inhibitors; the important significance of the present invention is to provide the first dual-target inhibitor of SHP2 and CDK4 / 6, providing a basis for later solving the problem of drug resistance of kinases and phosphatases. At the same time, the embodiments of the present invention confirm that SHP2 inhibitors and CDK4 / 6 inhibitors have synergistic effects, providing theoretical support for the subsequent development of SHP2 dual-target inhibitors.

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and specifically relates to a synthesis of a dual-target inhibitor containing SHP2 and CDK4 / 6, as well as a preparation method and application thereof. Background Art

[0002] SHP2 is a ubiquitous non-receptor protein tyrosine phosphatase with two N-terminal Src homology 2 domains (N-SH2 and C-SH2), a catalytic domain (PTP), and a C-terminal tail. These two SH2 domains control SHP2's subcellular localization and functional regulation. As a downstream signaling molecule for growth factors such as platelet-derived growth factor (PDGF), epidermal growth factor (EGF), fibroblast growth factor (FGF), interleukin-3 (IL-3), leukemia inhibitory factor (LIF), and interferon-α (INF-α), SHP2 participates in multiple signaling pathways, including the RAS / MARK pathway, the PI3K / AKT pathway, the JAK / STAT pathway, and the JNK pathway. Studies have shown that SHP2 mutations or overactivation can lead to the development of Noonan syndrome, juvenile myelomonocytic leukemia, myelodysplastic syndrome, B-cell acute lymphoblastic leukemia, and solid tumors (lung cancer, colon cancer, neuroblastoma, melanoma, and liver cancer).

[0003] CDKs are a class of serine / threonine protein kinases. Among them, CDK4 / 6 are core regulators of the cell cycle, controlling transitions between cell cycle phases and serving as important targets for anti-tumor drugs. CDK4 / 6 inhibitors can exert anti-tumor activity by inhibiting the transition of tumor cell cycles from G1 to S phase, activating anti-tumor immune responses, and influencing the tumor microenvironment. Clinically, they have demonstrated promising efficacy against certain tumor types. However, prolonged treatment with CDK4 / 6 inhibitors can easily lead to tumor resistance, resulting in the inhibitors losing their efficacy.

[0004] Studies have shown that the combination of SHP2 inhibitors and CDK4 / 6 inhibitors has a strong synergistic effect. This combination has entered Phase I clinical trials for the treatment of malignant tumors such as head and neck cancer, non-small cell lung cancer, and colon cancer.

[0005] The present invention prepares a new class of SHP2 and CDK4 / 6 dual-target inhibitors through pharmacophore fusion and rational drug design, providing a basis for the later solution to the problem of kinase and phosphatase resistance. Summary of the Invention

[0006] One of the technical problems to be solved by the present invention is the poor clinical efficacy and drug resistance that often occur with long-term use of SHP2 and CDK4 / 6 inhibitors. Therefore, a novel dual-target inhibitory compound containing SHP2 and CDK4 / 6 is provided to facilitate subsequent drug development.

[0007] The solutions to the above technical problems are as follows:

[0008] 1. A compound of formula I, and its pharmaceutically acceptable salts, enantiomers, diastereomers, tautomers, solvates, polymorphs or prodrugs

[0009]

[0010] Ring A is selected from C3-C6 cycloalkyl, C 6- C 10 Aromatic ring, C 6- C 10 Heteroaromatic ring, monocyclic heterocyclic ring, spirocyclic ring, bridged ring, and fused ring; wherein the cycloalkyl, aromatic ring, heteroaromatic ring, monocyclic heterocyclic ring, spirocyclic ring, bridged ring, and fused ring may optionally contain N, O, and S(O) m wherein m is selected from 0, 1 and 2; wherein the cycloalkyl, aromatic ring, heteroaromatic ring, carbocyclic ring, monoheterocyclic ring, spirocyclic ring, bridged ring and cyclic ring may be optionally substituted with one or more substituents;

[0011] Ring B is selected from aromatic rings, heteroaromatic rings, carbocyclic rings, and heterocyclic rings; wherein the aromatic rings, heteroaromatic rings, carbocyclic rings, and heterocyclic rings may optionally contain N, O, and S(O) m wherein m is selected from 0, 1 and 2; wherein the aromatic ring, heteroaromatic ring, carbocyclic ring, heterocyclic ring may be optionally substituted with one or more substituents;

[0012] L 1 For chemical bonds, NR a , C1-C2 carbon chain, O, S(O) m , wherein m is selected from 0, 1 and 2;

[0013] L 2 For chemical bonds, NR a , C1-C2 carbon chain, O, S(O) m , wherein m is selected from 0, 1 and 2;

[0014] Y 1 N or CR 3 , where R 3 is selected from hydrogen, a deuterium atom, a halogen, an amino group, a hydroxyl group, a cyano group, a nitro group, a carboxyl group, a sulfonic acid group, a C1-C6 alkyl group, a C1-C6 deuterated alkyl group, a C1-C6 alkoxy group, a C2-C6 alkenyl group, a C2-C6 alkynyl group or a C3-C6 cycloalkyl group;

[0015] Y 2 N or CR 4 , where R 4 is selected from hydrogen, a deuterium atom, a halogen, an amino group, a hydroxyl group, a cyano group, a nitro group, a carboxyl group, a sulfonic acid group, a C1-C6 alkyl group, a C1-C6 deuterated alkyl group, a C1-C6 alkoxy group, a C2-C6 alkenyl group, a C2-C6 alkynyl group or a C3-C6 cycloalkyl group;

[0016] R 1 、R 2 They are hydrogen, deuterium atoms, halogen, amino, hydroxyl, cyano, nitro, carboxyl, sulfonic acid, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C6-C 10 Heteroaryl; wherein the amino, hydroxy, aryl, heteroaryl, cycloalkyl group may be optionally substituted with one or more substituents;

[0017] Or, R 1 and R 2 Together with the carbon atoms to which they are attached, they form a 3- to 12-membered monocyclic or polycyclic heterocyclic ring, wherein the monocyclic or polycyclic heterocyclic ring may optionally contain a radical selected from N, O and S(O) m wherein m is selected from 0, 1 and 2; wherein the monocyclic or polycyclic heterocyclic ring may be unsubstituted or substituted with one or more substituents;

[0018] Or, R 1 and R 2 Together with the carbon atoms to which they are all attached, they form

[0019]

[0020] Wherein W is absent or selected from CR 6 R 7 、O、NR b 、S(O) m ; wherein m is selected from 0, 1 and 2;

[0021] The C ring is absent or selected from a C3-C7 membered monocyclic ring or a C5-C 12 Meta-polycyclic;

[0022] R a 、R b Each independently selected from hydrogen, deuterium atoms, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C6-C 10 heteroaryl;

[0023] R 4 、R 5 、R 6 、R 7Each is independently selected from hydrogen, deuterium atoms, halogen, amino, hydroxyl, cyano, nitro, carboxyl, sulfonic acid, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C6-C 10 Heteroaryl; wherein the amino, hydroxy, aryl, heteroaryl, cycloalkyl group may be optionally substituted with one or more substituents;

[0024] or R 4 and R 5 Together with the carbon atoms to which they are connected, they form CO, C=NH, C3-C 12 Membered heterocyclic ring or C3-C8 cycloalkyl;

[0025] 2. A compound of formula II, and its pharmaceutically acceptable salts, enantiomers, diastereomers, tautomers, solvates, polymorphs or prodrugs

[0026]

[0027] Ring A is selected from C3-C6 cycloalkyl, C 6- C 10 Aromatic ring, C 6- C 10 Heteroaromatic ring, monocyclic heterocyclic ring, spirocyclic ring, bridged ring, and fused ring; wherein the cycloalkyl, aromatic ring, heteroaromatic ring, monocyclic heterocyclic ring, spirocyclic ring, bridged ring, and fused ring may optionally contain N, O, and S(O) m wherein m is selected from 0, 1 and 2; wherein the cycloalkyl, aromatic ring, heteroaromatic ring, carbocyclic ring, monoheterocyclic ring, spirocyclic ring, bridged ring and cyclic ring may be optionally substituted with one or more substituents;

[0028] Ring B is selected from aromatic rings, heteroaromatic rings, carbocyclic rings, and heterocyclic rings; wherein the aromatic rings, heteroaromatic rings, carbocyclic rings, and heterocyclic rings may optionally contain N, O, and S(O) m wherein m is selected from 0, 1 and 2; wherein the aromatic ring, heteroaromatic ring, carbocyclic ring, heterocyclic ring may be optionally substituted with one or more substituents;

[0029] L 1 For chemical bonds, NR a , C1-C2 carbon chain, O, S(O) m , wherein m is selected from 0, 1 and 2;

[0030] L 2 For chemical bonds, NR a , C1-C2 carbon chain, O, S(O) m , wherein m is selected from 0, 1 and 2;

[0031] R 1 、R 2They are hydrogen, deuterium atoms, halogen, amino, hydroxyl, cyano, nitro, carboxyl, sulfonic acid, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C6-C 10 Heteroaryl; wherein the amino, hydroxy, aryl, heteroaryl, cycloalkyl group may be optionally substituted with one or more substituents;

[0032] Or, R 1 and R 2 Together with the carbon atoms to which they are attached, they form a 3- to 12-membered monocyclic or polycyclic heterocyclic ring, wherein the monocyclic or polycyclic heterocyclic ring may optionally contain a radical selected from N, O and S(O) m wherein m is selected from 0, 1 and 2; wherein the monocyclic or polycyclic heterocyclic ring may be unsubstituted or substituted with one or more substituents;

[0033] Or, R 1 and R 2 Together with the carbon atoms to which they are all attached, they form

[0034]

[0035] Wherein W is absent or selected from CR 6 R 7 、O、NR b 、S(O) m ; wherein m is selected from 0, 1 and 2;

[0036] The C ring is absent or selected from a C3-C7 membered monocyclic ring or a C5-C 12 Meta-polycyclic;

[0037] R a 、R b Each independently selected from hydrogen, deuterium atoms, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C6-C 10 heteroaryl;

[0038] R 4 、R 5 、R 6 、R 7 Each is independently selected from hydrogen, deuterium atoms, halogen, amino, hydroxyl, cyano, nitro, carboxyl, sulfonic acid, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C6-C 10 Heteroaryl; wherein the amino, hydroxy, aryl, heteroaryl, cycloalkyl group may be optionally substituted with one or more substituents;

[0039] or R 4 and R 5Together with the carbon atoms to which they are connected, they form CO, C=NH, C3-C 12 Membered heterocyclic ring or C3-C8 cycloalkyl;

[0040] 3. A compound of formula III, and its pharmaceutically acceptable salts, enantiomers, diastereomers, tautomers, solvates, polymorphs or prodrugs

[0041]

[0042] Ring A is selected from hydrogen,

[0043] Ring B is selected from aromatic rings, heteroaromatic rings, carbocyclic rings, and heterocyclic rings; wherein the aromatic rings, heteroaromatic rings, carbocyclic rings, and heterocyclic rings may optionally contain N, O, and S(O) m wherein m is selected from 0, 1 and 2; wherein the aromatic ring, heteroaromatic ring, carbocyclic ring, heterocyclic ring may be optionally substituted with one or more substituents;

[0044] L 2 For chemical bonds, NR a , C1-C2 carbon chain, O, S(O) m , wherein m is selected from 0, 1 and 2;

[0045] R 1 、R 2 They are hydrogen, deuterium atoms, halogen, amino, hydroxyl, cyano, nitro, carboxyl, sulfonic acid, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C6-C 10 Heteroaryl; wherein the amino, hydroxy, aryl, heteroaryl, cycloalkyl group may be optionally substituted with one or more substituents;

[0046] Or, R 1 and R 2 Together with the carbon atoms to which they are attached, they form a 3- to 12-membered monocyclic or polycyclic heterocyclic ring, wherein the monocyclic or polycyclic heterocyclic ring may optionally contain a radical selected from N, O and S(O) m wherein m is selected from 0, 1 and 2; wherein the monocyclic or polycyclic heterocyclic ring may be unsubstituted or substituted with one or more substituents;

[0047] Or, R 1 and R 2 Together with the carbon atoms to which they are all attached, they form

[0048]

[0049] Wherein W is absent or selected from CR 6 R 7 、O、NRb 、S(O) m ; wherein m is selected from 0, 1 and 2;

[0050] The C ring is absent or selected from a C3-C7 membered monocyclic ring or a C5-C 12 Meta-polycyclic;

[0051] R a 、R b Each independently selected from hydrogen, deuterium atoms, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C6-C 10 heteroaryl;

[0052] R 4 、R 5 、R 6 、R 7 Each is independently selected from hydrogen, deuterium atoms, halogen, amino, hydroxyl, cyano, nitro, carboxyl, sulfonic acid, C1-C6 alkyl, C3-C6 cycloalkyl, C6-C 10 Aryl, C6-C 10 Heteroaryl; wherein the amino, hydroxy, aryl, heteroaryl, cycloalkyl group may be optionally substituted with one or more substituents;

[0053] or R 4 and R 5 Together with the carbon atoms to which they are connected, they form CO, C=NH, C3-C 12 Membered heterocyclic ring or C3-C8 cycloalkyl;

[0054] A pharmaceutical composition characterized by containing compounds I-III and pharmaceutically acceptable excipients.

