Compounds as KIF18A inhibitors and uses thereof

Abstract

The invention relates to a compound serving as a KIF18A inhibitor, a pharmaceutical composition and application of the compound and the pharmaceutical composition. Specifically, the invention provides compounds as shown in formulas 1-3, a pharmaceutical composition and application of the compounds in preparation of drugs for preventing and / or treating diseases related to the activity or expression quantity of KIF18A. The compounds provided by the invention have good KIF18A inhibitory activity, and can be used for preparing drugs for preventing and / or treating diseases related to the activity or expression quantity of KIF18A. The compound can be used for treating bladder cancer, kidney cancer, ovarian cancer, leukemia, B-cell lymphoma, T-cell lymphoma and other diseases.

Description

Technical Field

[0001] The present invention relates to a class of compounds having KIF18A inhibitory activity, and uses thereof as KIF18A inhibitors and for treating cancer. Background Art

[0002] The kinesin family, a superfamily of microtubule-associated molecular motor proteins, plays a central role in numerous physiological processes in eukaryotic cells, including organelle transport, spindle assembly, chromosome segregation, and intracellular signaling. Among them, KIF18A (kinesin-like protein 18A), a member of the kinesin-8 subfamily, is an ATP-dependent motor protein localized to the plus ends of mitotic spindle microtubules. Its unique molecular structure, comprising an N-terminal motor domain, a central rod domain, and a C-terminal tail domain, precisely regulates mitotic progression through mechanisms such as microtubule dynamics inhibition, chromosome oscillation regulation, and spindle length maintenance.

[0003] In recent years, an increasing number of studies have shown that KIF18A plays a key role in the occurrence and development of tumors: in a variety of solid tumors such as breast cancer, lung cancer, and colorectal cancer, the KIF18A gene is abnormally highly expressed and is significantly correlated with tumor grade, lymph node metastasis, and poor patient prognosis; its overexpression can not only accelerate the mitotic process of tumor cells and enhance cell proliferation ability, but also promote the migration and invasion of tumor cells by regulating cytoskeleton dynamics; in addition, abnormal expression of KIF18A can also lead to chromosome segregation errors, aggravate the genomic instability of tumor cells, and induce tumor cells to develop resistance to chemotherapy drugs such as taxanes.

[0004] Given the central role of KIF18A in tumor cell proliferation and survival, the development of highly effective inhibitors has become a cutting-edge focus in anti-tumor drug research and development. Amgen's AMG-650, the world's first KIF18A inhibitor to enter clinical trials, is being evaluated for triple-negative breast cancer and platinum-resistant high-grade serous ovarian cancer and has been granted FDA Fast Track designation. AMG-650 binds with high affinity to the ATP-binding pocket of KIF18A, inhibiting its microtubule-depolymerizing activity and demonstrating significant anti-tumor activity in animal models.

[0005]

[0006] Currently, there are no approved drugs based on the KIF18A inhibition pathway in the world. Therefore, developing a new KIF18A inhibitor compound to provide a new solution for cancer treatment has important clinical significance and market potential. Summary of the Invention

[0007] The present invention provides a novel KIF18A inhibitor and its use in preparing a medicament for preventing or treating diseases related to KIF18A activity or expression.

[0008] The technical solutions of the present invention are as follows:

[0009] The present invention provides a KIF18A inhibitor selected from the following compounds, pharmaceutically acceptable salts or stereoisomers thereof:

[0010] .

[0011] The present invention also provides a pharmaceutical composition comprising any of the aforementioned compounds, a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier. The pharmaceutical composition can be prepared into various pharmaceutically acceptable dosage forms, such as tablets, capsules, oral solutions, granules, injections, or various sustained-release formulations. The pharmaceutical composition can be administered orally or parenterally (e.g., intravenously, subcutaneously, or topically). The dosage can be adjusted appropriately based on the patient's age, gender, and disease type; the typical daily dose is approximately 1-200 mg.

