Compounds as KIF18A inhibitors and their uses

Abstract

This invention relates to compounds, pharmaceutical compositions, and their applications as KIF18A inhibitors. Specifically, this invention provides compounds represented by Formulas 1-3, pharmaceutical compositions, and their applications in the preparation of medicaments for the prevention and / or treatment of diseases related to KIF18A activity or expression levels. The compounds provided by this invention exhibit good KIF18A inhibitory activity and can be used to treat diseases such as bladder cancer, kidney cancer, ovarian cancer, leukemia, B-cell lymphoma, and T-cell lymphoma.

Description

Technical Field

[0001] This invention relates to a class of compounds with KIF18A inhibitory activity, and their use as KIF18A inhibitors and for the treatment of cancer. Background Technology

[0002] The kinesin family, a superfamily of microtubule-associated motor proteins, plays a central role in various physiological processes in eukaryotic cells, including organelle transport, spindle assembly, chromosome segregation, and intracellular signal transduction. KIF18A (kinesin-like protein 18A), a member of the kinesin-8 subfamily, is an ATP-dependent motor protein located at the positive end of microtubules in the mitotic spindle. Its unique molecular structure includes an N-terminal motor domain, a central rod domain, and a C-terminal tail domain, enabling it to precisely regulate the process of mitosis through mechanisms such as microtubule dynamic 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 tumorigenesis and development: the KIF18A gene is abnormally overexpressed in various solid tumors such as breast cancer, lung cancer, and colorectal cancer, and is significantly associated 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, but also promote the migration and invasion of tumor cells by regulating the dynamics of the cytoskeleton; in addition, abnormal expression of KIF18A can lead to chromosome segregation errors, exacerbate the instability of tumor cell genome, and thus induce tumor cells to develop resistance to chemotherapeutic drugs such as taxanes.

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

[0005]

[0006] Currently, no drugs based on the KIF18A inhibition pathway have been approved for marketing globally. Therefore, developing a new KIF18A inhibitor compound to provide a new solution for cancer treatment has significant clinical implications and market potential. Summary of the Invention

[0007] This invention provides a novel KIF18A inhibitor and its use in the preparation of medicaments for the prevention or treatment of diseases related to KIF18A activity or expression levels.

[0008] The technical solution of the present invention is as follows:

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

[0010] .

[0011] This invention also provides a pharmaceutical composition comprising any of the above-described compounds, a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable carrier. The pharmaceutical composition can be formulated into various pharmaceutically acceptable dosage forms, such as tablets, capsules, oral liquids, 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 appropriately adjusted according to the patient's age, sex, and disease type, generally with a daily dose of approximately 1-200 mg.

[0012] In another aspect of the invention, the invention provides the use of any of the above-described compounds, their pharmaceutically acceptable salts, stereoisomers, or pharmaceutical compositions, in the preparation of medicaments for the prevention and / or treatment of diseases related to KIF18A activity or expression levels. The disease is cancer; the cancer is a solid tumor or a hematopoietic tumor, the solid tumor being 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 carcinoma and small cell lung cancer), head and neck cancer, thyroid cancer (including follicular thyroid carcinoma), brain cancer, skin cancer, melanoma, keratoacanthoma, xeroderma pigmentosum, breast cancer, osteosarcoma, teratoma, seminoma, Kaposi's sarcoma, fibrosarcoma, rhabdomyosarcoma, astrocytoma, neuroblastoma, glioma, or schwannoma; the hematopoietic tumor is leukemia (including acute lymphoblastic leukemia and acute lymphoblastic leukemia), B-cell lymphoma, T-cell lymphoma, etc. Pilocytic lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, pilocytic lymphoma, Burkert lymphoma, acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome, or promyelocytic leukemia.

[0013] The compounds provided by this invention have good KIF18A inhibitory activity and can be used to treat / or prevent KIF18A-mediated diseases. Detailed Implementation

[0014] The present invention will be further illustrated below with reference to specific embodiments and test examples, but this does not limit the scope of the invention in any way.

[0015] Example 1: Synthesis of Compound 1

[0016]

[0017] Synthesis route:

[0018]

