Pharmaceutical combinations of kif18a inhibitors and tubulin inhibitors and uses thereof

Abstract

The present application provides a pharmaceutical combination comprising a kinesin family member 18A (KIF18A) inhibitor and a tubulin inhibitor, a combination product or a pharmaceutical composition comprising the same, and use of the pharmaceutical combination, the combination product or the pharmaceutical composition for treating a tumor. Formula (K).

Description

Drug combination and use of KIF18A inhibitor and tubulin inhibitor

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of and priority to the following four Chinese invention patent applications, the entire contents of which are hereby incorporated by reference in their entirety:

[0003] Patent application No. 202410288354.4 filed with the State Intellectual Property Office of China on March 13, 2024;

[0004] Patent application No. 202411016438.9 filed with the State Intellectual Property Office of China on July 27, 2024;

[0005] Patent application No. 202411043089.X filed with the State Intellectual Property Office of China on July 31, 2024; and patent application No. 202411138467.2 filed with the State Intellectual Property Office of China on August 19, 2024. Technical Field

[0006] The present disclosure belongs to the field of medicine and relates to a drug combination comprising a kinesin family member 18A (KIF18A) inhibitor and a tubulin inhibitor, a combination product or a pharmaceutical composition comprising the drug combination, and the use of the drug combination, the combination product or the pharmaceutical composition for treating tumors. Background Art

[0007] Cancer is often characterized by unregulated cell proliferation. Damage to one or more genes in the cell proliferation pathway can cause the loss of normal regulation of cell proliferation. These dysregulated genes can encode various tumor suppressors or oncogene proteins, leading to unchecked cell cycle progression and cell proliferation. Various kinases and kinesins have been identified as playing a key role in the cell cycle, mitosis regulation and progression of normal and cancer cells.

[0008] Kinesins are molecular motors that play a vital role in cell division and the transport of intracellular vesicles and organelles. Mitotic kinesins play a role in multiple processes, including spindle assembly, chromosome segregation, and centrosome separation. Human kinesins are classified into 14 subfamilies based on sequence homology within the so-called "motor domain," whose ATPase activity drives unidirectional movement along microtubules. The non-motor domains of these proteins are responsible for attaching to their "cargo," which can be a variety of membranous organelles, signal transduction scaffolds, and chromosomes. Kinesins utilize the energy from ATP hydrolysis to move their "cargo" along polarized microtubules. For this reason, kinesins are often referred to as "plus-end" or "minus-end"-directed motor proteins. The KIF18A gene belongs to the kinesin-8 subfamily and is a plus-end-directed motor protein. KIF18A is believed to influence the movement of the plus ends of centromeric microtubules to control correct chromosome positioning and maintain spindle tension. Ablation of human KIF18A in HeLa cervical cancer cells results in longer spindles, increased chromosome oscillations during metaphase, and activation of the spindle assembly checkpoint (SAC). KIF18A is overexpressed in multiple types of cancer. In cancer cells with chromosomal instability (CIN), KIF18A gene deletion, knockout, or inhibition affects the mitotic spindle. In particular, it has been found that inhibition of KIF18A can activate the SAC, induce mitotic arrest, and ultimately lead to mitotic catastrophe in cancer cells or mitotic slippage during interphase, resulting in apoptosis of cancer cells. Therefore, KIF18A may be a viable target for cancer therapy.

[0009] The spindle assembly checkpoint is a surveillance mechanism that prevents centromeres from exiting mitosis in the presence of unattached or incorrectly attached chromosomes. This prevents the addition or deletion of entire chromosomes and their deleterious effects on cell physiology, and promotes accurate chromosome segregation during mitosis. Microtubule dynamics are crucial for successful mitosis, particularly for the proper function of the spindle during mitosis. Microtubule inhibitors disrupt microtubule dynamics, arresting the cell cycle in the G2 / M phase and ultimately leading to apoptosis. Microtubule inhibitors can be divided into two categories: microtubule stabilizers and microtubule destabilizers. Microtubule stabilizers, such as taxanes and epothilones, act by promoting microtubule polymerization and increasing the mass of microtubule polymers in cells. Microtubule destabilizers, such as vinca alkaloids, reduce the mass of microtubule polymers by depolymerizing microtubules or inhibiting their polymerization. Microtubule inhibitors have been used to treat a variety of solid tumors and hematological malignancies. However, intrinsic or acquired drug resistance limits the use of microtubule inhibitors. Summary of the Invention

[0010] In a first aspect, the present disclosure provides a drug combination comprising at least one KIF18A inhibitor and at least one tubulin inhibitor.

[0011] The present disclosure discloses the above-mentioned drug combination, wherein the tubulin inhibitor is selected from small molecule cytotoxic drugs and antibody-cytotoxic drug conjugates (ADCs). In some embodiments, the payloads of the small molecule cytotoxic drug and the antibody-cytotoxic drug conjugate (ADC) are each independently selected from dolastatins, auristatins, maytansines, tubulysins, and cryptomycins. In some embodiments, the payloads of the small molecule cytotoxic drug and the antibody-cytotoxic drug conjugate (ADC) are each independently selected from auristatins and maytansines. In some embodiments, the payloads of the small molecule cytotoxic drug and the antibody-cytotoxic drug conjugate (ADC) are auristatins. In some embodiments, the small molecule cytotoxic drug is selected from colchicine, vincristine, vinblastine, vindesine, vinblastine, vinorelbine, eribulin, methyl auristatin E (MMAE), methyl auristatin F (MMAF), paclitaxel, docetaxel (docetaxel), maytansine, maytansine DM1, maytansine DM3, and maytansine DM4. In some embodiments, the small molecule cytotoxic drug is selected from vincristine, vinblastine, vinorelbine, eribulin, MMAE, MMAF, and maytansine DM1. In some embodiments, the small molecule cytotoxic drug is selected from eribulin, MMAE, MMAF, and maytansine DM1. In some embodiments, the small molecule cytotoxic drug is selected from MMAE and MMAF. In some embodiments, the payload of the antibody-cytotoxic drug conjugate (ADC) is selected from colchicine, vincristine, vinblastine, vindesine, vinblastine, vinorelbine, eribulin, MMAE, MMAF, paclitaxel, docetaxel, maytansine, maytansine DM1, maytansine DM3 and maytansine DM4. In some embodiments, the payload of the antibody-cytotoxic drug conjugate (ADC) is selected from vincristine, vinblastine, vinorelbine, eribulin, MMAE, MMAF and maytansine DM1. In some embodiments, the payload of the antibody-cytotoxic drug conjugate (ADC) is selected from eribulin, MMAE, MMAF and maytansine DM1. In some embodiments, the payload of the antibody-cytotoxic drug conjugate (ADC) is selected from MMAE and MMAF.

[0012] The present disclosure discloses any of the above-mentioned drug combinations, wherein the KIF18A inhibitor is selected from the compound represented by formula (K) or a pharmaceutically acceptable salt thereof, the compound represented by formula (L) or a pharmaceutically acceptable salt thereof, the compound represented by formula (M) or a pharmaceutically acceptable salt thereof, the compound represented by formula (N) or a pharmaceutically acceptable salt thereof, the compound represented by formula (P) or a pharmaceutically acceptable salt thereof, wherein:

[0013] The formula (K) is selected from the following structures:

[0014] A is a monocyclic or bicyclic ring selected from a 4-10 membered carbocyclic ring or a 4-10 membered heterocyclic ring, wherein the 4-10 membered carbocyclic ring or the 4-10 membered heterocyclic ring is optionally substituted by one or more R b Replace, each R b Independently selected from halogen, OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2, the C1-C6 alkyl is optionally substituted by one or more groups independently selected from halogen, cyano, OH, NH2 or O(C1-C4 alkyl);

[0015] X1, X2, X3, X4 and X5 are each independently CR a or nitrogen, where R a are each independently selected from absent, H or halogen;

[0016] R 1 is absent, H, C1-C6 alkyl, cyano, C1-C6 alkoxy, (CH2) r -C(O)R d 、Halogen、OH、NH2、(CH2) r -C3-C 10 Cycloalkyl, (CH2) r -C5-C 10 Cycloalkenyl, (CH2) r -4-10 membered heterocyclic group, (CH2) r -C6-C 10 Aryl or (CH2) r -5-10 membered heteroaryl, wherein r is 0, 1, 2 or 3, the C1-C6 alkyl, C1-C6 alkoxy, NH2, C3-C 10 Cycloalkyl, C5-C 10 Cycloalkenyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted with one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0017] R 2 is H, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy, C(O)R d , halogen, C1-C6 haloalkyl, C1-C6 haloalkoxy, NH2, C3-C6 cycloalkyl, 4-7 heterocyclyl, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0018] R d is selected from OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0019] Ring E is absent, or is selected from C5-C6 cycloalkenyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl, wherein the C5-C6 cycloalkenyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl is optionally substituted with one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl), or N(C1-C4 alkyl);