[0012] In another aspect, the present invention provides the use of any one of the above compounds, pharmaceutically acceptable salts, stereoisomers, or pharmaceutical compositions thereof in the preparation of a medicament for preventing and / or treating a disease associated with KIF18A activity or expression. The disease is cancer; the cancer is a solid tumor or a blood-derived tumor, the solid tumor is bladder cancer, kidney cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, colon cancer, liver cancer, esophageal cancer, gallbladder cancer, pancreatic cancer, gastric cancer, lung cancer (including squamous cell lung cancer, small cell lung cancer), head and neck cancer, thyroid cancer (including follicular thyroid cancer), brain cancer, skin cancer, melanoma, keratoacanthoma, xeroderma pigmentosum, breast cancer, osteosarcoma, teratoma, seminoma, Kaposi's sarcoma, fibrosarcoma, rhabdomyosarcoma, astrocytoma, neuroblastoma, glioma, or neurilemmoma; the blood-derived tumor is leukemia (including acute lymphoblastic leukemia, acute lymphoblastic leukemia), B cell lymphoma, T cell lymphoma, Myeloid leukemia, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, Burkett lymphoma, acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome, or promyelocytic leukemia.

[0013] The compounds provided by the present invention have good KIF18A inhibitory activity and can be used to treat / or prevent KIF18A-mediated diseases. DETAILED DESCRIPTION

[0014] The present invention is further described below with reference to specific embodiments and test examples, but they are not intended to limit the scope of the present invention in any form.

[0015] Example 1 Synthesis of Compound 1

[0016]

[0017] Synthesis route:

[0018]

[0019] Step 1: Synthesis of compound 1-2

[0020] Compound 1-1 (2 g, 5.60 mmol, 1 eq.), N,N-diisopropylethylamine (4.34 g, 33.60 mmol, 5.85 mL, 6 eq.), and N,O-dimethylhydroxylamine hydrochloride (900.40 mg, 6.72 mmol, 1.2 eq.) were dissolved in dimethyl sulfoxide (30 mL). n-Butylphosphonic anhydride (50% ethyl acetate solution) (4.84 g, 6.72 mmol, 50% purity, 1.2 eq.) was added with stirring at 20°C. The reaction mixture was then stirred at 25°C for 1.0 hour. LCMS confirmed the complete reaction of the starting material. The reaction mixture was quenched with water (80 mL) and extracted with ethyl acetate (75 mL × 2). The organic phase was washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate: 1 / 0 to 5 / 1) to yield compound 1-2 (1.7 g, 3.82 mmol, 68.27% yield) as a pale yellow oil. (ESI) m / z = 401.0 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 7.42 - 7.34 (m, 2H), 6.98 (br d, J = 7.6 Hz, 1H), 3.98 - 3.43 (m,3H), 3.40 - 2.91 (m, 7H), 1.46 (br s, 4H), 0.33 (s, 4H).

[0021] Step 2: Synthesis of Compounds 1-4

[0022] Compound 1-3 (5 g, 15.71 mmol, 1 eq.) was dissolved in tetrahydrofuran (20 mL). A 2 M solution of lithium diisopropylamine in tetrahydrofuran (2 M, 11.78 mL, 1.5 eq.) was added at -78°C under a nitrogen atmosphere. The reaction mixture was then stirred at -78°C for 1 hour. Compound 1-2 (6.29 g, 15.71 mmol, 1 eq.) was then dissolved in tetrahydrofuran (20 mL). This mixture was slowly added to the reaction mixture at -78°C under a nitrogen atmosphere. The mixture was then slowly warmed to room temperature and stirred for 0.5 hour. LCMS monitoring of the reaction was performed. The reaction mixture was quenched with saturated aqueous ammonium chloride (20 mL) at 0°C and extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: technical n-hexane / ethyl acetate: 20 / 1) to obtain compound 1-4 (3.52 g, 4.98 mmol, 25.16% yield) as a yellow gum. (ESI) m / z = 657.1 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 7.51 (s, 1H), 7.48 - 7.43(m, 1H), 7.14 (d, J = 8.0 Hz, 1H), 6.91 (s, 1H), 6.08 (s, 2H), 3.67 - 3.60(m, 2H), 3.06 - 3.00 (m, 4H), 1.21 - 1.15 (m, 4H), 0.97 - 0.92 (m, 2H), 0.24 (s, 4H), -0.02 (s, 9H).