[0019] Step 1: Synthesis of Compounds 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). Butylphosphine 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 h. The reaction mixture was monitored by LCMS to ensure complete reaction of the starting materials. The reaction mixture was quenched with water (80 mL), extracted with ethyl acetate (75 mL × 2), 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 the crude product. The crude product was purified by silica gel column chromatography (elution: petroleum ether / ethyl acetate: 1 / 0 to 5 / 1) to give compounds 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] Compounds 1-3 (5 g, 15.71 mmol, 1 eq.) were dissolved in tetrahydrofuran (20 mL), and a 2M solution of lithium diisopropylamine in tetrahydrofuran (2 M, 11.78 mL, 1.5 eq.) was added under stirring at -78 °C under a nitrogen atmosphere. The reaction mixture was then stirred at -78 °C for 1 hour. Compounds 1-2 (6.29 g, 15.71 mmol, 1 eq.) were then dissolved in tetrahydrofuran (20 mL), and this mixture was slowly added to the above reaction mixture under a nitrogen atmosphere at -78 °C, followed by slow warming to room temperature and stirring for 0.5 hours. The reaction was monitored by LCMS. The reaction mixture was quenched at 0 °C with a saturated ammonium chloride aqueous solution (20 mL), 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 the crude product. The crude product was purified by silica gel column chromatography (eluent: industrial hexane / ethyl acetate: 20 / 1) to give compounds 1-4 (3.52 g, 4.98 mmol, 25.16% yield), as a yellow gel. (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] Compounds 1-4 (3.52 g, 5.35 mmol, 1 eq.) and ferric acetylacetone (189.09 mg, 535.41 μmol, 0.1 eq.) were dissolved in tetrahydrofuran (35 mL). A 3 M methylmagnesium bromide tetrahydrofuran solution (3 M, 2.14 mL, 1.2 eq.) was added under nitrogen atmosphere at 0 °C with stirring. The reaction mixture was then stirred at 0 °C for 1 hour. LC-MS was used to monitor the reaction until complete. 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 give crude compound 1-5 (3.26 g, crude), which was a brown oil. (ESI) m / z = 637.2 [M+H] + .

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

[0026] Compounds 1-5 (3.26 g, 5.12 mmol, 1 eq.), compound 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. The reaction mixture was monitored by LCMS to ensure 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), 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 give crude compound 1-6 (4.21 g, crude), which was a brown oil. (ESI) m / z = 722.3 [M+H] + .

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

[0028] Compounds 1-6 (2.1 g, 2.91 mmol, 1 eq.) were dissolved in dichloromethane (25 mL), and trifluoroacetic acid (7.68 g, 67.31 mmol, 5 mL, 23.13 eq.) was added at room temperature. The reaction mixture was then stirred at 40 °C for 1 hour, concentrated, and dissolved in acetonitrile (20 mL). 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. The reaction was monitored by LCMS to ensure complete reaction of the starting materials. The crude product was concentrated and then prepared by 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). After lyophilization, compounds 1-7 (0.4 g, 662.80 μmol, 22.88% yield) were obtained as a yellow solid. (ESI) m / z = 255.2 [M+H] + .

[0029] 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.

[0030] Step 6: Synthesis of Compound 1

[0031] Compounds 2-hydroxy-1-sulfonamide (160.82 mg, 1.29 mmol, 2 eq.) and 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. Compounds 1-7 (380 mg, 642.51 μmol, 1 eq.) were then added to the reaction mixture, and the mixture was back-stirred at 130 °C for 2 hours. LC-MS analysis confirmed complete reaction of the starting materials. The reaction mixture was cooled to room temperature, and 10 mL of water was added. The mixture was extracted with ethyl acetate (30 mL × 3), and the organic phase was washed with saturated brine (30 mL × 2). The organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was then prepared by reverse-phase 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) and lyophilized to obtain compound 1 (15 mg, 24.89 μmol, yield 3.87%, purity 97.67%) 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.

[0032] Example 2: Synthesis of Compound 2

[0033]

[0034] Synthesis route:

[0035]

[0036] Step 1: Synthesis of Compound 2-2

[0037] Compound 2-1 (2 g, 5.84 mmol, 1 eq.) and ferric acetylacetone (206.41 mg, 584.45 μmol, 0.1 eq.) were dissolved in tetrahydrofuran (20 mL), and 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. The reaction mixture was monitored by LCMS to ensure complete reaction of the starting material. The reaction mixture was quenched at 0 °C with saturated ammonium chloride aqueous solution (20 mL), 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 give crude compound 2-2 as a red solid (1.88 g, 5.26 mmol, yield 89.97%, purity 90%). 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).

[0038] Step 2: Synthesis of compounds 2-3

[0039] Compound 2-2 (1.7 g, 5.28 mmol, 1 eq.) and compound 4,4-difluorohexahydropyridine (999.07 mg, 6.34 mmol, 1.2 eq.) were dissolved in dimethyl sulfoxide (20 mL), and 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 h. The reaction mixture was monitored by LCMS to ensure complete reaction. The reaction mixture was cooled to room temperature, and water (20 mL) was added. The mixture 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 the crude product. The crude product was purified by silica gel column chromatography (elution: petroleum ether / ethyl acetate = 5 / 1) to give a white solid compound 2-3 (1.85 g, 4.40 mmol, yield 81.71%, purity 96.74%). (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.