[0020] and R 3 Selected from C1-C6 alkyl optionally substituted by one or more halogen or OH;

[0021] The formula (L) is selected from the following structures:

[0022] R 4 、R 5 Each is independently selected from H, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclyl;

[0023] R 6 Selected from C1-C6 alkyl optionally substituted by one or more halogen or OH;

[0024] Ring B is selected from C5-C7 cycloalkyl or 4-7 membered heterocyclic group;

[0025] Each R 7 The groups are independently selected from halogen, oxo, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy, C1-C6 haloalkyl or C1-C6 haloalkoxy, or two adjacent R 7 The group together with the carbon atom to which it is attached forms a C3-C6 cycloalkyl group, or two geminal R 7 The group together with the carbon atom to which it is attached forms a C3-C6 cycloalkyl group;

[0026] m is 0, 1, 2, 3 or 4;

[0027] The formula (M) is selected from the following structures:

[0028] Each R 8 The groups are independently selected from halogen, oxo, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy, C1-C6 haloalkyl or C1-C6 haloalkoxy,

[0029] n is 0, 1, 2, 3 or 4;

[0030] Y1, Y2, Y3 are each independently selected from C, CH or N;

[0031] Indicates a single bond or a double bond;

[0032] Ring C is selected from 5-6 membered heteroaryl, phenyl or 5-6 membered heterocyclyl, wherein the 5-6 membered heteroaryl, phenyl or 5-6 membered heterocyclyl is optionally substituted with one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl);

[0033] R 9 is selected from 4-10 membered heterocyclyl, wherein the 4-10 membered heterocyclyl is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl);

[0034] R 10 Selected from C1-C6 alkyl optionally substituted by one or more halogen or OH;

[0035] The formula (N) is selected from the following structures:

[0036] R 11 Selected from C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group, the C3-C 10 Cycloalkyl or 4-10 membered heterocyclyl is optionally substituted with one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl);

[0037] R 12is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0038] Z1 and Z2 are each independently selected from CR 14 or N, the R 14 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0039] Ring D is absent, or is selected from C5-C6 cycloalkenyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl, wherein the C5-C6 cycloalkenyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl is optionally substituted with one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl), or N(C1-C4 alkyl);

[0040] R 13 Selected from C1-C6 alkyl optionally substituted by one or more halogen or OH;

[0041] The formula (P) is selected from the following structures:

[0042] R 18 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0043] Z4 and Z5 are each independently selected from CR 22 or N, the R 22 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0044] R 19 Selected from H, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl or C1-C6 haloalkyl;

[0045] R 20 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2, or R 19 、R 20 and the atoms to which they are attached together form a 4-7 membered heterocyclyl, which is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl);

[0046] L1 is selected from (L2) a -(L3) b -(L4) c -(L5) d , L2 and L4 are each independently selected from CH2 or L3 and L5 are each independently selected from O, NH or NH(C1-C4 alkyl), a is selected from 0, 1, 2 or 3, c is selected from 1, 2, 3 or 4, b and d are each independently selected from 0 or 1, said (L2) a -(L3) b -(L4) c -(L5) d Optionally substituted with one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl);

[0047] R 21 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0048] R 23 is selected from C1-C6 alkyl groups optionally substituted by one or more halogen or OH.

[0049] In some embodiments, ring E in formula (K) is absent, or ring E is a C5-C6 cycloalkenyl group, which is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0050] In some embodiments, in the formula (K), R 1 is absent, H, C1-C6 alkyl, cyano, C1-C6 alkoxy, (CH2) r -C(O)R d , halogen, OH, (CH2) r -C3-C 10 Cycloalkyl, (CH2) r -C5-C 10 Cycloalkenyl, (CH2) r -4-10 membered heterocyclic group, (CH2) r -C6-C 10 Aryl or (CH2) r -5-10 membered heteroaryl, wherein r is 0, 1, 2 or 3, the C1-C6 alkyl, C1-C6 alkoxy, C3-C 10 Cycloalkyl, C5-C 10 Cycloalkenyl, 4-10 membered heterocyclic group, C6-C 10 The aryl or 5-10 membered heteroaryl is optionally substituted by one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0051] In some embodiments, in the formula (K), R 2 is H, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy, C(O)R d , halogen, C1-C6 haloalkyl or C1-C6 haloalkoxy.

[0052] In some embodiments, the compound represented by formula (K) or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (Ka) or a pharmaceutically acceptable salt thereof:

[0053] A is a monocyclic or bicyclic ring selected from a 4-10 membered carbocyclic ring or a 4-10 membered heterocyclic ring, wherein the 4-10 membered carbocyclic ring or the 4-10 membered heterocyclic ring is optionally substituted by one or more R b Replace, each R bIndependently selected from halogen, OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2, the C1-C6 alkyl is optionally substituted by one or more groups independently selected from halogen, cyano, OH, NH2 or O(C1-C4 alkyl);

[0054] X1, X2, X3, X4 and X5 are each independently CR a or nitrogen, where R a are each independently selected from absent, H or halogen;

[0055] R 1 is absent, H, C1-C6 alkyl, cyano, C1-C6 alkoxy, (CH2) r -C(O)R d , halogen, OH, (CH2) r -C3-C 10 Cycloalkyl, (CH2) r -C5-C 10 Cycloalkenyl, (CH2) r -4-10 membered heterocyclic group, (CH2) r -C6-C 10 Aryl or (CH2) r -5-10 membered heteroaryl, wherein r is 0, 1, 2 or 3, the C1-C6 alkyl, C1-C6 alkoxy, C3-C 10 Cycloalkyl, C5-C 10 Cycloalkenyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted with one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0056] R 2 is H, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy, C(O)R d , halogen, C1-C6 haloalkyl or C1-C6 haloalkoxy;

[0057] R d is selected from OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; and

[0058] R 3 is selected from C1-C6 alkyl groups optionally substituted by one or more halogen or OH.

[0059] In some embodiments, the ring A in formula (K) or formula (Ka) is a monocyclic or bicyclic ring selected from a 4-10 membered carbocyclic ring or a 4-10 membered heterocyclic ring, wherein the 4-10 membered carbocyclic ring or the 4-10 membered heterocyclic ring is optionally substituted by one or more R b Replace, each R b independently selected from F or C1-C4 alkyl, said C1-C4 alkyl being optionally substituted with one or more groups independently selected from F or O(C1-C4 alkyl).

[0060] In some embodiments, Ring A in Formula (K) or Formula (Ka) is a monocyclic or bicyclic ring selected from a 4-10 membered carbocyclic ring or a 4-10 membered heterocyclic ring, wherein the 4-10 membered carbocyclic ring or the 4-10 membered heterocyclic ring is optionally substituted with one or more halogens.

[0061] In some embodiments, X1, X2, X3, X4 and X5 in the formula (K) are each independently CR a or N, where R a Select from does not exist.

[0062] In some embodiments, X1 and X4 in Formula (K) or Formula (Ka) are C, X5 is N, and one of X2 and X3 is C and the other is N.

[0063] In some embodiments, R in Formula (K) or Formula (Ka) 1 is absent, H, OH, C1-C6 alkyl, halogen, cyano, C1-C6 alkoxy or C(O)R d , the C1-C6 alkyl group is optionally substituted by one or more halogens, R d Selected from OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0064] In some embodiments, R in Formula (K) or Formula (Ka) 1 is H or cyano.

[0065] In some embodiments, R in Formula (K) or Formula (Ka) 2 is H, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy or C(O)R d , R d Selected from OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0066] In some embodiments, R in Formula (K) or Formula (Ka) 2 For H.

[0067] In some embodiments, R in Formula (K) or Formula (Ka) 3 is selected from C1-C4 alkyl groups optionally substituted by one or more halogen or OH.

[0068] In some embodiments, R in Formula (K) or Formula (Ka) 3 is selected from C1-C4 alkyl groups optionally substituted by one or more OH groups.

[0069] In some embodiments, R in Formula (K) or Formula (Ka) 3 Selected from

[0070] In some embodiments, the compound represented by formula (K) or formula (Ka) or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (K-1) or formula (Ka-1) or a pharmaceutically acceptable salt thereof:

[0071] wherein the rings A and R in formula (K-1) or formula (Ka-1) 1 、R 2 、R 3 As defined above, one of X2 and X3 is C, and the other is N. Ring E in formula (K-1) is as defined above.