[0023] Step 3: Synthesis of Compounds 1-5

[0024] Compound 1-4 (3.52 g, 5.35 mmol, 1 eq.) and iron acetylacetonate (189.09 mg, 535.41 μmol, 0.1 eq.) were dissolved in tetrahydrofuran (35 mL). 3M methylmagnesium bromide in tetrahydrofuran (3 M, 2.14 mL, 1.2 eq.) was added under stirring at 0°C under a nitrogen atmosphere. The reaction mixture was then stirred at 0°C for 1 hour. LCMS confirmed the completion of the reaction. Water (20 mL) was added to the reaction mixture at 0°C, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield crude compound 1-5 (3.26 g, crude) as a brown oil. (ESI) m / z = 637.2 [M+H] + .

[0025] Step 4: Synthesis of Compounds 1-6

[0026] Compound 1-5 (3.26 g, 5.12 mmol, 1 eq.), 4,4-difluorohexahydropyridine hydrochloride (1.61 g, 10.24 mmol, 2 eq.), and N,N-diisopropylethylamine (1.32 g, 10.24 mmol, 1.78 mL, 2 eq.) were dissolved in dimethyl sulfoxide (40 mL). The reaction mixture was stirred at 100°C for 1 hour. LCMS confirmed the complete reaction. The reaction mixture was cooled to room temperature, and water (40 mL) was added. The mixture was extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain crude compound 1-6 (4.21 g, crude) as a brown oil. (ESI) m / z = 722.3 [M+H] + .

[0027] Step 5: Synthesis of Compounds 1-7

[0028] Compound 1-6 (2.1 g, 2.91 mmol, 1 eq.) was dissolved in dichloromethane (25 mL). Trifluoroacetic acid (7.68 g, 67.31 mmol, 5 mL, 23.13 eq.) was added at room temperature. The reaction mixture was stirred at 40°C for 1 hour. The concentrate was then concentrated and dissolved in acetonitrile (20 mL). Aqueous ammonia (6.55 g, 43.00 mmol, 7.20 mL, 23% purity, 14.85 eq.) was added at 20°C and the reaction mixture was stirred at 20°C for 1 hour. LCMS confirmed the completion of the reaction. The crude product was concentrated and lyophilized via reverse-phase HPLC (column: Waters xbridge 150 × 25 mm × 10 μm; mobile phase: [H2O(0.1%TFA)-ACN]; gradient: 0%-100% B over 15.0 min) to afford compound 1-7 (0.4 g, 662.80 μmol, 22.88% yield) as a yellow solid. (ESI) m / z = 255.2 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 9.18 - 9.00 (m, 1H), 7.46 - 7.42 (m, 1H), 7.41 - 7.35 (m, 1H), 7.14 - 7.10 (m, 1H), 6.90 - 6.83 (m, 1H), 4.10 - 4.05(m, 4H), 3.08 - 3.03 (m, 4H), 2.59 - 2.52 (m, 3H), 2.08 (s, 4H), 1.27 - 1.24(m, 4H), 0.24 (s, 4H). 19 F NMR (400 MHz, CDCl3) δ -97.049.

[0029] Step 6: Synthesis of Compound 1

[0030] Compound 2-hydroxy-1-sulfonamide (160.82 mg, 1.29 mmol, 2 eq.), sarcosine (114.49 mg, 1.29 mmol, 2 eq.), potassium phosphate (4681.91 mg, 3.21 mmol, 5 eq.), and cuprous iodide (122.37 mg, 642.51 μmol, 1 eq.) were dissolved in N,N-dimethylformamide (5 mL). The reaction mixture was stirred at 50°C under a nitrogen atmosphere for 5 minutes. Compound 1-7 (380 mg, 642.51 μmol, 1 eq.) was then added to the reaction mixture, which was then stirred at 130°C for 2 hours. LCMS confirmed the complete reaction. The reaction mixture was cooled to room temperature and added with 10 mL of water. The mixture was extracted with ethyl acetate (30 mL × 3). The organic phase was washed with saturated brine (30 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield the crude product, which was then lyophilized by reverse-phase preparative HPLC (Prep-HPLC: column: Phenomenex luna 150 × 25 mm × 10 μm; mobile phase: [H2O(0.225% FA)-ACN]; gradient: 40%-70% B over 9.0 min) to afford compound 1 (15 mg, 24.89 μmol, 3.87% yield, 97.67% purity) as a yellow solid. (ESI) m / z = 589.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d6) δ11.94 (s, 1H), 9.97 (s, 1H), 7.30 (d, J = 8.0 Hz, 1H), 6.95 - 6.93 (m, 2H), 6.85 (dd, J = 2.0, 8.4 Hz, 1H), 4.97 (t, J = 6.0 Hz, 1H), 3.96 (t, J = 4.0Hz, 4H), 3.78 (q, J = 6.4 Hz, 2H), 3.37 - 3.34 (m, 2H), 3.33 - 3.33 (m, 2H), 2.95 (t, J = 4.0 Hz, 4H), 2.52 (s, 3H), 2.05 - 1.96 (m, 4H), 1.13 (t, J = 5.2Hz, 4H), 0.20 (s, 4H). 19 F NMR (400 MHz, DMSO-d6) δ -94.967.