[0040] Step 3: Synthesis of compounds 2-4

[0041] Compound 2-3 (0.5 g, 1.23 mmol, 1 eq.) was dissolved in a mixed solvent of methanol (5 mL) and tetrahydrofuran (10 mL), and 5 mL of sodium hydroxide aqueous solution (2 mol / mL) was slowly added at room temperature. The reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was monitored by LCMS to ensure complete reaction. The reaction mixture was directly 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 give compound 2-4 (0.37 g, 1.47 mmol, yield 99.36%, purity 100%) 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.

[0042] Step 4: Synthesis of compounds 2-5

[0043] Compound 2-4 (0.27 g, 1.07 mmol, 1 eq.) was dissolved in a mixed solvent of ethyl acetate (5 mL) and glacial acetic acid (0.81 mL), and 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 (15 Psi) atmosphere for 12 hours. The reaction mixture was monitored by LCMS to ensure complete reaction of the starting materials. The reaction mixture was cooled to room temperature, filtered, and washed with ethyl acetate (8 mL × 2). The filtrate was concentrated to obtain the crude product, which was dissolved in (5 mL) water, and the pH was adjusted to 8-9 with a saturated sodium bicarbonate aqueous solution. The solution 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 the crude white solid compound 2-5 (262 mg, crude). The crude product was used directly in the next reaction. (ESI) m / z = 255.2 [M+H] + .

[0044] Step 5: Synthesis of compounds 2-6

[0045] Compounds 2-5 (50 mg, 196.63 μmol, 1 eq.) and 4-iodo-2-(6-azaspiro[2.5]oct-6-yl)benzoic acid (70.24 mg, 196.63 μmol, 1 eq.) were dissolved in dichloromethane (1 mL), followed by the addition of 2-chloro-1-methyliodopyridine (60.28 mg, 235.96 μmol, 1.2 eq.) and N,N-diisopropylethylamine (50.83 mg, 393.27 μmol, 2 eq.). The mixture was back-stirred at room temperature for 12 hours. The reaction proceeds were determined to be complete by LCMS. The reaction solution was concentrated to obtain a crude product, which was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 3 / 1) to give a white solid compound 2-6 (71 mg, 110.07 μmol, yield 46.65%, purity 92%). (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.

[0046] Step 6: Synthesis of Compound 2

[0047] Compounds 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), followed by the addition of 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.). The reaction mixture was stirred at 130 °C for 2.0 h under nitrogen protection. The reaction was detected by LCMS. The reaction mixture was cooled to room temperature, quenched with saturated brine (20 mL), extracted with ethyl acetate (30 mL × 2), and 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 (elution: n-hexane / ethyl acetate = 1 / 0 to 3 / 7) to give a grayish-white solid compound 2 (4.7 mg, 7.64 μmol, yield 11.34%, purity 96.08%). (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).

[0048] Example 3: Synthesis of Compound 3

[0049]

[0050] Synthesis route:

[0051]

[0052] Step 1: Synthesis of compound 3-2

[0053] 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), and then heated to 50 °C. 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 first stirred at 50 °C for 2.5 h, and then at 60 °C for 1.0 h. The reaction mixture was monitored by LC-MS to ensure complete reaction. The reaction mixture was concentrated to give a crude product. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to give 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).

[0054] Step 2: Synthesis of compound 3-3

[0055] 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 acetylacetone (258.62 mg, 732.28 μmol, 0.1 eq.) in THF (20 mL). The reaction mixture was stirred at 20 °C for 2.0 h. The reaction was monitored by LCMS. At 0 °C, the reaction mixture was quenched with saturated ammonium chloride solution, then 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 (elution: petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to give compound 3-3 (1.28 g, 5.07 mmol, yield 69.17%) as a pale 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).

[0056] Step 3: Synthesis of compounds 3-4

[0057] 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 h. The reaction mixture was monitored by LCMS to ensure complete reaction. The reaction mixture was cooled to room temperature, quenched with water (60 mL), extracted with ethyl acetate (75 mL × 2), 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 the crude product. The crude product was purified by silica gel column chromatography (elution: petroleum ether / ethyl acetate = 1 / 0 to 1 / 1) to give compounds 3-4 (710 mg, 2.10 mmol, yield 41.55%) 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).

[0058] Step 4: Synthesis of compounds 3-5

[0059] Under nitrogen protection at -65°C, a 2M tetrahydrofuran-heptane mixture (2 M, 1.11 mL, 1.5 eq.) of lithium diisopropylamine was slowly added dropwise to a tetrahydrofuran (10 mL) solution of compounds 3-4 (500 mg, 1.48 mmol, 1 eq.), and the reaction mixture was stirred at -65°C for 0.5 h. Then, at -65°C, 2.5 mL of tetrahydrofuran of compound 4-iodo-N-methoxy-N-methyl-2-(6-azaspiro[2.5]octane-6-yl)benzamide (652.52 mg, 1.63 mmol, 1.1 eq.) was added to the reaction mixture with stirring. The reaction mixture was then stirred at -65°C for 2 h. The reaction of the starting materials was monitored by LCMS to ensure complete reaction. The reaction solution was quenched with saturated ammonium chloride solution at 0°C, followed by extraction 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 give compound 3-5 (525 mg, 373 μmol, yield 24.87%) 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).