[0072] In some embodiments, the compound represented by formula (K) or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (O) or a pharmaceutically acceptable salt thereof:

[0073] R 15 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0074] Z3 is selected from CR 17 or N, the R 17 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2;

[0075] Ring E is selected from C5-C6 cycloalkenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl, wherein the C5-C6 cycloalkenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl is optionally substituted with one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl);

[0076] Ring F is selected from C5-C6 cycloalkenyl, 4-7 membered heterocyclic group, C8-C 14 Bicyclic or tricyclic cycloalkenyl, 8-14 membered bicyclic or tricyclic heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl, the C5-C6 cycloalkenyl, 4-7 membered heterocyclic group, C8-C 14 Bicyclic or tricyclic cycloalkenyl, 8-14 membered bicyclic or tricyclic heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted with one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl);

[0077] R 16 is selected from C1-C6 alkyl groups optionally substituted by one or more halogen or OH.

[0078] In some embodiments, R in formula (O) 15 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

[0079] In some embodiments, Z3 in the formula (O) is selected from CR 17 or N, the R 17 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

[0080] In some embodiments, ring E in formula (O) is selected from C5-C6 cycloalkenyl, and the C5-C6 cycloalkenyl is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0081] In some embodiments, the ring F in the formula (O) is selected from C5-C6 cycloalkenyl, 4-7 membered heterocyclyl, C8-C10 Bicyclic cycloalkenyl, C 11 -C 14 tricyclic cycloalkenyl, 8-10 membered bicyclic heterocyclyl, 11-14 membered tricyclic heterocyclyl, C6-C 10 Aryl or 5-10 membered heteroaryl, the C5-C6 cycloalkenyl, 4-7 membered heterocyclic group, C8-C 10 Bicyclic cycloalkenyl, C 11 -C 14 tricyclic cycloalkenyl, 8-10 membered bicyclic heterocyclyl, 11-14 membered tricyclic heterocyclyl, C6-C 10 The aryl or 5-10 membered heteroaryl is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0082] In some embodiments, the compound of formula (K) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0083] In some embodiments, R in the formula (L) 4 、R 5 Each is independently selected from H or C1-C6 alkyl.

[0084] In some embodiments, the structural unit in formula (L) Selected from wherein * indicates the point of attachment to the rest of formula (L).

[0085] In some embodiments, R in the formula (L) 6 Selected from C1-C6 alkyl.

[0086] In some embodiments, the compound of formula (L) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0087] In some embodiments, ring C in the formula (M) is selected from a 5-6 membered heteroaryl or a phenyl group, and the 5-6 membered heteroaryl or phenyl group is optionally substituted by one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl)2, and Y1, Y2, and Y3 in the formula (M) are each independently selected from C or N.

[0088] In some embodiments, Y1 in the formula (M) is selected from CH or N, and Y2 and Y3 in the formula (M) are each independently selected from C or N.

[0089] In some embodiments, each R in Formula (M) 8 The groups are independently selected from halogen; preferably, the halogen is F.

[0090] In some embodiments, Ring C in Formula (M) is selected from 5-6 membered heteroaryl, phenyl or 5-6 membered heterocyclyl, and the 5-6 membered heteroaryl, phenyl or 5-6 membered heterocyclyl is optionally substituted with one or more halogens; preferably, the halogen is F.

[0091] In some embodiments, Ring C in Formula (M) is selected from 5-6 membered heteroaryl or phenyl, and the 5-6 membered heteroaryl or phenyl is optionally substituted with one or more halogens; preferably, the halogen is F.

[0092] In some embodiments, R in the formula (M) 9 Selected from

[0093] In some embodiments, R in the formula (M) 10 is selected from C1-C6 alkyl groups optionally substituted with one or more OH groups.

[0094] In some embodiments, the compound of formula (M) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0095] In some embodiments, R in formula (N) 11 Selected from C5-C7 cycloalkyl or 4-7 membered heterocyclyl, wherein the C5-C7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0096] In some embodiments, R in formula (N) 11 Selected from 5-6 membered heterocyclyl, wherein the 5-6 membered heterocyclyl is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0097] In some embodiments, R in formula (N) 11 is selected from 5-6 membered heterocyclyl, wherein the 5-6 membered heterocyclyl is optionally substituted by one or more halogens.

[0098] In some embodiments, R in formula (N) 12 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

[0099] In some embodiments, Z1 and Z2 in the formula (N) are each independently selected from CR 14 or N, the R 14 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

[0100] In some embodiments, ring D in formula (N) is selected from C5-C6 cycloalkenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl, and the C5-C6 cycloalkenyl, 4-7 membered heterocyclyl or 5-6 membered heteroaryl is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0101] In some embodiments, ring D in formula (N) is absent, or ring D is C5-C6 cycloalkenyl, which is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0102] In some embodiments, ring D in formula (N) is C5-C6 cycloalkenyl, which is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0103] In some embodiments, the compound of formula (N) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0104] In some embodiments, R in Formula (P)18 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

[0105] In some embodiments, Z4 and Z5 in the formula (P) are each independently selected from CR 22 or N, the R 22 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

[0106] In some embodiments, R in Formula (P) 19 is selected from H, C1-C6 alkyl or C1-C6 haloalkyl, R in the formula (P) 20 is selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl, or R 19 、R 20 and the atoms to which they are attached together form a 5-6 membered heterocyclic group, which is optionally substituted by one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0107] In some embodiments, L1 in the formula (P) is selected from (CH2) a -O-(CH2) c , a is selected from 1 or 2, c is selected from 1 or 2, the (CH2) a -O-(CH2) c Optionally substituted by one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

[0108] In some embodiments, R in Formula (P) 21 Selected from H, C1-C6 alkyl or C1-C6 haloalkyl.

[0109] In some embodiments, the compound of formula (P) or a pharmaceutically acceptable salt thereof is selected from the following compounds or a pharmaceutically acceptable salt thereof:

[0110] In some embodiments, the KIF18A inhibitor is selected from the following compounds or pharmaceutically acceptable salts thereof:

[0111] The present disclosure provides a drug combination comprising at least one KIF18A inhibitor and at least one tubulin inhibitor, wherein the KIF18A inhibitor is a compound represented by formula (K) or formula (Ka) as defined in any of the above items, or a pharmaceutically acceptable salt thereof.

[0112] The present disclosure provides a drug combination comprising at least one KIF18A inhibitor and at least one tubulin inhibitor, wherein the tubulin inhibitor is any one of the above-defined tubulin inhibitors.

[0113] The present disclosure provides a drug combination comprising at least one KIF18A inhibitor and at least one tubulin inhibitor, wherein the KIF18A inhibitor is a compound represented by formula (K) or formula (Ka) as defined in any of the foregoing items, or a pharmaceutically acceptable salt thereof, and the tubulin inhibitor is a tubulin inhibitor as defined in any of the foregoing items.

[0114] In a second aspect, the present disclosure further provides a pharmaceutical composition comprising any one of the above-mentioned drug combinations and at least one pharmaceutically acceptable excipient.

[0115] In a third aspect, the present disclosure provides a combination product comprising a first pharmaceutical composition and a second pharmaceutical composition, wherein the first pharmaceutical composition comprises at least one of the aforementioned KIF18A inhibitors and a pharmaceutically acceptable excipient, and the second pharmaceutical composition comprises at least one of the aforementioned microtubule inhibitors and a pharmaceutically acceptable excipient.

[0116] In some embodiments, the present disclosure provides a combination product comprising a first pharmaceutical composition and a second pharmaceutical composition, wherein the first pharmaceutical composition comprises at least one compound of the aforementioned formula (K) or formula (Ka) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, and the second pharmaceutical composition comprises at least one aforementioned microtubule inhibitor and a pharmaceutically acceptable excipient.

[0117] In a fourth aspect, the present disclosure relates to the use of any of the aforementioned drug combinations, pharmaceutical compositions or combination products in the preparation of anti-tumor drugs.

[0118] In another aspect, the present disclosure relates to use of any of the aforementioned pharmaceutical combinations, pharmaceutical compositions or combination products in anti-tumor treatment.

[0119] In another aspect, the present disclosure relates to any one of the aforementioned pharmaceutical combinations, pharmaceutical compositions or combination products for anti-tumor use.

[0120] In another aspect, the present disclosure relates to an anti-tumor method, which comprises administering a therapeutically effective amount of any one of the aforementioned pharmaceutical combinations, pharmaceutical compositions, or combined products to a patient in need thereof.

[0121] In another aspect, the present disclosure relates to a use of the aforementioned KIF18A inhibitor in combination with at least one aforementioned tubulin inhibitor in the preparation of an anti-tumor drug.

[0122] On the other hand, the present disclosure relates to the use of the aforementioned compound of formula (K) or formula (Ka) or a pharmaceutically acceptable salt thereof in combination with at least one aforementioned microtubule inhibitor in the preparation of an anti-tumor drug.

[0123] In some embodiments, the tumor is selected from ovarian cancer, non-small cell lung cancer, colon cancer, or cervical cancer.

[0124] In some embodiments, the tumor is selected from chromosomal instability positive (CIN+) and / or chromosomal instability negative (CIN-) tumors.