[0031] Example 2 Synthesis of Compound 2

[0032]

[0033] Synthesis route:

[0034]

[0035] Step 1: Synthesis of compound 2-2

[0036] Compound 2-1 (2 g, 5.84 mmol, 1 eq.) and ferric acetylacetonate (206.41 mg, 584.45 μmol, 0.1 eq.) were dissolved in tetrahydrofuran (20 mL). Methylmagnesium bromide (3 M, 2.92 mL, 1.5 eq.) was added under stirring at 0°C under a nitrogen atmosphere. The reaction mixture was then stirred at 0°C for 1 hour. LCMS confirmed the complete reaction. The reaction mixture was quenched with saturated aqueous ammonium chloride (20 mL) at 0°C and extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain crude compound 2-2 (1.88 g, 5.26 mmol, 89.97% yield, 90% purity) as a red solid. The crude product was used directly in the next reaction. (ESI)m / z = 322.1 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 8.12 (d, J = 8.4 Hz, 2H), 7.68(d, J = 4.0 Hz, 1H), 7.35 (d, J = 8.4 Hz, 2H), 6.62 (d, J = 3.6 Hz, 1H), 2.67(s, 3H), 2.42 (s, 3H).

[0037] Step 2: Synthesis of compound 2-3

[0038] Compound 2-2 (1.7 g, 5.28 mmol, 1 eq.) and 4,4-difluorohexahydropyridine (999.07 mg, 6.34 mmol, 1.2 eq.) were dissolved in dimethyl sulfoxide (20 mL). N,N-diisopropylethylamine (2.05 g, 15.85 mmol, 2.76 mL, 3 eq.) was added with stirring at 20°C. The reaction mixture was then stirred at 120°C for 1.5 hours. LCMS monitoring confirmed the complete reaction of the starting material. The reaction mixture was cooled to room temperature, water (20 mL) was added, and extraction with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1) to afford compound 2-3 as a white solid (1.85 g, 4.40 mmol, 81.71% yield, 96.74% purity). (ESI) m / z = 407.0 [M+H] + . 1 HNMR (400 MHz, CDCl3) δ 8.00 (d, J = 8.4 Hz, 2H), 7.35 (d, J = 4.0 Hz, 1H), 7.29 (d, J = 8.4 Hz, 2H), 6.44 (d, J = 4.0 Hz, 1H), 4.04 (t, J = 5.6 Hz, 4H), 2.47 (s, 3H), 2.41 (s, 3H), 2.01 - 1.92 (m, 4H). 19 F NMR (400 MHz, CDCl3) δ-96.700.

[0039] Step 3: Synthesis of Compound 2-4

[0040] Compound 2-3 (0.5 g, 1.23 mmol, 1 eq.) was dissolved in a mixture of methanol (5 mL) and tetrahydrofuran (10 mL). 5 mL of 2 mol / mL sodium hydroxide solution was slowly added at room temperature, and the reaction mixture was stirred for 12 hours. LCMS confirmed the completion of the reaction. The reaction mixture was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1) to obtain compound 2-4 (0.37 g, 1.47 mmol, 99.36% yield, 100% purity) as a white solid. (ESI) m / z = 253.1 [M+H] + . 1H NMR (400 MHz, CDCl3)δ 8.71 (s, 1H), 6.90 (q, J = 2.4Hz, 1H), 6.39 (q, J = 2.0Hz, 1H), 4.01 (t, J= 5.6Hz, 1H), 2.58(s, 3H), 2.07 - 1.97 (m, 4H). 19 F NMR (400 MHz, CDCl3) δ = -96.695.