[0055] The pharmaceutical composition is characterized in that the pharmaceutical composition is prepared into tablets, capsules, injections or freeze-dried powders.

[0056] The aromatic spirocyclic compound and the pharmaceutical composition are used in the preparation of anti-tumor drugs, as prodrugs of anti-tumor drugs or as intermediates of anti-tumor drugs.

[0057] Design ideas:

[0058]

[0059] In dual-target drug design, the pharmacophore fusion approach often avoids issues such as high molecular weight and poor metabolism. In the design of a dual-target inhibitor for SHP2 / CDK4, we observed that both compounds possess pyridine rings and that modifications are located at the edge of the solvent region, leading to the innovative merging of their pharmacophores and linkers. Because this fusion-based dual-target inhibitor presents challenges in balancing its activity across its two targets, we briefly explored the effects of different pharmacophores and linkers on inhibitory activity, resulting in compounds 1-26.

[0060] SAR analysis found that compounds with two pharmacophores linked via the para position of the pyridine ring can maintain CDK4 activity but reduce SHP2 activity; compounds with two pharmacophores linked via the meta position of the pyridine ring can maintain SHP2 activity but reduce CDK activity; the overall pharmacophore effect of Abemaciclib is better than that of Ribociclib; in summary, we explored the SAR of SHP2 / CDK4 dual-target inhibitors by synthesizing 26 compounds and obtained multiple compounds with dual-target inhibitory activity.

[0061] Beneficial effects

[0062] 1. This invention prepares a new class of SHP2 and CDK4 / 6 dual-target inhibitors based on pharmacophore fusion and rational drug design to address the current problems of SHP2 inhibitors and CDK4 / 6 inhibitors such as drug resistance and poor efficacy.

[0063] 2. In the in vitro enzyme activity and anti-proliferative activity tests, compounds 10, 14, 19, 23, 25, and 26 showed good dual-target inhibitory activity; in the cell cycle experiment, compound 14 showed good cycle G0 / G1 phase arrest effect.

[0064] 3. The specific embodiment structure is as follows:

[0065]

[0066]

[0067] BRIEF DESCRIPTION OF THE DRAWINGS

[0068] Figure 1 Effects of compound 14 on the cell cycle of MDA-MB-231 cells; A is control, B is TNO155, C is Abemaciclib, D is TNO155+Abemaciclib, E is Cmpd14 (1 μM), and F is Cmpd14 (6 μM)

[0069] Figure 2Effect of compound 14 on the cell cycle of MDA-MB-468 cells; A is control, B is TNO155, C is Abemaciclib, D is TNO155+Abemaciclib, E is Cmpd 6 (1 μM), and F is Cmpd6 (6 μM). DETAILED DESCRIPTION

[0070] Synthesis of intermediate 5-chloro-2-pyrazinethiosodium (II-1):

[0071]

[0072] Step 1: Synthesis of compound II-1-1

[0073] At room temperature, 2,5-dichloropyrazine (5.00 g, 33.6 mmol), ethyl 3-mercaptopropionate (4.73 g, 1.05 eq.), and potassium carbonate (4.64 g, 33.6 mmol) were placed in a 100 mL single-necked flask. DMF (42 mL) was added and the reaction was allowed to proceed for 4 h. TLC plates were used to monitor the complete conversion of the starting materials. Ethyl acetate was added and the mixture was washed several times with saturated sodium chloride solution. The organic phase was concentrated and purified by column chromatography to obtain compound II-1-1 (7.78 g, 94% yield).

[0074] Step 2: Synthesis of Compound II-1

[0075] Compound II-1-1 (7.38 g, 30.0 mmol) was dissolved in THF and placed at 0°C. Sodium ethoxide (2.25 g, 33.0 mmol, 1.1 eq.) was added in batches. The mixture was checked at room temperature until the reaction of the raw materials was complete. The solid was filtered to obtain compound II-1 (5.13 g, crude product).

[0076] Synthesis of intermediate (III-1):

[0077]

[0078] Step 1: Synthesis of compound III-1-1

[0079] Ethyl N-tert-butyloxycarbonyl-4-carboxylate piperidine (19.00 g, 73.9 mmol) was dissolved in THF and placed at -70°C. LDA (1.28 eq.) was slowly added dropwise. The reaction was allowed to proceed for 1.5 h. Benzyl bromide (1.1 eq.) was added and the temperature was maintained for 2 h. After monitoring by TLC, complete conversion of the starting material was achieved. Saturated ammonium chloride was slowly added dropwise to quench the reaction. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to afford compound III-1-1 (15.10 g, 58% yield).

[0080] Step 2: Synthesis of compound III-1-2

[0081] Compound III-1-1 (12.00 g, 34.6 mmol) and PPA (1.35 eq.) were placed in a 50 mL single-necked bottle and reacted at 150° C. for 3 h. After monitoring by TLC plate until the conversion of the raw materials was complete, the reaction system was slowly adjusted to alkaline with sodium hydroxide (2 M) at low temperature, and BOC anhydride (12.10 g, 55.4 mmol, 1.6 eq.) was added. The reaction was allowed to proceed overnight. After monitoring by TLC plate until the conversion of the raw materials was complete, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound III-1-2 (3.95 g, yield 38%).

[0082] Step 3: Synthesis of compound III-1-3

[0083] Compound III-1-2 (4.80 g, 15.9 mmol), tert-butylsulfonamide (4.05 g, 33.4 mmol, 2.1 eq.), and tetraethyl titanate (24 mL) were placed in a 100 mL single-necked flask and reacted at 120°C for 2 h. TLC plate monitoring was performed until complete conversion of the starting material was achieved. The reaction was then placed at -50°C and BH3-THF (614.4 mg, 44.5 mmol, 2.8 eq.) was slowly added. After addition, the reaction was continued at room temperature for 1 h. TLC plate monitoring was performed until complete conversion of the starting material was achieved. Saturated ammonium chloride was slowly added dropwise at -50°C to quench the reaction. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound III-1-3 (3.10 g, 48% yield).

[0084] Step 4: Synthesis of Compound III-1

[0085] Dissolve III-1-3 (3.1 g, 7.64 mmol) in DCM (30 mL) and slowly add trifluoroacetic acid (9 eq.). Allow to react overnight at room temperature. After monitoring the reaction for completion, adjust the base with aqueous sodium carbonate solution. Extract with DCM, dry, and concentrate to obtain compound III-1 (2.30 g, 98% yield).

[0086] Synthesis of intermediate (III-2):

[0087]

[0088] Step 1: Synthesis of compound (S)-ethyl 2-((tert-butyldimethylsilyl)oxy)propionate

[0089] Compound (S)-ethyl 2-hydroxypropionate (40 g, 338.6 mmol), TBSCI (76.6 g, 507.9 mmol, 1.5 eq.), and imidazole (461 g, 677.2 mmol, 2.0 eq.) were dissolved in DCM and reacted overnight at room temperature. TLC plates were used to monitor complete conversion of the starting materials. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound (S)-ethyl 2-((tert-butyldimethylsilyl)oxy)propionate (90.00 g, crude).

[0090] Step 2: Synthesis of compound (S)-2-(tert-butyldimethylsilyl)oxy)propanal

[0091] DIBAL-H (361.9 mL, 361.9 mmol, 1.3 eq.) was slowly added dropwise to a hexane (650 mL) solution of ethyl (S)-2-((tert-butyldimethylsilyl)oxy)propanoate (70.7 g, 304.4 mmol) at -78°C. The mixture was allowed to react for 1 hour and then placed at 0°C for 30 minutes. After monitoring the reaction for completion, the reaction mixture was stirred at -78°C and quenched by the slow addition of potassium sodium tartrate (aq.). The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound (S)-2-(tert-butyldimethylsilyl)oxy)propanal (38.6 g, 41% yield over two steps).

[0092] Step 3: Synthesis of compound III-2-1

[0093] Ethyl N-tert-butyloxycarbonyl-4-piperidincarboxylate (101.5 g, 394.5 mmol, 1.0 eq.) was dissolved in THF (742 mL). LDA (295.9 mL, 591.8 mmol, 1.5 eq.) was slowly added dropwise at 0°C. (S)-2-(tert-butyldimethylsilyloxy)propanal (74.3 g, 394.5 mmol) was then added portionwise. The mixture was allowed to react at room temperature for 1.5 h. The reaction was monitored by TLC until complete conversion of the starting material was achieved. The mixture was quenched, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to afford compound III-2-1 (163.4 g, 93% yield).

[0094] Step 4: Synthesis of compound III-2-2

[0095] LiBH4 (12 g, 550.2 mmol, 1.5 eq.) was added portionwise to a solution of compound III-2-1 (163.35 g, 366.8 mmol) in THF (700 mL) at 0°C. The mixture was allowed to react overnight at room temperature. After monitoring the reaction for completion, the reaction mixture was stirred at 0°C and quenched by slowly adding NH4Cl (aq.). The mixture was extracted with DCM, dried over anhydrous sodium sulfate, and concentrated to afford compound III-2-2 (103.7 g, 70% yield).

[0096] Step 5: Synthesis of compound III-2-3

[0097] Compound III-2-2 (103.7 g, 257.1 mmol) and TBAF (282.9 mL, 282.9 mmol, 1.1 eq.) were dissolved in THF (500 mL) and reacted at room temperature for 2 h. TLC plates were used to monitor complete conversion of the starting material. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound III-2-3 (104.7 g, crude).

[0098] Step 6: Synthesis of compound III-2-4

[0099] Compound III-2-3 (104.7 g, 363.3 mmol) and TsCI (55.4 g, 290.6 mmol, 0.8 eq.) were dissolved in THF (600 mL). Sodium hydroxide (30.5 g, 127.2 mmol, 2.0 eq.) was slowly added at 0°C and reacted for 1 h. The reaction was monitored by TLC plate until the conversion of the starting material was complete. The mixture was quenched, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound III-2-4 (4.3.9 g, 63% yield).

[0100] Step 7: Synthesis of compound III-2-5

[0101] Compound III-2-4 (43.9 g, 161.9 mmol) was dissolved in DCM at 0°C and Dess-Martin periodinane (89.3 g, 1.3 mmol) was added for 2 h. The reaction was monitored by TLC until complete conversion of the starting material was complete. The mixture was quenched, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to afford compound III-2-5 (41.3 g, crude).

[0102] Step 8: Synthesis of compound III-2-6

[0103] Compound III-2-5 (41.3 g, 153.3 mmol), tert-butylsulfonamide (37.2 g, 306.6 mmol, 2.0 eq.), and tetraethanol titanate (139.9 g, 613.2 mmol, 4.0 eq.) were dissolved in THF (300 mL) and placed in a 100 mL single-necked bottle. The mixture was reacted at 90°C for 21 h. The reaction was monitored by TLC plate until the conversion of the raw materials was complete. LiBH4 (6.7 mg, 306.6 mmol, 2.0 eq.) was added at -4°C and dissolved in methanol. The mixture was reacted at room temperature for 1 h. The reaction was monitored by TLC plate until the conversion of the raw materials was complete. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound III-2-6 (34.6 g, crude).