[0125] In some embodiments of the present disclosure, the compound of formula (K) or formula (Ka) or a pharmaceutically acceptable salt thereof in the use or treatment method can be administered at a frequency of 3 times a day (tid), 2 times a day (bid) or once a day (qd); the daily dosage is 0.01 to 100 mg / kg body weight, preferably 0.05 to 50 mg / kg body weight, and more preferably 0.1 to 30 mg / kg body weight.

[0126] In some embodiments of the present disclosure, in the uses or methods of treatment, the tubulin inhibitor can be administered at a frequency of 3 times a day (tid), 2 times a day (bid), once a day (qd), once a week (q1w), once every 2 weeks (q2w), once every 3 weeks (q3w), or once every 4 weeks (q4w); the daily dosage is 0.01 to 100 mg / kg body weight, preferably 0.05 to 50 mg / kg body weight, and more preferably 0.1 to 30 mg / kg body weight.

[0127] In some embodiments of the present disclosure, the compound represented by Formula (L), Formula (M), Formula (N) or Formula (P) or a pharmaceutically acceptable salt thereof in the use or treatment method can be administered at a frequency of 3 times a day (tid), 2 times a day (bid) or once a day (qd); the daily dosage is 0.01 to 100 mg / kg body weight, preferably 0.05 to 50 mg / kg body weight, and more preferably 0.1 to 30 mg / kg body weight.

[0128] Technical Effects

[0129] Compared with the administration of any drug in the combination alone, the combination of the KIF18A inhibitor of the present disclosure and the tubulin inhibitor produces better therapeutic effects in reducing tumor growth or even eliminating tumors, showing an excellent anti-tumor synergistic effect.

[0130] Definitions and Explanations of Terms

[0131] Unless otherwise indicated, the definitions of groups and terms in this disclosure and claims, including definitions provided as examples, exemplary definitions, preferred definitions, definitions in tables, and definitions of specific compounds in the Examples, may be arbitrarily combined and coupled with one another. The resulting group definitions and compound structures shall fall within the scope of this disclosure.

[0132] In this article Indicates the attachment site.

[0133] The diagrammatic representations of racemates or enantiomerically pure compounds herein are from Maehr, J. Chem. Ed. 1985, 62: 114-120. Unless otherwise indicated, the solid and imaginary bonds are wedge-shaped. To express the absolute configuration of a stereocenter, use direct real bonds and direct virtual bonds. Indicates the relative configuration of a stereocenter (such as the cis-trans configuration of an alicyclic compound).

[0134] The term "tautomer" refers to functional group isomers resulting from the rapid shift of an atom in a molecule between two positions. The compounds of the present disclosure may exhibit tautomerism. Tautomeric compounds may exist as two or more interconvertible species. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer usually result in a mixture whose physical and chemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates, while in phenols, the enol form predominates. The present disclosure encompasses all tautomeric forms of the compounds.

[0135] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0136] The compounds of the present invention may have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms or asymmetric double bonds, so that the compounds of the present invention may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis and trans isomers, E and Z geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures or other mixtures thereof, such as mixtures enriched in enantiomers or diastereomers, all of which are within the definition of the compounds of the present invention and mixtures thereof. Additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms or asymmetric phosphorus atoms may be present in substituents such as alkyl groups, and all of which are within the definition of the compounds of the present invention and mixtures thereof. Compounds of the present disclosure containing an asymmetric atom can be isolated in optically pure or racemic forms. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or reagents.

[0137] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, as long as the valence of the particular atom is normal and the compound after the substitution is stable.

[0138] The term "optional" or "optionally" refers to that the event or situation described subsequently may or may not occur, and the description includes that the event or situation occurs and that the event or situation does not occur. For example, an ethyl group is "optionally" substituted with halogen, meaning that the ethyl group may be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl, etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2, etc.), or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3, etc.). It will be appreciated by those skilled in the art that for any group comprising one or more substituents, any sterically impossible and / or incomposable replacement or substitution pattern will not be introduced.

[0139] When any variable (such as R a 、R b ) appears more than once in the composition or structure of a compound, its definition is independent in each case. For example, if a group is represented by two R b is replaced, then each R b There are independent options.

[0140] The term "alkyl" refers to a group of the formula C n H 2n+1The term "C1-C6 alkyl" may be understood to mean an alkyl group having 1 to 6 carbon atoms, and specific examples include but are not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C4 alkyl" may be understood to mean a straight or branched saturated alkyl group having 1 to 4 carbon atoms. The "C1-C6 alkyl" may include "C1-C4 alkyl".

[0141] The term "alkoxy" refers to a group generated by losing a hydrogen atom from a hydroxyl group of a straight-chain or branched alcohol, and can be understood as "alkyloxy" or "alkyl-O-".

[0142] The term "cycloalkyl" refers to a fully saturated carbocyclic group that exists in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbocyclic ring is typically a 3- to 10-membered ring. The term "C3-C 10 "Cycloalkyl" is understood to mean a saturated monocyclic, bicyclic, spirocyclic or bridged ring group having 3 to 10 carbon atoms. Specific examples of the cycloalkyl group include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decyl, and the like. The term "C3-C 10 "Cycloalkyl" may include "C3-C6 cycloalkyl" and "C5-C7 cycloalkyl". The term "C3-C6 cycloalkyl" may be understood to mean a saturated monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms, specific examples of which include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; the term "C5-C7 cycloalkyl" may be understood to mean a saturated monocyclic or bicyclic hydrocarbon ring having 5 to 7 carbon atoms.

[0143] The term "C5-C 10 "Cycloalkenyl" refers to a non-aromatic carbocyclic group that is not fully saturated and has at least one carbon-carbon double bond and exists in the form of a monocyclic, condensed, bridged or spirocyclic ring, and has 5 to 10 ring carbon atoms. The term "C5-C6 cycloalkenyl" refers to a cycloalkenyl group with 5 or 6 ring carbon atoms. "C8-C 14 "Bicyclic or tricyclic cycloalkenyl" refers to a bicyclic or tricyclic cycloalkenyl group having 8, 9, 10, 11, 12, 13 or 14 ring carbon atoms. "C8-C 14 Bicyclic or tricyclic cycloalkenyl" may contain "C8-C 10 Bicyclic cycloalkenyl" and "C 11 -C 14 "tricyclic cycloalkenyl".

[0144] The term "carbocycle" refers to a cyclic structure composed of carbon atoms, wherein the carbon atoms are linked together by covalent bonds to form a monocyclic, fused, spirocyclic or bridged ring.

[0145] The term "heterocyclyl" refers to a fully saturated or partially saturated (heteroaromatic as a whole that is not aromatic) monocyclic, fused, spiro or bridged ring group, which contains 1 to 5 heteroatoms or heteroatomic groups (i.e., heteroatom-containing groups) in its ring atoms, wherein the "heteroatoms or heteroatomic groups" include, but are not limited to, nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(=O)2-, -S(=O)-, -P(=O)2-, -P(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH- or -NHC(=O)NH-. The term "4-10 membered heterocyclyl" refers to a heterocyclyl group having 4, 5, 6, 7, 8, 9 or 10 ring atoms, and containing 1 to 5 heteroatoms or heteroatomic groups independently selected from the above-mentioned heteroatoms in its ring atoms. The term “4-10 membered heterocyclyl” may include a “4-7 membered heterocyclyl”. The term “4-7 membered heterocyclyl” refers to a heterocyclyl group having 4, 5, 6 or 7 ring atoms, and containing 1, 2, 3, 4 or 5 heteroatoms or heteroatom groups independently selected from the above-mentioned heteroatoms in the ring atoms. The term “8-14 membered bicyclic or tricyclic heterocyclyl” refers to a bicyclic or tricyclic heterocyclyl group having 8, 9, 10, 11, 12, 13 or 14 ring atoms, and containing 1 to 8 heteroatoms or heteroatom groups independently selected from the above-mentioned heteroatoms in the ring atoms. The term “8-14 membered bicyclic or tricyclic heterocyclyl” may include an “8-10 membered bicyclic heterocyclyl” and an “11-14 membered tricyclic heterocyclyl”. In particular, the heterocyclic group may include but is not limited to: specific examples of 4-membered heterocyclic groups include but are not limited to azetidinyl or oxetanyl; specific examples of 5-membered heterocyclic groups include but are not limited to tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl or 2,5-dihydro-1H-pyrrolyl; specific examples of 6-membered heterocyclic groups include but are not limited to tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridinyl or 4H-[1,3,4]thiadiazinyl; specific examples of 7-membered heterocyclic groups include but are not limited to diazepanyl. The heterocyclic group may also be a bicyclic group, wherein specific examples of 5,5-membered bicyclic groups include, but are not limited to, hexahydrocyclopenta[c]pyrrol-2(1H)-yl; specific examples of 5,6-membered bicyclic groups include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl, or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Although some bicyclic heterocyclic groups in the present disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclic group as a whole is still non-aromatic.