[0041] Step 4: Synthesis of Compound 2-5

[0042] Compound 2-4 (0.27 g, 1.07 mmol, 1 eq.) was dissolved in a mixture of ethyl acetate (5 mL) and glacial acetic acid (0.81 mL). 10% wet palladium on carbon (270 mg) was added under an argon atmosphere. The reaction mixture was then stirred at 25°C under a hydrogen atmosphere (15 psi) for 12 hours. LCMS confirmed the completion of the reaction. The reaction mixture was cooled to room temperature, filtered, and rinsed with ethyl acetate (8 mL × 2). The filtrate was concentrated to obtain the crude product, which was then dissolved in water (5 mL) and the pH adjusted to 8–9 with saturated sodium bicarbonate solution. The product was extracted with ethyl acetate (50 mL × 3), and the organic phase was washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude white solid compound 2-5 (262 mg, crude). The crude product was used directly in the next step. (ESI) m / z = 255.2 [M+H] + .

[0043] Step 5: Synthesis of Compounds 2-6

[0044] Compound 2-5 (50 mg, 196.63 μmol, 1 eq.) and 4-iodo-2-(6-azaspiro[2.5]octan-6-yl)benzoic acid (70.24 mg, 196.63 μmol, 1 eq.) were dissolved in dichloromethane (1 mL). 2-Chloro-1-methylpyridine iodide (60.28 mg, 235.96 μmol, 1.2 eq.) and N,N-diisopropylethylamine (50.83 mg, 393.27 μmol, 2 eq.) were added, respectively. The mixture was stirred at room temperature for 12 hours. LCMS analysis indicated that the reaction was complete. The reaction mixture was concentrated to obtain a crude product, which was then purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1) to afford compound 2-6 as a white solid (71 mg, 110.07 μmol, 46.65% yield, 92% purity). (ESI) m / z = 594.2 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 7.47 - 7.42 (m, 1H), 7.37 - 7.33 (m, 1H), 7.06 -7.03 (m, 1H), 4.94 - 4.62 (m, 4H), 4.28 - 4.14 (m, 2H), 3.54 - 3.25 (m, 4H),3.10 - 2.92 (m, 4H), 2.72 - 2.59 (m, 2H), 2.24 (s, 3H), 1.87 - 1.73 (m, 4H),0.25 (s, 4H). 19 F NMR (400 MHz, CDCl3) δ -97.232.

[0045] Step 6: Synthesis of Compound 2

[0046] Compound 2-6 (40 mg, 67.40 μmol, 1 eq.) and 2-hydroxy-1-sulfonamide (16.87 mg, 134.81 μmol, 2 eq.) were dissolved in N,N-dimethylformamide (2 mL). Anhydrous potassium phosphate (71.54 mg, 337.01 μmol, 5 eq.), cuprous iodide (12.84 mg, 67.40 μmol, 1 eq.), and sarcosine (12.01 mg, 134.81 μmol, 2 eq.) were then added. Under nitrogen, the reaction mixture was stirred at 130°C for 2.0 hours. LCMS was used to monitor the reaction. The reaction mixture was cooled to room temperature and quenched with saturated brine (20 mL). The mixture was extracted with ethyl acetate (30 mL × 2), and the organic phase was washed with saturated brine (20 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield the crude product. The crude product was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 0 to 3 / 7) to afford compound 2 as an off-white solid (4.7 mg, 7.64 μmol, 11.34% yield, 96.08% purity). (ESI) m / z = 591.3 [M+H] + . 19 F NMR (377 MHz, CD3SOCD3) δ -94.99 (br s, 2F). 1 H NMR(400 MHz, CD3SOCD3) δ 9.89 (s, 1H), 7.19 (d, J = 8.0 Hz, 1H), 6.91 - 6.84 (m,2H), 4.13 - 4.03 (m, 2H), 3.77 (t, J = 6.4 Hz, 2H), 3.31 - 3.29 (m, 4H), 2.97- 2.87 (m, 4H), 2.61 - 2.56 (m, 2H), 2.19 (s, 3H), 1.77 - 1.66 (m, 4H), 1.30- 1.17 (m, 4H), 1.05 - 0.95 (m, 2H), 0.23 (br s, 4H).