[0104] Step 9: Synthesis of Intermediate III-2

[0105] Dissolve III-2-6 (1 g, 2.67 mmol) in DCM (10 mL) and slowly add trifluoroacetic acid (2 mL). Allow to react overnight at room temperature. After monitoring the reaction for completion, adjust the base with aqueous sodium carbonate solution. Extract with ethyl acetate, dry, and concentrate to obtain compound III-2 (150.0 mg, 20% yield for all three steps).

[0106] Synthesis of intermediate (I-1):

[0107]

[0108] Step 1: Synthesis of compound I-1-2

[0109] Compound I-1-1 (17.60 g, 63.6 mmol), propionaldehyde diethyl acetal (12.20 mg, 95.4 mmol, 1.5 eq.), Pd(dppf)Cl2 (930.1 mg, 0.02 eq.), cuprous iodide (242.3 mg, 0.02 eq.), and triethylamine (1.5 eq.) were placed in a sealed tube. Anhydrous THF (176 mL) was added under nitrogen protection and reacted at 60°C for 48 h. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound I-1-2 (6.61 g, 32% yield).

[0110] Step 2: Synthesis of compound I-1-3

[0111] Compound I-1-2 (6.30 g, 19.6 mmol) and TBAF (25.63 g, 98.0 mmol, 5.0 eq.) were dissolved in 2 mL of THF and reacted at 70°C overnight. After monitoring the completion of the reaction, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound I-1-3 (8.50 g, crude).

[0112] Step 3: Synthesis of compound I-1-4

[0113] At room temperature, I-1-3 (8.50 g, 26.3 mmol) was dissolved in 2.0 mL of dioxane, and concentrated hydrochloric acid (0.76 mL) was slowly added dropwise and reacted for 20 min. After monitoring the completion of the reaction, the mixture was extracted with EA, dried and concentrated to obtain compound I-1-4 (3.10 g, two-step yield 63%).

[0114] Step 4: Synthesis of compound I-1-5

[0115] At room temperature, I-1-4 (3.10 g, 12.4 mmol) was dissolved in 1.0 mL DMF, and potassium bisulfate (760.0 mg, 5.58 mmol, 0.45 eq.) was added and reacted for 6 h. The reaction was monitored by TLC plate until the conversion of the raw material was complete. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound I-1-5 (5.20 g, crude).

[0116] Step 5: Synthesis of compound I-1-6

[0117] Compound I-1-5 (5.20 g, 19.6 mmol), HBTU (7.43 g, 19.6 mmol, 1.0 eq.), and DIPEA (7.60 g, 58.8 mmol, 3.0 eq.) were added to a sealed tube, and DMF (1.7 mL) was added under nitrogen protection. The reaction was carried out for 30 min and monitored by TLC plate until the conversion of the raw materials was complete. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound I-1-6 (1.50 g, two-step yield 31%).

[0118] Example 1:

[0119]

[0120] Step 1: Synthesis of compound 1-1

[0121] At room temperature, 2,5-dichloropyrazine (1.50 g, 10.08 mmol), ethyl 3-mercaptopropionate (1.35 g, 10.08 mmol, 1.0 eq.), and potassium carbonate (1.53 g, 11.1 mmol, 1.1 eq.) were placed in a 50 mL single-necked flask. DMF (15 mL) was added and the reaction was allowed to proceed for 3 h. TLC plate monitoring was performed until the conversion of the starting materials was complete. Ethyl acetate was added and the mixture was washed several times with saturated sodium chloride solution. The organic phases were combined, concentrated, and purified by column chromatography to obtain compound 1-1 (2.51 g, 76% yield).

[0122] Step 2: Synthesis of compound 1-2

[0123] Intermediate 1-1 (1.82 g, 7.37 mmol) and 4-methyl-4-(N-tert-butyloxycarbonyl)aminopiperidine (1.74 g, 8.1 mmol, 1.1 eq.) were dissolved in NMP and stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 1-2 (969.2 mg, 31% yield).

[0124] Step 3: Synthesis of compound 1-3

[0125] 1-2 (969.2 mg, 2.28 mmol) was dissolved in ethanol, and sodium ethoxide (170.8 mg, 2.51 mmol, 1.1 eq.) was added in batches. The reaction was monitored at room temperature until the raw material reaction was complete. The solid was filtered to obtain compound 1-3 (674.1 mg, yield 85%).

[0126] Step 4: Synthesis of Compound 1-4

[0127] Intermediate I-1 (99.6 mg, 0.34 mmol), compound 1-3 (118.3 mg, 0.34 mmol), Pd2(dba)3 (6.2 mg, 2 mol%), XantPhos (23.1 mg, 4 mol%), and DIPEA (87.9 mg, 0.68 mmol, 2.0 eq.) were placed in a sealed tube. Under nitrogen protection, anhydrous dioxane (2 mL) was added and reacted at 90°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 1-4 (40.4 mg, 21% yield).

[0128] Step 5: Synthesis of compound 1

[0129] Compound 1-4 (20.2 mg, 0.035 mmol) was dissolved in EA (1 mL), and EA / HCl (1 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 1 (14.2 mg, 87% yield). 1 H NMR(300MHz,Chloroform-d)δ8.75(s,1H),8.35(d,J=1.4Hz,1H),8.21(d,J=1.4Hz,1H),6.43(s,1H),4.56(p,J=8.8Hz ,1H),3.85–3.63(m,4H),3.10(d,J=17.1Hz,6H),2.26–2.10(m,2H),1.94–1.83(m,2H),1.73–1.40(m,8H),1.24(s,3H).

[0130] Example 2:

[0131]

[0132] Step 1: Synthesis of compound 2-2

[0133] Compound 1-3 (240.0 mg, 0.69 mmol), 2-amino-5-iodopyridine (151.8 mg, 0.69 mmol, 1.0 eq.), Pd2(dba)3 (12.6 mg, 2 mol%), XantPhos (16.0 mg, 4 mol%), and DIPEA (178.4 mg, 1.4 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (3 mL) was added under nitrogen protection and reacted at 105°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 2-2 (49.8 mg, 17% yield).

[0134] Step 2: Synthesis of compound 2-3

[0135] Compound 2-2 (89.9 mg, 0.22 mmol), intermediate I-1 (70.3 mg, 0.24 mmol, 1.1 eq.), Pd2(dba)3 (4.0 mg, 2 mol%), BINAP (8.2 mg, 6 mol%), and DIPEA (178.4 mg, 1.4 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (1 mL) was added under nitrogen protection and reacted at 105°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 2-3 (90.2 mg, 61% yield).

[0136] Step 3: Synthesis of compound 2

[0137] Compound 2-3 (62.0 mg, 0.09 mmol) was dissolved in EA (1 mL), and EA / HCl (1 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 2-4 (35.2 mg, 68% yield). 1H NMR(300MHz,Chloroform-d)δ8.72(s,1H),8.49(d,J=8.8Hz,1H),8.41(d,J=2.3 Hz,1H),8.07(d,J=1.4Hz,2H),8.04(d,J=1.4Hz,1H),7.83(dd,J=8.8,2.3Hz,1H ),6.46(s,1H),4.79(p,J=9.4Hz,1H),3.71–3.52(m,4H),3.16(s,6H),2.64–2.4 7(m,2H),2.16–1.96(m,4H),1.77–1.69(m,2H),1.69–1.46(m,4H),1.20(s,3H).

[0138] Example 3:

[0139]

[0140] Step 1: Synthesis of compound 3-1

[0141] 3-Chloro-4-iodoaniline (700.0 mg, 2.8 mmol), 5-chloro-2-pyrazine sodium sulfide (472.1 mg, 2.8 mmol, 1.0 eq.), Pd2(dba)3 (18.3 mg, 2 mol%), XantPhos (23.1 mg, 4 mol%), and DIPEA (724.1 mg, 5.6 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (10 mL) was added under nitrogen protection and reacted overnight at 100°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 3-1 (226.1 mg, 30% yield).

[0142] Step 2: Synthesis of compound 3-3

[0143] Compound 3-1 (150.0 mg, 0.55 mmol) and compound 3-2 (337.2 mg, 1.1 mmol, 2.0 eq.) were dissolved in NMP and stirred at 100°C overnight. After monitoring the reaction completion, the mixture was diluted with ethyl acetate and washed with saturated brine five times. The organic phase was concentrated and separated by column chromatography to obtain compound 3-3 (116.6 mg, 39% yield).

[0144] Step 3: Synthesis of compound 3-4

[0145] Compound 3-4 (100.0 mg, 0.18 mmol), intermediate I-1 (1.1 eq.), Pd2(dba)3 (18.3 mg, 2 mol%), BINAP (6.7 mg, 6 mol%), and DIPEA (46.5 mg, 0.36 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (1 mL) was added under nitrogen protection and reacted at 100°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 3-4 (67.0 mg, 47% yield).

[0146] Step 4: Synthesis of compound 3

[0147] Compound 3-4 (60.0 mg, 0.075 mmol) was dissolved in EA (1 mL), and EA / HCl (1 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 3 (42.2 mg, 81% yield). 1 H NMR(300MHz,Chloroform-d)δ8.65(s,1H),8.25(d,J=2.3Hz,1H),8.11(d,J=1.4Hz,1H),8 .08(d,J=1.4Hz,1H),7.37–7.27(m,3H),7.25–7.18(m,4H),6.44(s,1H),4.74(p,J=9.2Hz, 1H),4.21–4.09(m,2H),3.98(s,1H),3.27–3.13(m,8H),3.08(d,J=15.7Hz,1H),2.72(d,J =15.7Hz,1H),2.67–2.52(m,2H),2.17–2.00(m,4H),1.91–1.62(m,5H),1.42–1.33(m,1H).

[0148] Example 4:

[0149]

[0150] Step 1: Synthesis of compound 4-2

[0151] Compound 4-1 (100.0 mg, 0.22 mmol), intermediate I-1 (67.6 mg, 0.23 mmol, 1.05 eq.), Pd2(dba)3 (36.6 mg, 4 mol%), BINAP (74.7 mg, 12 mol%), and Cs2CO3 (143.4 mg, 0.44 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (1 mL) was added under nitrogen protection and reacted at 100°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 4-2 (41.0 mg, 26% yield).

[0152] Step 2: Synthesis of compound 4

[0153] Compound 4-2 (41.0 mg, 0.060 mmol) was dissolved in EA (1 mL), and EA / HCl (1 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 4 (21.0 mg, 60% yield). 1 H NMR (300MHz, Chloroform-d) δ8.81 (s, 1H), 8.31 (d, J = 1.4Hz, 1H), 8.27 (d, J = 1. 4Hz,1H),8.06(d,J=5.4Hz,1H),7.74(s,1H),6.47(s,1H),6.35(d,J=5.4Hz,1H) ,4.78(p,J=8.7Hz,1H),3.92–3.80(m,2H),3.78–3.65(m,2H),3.17(s,3H),3.1 4(s,3H),2.53–2.36(m,2H),2.14–1.92(m,4H),1.76–1.59(m,6H),1.27(s,3H).

[0154] Example 5:

[0155]

[0156] Step 1: Synthesis of compound 5-2

[0157] Intermediate III-1 (300.0 mg, 1.10 mmol) and compound 5-1 (450.6 mg, 1.65 mmol, 1.5 eq.) were dissolved in NMP, and DIPEA (10 mL) was added. The mixture was stirred at 100°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 5-2 (444.0 mg, 74% yield).

[0158] Step 2: Synthesis of compound 5-3

[0159] Compound 5-2 (150.0 mg, 0.28 mmol), intermediate I-1 (84.9 mg, 0.29 mmol, 1.05 eq.), Pd2(dba)3 (10.3 mg, 4 mol%), BINAP (20.9 mg, 12 mol%), and Cs2CO3 (182.4 mg, 0.56 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (1 mL) was added under nitrogen protection and reacted at 100°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 5-3.