[0146] The term "aryl" refers to an all-carbon monocyclic or fused polycyclic aromatic ring group having a conjugated π electron system. The aryl group may have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. The term "C6-C 10 "Aryl" is understood to mean an aromatic group having 6 to 10 carbon atoms. For example, a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 "aryl"), for example tetrahydronaphthyl, dihydronaphthyl or naphthyl.

[0147] The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system with aromatic character, which contains at least one ring atom selected from N, O, S, and the remaining ring atoms are C. The term "5-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems: which have 5, 6, 7, 8, 9 or 10 ring atoms, such as 5 or 6 or 9 or 10 ring atoms, and which contain 1 to 5, such as 1 to 3 heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl, and the like, and benzo derivatives thereof, such as benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl, and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl, and the like, and benzo derivatives thereof, such as quinolyl, quinazolinyl or isoquinolyl, and the like; or acinyl, indolizinyl, purinyl, and the like, and benzo derivatives thereof; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, and the like. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms and containing 1-3, for example 1-2, heteroatoms independently selected from N, O and S.

[0148] When it appears inside a ring, it indicates that the ring is aromatic, for example It refers to the benzene ring.

[0149] The term "halo" or "halogen" refers to fluorine, chlorine, bromine or iodine.

[0150] The term "payload" refers to the therapeutic compound (such as a cytotoxic drug) carried in the ADC. By combining the payload with the antibody, the ADC can achieve more precise treatment and reduce damage to normal cells.

[0151] The term "drug combination" refers to a combination of two or more active ingredients or pharmaceutically acceptable salts thereof. In some embodiments of the present disclosure, the active ingredients or pharmaceutically acceptable salts thereof in the drug combination may be administered simultaneously. In some embodiments of the present disclosure, the active ingredients or pharmaceutically acceptable salts thereof in the drug combination may also be administered separately or sequentially.

[0152] The term "pharmaceutically acceptable salts" refers to salts of pharmaceutically acceptable acids or bases, including salts of inorganic acids and bases, organic acids and bases, such as succinate.

[0153] The term "pharmaceutical composition" refers to a mixture of one or more active ingredients of the present disclosure and pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound of the present disclosure or a pharmaceutical combination thereof to a subject.

[0154] The term "treating" means administering a compound or formulation of the present disclosure to improve or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0155] (i) inhibiting a disease or disease state, i.e., arresting its development;

[0156] (ii) ameliorating the disease or condition, i.e., causing regression of the disease or condition.

[0157] The term "therapeutically effective amount" means an amount of a compound of the present disclosure that (i) treats a particular disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) delays the onset of one or more symptoms of a particular disease, condition, or disorder as described herein. The amount of a compound of the present disclosure that constitutes a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by one skilled in the art based on their own knowledge and this disclosure.

[0158] As used herein, the terms "subject" or "patient" are used interchangeably. In some embodiments, the term "subject" or "patient" is a mammal. In some embodiments, the subject or patient is a mouse. In some embodiments, the subject or patient is a human.

[0159] The term "administering" refers to physically introducing a composition comprising a therapeutic agent into a subject using any of a variety of methods and delivery systems known to those skilled in the art. Routes of administration of KIF18A inhibitors and tubulin inhibitors include, but are not limited to, oral, parenteral, intravenous, transdermal, sublingual, intramuscular, and subcutaneous administration. In certain specific embodiments, the KIF18A inhibitor and tubulin inhibitor are administered orally.

[0160] In the pharmaceutical combination or composition of the present disclosure, the KIF18A inhibitor and the tubulin inhibitor may be in separate or single formulations. When the KIF18A inhibitor and the tubulin inhibitor are in separate formulations, the KIF18A inhibitor and the tubulin inhibitor may be administered simultaneously, separately or sequentially.

[0161] The term "tubulin inhibitors" refers to a class of compounds / drugs that exert therapeutic effects by inhibiting the formation, stability or function of intracellular microtubules. Tubulin inhibitors include both small molecule compounds / drugs and biomacromolecule compounds / drugs.

[0162] The term "pharmaceutically acceptable excipient" refers to an excipient that is non-irritating to organisms and does not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art and include, for example, carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0163] The words “comprise,” “comprise,” or “comprises,” and variations thereof such as comprises or comprising, should be construed in an open, non-exclusive sense, ie, “including but not limited to.”

[0164] In the present disclosure, “A is a monocyclic or bicyclic ring selected from a 4-10 membered carbocyclic ring or a 4-10 membered heterocyclic ring” means that ring A is a monocyclic or bicyclic 4-10 membered carbocyclic ring, or ring A is a monocyclic or bicyclic 4-10 membered heterocyclic ring.

[0165] The compounds disclosed herein can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments described in PCT / CN2023 / 117694, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitution methods well known to those skilled in the art.

[0166] The present application also includes isotopically labeled compounds of the present application that are identical to those described herein, but in which one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present application include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O.18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0167] Certain isotope-labeled compounds of the present application (e.g. 3 H and 14 C-labeled) can be used in compound and / or substrate tissue distribution assays. 3 H) and carbon-14 (i.e. 14 C) isotopes are particularly preferred due to their ease of preparation and detectability. Positron emitting isotopes, such as 15 O. 13 N. 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the present application can generally be prepared by following procedures analogous to those disclosed in the Schemes and / or Examples below, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0168] In addition, the use of heavier isotopes such as deuterium (i.e. 2 H)) substitution may offer certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and may therefore be preferred in certain circumstances, wherein deuterium substitution may be partial or complete, partial deuterium substitution meaning that at least one hydrogen is replaced by at least one deuterium.

[0169] The pharmaceutical compositions of the present application can be prepared by combining the compounds of the present application with suitable pharmaceutically acceptable excipients, and can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres and aerosols, etc.

[0170] Typical routes of administration of the compounds of the present invention, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0171] The pharmaceutical composition of the present application can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, sugar-coated pill making methods, grinding methods, emulsification methods, freeze-drying methods, etc.

[0172] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present application to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, slurries, suspensions, and the like for oral administration to a patient.

[0173] Solid oral compositions can be prepared by conventional mixing, filling, or tableting methods. For example, they can be prepared by mixing the active compound with a solid excipient, optionally grinding the resulting mixture, adding other suitable excipients as needed, and then granulating the mixture to obtain a tablet or dragee core. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants, sweeteners, or flavoring agents.

[0174] The pharmaceutical composition may also be suitable for parenteral administration, such as sterile solutions, suspensions or lyophilized products in appropriate unit dosage forms. BRIEF DESCRIPTION OF THE DRAWINGS

[0175] Figure 1 shows that the combination of compound 1 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H358 cells.

[0176] Figure 2 shows that the combination of compound 1 and vincristine has a synergistic inhibitory effect on OVCAR3 cell proliferation.

[0177] Figure 3 shows that the combination of compound 1 and maytansine DM1 has a synergistic inhibitory effect on Hela cell proliferation.

[0178] Figure 4 shows that the combination of compound 1 and eribulin has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0179] Figure 5 shows that the combination of compound 4 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H358 cells.

[0180] Figure 6 shows that the combination of compound 4 and MMAE has a synergistic inhibitory effect on Hela cell proliferation.

[0181] FIG7 shows that the combination of compound 4 and vincristine has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0182] FIG8 shows that the combination of compound 4 and vinblastine has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0183] FIG9 shows that the combination of compound 4 and vinorelbine has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0184] FIG10 shows that the combination of compound 4 and MMAE has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0185] FIG11 shows that the combination of compound 4 and MMAF has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0186] FIG12 shows that the combination of compound 4 and eribulin has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0187] FIG13 shows that the combination of compound 4 and maytansine DM1 has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0188] FIG14 shows that the combination of compound BTB-1 and vincristine has no synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0189] FIG15 shows that the combination of compound 5 and docetaxel has no synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0190] FIG16 shows that the combination of compound 5 and MMAE has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0191] FIG17 shows that the combination of compound 5 and MMAF has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0192] FIG18 shows that the combination of compound 5 and eribulin has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0193] FIG19 shows that the combination of compound 5 and maytansine DM1 has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0194] FIG20 shows that the combination of BTB-1 and vincristine has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0195] FIG21 shows that the combination of Compound 1 and docetaxel has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0196] FIG22 shows that the combination of Compound 1 and vincristine has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0197] FIG23 shows that the combination of compound 1 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0198] FIG24 shows that the combination of compound 4 and docetaxel has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0199] FIG25 shows that the combination of compound 4 and vincristine has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0200] FIG26 shows that the combination of compound 4 and MMAF has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0201] FIG27 shows that the combination of compound 5 and docetaxel has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0202] FIG28 shows that the combination of compound 5 and vincristine has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0203] FIG29 shows that the combination of compound 5 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0204] FIG30 shows that the combination of compound 5 and MMAF has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0205] FIG31 shows that the combination of compound 6 and docetaxel has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0206] FIG32 shows that the combination of compound 6 and vincristine has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0207] FIG33 shows that the combination of compound 6 and MMAF has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0208] FIG34 shows that the combination of compound 7 and docetaxel has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0209] FIG35 shows that the combination of compound 7 and vincristine has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0210] FIG36 shows that the combination of compound 7 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0211] FIG37 shows that the combination of compound 9 and docetaxel has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0212] FIG38 shows that the combination of compound 9 and vincristine has no synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0213] FIG39 shows that the combination of compound 9 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0214] FIG40 shows that the combination of compound 4 and Ladiratuzumab-MMAE has a synergistic inhibitory effect on the proliferation of OVCAR3 cells.