[0047] Example 3 Synthesis of Compound 3

[0048]

[0049] Synthesis route:

[0050]

[0051] Step 1: Synthesis of compound 3-2

[0052] Compound 3-1 (5 g, 26.45 mmol, 1 eq.) and p-toluenesulfonic acid monohydrate (251.61 mg, 1.32 mmol, 0.05 eq.) were dissolved in ethyl acetate (75 mL). The mixture was then heated to 50°C and 2,3-dihydropyran (3.34 g, 39.68 mmol, 3.63 mL, 1.5 eq.) was added with stirring at 50°C. The reaction mixture was stirred at 50°C for 2.5 hours and then at 60°C for 1.0 hour. LCMS confirmed the completion of the reaction. The reaction mixture was concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to obtain compound 3-2 (5.6 g, 20.50 mmol, 77.51% yield) as a white solid. (ESI) m / z = 189.0 [M-THP+H] + . 1 H NMR (400MHz, CDCl3) δ 8.33 (s, 1H), 5.75 (dd, J = 2.4, 10.4 Hz, 1H), 4.22 - 4.15 (m,1H), 3.80 - 3.74 (m, 1H), 2.21 - 2.13 (m, 1H), 2.12 - 2.05 (m, 1H), 2.02 -1.90 (m, 1H), 1.85 - 1.71 (m, 2H), 1.70 - 1.64 (m, 1H).

[0053] Step 2: Synthesis of compound 3-3

[0054] At 0°C, 3 mol / L methylmagnesium bromide (3 M, 3.66 mL, 1.5 eq.) was slowly added dropwise to a mixture of compound 3-2 (2 g, 7.32 mmol, 1 eq.) and ferric acetylacetonate (258.62 mg, 732.28 μmol, 0.1 eq.) in THF (20 mL). The reaction was stirred at 20°C for 2.0 hours. The reaction was monitored by LCMS. The reaction was quenched with saturated ammonium chloride solution at 0°C and extracted with ethyl acetate (75 mL × 2). The organic phase was washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to obtain compound 3-3 (1.28 g, 5.07 mmol, yield 69.17%) as a light yellow solid.1 H NMR (400 MHz, CDCl3) δ 8.24 (s, 1H), 5.77 (dd, J = 2.4, 10.8Hz, 1H), 4.21 - 4.15 (m, 1H), 3.83 - 3.75 (m, 1H), 2.85 (s, 3H), 2.18 - 2.12(m, 1H), 2.11 - 2.06 (m, 1H), 2.03 - 1.93 (m, 1H), 1.81 - 1.76 (m, 1H), 1.70- 1.65 (m, 2H).

[0055] Step 3: Synthesis of compound 3-4

[0056] Compound 3-3 (1.28 g, 5.07 mmol, 1 eq.) and 4,4-difluorohexahydropyridine (1.20 g, 7.60 mmol, 1.5 eq., HCl) were dissolved in dimethyl sulfoxide (15 mL). N,N-diisopropylethylamine (1.96 g, 15.20 mmol, 2.65 mL, 3 eq.) was added with stirring. The reaction mixture was then stirred at 130°C for 2.0 hours. LCMS monitoring indicated that the reaction was complete. The reaction mixture was cooled to room temperature, quenched with water (60 mL), and extracted with ethyl acetate (75 mL × 2). The organic phase was washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to yield the crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to yield compound 3-4 (710 mg, 2.10 mmol, 41.55% yield) as a pale yellow solid. (ESI) m / z = 338.2 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 7.91 (s, 1H), 5.63 - 5.57 (m, 1H), 4.07 - 3.99 (m,4H), 3.80 - 3.73 (m, 1H), 2.66 (s, 3H), 2.08 - 1.96 (m, 7H), 1.82 - 1.70 (m,3H), 1.69 - 1.63 (m, 1H). 19 F NMR (376 MHz, CDCl3) δ -96.91 (br s, 2F).