[0160] Step 3: Synthesis of compound 5

[0161] Compound 5-3 was dissolved in EA (1 mL), and EA.HCl (1 mL) was added. The mixture was allowed to react overnight at room temperature. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with DCM / MeOH (5:1). The mixture was dried and concentrated to obtain compound 5 (29.0 mg, 20% yield). 1 H NMR(300MHz,Chloroform-d)δ8.79(s,1H),8.30(d,J=1.2Hz,1H),8.27(d,J=1.2Hz,1H),8.05 (d,J=5.4Hz,1H),7.74(s,1H),7.38–7.31(m,1H),7.26–7.20(m,3H),6.45(s,1H),6.35(d,J= 5.4Hz,1H),4.76(p,J=8.8Hz,1H),4.38–4.19(m,2H),4.02(s,1H),3.38–3.22(m,2H),3.20–3 .06(m,7H),2.77(d,J=15.7Hz,1H),2.54–2.36(m,2H),2.13–1.75(m,9H),1.48–1.38(m,1H).

[0162] Examples 6 and 7:

[0163]

[0164] Step 1: Synthesis of compound 6-2

[0165] Compound 6-1 (191.0 mg, 0.66 mmol) was dissolved in DCM (3 mL), trifluoroacetic acid (1 mL) was added, and the mixture was allowed to react at room temperature for 3 h. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with DCM / MeOH (5:1). The mixture was dried and concentrated to obtain compound 6-2 (137.1 mg).

[0166] Step 2: Synthesis of compound 6-3

[0167] Compound 6-2 (137.1 mg, 0.51 mmol), intermediate 5-1 (120.9 mg, 0.51 mmol, 1.0 eq.), and cesium carbonate (498.5 mg, 1.53 mmol, 3.0 eq.) were dissolved in DMSO (3 mL) and stirred at 90°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 6-3 (66.7 mg, 20% yield).

[0168] Step 3: Synthesis of compounds 6-4 and 6-5

[0169] Compound 6-3 (66.7 mg, 0.13 mmol), intermediate I-1 (58.6 mg, 0.20 mmol, 1.5 eq.), Pd2(dba)3 (2.4 mg, 2 mol%), BINAP (4.9 mg, 6 mol%), and DIPEA (33.6 mg, 0.26 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (1 mL) was added under nitrogen protection and reacted at 100°C overnight. After monitoring the reaction completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compounds 6-4 (24.6 mg, 24% yield) and 6-5 (18.9 mg).

[0170] Step 4: Synthesis of compounds 6 and 7

[0171] Compound 6-4 (24.6 mg, 0.032 mmol) was dissolved in EA (1 mL), and EA-HCl (1 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with DCM / MeOH (5:1). The mixture was dried and concentrated to obtain compound 6 (14.5 mg, 68% yield). 1 H NMR(300MHz,Chloroform-d)δ8.78(s,1H),8.26(d,J=1.4Hz,1H),8.24(d,J=1.4Hz,1H) ,8.02(d,J=5.5Hz,2H),7.46–7.21(m,4H),6.46(s,1H),6.35(d,J=5.5Hz,2H),5.05(d, J=15.2Hz,1H),4.86–4.67(m,2H),4.49–4.40(m,1H),4.27–4.03(m,2H),3.14(s,3H),3 .12(s,3H),2.53–2.33(m,2H),2.31–2.16(m,1H),2.04–1.86(m,5H),1.71–1.48(m,2H).

[0172] Compound 6-5 (18.9 mg, 0.019 mmol) was reacted according to the above steps to obtain compound 7 (11.4 mg, yield 70%). 1 H NMR(300MHz,Chloroform-d)δ8.85(s,2H),8.25(s,2H),8.11(d,J=5.3Hz,1H),7.45 –7.36(m,2H),7.33–7.18(m,2H),6.60(d,J=5.3Hz,1H),6.46(s,2H),5.05(d,J=15. 1Hz,1H),4.74(d,J=15.1Hz,1H),4.69–4.59(m,2H),4.54–4.44(m,1H),4.21–4.06( m,2H),3.21–2.98(m,12H),2.30–2.07(m,6H),2.02–1.82(m,6H),1.51–1.27(m,6H).

[0173] Example 8:

[0174]

[0175] Step 1: Synthesis of compound 8-2

[0176] Compound 8-1 (200.0 mg, 0.63 mmol), compound 5-1 (172.0 mg, 0.63 mmol, 1.0 eq.), and cesium carbonate (310 mg, 0.95 mmol, 1.5 eq.) were dissolved in DMSO (2 mL) and stirred at 90°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 8-2 (168.1 mg, 50% yield).

[0177] Step 2: Synthesis of compound 8-3

[0178] Compound 8-2 (1500 mg, 0.27 mmol), intermediate I-1 (83.0 mg, 0.28 mmol, 1.05 eq.), Pd2(dba)3 (5 mg, 2 mol%), BINAP (11 mg, 6 mol%), and Cs2CO3 (176 mg, 0.54 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1 mL) was added under nitrogen protection and reacted at 110°C overnight. After monitoring the reaction for completion, the product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 8-3 (64 mg, 30% yield).

[0179] Step 3:

[0180] Compound 8-3 (60 mg, 0.07 mmol) was dissolved in EA (1.0 mL), and EA.HCl (1.0 mL) was added. The mixture was reacted at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 8 (11 mg, yield 21%). 1 HNMR(300MHz,Chloroform-d)δ8.79(s,1H),8.32(d,J=1.4Hz,1H),8.26(d,J=1.4Hz,1H),8.04(d, J=5.3Hz,1H),7.72(s,1H),7.43–7.35(m,1H),7.26–7.20(m,3H),6.44(s,1H),6.34(d,J=5.3Hz,1 H),4.76(p,J=8.8Hz,1H),4.32(d,J=4.8Hz,1H),3.87–3.71(m,4H),3.13(s,6H),3.07–2.88(m,3H ),2.87–2.76(m,2H),2.76–2.62(m,2H),2.55–2.36(m,2H),2.13–1.88(m,4H),1.61–1.52(m,2H).

[0181] Example 9:

[0182]

[0183] Step 1: Synthesis of compound 6-(N-tert-butyloxycarbonyl)amino-2-chloro-3-iodopyridine

[0184] 5-Amino-2-chloro-3-iodopyridine (733.0 mg, 2.88 mmol) was dissolved in DCM, and triethylamine (349.1 mg, 3.46 mmol, 1.2 eq.) and DMAP (17.6 mg, 0.14 mmol, 0.05 eq.) were added. BOC anhydride (690.6 mg, 3.17 mmol, 1.1 eq.) was added at 0°C and allowed to react at room temperature for 3 h. After monitoring the reaction for completion, the mixture was washed several times with saturated sodium chloride solution. The organic phase was concentrated, dried over anhydrous sodium sulfate, and then concentrated. The resulting mixture was separated by column chromatography to afford 6-(N-tert-butoxycarbonyl)amino-2-chloro-3-iodopyridine (513.2 mg, 50% yield).

[0185] Step 2: Synthesis of compound 9-1

[0186] 6-(N-tert-Butoxycarbonyl)amino-2-chloro-3-iodopyridine (1.28 g, 3.62 mmol), 5-chloro-2-pyrazine sodium sulfide (669.0 mg, 3.98 mmol, 1.1 eq.), Pd2(dba)3 (66.2 mg, 2 mol%), Xantphos (125.5 mg, 6 mol%), and DIPEA (934.0 mg, 7.24 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and the mixture was reacted overnight at 110°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 9-1 (0.85 g, 65% yield).

[0187] Step 3: Synthesis of compound 9-2

[0188] Compound 9-1 (0.80 g, 2.15 mmol) was dissolved in EA (1.0 mL) and EA-HCl (1.0 mL) was added. The mixture was allowed to react overnight at room temperature. After monitoring the reaction for completion, sodium carbonate aqueous solution was added to adjust the base. The mixture was extracted with DCM / MeOH (5:1) and dried and concentrated to afford compound 9-2 (0.55 g, 94% yield).

[0189] Step 4: Synthesis of compound 9-3

[0190] Intermediate III-1 (80.0 mg, 0.29 mmol) and compound 9-2 (99.0 mg, 0.32 mmol, 1.0 eq.) were dissolved in 1.0 ml of NMP, and DIPEA (10 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 9-3 (128.0 mg, 80% yield).

[0191] Step 5: Synthesis of compound 9-4

[0192] Compound 9-4 (100 mg, 0.18 mmol), intermediate I-1 (56.6 mg, 0.19 mmol, 1.05 eq.), Pd2(dba)3 (7 mg, 2 mol%), BINAP (14 mg, 5 mol%), and cesium carbonate (120 mg, 0.37 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (2.0 mL) was added under nitrogen protection and reacted at 110°C overnight. After monitoring the reaction for completion, the product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 9-4 (53.3 mg, 37% yield).

[0193] Step 6: Synthesis of compound 9

[0194] Compound 9-4 (33.0 mg, mmol) was dissolved in EA (1.0 mL), and EA-HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with DCM / MeOH (5:1). The mixture was dried and concentrated to obtain compound 9 (24.1 mg, 83% yield). 1 H NMR(300MHz,Chloroform-d)δ8.83(s,1H),8.45(s,1H),8.42(d,J=8.6Hz,1H),8.15(d,J=1.3Hz,1H ),8.09(d,J=1.3Hz,1H),7.78(d,J=8.6Hz,1H),7.39–7.29(m,1H),7.26–7.17(m,3H),6.48(s,1H), 4.79(p,J=9.0Hz,1H),4.25–4.09(m,2H),3.98(s,1H),3.29–3.12(m,8H),3.08(d,J=15.6Hz,1H),2 .73(d,J=15.6Hz,1H),2.63–2.44(m,2H),2.17–1.94(m,4H),1.92–1.61(m,5H),1.42–1.33(m,1H).

[0195] Example 10:

[0196]

[0197] Step 1: Synthesis of compound 10-1

[0198] Compound 8-2 (150 mg, 0.28 mmol), compound 11-5 (98 mg, 0.3 mmol, 1.1 eq.), Pd2(dba)3 (11 mg, 4 mol%), Xantphos (22 mg, 12 mol%), and cesium carbonate (180 mg, 0.55 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (2.0 mL) was added under nitrogen protection and reacted at 110°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 10-1 (132 mg, 57% yield).

[0199] Step 2: Synthesis of compound 10

[0200] Compound 10-1 (102.0 mg, 0.12 mmol) was dissolved in EA (1.0 mL), and EA-HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with DCM / MeOH (5:1). The mixture was dried and concentrated to obtain compound 10 (74.7 mg, 57% yield). 1H NMR(300MHz,Chloroform-d)δ8.50(d,J=3.7Hz,1H),8.32(d,J=1.0Hz,1H),8.27(s,1H ),8.19(d,J=1.0Hz,1H),8.12(d,J=5.4Hz,1H),7.97(s,1H),7.82(d,J=11.9Hz,1H),7. 53(d,J=6.6Hz,1H),7.32(m,3H),6.40(d,J=5.4Hz,1H),4.73(p,J=7.0Hz,1H),4.30–4. 06(m,3H),3.43–3.14(m,3H),2.89(d,J=16.2Hz,1H),2.68(s,3H),1.97–1.50(m,10H).

[0201] Example 11:

[0202]

[0203] Step 1: Synthesis of compound 6-amino-2-cyanopyridine

[0204] 2-Bromo-6-aminopyridine (5.00 g, 28.9 mmol), zinc cyanide (3.97 g, 33.8 mmol, 1.17 eq.), and tetrakistriphenylphosphine palladium (668.0 mg, 2 mol%) were placed in a 50 mL single-necked bottle. DMF (mL) was added under nitrogen protection and the mixture was allowed to react at 100°C for 2 h. After monitoring the reaction completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 6-amino-2-cyanopyridine (2.22 g, 64% yield).

[0205] Step 2: Synthesis of compound 3-bromo-6-amino-2-cyanopyridine

[0206] 6-Amino-2-cyanopyridine (2.22 g, 18.63 mmol), tetrabutylammonium tribromide (10.33 g, 21.43 mmol), methanol (15.0 mL), and DCM (15.0 mL) were placed in a 100 mL single-necked flask and allowed to react overnight at room temperature. After monitoring the reaction for completion, the product was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to afford 3-bromo-6-amino-2-cyanopyridine (1.92 g, 52% yield).