[0215] FIG41 shows that the combination of compound 4 and Ladiratuzumab-MMAE has a synergistic inhibitory effect on HT29 cell proliferation.

[0216] Figure 42 shows that the combination of Compound 4 and Ladiratuzumab-MMAE has a synergistic effect on the in vivo efficacy in the OVCAR3 model. DETAILED DESCRIPTION

[0217] The invention is described in detail below by way of examples, but this is not intended to limit the present disclosure in any way. While the present disclosure has been described in detail herein, including specific embodiments thereof, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments without departing from the spirit and scope of the present disclosure. All reagents used in the present disclosure are commercially available and can be used without further purification.

[0218] Example 1 Preparation of KIF18A Inhibitor

[0219] The following compounds 1, 2, 3, and 7 were prepared according to the preparation methods described in Examples 64, 110, 86, and 28 of PCT patent application PCT / CN2023 / 117694 (filing date: September 8, 2023), respectively.

[0220] Compound 4 The synthesis of PCT patent application PCT / CN2023 / 117694 (filing date: September 8, 2023) Example 64, and the starting material Replace with Obtain a mixture Compound 4 was obtained by HPLC preparative purification (acetonitrile: water, 20-85%, 0.1% ammonia water).

[0221] 1 H NMR (400MHz, DMSO-d6) δ13.02 (s, 1H), 10.21 (s, 1H), 8.06 (d, J = 8.6Hz, 1H), 7.88 (dd,J=7.6,1.2Hz,1H),7.40–7.30(m,2H),7.25(dd,J=8.9,7.5Hz,1H),7.12(dd, J=8.6,2.1Hz,1H),4.94(s,1H),3.77(t,J=6.5Hz,2H),3.37(t,J=6.5Hz,2H),3. 10–2.96(m,6H),2.92–2.81(m,2H),2.30–2.12(m,4H),1.76(s,4H),0.33(s,4H).

[0222] MS m / z(ESI):574.1[M+H] + .

[0223] Compound 5 It is the disclosed compound AMG650, which was prepared according to Example 4 of patent application WO2020132648A1.

[0224] Compound 6 Prepared with reference to patent application WO2023028564A1.

[0225] Compound 8 and compound 9 They were prepared with reference to Examples 9 and 15 of patent application WO2024067675A1, respectively.

[0226] Compound 10 and compound 11 Prepared with reference to Examples 36 and 81 of patent application WO2024153217A1 respectively.

[0227] Example 2: Preparation of Antibody-Drug Conjugate Ladiratuzumab-MMAE

[0228] 2.1 Construction and production of anti-human LIV-1 monoclonal antibodies

[0229] The sequence of the anti-human LIV-1 monoclonal antibody Ladiratuzumab is shown in Table 1 below (antibody sequence source US20200165335A). The nucleic acid sequences encoding the antibody VH and VL were recombined into the expression vector pTT5 with a signal peptide and heavy chain constant region (sequence) / light chain constant region sequence to obtain a recombinant plasmid expressing Ladiratuzumab. After sequencing verification, the plasmid was extracted. The plasmid and transfection reagent PEI (Polysciences, catalog number: 24765-1) were added to OPTI-MEM (Gibco, catalog number: 11058021) and mixed well. After standing for 15 minutes, the cells were added to Expi293 cells (manufacturer: Thermofisher, catalog number: A14527) and cultured in a shaker at 5% CO2, 120 rpm, and 37°C. On the second day of transfection, OPM-293 ProFeed (Shanghai Aopuma, Catalog No.: F081918-001) and 6 g / L glucose (Manufacturer: Sigma, Catalog No.: G7528) were added. On the sixth day of transfection, the cell supernatant was collected.

[0230] The sequence information of the anti-human LIV-1 antibody Ladiratuzumab is as follows:

[0231] 2.2 Antibody Purification

[0232] Purify the above-mentioned antibodies from cell culture supernatant using Protein A affinity chromatography (GE, Catalog No. 17549802). Wash the Protein A affinity column with 6M guanidine hydrochloride for 3-5 column volumes, followed by 3-5 column volumes with purified water. Equilibrate the column with a buffer system such as 1× PBS (pH 7.4) for 3-5 column volumes. Load the cell supernatant for binding at a low flow rate, controlling the flow rate to achieve a retention time of approximately 1 minute or longer. After binding, wash the column with 1× PBS (pH 7.4) for 3-5 column volumes until UV absorbance returns to baseline. Elute the sample with 0.02M citrate buffer, pH 3.5. Collect the elution peak under UV monitoring. Adjust the pH of the eluted product to 5-6 using 1M Tris-HCl (pH 8.0, Sinopharm, Catalog No. 30188336) for storage. The eluted product can be subjected to solvent exchange using methods well known to those skilled in the art, such as ultrafiltration and concentration using ultrafiltration tubes and solvent exchange into the desired buffer system, or using molecular exclusion chromatography, such as G-25 desalting, to replace the desired buffer system, or using a high-resolution molecular exclusion column such as Superdex 200 to remove aggregates from the eluted product to improve sample purity. After purification, proteins that meet purity requirements are sterile filtered using a 0.22 μm filter (Millipore, Catalog No. SLGVR13SL). After passing SEC-HPLC purity testing, the samples are aliquoted and stored at -80°C until ready for use.

[0233] 2.3 Preparation of Antibody-Drug Conjugates

[0234] 2.3.1 Sources of Drug-Linker Compounds

[0235] The structure of drug-linker 1 is shown below, which was purchased from MedChemExpress (MCE) with the product number HY-15575.

[0236] 2.3.2 Antibody and Drug-Linker Conjugation

[0237] The prepared antibody was dialyzed into 20 mM phosphate buffer, 150 mM NaCl, and 1 mM EDTA (pH 6.5). A 2.4-fold volume of 10 mM tris(2-carboxyethyl)phosphine solution (TCEP, Thermo Scientific #77720) was added to the antibody solution, and the mixture was reduced on a thermostatic metal shaker at 37°C for 3 hours. A 10-fold molar equivalent of the aforementioned drug-linker 1 compound (purchased from MCE, Cat. HY-15575) (dissolved in DMSO to 20 mg / ml) was added to the reaction system, and the reaction mixture was conjugated at 25°C for 12 hours. The reaction product was desalted on a G25 column and exchanged into phosphate-buffered saline (PBS) buffer to remove unreacted free small molecule toxins, yielding the antibody-drug conjugate. The ADC product was analyzed for purity and DAR value using SEC and LC-MS.

[0238] SEC purity analysis: SEC-HPLC method was used to analyze the protein sample to be tested, to characterize the molecular size homogeneity of the recombinant protein, and to measure the purity of the recombinant protein. The HPLC used in this method is an Agilent 1260, and the chromatographic column is a TSKgel G3000SWXL from Tosoh Bioscience, with a mobile phase of 200mM phosphate buffer pH 7.0 containing 10% isopropanol, a detection temperature of 25°C, a flow velocity of 0.5mL / min, a detection wavelength of 280nm, and a target protein loading of 50μg, with an analysis time of 40 minutes. To SEC-HPLC data, manual integration was used to analyze the chromatogram, and protein purity was calculated according to the area normalization method. The main peak was considered to be a monomer, the chromatographic peak before the main peak was called an aggregate, and the chromatographic peak after the main peak was called a fragmentation body.

[0239] DAR value determination: The DAR value of the ADC molecule was measured using ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS). First, the ADC molecule to be tested was treated with PNGase F (NEB#P0705L) to remove the N-glycan modification, and then treated with dithiothreitol (DTT, Sigma#646563) and incubated at 37°C for 1 hour to reduce it to light and heavy chains. Then, it was analyzed using a Thermo Vanquish UHPLC-Q Exactive Plus mass spectrometry system. 2 μg of protein was injected onto a Waters ACQUITY Protein BEH molecular exclusion chromatography column. The mobile phase was an aqueous solution containing 0.1% formic acid, 0.05% TFA, and 25% acetonitrile. The flow rate was 0.2 mL / min and the analysis time was 30 min. The mass spectrometer was a Thermo Q Exactive Plus. The main mass spectrometry parameters were as follows: spray voltage 3.8 kV, capillary heating temperature 300°C, sheath gas flow rate 35 arb, parent ion scan range 800-3000, etc. Finally, the mass spectrometry data were analyzed using the Biopharma Finder software. 4.1. Deconvolution processing is performed using the Respect algorithm to calculate the molecular weight information of the light and heavy chain mass spectrum peaks and the mass spectrum response signals of each component, respectively, and thus calculate the DAR value of the ADC sample to be tested.