[0057] Step 4: Synthesis of Compound 3-5

[0058] Under nitrogen at -65°C, a 2 M solution of lithium diisopropylamine in tetrahydrofuran / n-heptane (2 M, 1.11 mL, 1.5 eq.) was slowly added dropwise to a solution of compound 3-4 (500 mg, 1.48 mmol, 1 eq.) in tetrahydrofuran (10 mL). The reaction was stirred at -65°C for 0.5 hour. Then, a solution of 4-iodo-N-methoxy-N-methyl-2-(6-azaspiro[2.5]octan-6-yl)benzamide (652.52 mg, 1.63 mmol, 1.1 eq.) in tetrahydrofuran (2.5 mL) was added to the reaction mixture with stirring at -65°C. The reaction was then stirred at -65°C for 2 hours. LCMS monitoring indicated that the reaction was complete. The reaction mixture was quenched with saturated ammonium chloride solution at 0°C and extracted with ethyl acetate (50 mL × 2). The organic phase was washed with saturated brine (30 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 0 to 4 / 1) to obtain compound 3-5 (525 mg, 373 μmol, 24.87% yield) as a yellow solid. (ESI) m / z = 677.1 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ 7.52 (s, 1H), 7.43 (d, J = 8.0 Hz, 1H), 7.25 (s, 1H), 6.05 (br d, J= 9.6 Hz, 1H), 4.07 (br s, 4H), 3.64 - 3.56 (m, 1H), 3.04 (br d, J = 2.0 Hz,1H), 2.96 (br t, J = 5.2 Hz, 4H), 2.59 (s, 3H), 2.07 - 1.97 (m, 4H), 1.80 -1.68 (m, 2H), 1.58 (br s, 4H), 1.21 - 1.10 (m, 4H), 0.22 (s, 4H).

[0059] Step 5: Synthesis of Compounds 3-6

[0060] Compound 3-5 (525 mg, 776.01 μmol, 1 eq.) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (5 mL) was added with stirring. The reaction mixture was then stirred at 20°C for 1.0 hour. LCMS confirmed the completion of the reaction. The reaction mixture was concentrated to obtain a crude product, which was quenched with saturated sodium bicarbonate solution (20 mL) and extracted with ethyl acetate (50 mL × 2). The organic phase was washed with saturated brine (30 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 0 to 4 / 1) to obtain compound 3-6 (440 mg, 742.71 μmol, 95.71% yield) as a brown solid. (ESI) m / z = 593.2 [M+H] + . 19 FNMR (376 MHz, CDCl3) δ -97.13 (br s, 2F). 1 H NMR (400 MHz, CDCl3) δ 10.03 (brs, 1H), 7.52 (s, 1H), 7.44 (d, J = 1.6 Hz, 2H), 4.07 (br t, J = 5.6 Hz, 4H), 3.07 - 3.01 (m, 4H), 2.67 (s, 3H), 2.10 - 2.06 (m, 2H), 2.04 - 1.98 (m, 2H), 1.26 - 1.22 (m, 4H), 0.26 (s, 4H).

[0061] Step 6: Synthesis of compound 3

[0062] 2-Hydroxyethane-1-sulfonamide (97.17 mg, 776.47 μmol, 2 eq.) was dissolved in N,N-dimethylformamide (5 mL), followed by the addition of anhydrous potassium phosphate (412.05 mg, 1.94 mmol, 5 eq.), cuprous iodide (73.94 mg, 388.24 μmol, 1 eq.), and sarcosine (69.18 mg, 776.47 μmol, 2 eq.). Under nitrogen, the reaction mixture was stirred at 50°C for 5 minutes. Compound 3-6 (230 mg, 388.24 μmol, 1 eq.) was then added to the reaction mixture at 50°C. Finally, the reaction mixture was stirred at 130°C for 1.0 hour under nitrogen. LCMS confirmed the complete reaction. The reaction mixture was cooled to room temperature and filtered. The reaction mixture was then quenched with saturated brine (20 mL) and extracted with ethyl acetate (30 mL × 2). The organic phase was washed with saturated brine (20 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 0 to 0 / 1) to obtain the product. The product was then lyophilized by reverse phase preparative chromatography to obtain the target compound 3 (4.2 mg, 7.08 μmol, 1.82% yield, 99.38% purity) as a yellow solid. Reverse phase preparative chromatography (mobile phase: [H2O(0.1%FA)-ACN]; B%: 0-60%, 20 min). (ESI) m / z = 590.3 [M+H] + . 19 F NMR (377 MHz, DMSO-d6) δ-95.04 (br s, 2F). 1H NMR (400 MHz, CD3SOCD3) δ 13.44 (br s, 1H), 10.10 (br s,1H), 7.54 (d, J = 8.3 Hz, 1H), 7.01 (d, J = 1.6 Hz, 1H), 6.90 (dd, J = 1.6,8.4 Hz, 1H), 4.99 - 4.98 (m, 1H), 3.95 (br dd, J = 4.8, 6.0 Hz, 4H), 3.78 (brt, J = 6.4 Hz, 2H), 3.38 - 3.37 (m, 2H), 2.89 - 2.83 (m, 4H), 2.53 (br s,3H), 2.06 - 1.96 (m, 4H), 1.08 (br d, J = 4.54 Hz, 4H), 0.20 (s, 4H).