[0207] Step 3: Synthesis of compound 11-1

[0208] 3-Bromo-6-amino-2-cyanopyridine (1.92 g, 9.7 mmol), ethyl 3-mercaptopropionate (3.9 g, 29.1 mmol, 3 eq.), Pd(dba) (178 mg, 2 mol%), Xantphos (337 mg, 6 mol%), and DIPEA (3.4 mL, 19.4 mmol, 2.0 eq.) were placed in a sealed tube. Under nitrogen protection, anhydrous dioxane (1.0 mL) was added and the mixture was reacted at 105°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 11-1 (1.82 g, 75% yield).

[0209] Step 4: Synthesis of compound 11-2

[0210] Compound 11-1 (1.82 g, mmol), sodium ethoxide (641 mg, 9.4 mmol, 1.3 eq.), and ethanol (mL) were placed in a 100 mL single-necked flask and reacted overnight at room temperature. After monitoring the reaction completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 11-2 (1.67 g, crude).

[0211] Step 5: Synthesis of compound 11-3

[0212] Compound 11-2 (1.54 g, 8.9 mmol), 2,5-dichloropyrazine (1.3 g, 8.9 mmol, 1.0 eq.), and potassium carbonate (9.4 g, 9.8 mmol, 1.1 eq.) were dissolved in 10 mL of DMF and reacted at room temperature for 12 h. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to afford compound 11-3 (510.0 mg, 22% yield).

[0213] Step 6: Synthesis of compound 11-4

[0214] Compound III-3 (200.0 mg, 0.76 mmol) and tert-butyl (4-methylpiperidinyl)carboxamide (162.9 mg, 0.84 mmol, 1.1 eq.) were dissolved in 3.0 mL of NMP, and DIPEA (8.0 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 11-4 (254.0 mg, 76% yield).

[0215] Step 7: Synthesis of compound 11-5

[0216] Compound 11-4 (200.0 mg, 0.45 mmol, 0.45 eq), compound 11-5 (322.7 mg, 1.0 mmol, 1.0 eq.), Pd2(dba)3 (16.5 mg, 4 mol%), Xantphos (69.4 mg, 12 mol%), and cesium carbonate (651.6 mg, 2.0 mmol, 2.0 eq.) were placed in a sealed tube. Under nitrogen protection, anhydrous dioxane (4.0 mL) was added and the mixture was reacted at 110°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 11-5 (70.6 mg, 22% yield).

[0217] Step 7: Synthesis of compound 11

[0218] Compound 11-5 (65.0 mg, 0.09 mmol) was dissolved in EA (1.0 mL), and EA.HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 11 (38.0 mg, yield 67%). 1 H NMR (300MHz, Chloroform-d) δ8.65(d,J=9.0Hz,1H),8.50(d,J=3.6Hz,2H),8.24(d,J=1.3Hz,1H),8.15(d,J=1.0Hz,1H),8.07(d,J=1.3Hz,1H),7.85(d, J=9.0Hz,1H),7.75(d,J=11.6Hz,1H),4.74(p,J=7.0Hz,1H),3.78–3.55(m, 4H),2.70(s,3H),1.72(s,3H),1.69(s,3H),1.68–1.47(m,4H),1.21(s,3H).

[0219] Example 12:

[0220]

[0221] Step 1: Synthesis of compound 12-1

[0222] Intermediate III-2 (865.9 mg, 0.91 mmol, 1.2 eq.) and compound 11-3 (200.0 mg, 0.76 mmol) were dissolved in NMP (3.0 mL), and DIPEA (8.0 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 12-1 (162.2 mg, 43% yield).

[0223] Step 2: Synthesis of compound 12-2

[0224] Compound 12-1 (150.0 mg, 0.30 mmol), compound 11-5 (96.8 mg, 0.3 mmol, 1.0 eq.), Pd2(dba)3 (10.1 mg, 4 mol%), Xantphos (20.8 mg, 12 mol%), and cesium carbonate (195.5 mg, 0.6 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (2.0 mL) was added under nitrogen protection and reacted at 110°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 12-2 (123.0 mg, 52% yield).

[0225] Step 3: Synthesis of compound 12

[0226] Compound 12-2 (123.0 mg, 0.16 mmol) was dissolved in EA (1 mL), and EA / HCl (2 mL) was added. The mixture was allowed to react overnight at room temperature. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 12 (37.1 mg, yield 64%). 1 HNMR(300MHz,Chloroform-d)δ8.65(d,J=9.0Hz,1H),8.49(d,J=3.7Hz,1H),8.43(s,1H),8.24(d,J= 1.2Hz,1H),8.15(d,J=1.0Hz,1H),8.06(d,J=1.2Hz,1H),7.87(d,J=9.0Hz,1H),7.75(d,J=11.8Hz,1H ),4.74(p,J=7.0Hz,1H),4.24–4.13(m,1H),3.96–3.75(m,3H),3.68(d,J=8.8Hz,1H),3.50–3.26(m, 2H), 2.99(d,J=4.5Hz,1H),2.70(s,3H),1.93–1.80(m,1H),1.77–1.64(m,9H),1.23(d,J=6.4Hz,3H).

[0227] Example 13:

[0228]

[0229] Step 1: Synthesis of compound 13-1

[0230] 5-Nitro-2,3-dichloropyridine (500.0 mg, 2.59 mmol, 1.2 eq.) and 5-chloro-2-pyrazine sodium sulfide (524.0 mg, 3.11 mmol) were dissolved in NMP (6 mL). KCO (716.0 mg, 5.18 mmol, 2 eq.) was added and stirred at room temperature overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 13-3 (526.0 mg, 67% yield).

[0231] Step 2: Synthesis of compound 13-2

[0232] Compound 13-1 (400.0 mg, 1.32 mmol), iron powder (295.7 mg, 5.28 mmol, 4.0 eq.), acetic acid (4.0 mL), and water (4.0 mL) were placed in a 25 mL single-necked flask and reacted at room temperature for 12 h. After monitoring the reaction for completion, dilute hydrochloric acid was added dropwise to quench the reaction. The mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 13-2 (318.0 mg, 88% yield).

[0233] Step 3: Synthesis of compound 13-3

[0234] Intermediate III-2 (865.9 mg, 0.91 mmol, 1.2 eq.) and compound 13-2 (300.0 mg, 1.10 mmol) were dissolved in 4.0 mL of NMP, and DIPEA (10.0 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 13-3 (340.7 mg, 61% yield).

[0235] Step 4: Synthesis of compound 13-4

[0236] Compound 13-3 (108.0 mg, 0.33 mmol), compound 11-5 (106.5 mg, 0.33 mmol, 1.0 eq.), Pd2(dba)3 (12.1 mg, 4 mol%), Xantphos (22.9 mg, 12 mol%), and cesium carbonate (215.0 mg, 0.66 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (2.0 mL) was added under nitrogen protection and reacted overnight at 110°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 13-4 (144.0 mg, 54% yield).

[0237] Step 5: Synthesis of compound 13

[0238] Compound 13-4 (43.0 mg, 0.05 mmol) was dissolved in EA (1 mL), and EA / HCl (1 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 13 (13.3 mg, 35% yield). 1 HNMR(300MHz,Chloroform-d)δ8.46(d,J=2.4Hz,1H),8.37(d,J=3.7Hz,1H),8.30(d,J=2.4Hz,1H),8.28(d ,J=1.2Hz,1H),8.19(d,J=1.2Hz,1H),8.11(d,J=1.1Hz,1H),7.73(d,J=11.7Hz,1H),7.33(s,1H),4.74(p, J=6.9Hz,1H),4.26–4.14(m,1H),4.01–3.87(m,2H),3.82(d,J=8.8Hz,1H),3.70(d,J=8.8Hz,1H),3.54–3. 31(m,2H),3.00(d,J=4.5Hz,1H),2.70(s,3H),1.95–1.83(m,1H),1.80–1.64(m,9H),1.25(d,J=6.4Hz,3H).

[0239] Example 14:

[0240]

[0241] Step 1: Synthesis of compound 14-1

[0242] Intermediate III-2 (282.0 mg, 1.27 mmol) and compound 5-1 (278.6 mg, 1.02 mmol, 0.8 eq.) were dissolved in 4.0 mL of NMP, and DIPEA (10.0 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 14-1 (151 mg, 23% yield).

[0243] Step 2: Synthesis of compound 14-2

[0244] Compound 14-1 (131.0 mg, 0.26 mmol), compound 11-5 (83.9 mg, 0.26 mmol, 1.0 eq.), Pd2(dba)3 (9.5 mg, 4 mol%), Xantphos (18.1 mg, 12 mol%), and cesium carbonate (169.4 mg, 0.52 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (4.0 mL) was added under nitrogen protection and reacted overnight at 110°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 14-2 (59.6 mg, 70% yield).

[0245] Step 3: Synthesis of compound 14

[0246] Compound 14-2 (59.6 mg, 0.07 mmol) was dissolved in EA (1 mL), and EA-HCl (1 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with DCM / MeOH (5:1). The mixture was dried and concentrated to obtain compound 14 (39.2 mg, 64% yield). 1 H NMR(300MHz,Chloroform-d)δ8.50(d,J=3.7Hz,1H),8.31(d,J=1.3Hz,1H),8.25(d,J=1.4Hz,1H),8.20(d, J=1.3Hz,1H),8.10(d,J=5.3Hz,1H),7.93(s,1H),7.82(d,J=11.8Hz,1H),6.38(d,J=5.3Hz,1H),4.73(p,J =6.9Hz,1H),4.25–4.16(m,1H),4.05–3.90(m,2H),3.84(d,J=8.8Hz,1H),3.71(d,J=8.8Hz,1H),3.61–3.3 7(m,2H),3.02(d,J=4.6Hz,1H),2.69(s,3H),1.97–1.87(m,1H),1.82–1.67(m,9H),1.25(d,J=6.4Hz,3H).

[0247] Example 15:

[0248]

[0249] Step 1: Synthesis of compound 15-1

[0250] Intermediate III-2 (301.8 mg, 1.1 mmol, 2.5 eq.) and compound 9-2 (120.0 mg, 0.44 mmol) were dissolved in 3.0 mL of NMP, and DIPEA (1.0 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 15-1 (92.5 mg, 41% yield).

[0251] Step 2: Synthesis of compound 15-2

[0252] Compound 15-1 (80.0 mg, 0.16 mmol), compound 11-5 (51.6 mg, 0.16 mmol, 1 eq.), Pd2(dba)3 (2.9 mg, 2 mol%), Xantphos (5.6 mg, 6 mol%), and cesium carbonate (104.3 mg, 0.32 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted at 105°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 15-2 (71.0 mg, 56% yield).

[0253] Step 3: Synthesis of compound 15

[0254] Compound 15-2 (71.0 mg, 0.09 mmol) was dissolved in EA (1.0 mL), and EA-HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with DCM / MeOH (5:1). The mixture was dried and concentrated to obtain compound 14 (29.4 mg, 47% yield). 1 H NMR(300MHz,Chloroform-d)δ8.47(d,J=3.8Hz,1H),8.34(d,J=8.6Hz,1H),8.29(s,1H),8.17(d,J=1.4Hz, 1H),8.16(d,J=1.2Hz,1H),8.10(d,J=1.4Hz,1H),7.75(d,J=11.5Hz,1H),7.68(d,J=8.6Hz,1H),4.73(p,J= 6.9Hz,1H),4.26–4.11(m,1H),3.96–3.76(m,3H),3.69(d,J=8.7Hz,1H),3.49–3.27(m,2H),2.99(d,J=4.5H z,1H),2.70(s,3H),1.96–1.79(m,2H),1.71(s,3H),1.69(s,3H),1.38–1.27(m,2H),1.24(d,J=6.4Hz,3H).

[0255] Example 16:

[0256]

[0257] Step 1: Synthesis of compound 16-1

[0258] Compound 9-2 (150.0 mg, 0.55 mmol) and 4-methyl-4-(N-tert-butoxycarbonyl)aminopiperidine (201.4 mg, 0.94 mmol, 1.7 eq.) were dissolved in 1 mL of NMP, and DIPEA (1.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 16-1 (165.5 mg, 67% yield).