[0240] The DAR value and SEC purity of the antibody drug conjugate Ladiratuzumab-MMAE are shown in the following table:

[0241] Test Example 1: KIF18A enzyme activity assay

[0242] Based on ADP-Glo TM Kinase Assay: Detect the amount of ADP generated in the reaction to reflect KIF18A enzyme activity.

[0243] 1. Experimental instruments and materials

[0244] The human KIF18A (1-376aa) protease used in the experiment was expressed by Panchao Biotechnology and stored at -80°C (freeze-thaw no more than 5 times).

[0245] Detection kit (ADP-Glo TMThe Kinase Assay was purchased from Promega under the catalog number V9101 and stored at -30°C. The kit detects enzyme activity by measuring the ADP generated during the enzyme reaction. KIF18A enzyme reacts with ATP to generate ADP, and the remaining ATP in the reaction is consumed by the ADP-Glo ​​reagent. The ADP generated during the KIF18A enzyme reaction is reduced to ATP by the detection reagent. ATP is then converted to ATP in the Ultra-Glo TM Under the action of luciferase, it reacts with luciferin to emit light, and the luminescence signal is positively correlated with the KIF18A enzyme activity.

[0246] Other reagents and consumables required for the experiment are as follows

[0247] 2. Experimental steps

[0248] The compound was diluted 3-fold with a starting concentration of 10 μM, and 10 concentration points were added. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument obtained the gradient dilution sample concentration through different ratios. KIF18A enzyme reaction buffer was prepared: 15 nM Tris, 10 mM MgCl2, 0.01% Pluronic F-68, 1 μM Taxol, and 50 μg / mL Microtubule. KIF18A was mixed with the enzyme reaction buffer and added to a 384-well plate. After incubation at room temperature for 15 minutes, ATP (Km 70 μM) was added to the mixture. After incubation at room temperature for 15 minutes, 10 μL ADP-Glo ​​was added. TM Reagent was mixed with 10 μL of reaction mixture and incubated at room temperature for 40 minutes. Finally, 20 μL of ADP-Glo ​​was added. TM Detection Reagent, incubate at room temperature for 30 minutes. Use Envision plate reader (PerkinElmer, emission wavelength 400-700nm) to measure the chemiluminescence signal in each well. In the experiment, set up 1 column of wells without compound and enzyme (chemiluminescence value is used as [RLU] background ([RLU]background)), 1 column of wells with enzyme but no compound (chemiluminescence value is used as [RLU] enzyme ([RLU]enzyme)), the chemiluminescence value of the drug-added group is [RLU]cpd, and the inhibition rate of the compound on proliferation is calculated according to the following formula: Inhibition rate (%) = ([RLU]enzyme-[RLU]cpd) / ([RLU]enzyme–[RLU]background)×100%, the inhibitory activity IC of the compound on the enzyme 50The values ​​were calculated using a four-parameter logistic model. In the following formula, x represents the logarithm of the compound concentration; F(x) represents the effect size (the rate of enzyme inhibition at that concentration): F(x) = (A + ((BA) / (1 + ((C / x)^D)))). A, B, C, and D are the four parameters. IC was calculated using Xlfit. 50 The values ​​were further calculated as the compound concentration required for 50% enzyme inhibition in the best-fit curve. The KIF18A inhibitory activities of the disclosed compounds are shown in Table 1.

[0249] Table 1: KIF18A inhibitory activity of the disclosed compounds In the above table, “++++” indicates the inhibitory activity of the test compound on the enzyme IC 50 The range is <200nM.

[0250] Test Example 2: Proliferation inhibition test of OVCAR-3 cells by the KIF18A inhibitor disclosed herein

[0251] Cells and materials: Human ovarian cancer cell line OVCAR3 was purchased from ATCC (catalog number HTB-161 TM ), RPMI 1640 medium (Gibco#A1049101), penicillin-streptomycin (Gibco#15140122) and 0.25% Trypsin-EDTA (Gibco#25200056) were purchased from Gibco (USA), bovine insulin (Yisheng#40107ES60) was purchased from Yisheng, 384-well plates (Corning#CLS3765) were purchased from Corning (USA), and Cell-Titer Glo reagent (Promega#G7568) was purchased from Promega (USA).

[0252] Cell culture: OVCAR3 cells were cultured in RPMI 1640 complete medium (RPMI 1640 medium containing 20% ​​fetal bovine serum, 10 μg / mL bovine insulin, and 1% penicillin-streptomycin) at 37°C in a 5% CO2 incubator. Cells in the logarithmic growth phase were used for experiments.

[0253] Cell proliferation activity assay: Cell-Titer Glo reagent was used to detect the inhibitory effect of compounds on the proliferation of OVCAR3 cells. Compounds prepared in DMSO and pure DMSO (control) were added to a 384-well plate using an Echo instrument. Compounds were diluted three-fold starting at 30 μM for a total of 11 concentration points. The volume of compound or DMSO added was 100 nL.

[0254] OVCAR3 cells were digested and resuspended in RPMI 1640 complete medium. The cells were plated in a 384-well plate (1000 cells / 50 μL / well), mixed with the compound, and cultured in a 37°C, 5% CO2 incubator for 3 days. 25 μL of Cell-Titer Glo reagent was added to each well, mixed by vortexing, and incubated for 10 minutes. Cell-Titer Glo readings were measured using a Multimode Plate Reader (PerkinElmer, Model: SIC-PV-018).

[0255] A negative control group (Bottom) was set up. The negative control group consisted of the culture medium wells added with 0.2% DMSO, which was defined as 100% proliferation inhibition. The positive control group (Top) consisted of the OVCAR3 cell wells added with 0.2% DMSO.

[0256] Data Analysis:

[0257] The percentage of proliferation inhibition (% Inhibition) was calculated using the four-parameter formula: Y = Bottom + (Top-Bottom) / (1 + (IC 50 / X)^HillSlope) fitting curve to obtain the compound proliferation inhibition IC 50 .

[0258] Inhibition percentage: %Inhibition=(1-(Signal-Bottom) / (Top-Bottom))×100%.

[0259] Signal: Cell-Titer Glo reading of the wells with added compounds;

[0260] Bottom: Cell-Titer Glo reading of negative control well;

[0261] Top: Cell-Titer Glo readings of positive control wells.

[0262] Experimental results:

[0263] The inhibitory activity of the KIF18A inhibitor disclosed herein on OVCAR3 proliferation is shown in Table 2.

[0264] Table 2: Inhibitory activity of KIF18A inhibitors disclosed herein on OVCAR3 proliferation In the above table, “++++” indicates the IC value of the test compound for the inhibition of OVCAR3 proliferation. 50 The range is <50nM; "+++" indicates the IC value of the test compound for the inhibition of OVCAR3 proliferation. 50 Range 50nM <IC 50<100nM; “++” indicates the IC50 of the test compound against OVCAR3 proliferation 50 Range 100nM <IC 50 <200nM.

[0265] Test Example 3: Synergistic Killing Test of Cancer Cells by Combining KIF18A Inhibitors and Microtubule Inhibitors

[0266] 1. Experimental instruments and materials

[0267] The experimental instruments are as follows

[0268] Cell source: Human ovarian cancer cell line OVCAR3 was purchased from ATCC (catalog number HTB-161 TM ), human non-small cell lung cancer cell line NCI-H358 was purchased from ATCC (catalog number CRL-5807 TM ); human cervical cancer cell line Hela was purchased from Kebai (Cat. No. CBP60232); human non-small cell lung cancer cell line NCI-H292 was purchased from the Cell Bank of the Chinese Academy of Sciences (Cat. No. SCSP-582); compound BTB-1 was purchased from MCE (Cat. No. HY-101770).

[0269] 2. Experimental steps

[0270] Cell Culture: OVCAR3 cells were cultured in RPMI 1640 complete medium (RPMI 1640 medium supplemented with 20% fetal bovine serum, 10 μg / mL bovine insulin, and 1% penicillin-streptomycin); NCI-H358 and NCI-H292 cells were cultured in RPMI 1640 complete medium (RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin); HeLa cells were cultured in DMEM complete medium (RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin). All cells were cultured at 37°C in a 5% CO2 incubator. Cells were used in experiments only when they were in the logarithmic growth phase.