[0063] Experimental Example 1. KIF18A functional in vitro activity test

[0064] 1. Test of KIF18A enzyme inhibitory activity of the compounds of the present invention

[0065] 1.1 Test consumables

[0066]

[0067] 1.2 Test instruments

[0068]

[0069] 1.3 Test methods

[0070] (1) Perform serial dilutions of the compound using DMSO in a 384-well dilution plate. Use Echo to transfer 0.1 μL of the compound to a 384-well reaction microplate (OptiPlate 384), ensuring a final DMSO concentration of 1% (in duplicate).

[0071] (2) Add 5 μL of enzyme solution to each well of a 384-well reaction microplate and centrifuge at 1000 rpm for 1 minute.

[0072] (3) Incubate at 25°C for 10 minutes.

[0073] (4) The wells containing 1% DMSO and enzyme serve as the High Control, and the wells containing 1% DMSO and buffer serve as the Low Control. Add 5 μL of ATP solution to each well and centrifuge at 1000 rpm for 1 minute.

[0074] (5) Incubate at 25°C for 60 minutes.

[0075] (6) Add 5 μL of ADP-Glo ​​Reagent solution to each well and centrifuge at 1000 rpm for 1 minute.

[0076] (7) Incubate at 25°C for 40 minutes.

[0077] (8) Add 10 μL of ADP-Glo ​​Detection solution to each well and centrifuge at 1000 rpm for 1 minute.

[0078] (9) Incubate at 25°C for 40 minutes.

[0079] (10) Read the signal value of Relative Luminescence Unit on a BMG (PHERAstar FSX) microplate reader and calculate the inhibition percentage of the compound-treated wells.

[0080] (12) Fitting the four-parameter IC using XLfit 5.5.0 50 Curve and analysis, IC 50 is the concentration of compound that results in 50% inhibition.

[0081] Table 1 In vitro inhibitory activity test results of the compounds of the present invention on KIF18A enzyme

[0082]

[0083] The test results showed that compounds 1, 2, and 3 all exhibited inhibitory activity against KIF18A, and their inhibitory abilities were significantly superior to those of the positive control drug AMG650. Among them, compound 1 had the strongest inhibitory effect.

Claims

1. A KIF18A inhibitor compound, a pharmaceutically acceptable salt or stereoisomer thereof, characterized in that: The compound is selected from any one of the following compounds 1-3: 。 2. A pharmaceutical composition, characterized in that: The invention comprises the compound according to claim 1, a pharmaceutically acceptable salt or stereoisomer thereof and a pharmaceutically acceptable carrier.

3. Use of the compound according to claim 1, its pharmaceutically acceptable salt or stereoisomer, or the pharmaceutical composition according to claim 2 in the preparation of a medicament for preventing and / or treating diseases related to KIF18A activity or expression.

4. The use according to claim 3, characterized in that: The disease is cancer.

5. The use according to claim 4, characterized in that: The cancer is a solid tumor or a blood-derived tumor.

6. The use according to claim 5, characterized in that: The solid tumor is bladder cancer, kidney cancer, ovarian cancer, cervical cancer, endometrial cancer, prostate cancer, colon cancer, liver cancer, esophageal cancer, gallbladder cancer, pancreatic cancer, stomach cancer, lung cancer, head and neck cancer, thyroid cancer, brain cancer, skin cancer, melanoma, keratoacanthoma, xeroderma pigmentosum, breast cancer, osteosarcoma, teratoma, seminoma or Kaposi's sarcoma; the hematological tumor is leukemia, B cell lymphoma, T cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, hairy cell lymphoma, Burkett lymphoma, acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome or promyelocytic leukemia.

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