[0259] Step 2: Synthesis of compound 16-2

[0260] Compound 16-1 (120.0 mg, 0.27 mmol), intermediate I-1 (79.1 mg, 0.27 mmol, 1.0 eq.), Pd2(dba)3 (14.8 mg, 2 mol%), BINAP (8.4 mg, 5 mol%), and cesium carbonate (175.9 mg, 0.54 mmol, 2.0 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted at 100°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 16-2 (20.0 mg, 10% yield).

[0261] Step 3: Synthesis of compound 16

[0262] Compound 16-2 (20.0 mg, 0.03 mmol) was dissolved in EA (1.0 mL), and EA.HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 16 (14.1 mg, yield 77%). 1H NMR(300MHz,Chloroform-d)δ8.77(s,1H),8.40(d,J=8.6Hz,1H),8.21(s,1H),8.15(d,J=1.3Hz,1H),8.08(d,J=1.3Hz,1H),7.76(d,J=8.6Hz, 1H),6.47(s,1H),4.78(p,J=9.0Hz,1H),3.75–3.54(m,4H),3.16(s,6H) ,2.61–2.43(m,2H),2.16–1.95(m,4H),1.80–1.45(m,6H),1.21(s,3H).

[0263] Example 17:

[0264]

[0265] Step 1: Synthesis of compound 17-1

[0266] 6-Amino-3-chloro-4-bromopyridine (200.0 mg, 0.96 mmol), 5-chloro-2-mercaptopyrazine (155.4 mg, 1.06 mmol, 1.1 eq.), Pd2(dba)3 (17.6 mg, 2 mol%), Xantphos (33.3 mg, 6 mol%), and DIPEA (248.3 mg, 1.92 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted overnight at 95°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 17-1 (95.6 mg, 36% yield).

[0267] Step 2: Synthesis of compound 17-2

[0268] Compound 17-1 (95.6 mg, 0.35 mmol) and intermediate III-2 (107.0 mg, 0.39 mmol, 1.1 eq.) were dissolved in 1.0 ml of NMP, and DIPEA (1.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 17-2 (98.8 mg, 55% yield).

[0269] Step 3: Synthesis of compound 17-3

[0270] Compound 17-2 (95.0 mg, 0.19 mmol), 11-5 (67.8 mg, 0.21 mmol, 1.1 eq.), Pd2(dba)3 (3.5 mg, 2 mol%), Xantphos (6.6 mg, 6 mol%), and DIPEA (49.1 mg, 0.38 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted overnight at 95°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 17-3 (66.3 mg, 44% yield).

[0271] Step 3: Synthesis of compound 17

[0272] Compound 17-3 (61.0 mg, 0.076 mmol) was dissolved in EA (1.0 mL), and EA-HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 17 (38.2 mg, 73% yield). 1 H NMR(300MHz,Chloroform-d)δ8.26(d,J=1.3Hz,1H),8.17(d,J=3.6Hz,1H),8.15(s,1H),8.09 (d,J=1.3Hz,1H),8.04(s,1H),7.98(s,1H),7.73(s,1H),7.64(d,J=11.5Hz,1H),4.75(p,J=7 .0Hz,1H),4.24–4.15(m,1H),3.90–3.75(m,3H),3.68(d,J=8.8Hz,1H),3.45–3.24(m,2H),3. 00(d,J=4.5Hz,1H),2.70(s,3H),1.92–1.80(m,1H),1.75–1.61(m,9H),1.25(d,J=6.4Hz,3H).

[0273] Example 18:

[0274]

[0275] Step 1: Synthesis of compound 18-1

[0276] 2-Amino-5-bromopyrazine (400.0 mg, 2.30 mmol), sodium 5-chloro-2-pyrazinesulfide (426.6 mg, 2.53 mmol, 1.1 eq.), Pd2(dba)3 (42.1 mg, 2 mol%), Xantphos (79.8 mg, 6 mol%), and DIPEA (594.8 mg, 4.60 mmol, 2 eq.) were placed in a sealed tube. Under nitrogen protection, anhydrous dioxane (1.0 mL) was added and the mixture was reacted overnight at 90°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 18-1 (161.7 mg, 29% yield).

[0277] Step 2: Synthesis of compound 18-2

[0278] Compound 18-1 (120.0 mg, 0.50 mmol) and intermediate III-2 (150.9 mg, 0.55 mmol, 1.1 eq.) were dissolved in 1.0 mL of NMP, and DIPEA (1.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 18-2 (76.3 mg, 32% yield).

[0279] Step 3: Synthesis of compound 18-3

[0280] Compound 18-2 (76.3 mg, 0.16 mmol), compound 11-5 (58.1 mg, 0.18 mmol, 1.1 eq.), Pd2(dba)3 (2.9 mg, 2 mol%), BINAP (6.0 mg, 6 mol%), and sodium tert-butoxide (30.8 mg, 0.32 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted overnight at 90°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 18-3 (10.2 mg, 9% yield).

[0281] Step 4: Synthesis of compound 18

[0282] Compound 18-3 (10.2 mg, 0.014 mmol) was dissolved in EA (1.0 mL), and EA-HCl (1.0 mL) was added. The mixture was allowed to react overnight at room temperature. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 18 (7 mg, yield 75%). 1H NMR(300MHz,Chloroform-d)δ9.62(d,J=1.6Hz,1H),8.43(d,J=3.7Hz,1H),8.30(d,J=1.4Hz,1H),8.20 (d,J=1.5Hz,1H),8.16(d,J=1.4Hz,1H),8.14(d,J=1.5Hz,1H),8.01(s,1H),7.76(d,J=11.4Hz,1H),4. 73(p,J=5.9,4.8Hz,1H),4.23–4.13(m,1H),4.01–3.77(m,3H),3.70(d,J=8.8Hz,1H),3.52–3.30(m,2H ),3.01(d,J=4.5Hz,1H),2.69(s,3H),1.93–1.82(m,1H),1.70(m,J=6.9Hz,9H),1.24(d,J=6.4Hz,3H)..

[0283] Example 19:

[0284]

[0285] Step 1: Synthesis of compound 2-amino-4-chloro-3-cyanopyridine

[0286] 2-Amino-4-chloro-3-iodopyridine (850.0 mg, 3.32 mmol), zinc cyanide (234.8 mg, 2.0 mmol, 0.6 eq.), and Pd(PPh3)4 (191.8 mg, 5 mol%) were placed in a sealed tube. Anhydrous NMP (3.0 mL) was added under nitrogen protection and the mixture was reacted at 120°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 2-amino-4-chloro-3-cyanopyridine (432.2 mg, 85% yield).

[0287] Step 2: Synthesis of compound 19-1

[0288] 2-Amino-4-chloro-3-cyanopyridine (220.0 mg, 1.43 mmol), 5-chloro-2-mercaptopyrazine (252.2 mg, 1.72 mmol, 1.2 eq.), Pd2(dba)3 (26.2 mg, 2 mol%), Xantphos (49.6 mg, 6 mol%), and DIPEA (369.8 mg, 2.86 mmol, 2 eq.) were placed in a sealed tube. Under nitrogen protection, anhydrous dioxane (1.0 mL) was added and the mixture was reacted overnight at 95°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 19-1 (171.3 mg, 45% yield).

[0289] Step 3: Synthesis of compound 19-2

[0290] Compound 19-1 (107.0 mg, 0.41 mmol) and intermediate III-2 (123.5 mg, 0.45 mmol, 1.1 eq.) were dissolved in 1.0 mL of NMP, and DIPEA (1.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 19-2 (134.4 mg, 60% yield).

[0291] Step 4: Synthesis of compound 19-3

[0292] Compound 19-2 (100.0 mg, 0.20 mmol), compound 11-5 (71.0 mg, 0.22 mmol, 1.1 eq.), Pd2(dba)3 (3.7 mg, 2 mol%), Xantphos (6.9 mg, 6 mol%), and cesium carbonate (130.3 mg, 0.40 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted overnight at 95°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 19-3 (91.7 mg, 60% yield).

[0293] Step 5: Synthesis of compound 19

[0294] Compound 19-3 (80.0 mg, 0.10 mmol) was dissolved in EA (1.0 mL), and EA.HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 19 (8.9 mg, 13% yield). 1H NMR(300MHz,Chloroform-d)δ8.50(d,J=3.6Hz,1H),8.30(d,J=1.1Hz,1H),8.27(d,J=5.5Hz,1H),8.24(d, J=1.0Hz,1H),8.21(d,J=1.2Hz,1H),8.19(s,1H),7.76(d,J=11.7Hz,1H),6.55(d,J=5.5Hz,1H),4.75(p,J =6.9Hz,1H),4.26–4.15(m,1H),4.09–3.93(m,2H),3.85(d,J=8.8Hz,1H),3.71(d,J=8.8Hz,1H),3.59–3.3 6(m,2H),3.04(d,J=4.5Hz,1H),2.69(s,3H),1.99–1.88(m,1H),1.82–1.61(m,9H),1.26(d,J=6.2Hz,3H).

[0295] Example 20:

[0296]

[0297] Step 1: Synthesis of compound 20-1

[0298] 2-Amino-5-chloro-6-bromopyridine (250.0 mg, 1.20 mmol), 5-chloro-2-mercaptopyrazine (211.1 mg, 1.44 mmol, 1.2 eq.), CuI (45.7 mg, 0.2 eq.), 1,10-phenanthroline (86.5 mg, 0.4 eq.), and K3PO4 (509.5 mg, 2.40 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (2.0 mL) was added under nitrogen protection and reacted overnight at 110°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 20-1 (127.7 mg, 39% yield).

[0299] Step 2: Synthesis of compound 20-2

[0300] Compound 20-1 (102.0 mg, 0.37 mmol) and intermediate III-2 (112.5 mg, 0.41 mmol, 1.1 eq.) were dissolved in 1.0 ml of NMP, and DIPEA (1.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 20-2 (43.2 mg, 23% yield).

[0301] Step 3: Synthesis of compound 20-3

[0302] Compound 20-2 (43.2 mg, 0.085 mmol), compound 11-5 (30.3 mg, 0.094 mmol, 1.1 eq.), Pd2(dba)3 (1.6 mg, 2 mol%), Xantphos (3.0 mg, 6 mol%), and DIPEA (22.0 mg, 0.17 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted at 95°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain crude compound 20-3 (22.3 mg).

[0303] Step 4: Synthesis of compound 20

[0304] Compound 20-3 (22.3 mg, 0.028 mmol) was dissolved in EA (1.0 mL), and EA.HCl (1.0 mL) was added. The mixture was reacted at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 20 (9.7 mg, yield 16%). 1 H NMR(300MHz,Chloroform-d)δ8.37(d,J=3.7Hz,1H),8.29(d,J=1.3Hz,1H),8.22(d,J=1.3Hz,1H),8.1 4(d,J=8.7Hz,1H),8.10(d,J=1.0Hz,1H),7.76–7.67(m,2H),7.54(d,J=8.7Hz,1H),4.73(p,J=7.0Hz, 1H),4.27–4.16(m,1H),4.07–3.90(m,2H),3.84(d,J=8.8Hz,1H),3.73(d,J=8.8Hz,1H),3.58–3.36(m ,2H),3.03(d,J=4.6Hz,1H),2.69(s,3H),2.00–1.89(m,1H),1.84–1.67(m,9H),1.25(d,J=6.4Hz,3H).

[0305] Example 21:

[0306]

[0307] Step 1: Synthesis of compound 2-amino-4-chloro-5-iodopyridine

[0308] 2-Amino-4-chloropyridine (1.00 g, 7.78 mmol), NIS (1.92 g, 8.56 mmol, 1.1 eq.), and DMF (15.0 ml) were placed in a 50 ml single-necked flask and reacted at room temperature for 3 h. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain 2-amino-4-chloro-5-iodopyridine (1.10 g, 56% yield).