[0060] Step 5: Synthesis of compounds 3-6

[0061] 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 h. The reaction mixture was monitored by LCMS to ensure complete reaction. The reaction mixture was concentrated to obtain a crude product, which was quenched with saturated sodium bicarbonate solution (20 mL), extracted with ethyl acetate (50 mL × 2), and 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 give compound 3-6 (440 mg, 742.71 μmol, yield 95.71%) 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).

[0062] Step 6: Synthesis of Compound 3

[0063] 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.). The reaction mixture was stirred at 50 °C for 5 minutes under nitrogen protection. Then, compound 3-6 (230 mg, 388.24 μmol, 1 eq.) was added to the reaction mixture at 50 °C. Finally, the reaction mixture was stirred at 130 °C for 1 hour under nitrogen protection. LC-MS analysis confirmed complete reaction of the starting materials. The reaction solution was cooled to room temperature, filtered, and then quenched with saturated brine (20 mL). Extraction was performed 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 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 reversed-phase prepared by lyophilization to obtain the target compound 3 (4.2 mg, 7.08 μmol, yield 1.82%, purity 99.38%) as a yellow solid. Reverse-phase preparative chromatography (mobile phase: [H₂O(0.1%FA)-ACN]; B%: 0-60%, 20 min) showed an ESI result of 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).

[0064] Experimental Example 1. In vitro functional activity test of KIF18A

[0065] 1. Test of the inhibitory activity of the compound of the present invention against KIF18A enzyme

[0066] 1.1 Test Consumables

[0067]

[0068] 1.2 Test Instruments

[0069]

[0070] 1.3 Test Methods

[0071] (1) The compound was serially diluted with DMSO in a 384 dilution plate. 0.1 μL of the compound was transferred into a 384 reaction microplate (OptiPlate 384) using Echo, ensuring that the final concentration of DMSO was 1% (double replicate).

[0072] (2) Add 5 μL of enzyme solution to each well of the 384 reaction microplate and centrifuge at 1000 rpm for 1 minute.

[0073] (3) Incubate at 25℃ for 10 minutes.

[0074] (4) Wells containing 1% DMSO and enzyme were designated as High Control, and wells containing 1% DMSO and buffer were designated as Low Control. Add 5 μL of ATP solution to each well and centrifuge at 1000 rpm for 1 minute.

[0075] (5) Incubate at 25℃ for 60 minutes.

[0076] (6) Add 5 μL of ADP-Glo ​​Reagent solution to each well and centrifuge at 1000 rpm for 1 minute.

[0077] (7) Incubate at 25℃ for 40 minutes.

[0078] (8) Add 10 μL of ADP-Glo ​​Detection solution to each well and centrifuge at 1000 rpm for 1 minute.

[0079] (9) Incubate at 25℃ for 40 minutes.

[0080] (10) Read the signal value of the Relative Luminescence Unit on the BMG (PHERAstar FSX) microplate reader. Calculate the inhibition percentage of the wells treated with the compound.

[0081] (12) Fitting four-parameter IC using XLfit 5.5.0 50 Curve analysis, IC 50 This is the compound concentration corresponding to an inhibition rate of 50%.

[0082] Table 1. Results of in vitro inhibitory activity test of the compounds of the present invention against KIF18A enzyme.

[0083]

[0084] The experimental results showed that compounds 1, 2, and 3 all exhibited inhibitory activity against KIF18A, and their inhibitory abilities were significantly better than those of the positive control drug AMG650. Among them, compound 1 showed the strongest inhibitory effect.

Claims

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

3. The use of the compound of claim 1, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 2, in the preparation of a medicament for the prevention and / or treatment of diseases related to KIF18A activity or expression levels.

4. The application according to claim 3, characterized in that, The disease in question is cancer.

5. The application according to claim 4, characterized in that, The cancer is a solid tumor or a hematopoietic tumor.

6. The application according to claim 5, characterized in that, The solid tumors are 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, head and neck cancer, thyroid cancer, brain cancer, skin cancer, melanoma, breast cancer, osteosarcoma, teratoma, seminoma, or Kaposi's sarcoma; the hematogenous tumors are leukemia, B-cell lymphoma, T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, pilocellular lymphoma, Burkert lymphoma, acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome, or promyelocytic leukemia.

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