[0309] Step 2: Synthesis of compound 21-1

[0310] 2-Amino-4-chloro-5-iodopyridine (250.0 mg, 1.0 mmol), 5-chloro-2-mercaptopyrazine (161.3 mg, 1.1 mmol, 1.1 eq.), CuI (38.1 mg, 0.2 eq.), 1,10-phenanthroline (72.1 mg, 0.4 eq.), and K3PO4 (424.6 mg, 2.00 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (2.0 mL) was added under nitrogen protection and reacted overnight at 110°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 21-1 (145.2 mg, 53% yield).

[0311] Step 3: Synthesis of compound 21-2

[0312] Compound 21-1 (145.2 mg, 0.53 mmol) and intermediate III-2 (158.6 mg, 0.58 mmol, 1.1 eq.) were dissolved in 1.0 mL of NMP, and DIPEA (1.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 21-2 (115.0 mg, 23% yield).

[0313] Step 4: Synthesis of compound 21-3

[0314] Compound 21-2 (115.0 mg, 0.22 mmol), compound 11-5 (77.5 mg, 0.24 mmol, 1.1 eq.), Pd2(dba)3 (4.0 mg, 2 mol%), Xantphos (7.6 mg, 6 mol%), and DIPEA (56.9 mg, 0.44 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted at 95°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 21-3 (91.5 mg, 52% yield).

[0315] Step 4: Synthesis of compound 21

[0316] Compound 21-3 (75.0 mg, 0.094 mmol) was dissolved in EA (1.0 mL), and EA-HCl (1.0 mL) was added. The mixture was reacted at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 21 (18.0 mg, yield 28%). 1 H NMR(300MHz,Chloroform-d)δ8.66(s,1H),8.47(d,J=3.7Hz,1H),8.39(s,1H),8.38(s,1H),8.1 5(d,J=1.3Hz,1H),8.11(d,J=1.4Hz,1H),8.05(d,J=1.4Hz,1H),7.77(d,J=11.6Hz,1H),4.76(p ,J=6.9Hz,1H),4.23–4.13(m,1H),3.92–3.76(m,3H),3.68(d,J=8.8Hz,1H),3.46–3.24(m,2H), 2.98(d,J=4.5Hz,1H),2.70(s,3H),1.91–1.80(m,1H),1.75–1.61(m,9H),1.23(d,J=6.4Hz,3H).

[0317] Example 22:

[0318]

[0319] Step 1: Synthesis of compound 22-1

[0320] 2-Amino-4-chloropyrimidine (500.0 mg, 3.9 mmol) and 5-chloro-2-mercaptopyrazine (628.9 mg, 4.29 mmol, 1.1 eq.) were dissolved in 10.0 ml of NMP. Potassium carbonate (1.08 g, 7.80 mmol, 2 eq.) was added and stirred overnight at room temperature. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 22-1 (304.1 mg, 33% yield).

[0321] Step 2: Synthesis of compound 22-2

[0322] Compound 22-1 (120.0 mg, 0.50 mmol) and intermediate III-2 (175.6 mg, 0.64 mmol, 1.1 eq.) were dissolved in 1.0 ml of NMP, and DIPEA (1.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 22-2 (105.4 mg, 46% yield).

[0323] Step 3: Synthesis of compound 22-3

[0324] Compound 22-2 (105.4 mg, 0.22 mmol), compound 11-5 (77.5 mg, 0.24 mmol, 1.1 eq.), Pd2(dba)3 (4.0 mg, 2 mol%), Xantphos (7.6 mg, 6 mol%), and DIPEA (56.9 mg, 0.44 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted at 95°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 22-3 (47.1 mg, 37% yield).

[0325] Step 4: Synthesis of compound 22

[0326] Compound 22-3 (42.0 mg, 0.055 mmol) was dissolved in EA (1.0 mL), and EA-HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 22 (23.0 mg, yield 63%). 1 H NMR(300MHz,Chloroform-d)δ8.52(d,J=3.5Hz,1H),8.34(d,J=1.4Hz,1H),8.31(d,J=5.5Hz,1H),8.23 (d,J=1.3Hz,1H),8.21(d,J=1.4Hz,1H),7.84(d,J=11.7Hz,1H),6.60(d,J=5.5Hz,1H),4.76(p,J=7.0H z,1H),4.26–4.16(m,1H),4.07–3.92(m,2H),3.85(d,J=9.0Hz,1H),3.72(d,J=9.0Hz,1H),3.56–3.34( m,2H),3.05(d,J=4.6Hz,1H),2.70(s,3H),1.98–1.86(m,1H),1.82–1.68(m,9H),1.26(d,J=6.4Hz,3H).

[0327] Example 23:

[0328]

[0329] Step 1: Synthesis of compound 23-1

[0330] 3-Chloro-4-iodoaniline (300.0 mg, 1.20 mmol), 5-chloro-2-mercaptopyrazine (193.5 mg, 1.32 mmol, 1.1 eq.), CuI (45.7 mg, 0.2 eq.), 1,10-phenanthroline (86.5 mg, 0.4 eq.), and K3PO4 (331.7 mg, 2.40 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (2.0 mL) was added under nitrogen protection and reacted overnight at 110°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 23-1 (212.4 mg, 56% yield).

[0331] Step 2: Synthesis of compound 23-2

[0332] Compound 23-1 (120.0 mg, 0.44 mmol) and intermediate III-2 (131.7 mg, 0.48 mmol, 1.1 eq.) were dissolved in 1.0 ml of NMP, and DIPEA (1.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 23-2 (93.5 mg, 41% yield).

[0333] Step 3: Synthesis of compound 23-3

[0334] Compound 23-2 (80.0 mg, 0.16 mmol), compound 11-5 (58.1 mg, 0.18 mmol, 1.1 eq.), Pd2(dba)3 (2.9 mg, 2 mol%), Xantphos (5.6 mg, 6 mol%), and DIPEA (41.4 mg, 0.32 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted overnight at 95°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 23-3 (67.3 mg, 53% yield).

[0335] Step 4: Synthesis of compound 23

[0336] Compound 23-3 (60.0 mg, 0.075 mmol) was dissolved in EA (1.0 mL), and EA.HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 23 (42.6 mg, yield 82%). 1 H NMR(300MHz,Chloroform-d)δ8.37(d,J=3.8Hz,1H),8.15(d,J=1.2Hz,1H),8.11–8.07(m,2H),8.05( d,J=2.2Hz,1H),7.76(d,J=11.6Hz,1H),7.38(dd,J=8.6,2.2Hz,1H),7.32(d,J=8.6Hz,1H),7.28(s,1 H),4.75(p,J=7.0Hz,1H),4.23–4.14(m,1H),3.94–3.76(m,3H),3.68(d,J=8.8Hz,1H),3.47–3.24(m, 2H),2.99(d,J=4.5Hz,1H),2.70(s,3H),1.92–1.80(m,1H),1.76–1.67(m,9H),1.23(d,J=6.4Hz,3H).

[0337] Example 24:

[0338]

[0339] Step 1: Synthesis of compound 24-1

[0340] Compound 9-3 (106.0 mg, 0.2 mmol), compound 11-5 (58.0 mg, 0.18 mmol, 0.9 eq.), Pd2(dba)3 (2.9 mg, 2 mol%), Xantphos (5.6 mg, 6 mol%), and cesium carbonate (41.4 mg, 0.32 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted at 105°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 24-1 (88.6 mg, 61% yield).

[0341] Step 2: Synthesis of compound 24

[0342] Compound 24-1 (80.0 mg, 0.1 mmol) was dissolved in EA (1.0 mL), and EA.HCl (1.0 mL) was added. The mixture was allowed to react at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 24 (38.0 mg, 54% yield). 1H NMR(300MHz,Chloroform-d)δ8.48(d,J=3.7Hz,1H),8.34(d,J=8.6Hz,1H),8.31(s,1H),8.18(d,J=1.2Hz,1H) ,8.16(d,J=1.2Hz,1H),8.12(d,J=1.1Hz,1H),7.75(d,J=11.9Hz,1H),7.67(d,J=8.6Hz,1H),7.37–7.29(m,1H) ,7.26–7.16(m,3H),4.73(p,J=6.9Hz,1H),4.25–4.11(m,2H),3.99(s,1H),3.31–3.15(m,2H),3.09(d,J=15.6 Hz,1H),2.73(d,J=15.6Hz,1H),2.70(s,3H),1.93–1.75(m,2H),1.70(d,J=7.0Hz,6H),1.38(d,J=11.5Hz,2H).

[0343] Example 25:

[0344]

[0345] Step 1: Synthesis of 3-(tert-butylthio)-2-chloroaniline

[0346] 3-Fluoro-2-chloroaniline (1.00 g, 6.87 mmol), tert-butyl mercaptan (1.86 g, 20.61 mmol, 3.0 eq.), cesium carbonate (3.36 g, 10.31 mmol, 1.5 eq.), and DMSO (15.0 mL) were placed in a single-necked flask and reacted overnight at 120°C under nitrogen. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain a crude product of 3-(tert-butylthio)-2-chloroaniline (1.67 g).

[0347] Step 2: Synthesis of 2-chloro-3-mercaptoaniline

[0348] 3-(tert-Butylthio)-2-chloroaniline (1.67 g, 7.74 mmol) was dissolved in 15.0 mL of concentrated hydrochloric acid and allowed to react at 80°C overnight. After monitoring the reaction for completion, the mixture was adjusted to alkalinity with sodium carbonate, extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to afford 2-chloro-3-mercaptoaniline (577.5 mg, 47% yield over two steps).

[0349] Step 3: Synthesis of 25-1

[0350] 2-Chloro-3-mercaptoaniline (577.5 mg, 3.62 mmol) and 2,5-dichloropyrazine (593.0 mg, 3.98 mmol, 1.1 eq.) were dissolved in 4.0 ml of NMP. Potassium carbonate (1.00 g, 7.24 mmol, 2 eq.) was added and stirred overnight at room temperature. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 25-1 (112.5 mg, 11% yield).

[0351] Step 4: Synthesis of 25-2

[0352] Compound 25-1 (71.5.0 mg, 0.26 mmol) and intermediate III-2 (85.1 mg, 0.31 mmol, 1.2 eq.) were dissolved in 1.0 mL of NMP, and DIPEA (2.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 25-2 (71.7 mg, 54% yield).

[0353] Step 5: Synthesis of 25-3

[0354] Compound 25-2 (71.7 mg, 0.14 mmol), compound 11-5 (35.5 mg, 0.11 mmol, 0.8 eq.), Pd2(dba)3 (5.1 mg, 4 mol%), Xantphos (9.8 mg, 12 mol%), and cesium carbonate (91.3 mg, 0.28 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and reacted at 105°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 25-3 (50.0 mg, 46% yield).

[0355] Step 6: Synthesis of compound 25

[0356] Compound 25-3 (45.0 mg, 0.058 mmol) was dissolved in EA (1.0 mL), and EA.HCl (1.0 mL) was added. The mixture was reacted at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 25 (24.0 mg, yield 60%). 1H NMR(300MHz,Chloroform-d)δ8.48(dd,J=8.4,1.4Hz,1H),8.39(d,J=3.8Hz,1H),8.27–8.14(m,3H ),7.85–7.74(m,2H),7.17(t,J=8.1Hz,1H),6.73(dd,J=7.9,1.4Hz,1H),4.73(p,J=6.9Hz,1H),4. 26–4.14(m,1H),4.00–3.85(m,2H),3.82(d,J=8.8Hz,1H),3.70(d,J=8.8Hz,1H),3.53–3.28(m,2H ),3.00(d,J=4.5Hz,1H),2.69(s,3H),1.95–1.83(m,1H),1.79–1.66(m,9H),1.24(d,J=6.4Hz,3H).

[0357] Example 26:

[0358]

[0359] Step 1: Synthesis of compound 26-1

[0360] 2-Amino-4-iodopyridine (250.0 mg, 1.14 mmol), 5-chloro-2-pyrazine sodium sulfide (249.5 mg, 1.48 mmol, 1.3 eq.), Pd2(dba)3 (21.1 mg, 2 mol%), Xantphos (19.7 mg, 3 mol%), and DIPEA (294.1 mg, 2.28 mmol, 2 eq.) were placed in a sealed tube. Under nitrogen protection, anhydrous dioxane (3.0 mL) was added and reacted overnight at 95°C. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 26-1 (178.0 mg, 65% yield).

[0361] Step 2: Synthesis of compound 26-2

[0362] Compound 26-1 (120.0 mg, 0.50 mmol) and intermediate III-2 (164.6 mg, 0.60 mmol, 1.2 eq.) were dissolved in 1.0 ml of NMP, and DIPEA (2.5 mL) was added. The mixture was stirred at 95°C overnight. After monitoring the reaction for completion, the mixture was diluted with ethyl acetate and washed five times with saturated brine. The organic phase was concentrated and separated by column chromatography to obtain compound 26-2 (142.2 mg, 60% yield).

[0363] Step 3: Synthesis of compound 26-3

[0364] Compound 26-2 (130.0 mg, 0.27 mmol), compound 11-5 (74.2 mg, 0.23 mmol, 0.85 eq.), Pd2(dba)3 (10.1 mg, 4 mol%), Xantphos (18.5 mg, 12 mol%), and cesium carbonate (176.0 mg, 0.54 mmol, 2 eq.) were placed in a sealed tube. Anhydrous dioxane (1.0 mL) was added under nitrogen protection and the mixture was reacted at 105°C overnight. After monitoring the reaction for completion, the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated, and separated by column chromatography to obtain compound 26-3 (110.0 mg, 53% yield).

[0365] Step 4: Synthesis of compound 26

[0366] Compound 26-3 (90.0 mg, 0.12 mmol) was dissolved in EA (1.0 mL), and EA.HCl (1.0 mL) was added. The mixture was reacted at room temperature overnight. After monitoring the reaction completion, sodium carbonate aqueous solution was added to adjust the base, and the mixture was extracted with EA. The mixture was dried and concentrated to obtain compound 26 (65.0 mg, yield 82%). 1 H NMR(300MHz,Chloroform-d)δ8.37(d,J=3.7Hz,1H),8.35(dd,J=1.7,0.7Hz,1H),8.26(d,J=1.4Hz,1H ),8.21–8.04(m,3H),8.01(d,J=1.4Hz,1H),7.73(d,J=11.8Hz,1H),6.62(dd,J=5.4,1.7Hz,1H),4.77 (p,J=6.9Hz,1H),4.20(dd,J=6.5,4.6Hz,1H),3.97–3.78(m,3H),3.69(d,J=8.8Hz,1H),3.50–3.27(m ,2H),3.01(d,J=4.6Hz,1H),2.70(s,3H),1.94–1.82(m,1H),1.78–1.64(m,9H),1.25(d,J=6.4Hz,3H).

[0367] In vitro SHP2 enzyme activity test

[0368] The SHP2 enzyme activity of the compounds in the above examples was tested, and the specific operation was as follows:

[0369] 4.1 Compound Preparation

[0370] The compound was dissolved in 100% DMSO to prepare a 30 mM stock solution, which was stored in a -20°C refrigerator in the dark.

[0371] 4.2 SHP2 reaction process

[0372] (1) Prepare 1×Reaction Buffer.

[0373] (2) Preparation of compound concentration gradient: The starting concentration of the test compound was 30 μM, and the compound was diluted 3-fold to 10 concentrations in a single well. A 100% DMSO solution was diluted to 100-fold the final concentration in a 384 source plate. The compound was diluted 3-fold to 10 concentrations using a Precision RT-PCR system. 250 nL of the compound at 100-fold the final concentration was transferred to the destination 384 plate using an Echo 550 dispenser. 250 nL of DMSO was added to the positive control, and 250 nL of 1 mM SHP099 was added to the negative control.

[0374] (3) Prepare an activation peptide solution with 5 times the final concentration using 1× Reaction Buffer, add 5 μL to each reaction plate, and centrifuge at 1000 rpm for 1 min.

[0375] (4) Prepare an enzyme solution with a final concentration of 2.5 times using 1× Reaction Buffer, add 10 μL to each reaction plate, centrifuge at 1000 rpm for 1 min, and incubate at room temperature for 60 min.

[0376] (5) Prepare a substrate solution with a final concentration of 2.5 times using 1× Reaction Buffer, add 10 μL to each reaction plate, centrifuge at 1000 rpm for 1 min, and incubate at room temperature for 20 min.

[0377] (6) Read the Ex355 / Em460 fluorescence values using EnSight

[0378] 4.3 Data Analysis

[0379] Calculation formula

[0380]

[0381] Where: RFU: fluorescence value of the sample; Mean (NC): mean fluorescence value of the control wells containing 10 μM SHP099;

[0382] Mean (PC): Mean fluorescence value of positive control wells.

[0383] Fitting dose-effect curve

[0384] The log value of the concentration was used as the X-axis and the percentage inhibition rate was used as the Y-axis. The log (inhibitor) vs. response-Variable slope analysis software GraphPad Prism 5 was used to fit the dose-effect curve to obtain the IC50 value of each compound on the enzyme activity.

[0385] The calculation formula is Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) * HillSlope))

[0386] In vitro CDK4 enzyme activity test

[0387] 5.1 Compound Preparation

[0388] The compound was dissolved in 100% DMSO to prepare a 30 mM stock solution, which was stored in a -20°C refrigerator in the dark.

[0389] 5.2 Kinase reaction process

[0390] (1) Prepare 1× Kinase buffer.

[0391] (2) Preparation of compound concentration gradient: The test compound concentration was prepared as per customer specifications, starting at 30,000 nM. The compound was diluted in 100% DMSO solution to a 100-fold final concentration in a 384-source plate. The compound was diluted 3-fold to a total of 10 concentrations. 250 nL of the compound at 100-fold final concentration was transferred to the destination 384-well plate using an Echo 550 dispenser.

[0392] (3) Prepare a kinase solution with a final concentration of 2.5 times using 1× Kinase buffer.

[0393] (4) Add 10 μL of kinase solution at 2.5 times the final concentration to the compound wells and positive control wells respectively; add 10 μL of 1× Kinase buffer to the negative control wells.

[0394] (5) Centrifuge at 1000 rpm for 30 seconds, shake the reaction plate to mix, and incubate at room temperature for 10 minutes.

[0395] (6) Prepare a mixed solution of ATP and Kinase substrate 8 with a final concentration of 25 / 15 times using 1× Kinase buffer.

[0396] (7) Add 15 μL of a mixed solution of ATP and substrate at 5 / 3 times the final concentration to start the reaction.

[0397] (8) Centrifuge the 384-well plate at 1000 rpm for 30 seconds, shake to mix, and incubate at room temperature for 180 minutes.

[0398] (9) Add 30 μL of stop detection solution to stop the kinase reaction, centrifuge at 1000 rpm for 30 seconds, and shake to mix.

[0399] (10) Read the conversion rate using Caliper EZ Reader II.

[0400] 5.3 Data Analysis

[0401] Calculation formula

[0402]

[0403] Where: Conversion%_sample is the conversion rate reading of the sample; Conversion%_min is the mean value of the negative control wells, representing the conversion rate reading of the wells without enzyme activity; Conversion%_max is the mean value of the positive control wells, representing the conversion rate reading of the wells without compound inhibition.

[0404] The specific results are shown in the table:

[0405]

[0406]

[0407] In vitro antiproliferative activity of compounds

[0408] 1. Experimental steps

[0409] (1) Sterilize the PBS solution under high pressure and store it in a refrigerator at 4°C.

[0410] (2) Weigh trypsin and pancreatic digestion solution, add ultrapure water to fully dissolve, filter the liquid with a microporous filter, and store in a refrigerator at -20℃.

[0411] (3) Weigh the culture medium powder and NaHCO3 separately, add ultrapure water to fully dissolve, add 10% double antibody, filter with a microporous filter membrane to obtain the culture medium, store in a refrigerator at 4°C, and add 10% fetal bovine serum before use.

[0412] (4) Take out the MDA-MB-231, MDA-MB-468 or EMT6 cells from the liquid nitrogen tank and immediately place them in a 37-degree Celsius constant temperature water bath. Shake them to melt, then pour the cells into a culture flask and add culture medium (containing 10% fetal bovine serum) to dilute them. Transfer the diluted culture medium into a centrifuge tube, centrifuge at 1000r / min for 5 minutes, discard the supernatant, add fresh culture medium and mix thoroughly, transfer them into a culture flask and culture them in a 5% CO2, 37°C incubator. When the cells are attached to the wall and almost cover the bottom of the flask, start passage, add a small amount of fresh culture medium (containing 10% fetal bovine serum) to stop digestion, pour out the liquid in the culture flask, wash twice with PBS, add fresh culture medium and mix thoroughly, and divide them equally into two culture flasks for further culture.

[0413] (5) Take the logarithmic phase cells, pour out the old culture medium, add trypsin solution to digest for 3 minutes, add fresh culture medium containing 10% fetal bovine serum to stop digestion, transfer the solution to a centrifuge tube, centrifuge at 1000r / min for 5 minutes, and discard the supernatant. Add culture medium to prepare it into a cell suspension and count the cells. After counting, the cells are plated in a 96-well plate at a concentration of 5000-10000 cells per well. The 96-well plate with cells is placed in a 37°C, 5% CO2 incubator and cultured for 24 hours. Use culture medium to dilute the drug gradient to 90μmol / L, 30μmol / L, 10μmol / L, 3.3μmol / L, 1.1μmol / L, 0.37μmol / L, and then add them to the 96-well plate, 100μL per well, and set up three replicates for each concentration. The control group added the corresponding concentration of medium containing solvent, and the zero well added the same volume of blank medium. The cells were placed in a 5% CO2, 37°C incubator and incubated for 3 days. The medium was replaced every two days. 20 μL of MTT (5 mg / mL) was added to each well, mixed well, and incubated in a 5% CO2, 37°C incubator in the dark for 4 hours. The liquid in the 96-well plate was removed, and 150 μL of DMSO was added to each well. The cells were placed on a micro-oscillator and shaken to completely dissolve the crystals at the bottom. The 96-well plate was then placed in a microplate reader for detection, and the absorbance was measured at 490 nm.

[0414] Draw the curve and calculate the inhibition rate and IC of the drug on cells 50 .

[0415] Inhibition rate = [(average OD value of the control group - average OD value of the experimental group) / (average OD value of the control group - average OD value of the blank control group)] × 100%.

[0416] 3. Experimental results

[0417] The inhibitory activity of the compound on MDA-MB-231, MDA-MB-468, and EMT6 cell lines is as follows:

[0418]

[0419] Experimental conclusion: Compared with the administration of SHP2 and CDK4 / 6 inhibitors alone or in combination, the compounds of the embodiments of the present invention have better anti-proliferative effects on MDA-MB-231, MDA-MB-468 and EMT6 cells.

[0420] Compound cell cycle assay

[0421] MDA-MB-231 and MDA-MB-468 cell lines were seeded in six-well plates and treated with DMSO, TNO155, Abemaciclib, TNO155+Abemaciclib or different concentrations of compound 12 for 48 hours. The cells were washed once with PBS and 1mL of trypsin (0.25%) without EDTA was added. When the cells were rounded and some cells were suspended, PBS was added to stop digestion. Then, cells were harvested and fixed in 70% ethanol (500 μL) for 24 hours at 4°C. Cells were collected by washing with PBS and centrifuged at 1500rpm for 5 minutes, then stained with PI / RNase at room temperature in the dark for 30-60 minutes. After incubation, cells were analyzed by flow cytometry. All experiments were performed 3 times.

[0422] Experimental conclusion: Figure 1 and Figure 2 As shown, compared with the administration of SHP2 and CDK4 / 6 inhibitors alone or in combination, the compounds of the embodiments of the present invention have better G0 / G1 cycle arrest effects on MDA-MB-231 and MDA-MB-468 cells.

Claims

1. A compound, characterized in that The compound is any one of the following structural formulas:

2. A pharmaceutical composition, characterized in that Contains the compound as claimed in claim 1 and pharmaceutically acceptable excipients.

3. The pharmaceutical composition according to claim 2, wherein The pharmaceutical composition is prepared into tablets, capsules, injections or lyophilized powders.

4. Use of the compound according to claim 1 or the pharmaceutical composition according to claim 2 or 3 in the preparation of anti-tumor drugs.

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