[0271] Compound Dilution: For the NCI-H358 cell line, a KIF18A inhibitor (e.g., compounds 1 and 4 prepared in Example 1) was diluted 3-fold starting at 10 μM, with six dilution points. The compound and pure DMSO (control) were then dispensed into each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument used different ratios to obtain the sample concentrations of the gradient dilution. The compound methyl auristatin E was diluted 2-fold starting at 0.375 nM, with six dilution points. The compound and pure DMSO (control) were then dispensed into each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument used different ratios to obtain the sample concentrations of the gradient dilution.

[0272] For the OVCAR3 cell line, the KIF18A inhibitor was diluted at a starting concentration of 62.5 nM (e.g., compounds 1 and 4 prepared in Example 1) or 125 nM (e.g., compound 5 prepared in Example 1), and was serially diluted 2-fold to 6 concentration points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument obtained the gradient dilution sample concentration by different ratios. The compound BTB-1 was diluted at a starting concentration of 10,000 nM, and was serially diluted 3-fold to 6 concentration points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument obtained the gradient dilution sample concentration by different ratios. The compound methyl auristatin E was diluted 2-fold starting at 0.5 nM or 0.375 nM, with six dilution points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument used different ratios to obtain the sample concentrations of the gradient dilution. The compound methyl auristatin F was diluted 2-fold starting at 111.11 nM or 100 nM, with six dilution points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument used different ratios to obtain the sample concentrations of the gradient dilution. The compound vincristine (sulfate form) was diluted 3-fold at a starting concentration of 333.333 nM for 6 concentration points, or 5.0 nM for 6 concentration points, and the compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL, and the instrument used different ratios to obtain the concentration of the sample with a gradient dilution. The compound vinblastine was diluted 2-fold at a starting concentration of 1.0 nM for 6 concentration points, and the compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL, and the instrument used different ratios to obtain the concentration of the sample with a gradient dilution. Vinorelbine was diluted 2-fold starting at 50.0 nM to 6 concentration points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument obtained gradient dilution sample concentrations using different ratios.The compound eribulin was diluted three-fold starting at 10 nM or 0.2 nM, or diluted two-fold starting at 0.1 nM, over six concentration points. The compound and pure DMSO (control) were then added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument used different ratios to obtain the sample concentrations of the gradient dilution. Maytansine DM1 was diluted three-fold starting at 22 nM, or diluted two-fold starting at 12.5 nM, over six concentration points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument used different ratios to obtain the sample concentrations of the gradient dilution. Docetaxel was diluted 2-fold starting at 2.5 nM, with 6 dilution points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument obtained gradient dilution sample concentrations through different ratios.

[0273] For the Hela cell line, the KIF18A inhibitor (e.g., compounds 1 and 4 prepared in Example 1) was diluted 2-fold at a starting concentration of 62.5 nM, or 370.37 nM at a starting concentration, diluted 3-fold, and diluted to 6 concentration points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument obtained the gradient dilution sample concentration by different ratios. Maytansine DM1 was diluted 3-fold at a starting concentration of 37.037 nM, diluted 6 concentration points, and the compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument obtained the gradient dilution sample concentration by different ratios. The compound methyl auristatin E was diluted 2-fold starting at 0.375 nM, with 6 dilution points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument obtained gradient dilution sample concentrations through different ratios.

[0274] For the NCI-H292 cell line, a KIF18A inhibitor system (e.g., BTB-1, Compound 1, Compounds 4-7, and Compound 9 prepared in Example 1) was diluted 2-fold starting at a final concentration of 2500.0 nM, with six dilution points. The compound and pure DMSO (control) were then dispensed into each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL, and the instrument used different ratios to obtain a gradient dilution of sample concentrations. Vincristine and docetaxel were diluted 2-fold starting at a final concentration of 5.0 nM, with six dilution points. The compound and pure DMSO (control) were then dispensed into each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL, and the instrument used different ratios to obtain a gradient dilution of sample concentrations. The compound methyl auristatin E was diluted 2-fold starting at 0.75 nM for 6 concentration points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument used different ratios to obtain the sample concentrations of the gradient dilution. The compound methyl auristatin F was diluted 2-fold starting at 250.0 nM for 6 concentration points. The compound and pure DMSO (control) were added to each well of a 384-well plate using an Echo instrument. The total volume of the compound and DMSO was 100 nL. The instrument used different ratios to obtain the sample concentrations of the gradient dilution.

[0275] Cell proliferation activity assay: The combined use of a KIF18A inhibitor and a microtubule inhibitor was used to detect the inhibitory effects of KIF18A inhibitors on cancer cells (OVCAR3, Hela, NCI-H358, and NCI-H292). Cancer cells were digested and resuspended in the corresponding complete culture medium. The cells were plated in 384-well plates (1000 cells / 50 μL / well for OVCAR3, Hela, and NCI-H358 cells; 300 cells / 50 μL / well for NCI-H292 cells), mixed with the compounds, and cultured in a 37°C, 5% CO2 incubator for 7 days. 25 μL of Cell-Titer Glo reagent was added to each well, mixed thoroughly, and incubated for 10 minutes. Cell-Titer Glo readings were obtained using Envision.

[0276] The negative control group consisted of the culture medium wells added with 0.4% DMSO, which was defined as 100% proliferation inhibition; the positive control group consisted of the cancer cell (OVCAR3, Hela, NCI-H358 and NCI-H292) wells added with 0.4% DMSO.

[0277] Data Analysis:

[0278] % cell viability = (Cell-Titer Glo reading of compound wells - Cell-Titer Glo reading of negative control wells) / (Cell-Titer Glo reading of positive control wells - Cell-Titer Glo reading of negative control wells) x 100%.

[0279] Combenefit software was used to analyze the cell activity results of the two molecules in combination according to the Bliss model to obtain a Bliss score. A Bliss score greater than 10 indicates significant synergy between the two molecules.

[0280] Experimental results:

[0281] See Figures 1-39 and Table 3. Figure 1 shows that the combination of compound 1 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H358 cells; Figure 2 shows that the combination of compound 1 and vincristine has a synergistic inhibitory effect on the proliferation of OVCAR3 cells; Figure 3 shows that the combination of compound 1 and maytansine DM1 has a synergistic inhibitory effect on the proliferation of Hela cells; Figure 4 shows that the combination of compound 1 and eribulin has a synergistic inhibitory effect on the proliferation of OVCAR3 cells; Figure 5 shows that the combination of compound 4 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H358 cells; Figure 6 shows that the combination of compound 4 and MMAE has a synergistic inhibitory effect on the proliferation of Hela cells; Figure 7 shows that the combination of compound 4 and vincristine has a synergistic inhibitory effect on the proliferation of OVCAR3 cells Figure 8 shows that the combination of compound 4 and vinblastine has a synergistic inhibitory effect on the proliferation of OVCAR3 cells; Figure 9 shows that the combination of compound 4 and vinorelbine has a synergistic inhibitory effect on the proliferation of OVCAR3 cells; Figure 10 shows that the combination of compound 4 and MMAE has a synergistic inhibitory effect on the proliferation of OVCAR3 cells; Figure 11 shows that the combination of compound 4 and MMAF has a synergistic inhibitory effect on the proliferation of OVCAR3 cells; Figure 12 shows that the combination of compound 4 and eribulin has a synergistic inhibitory effect on the proliferation of OVCAR3 cells; Figure 13 shows that the combination of compound 4 and maytansine DM1 has a synergistic inhibitory effect on the proliferation of OVCAR3 cells; Figure 14 shows that the combination of compound BTB-1 and vincristine has a synergistic inhibitory effect on the proliferation of OVCAR3 cells. Figure 15 shows that the combination of compound 5 and docetaxel has no synergistic inhibitory effect on OVCAR3 cell proliferation; Figure 16 shows that the combination of compound 5 and MMAE has a synergistic inhibitory effect on OVCAR3 cell proliferation; Figure 17 shows that the combination of compound 5 and MMAF has a synergistic inhibitory effect on OVCAR3 cell proliferation; Figure 18 shows that the combination of compound 5 and eribulin has a synergistic inhibitory effect on OVCAR3 cell proliferation; Figure 19 shows that the combination of compound 5 and maytansine DM1 has a synergistic inhibitory effect on OVCAR3 cell proliferation; Figure 20 shows that the combination of BTB-1 and vincristine has no synergistic inhibitory effect on NCI-H292 cell proliferation; Figure 21 shows that compound 1 and docetaxel The combination had no synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 22 shows that the combination of compound 1 and vincristine had no synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 23 shows that the combination of compound 1 and MMAE had a synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 24 shows that the combination of compound 4 and docetaxel had no synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 25 shows that the combination of compound 4 and vincristine had no synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 26 shows that the combination of compound 4 and MMAF had a synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 27 shows that the combination of compound 5 and docetaxel had no synergistic inhibitory effect on the proliferation of NCI-H292 cells;Figure 28 shows that the combination of compound 5 and vincristine has no synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 29 shows that the combination of compound 5 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 30 shows that the combination of compound 5 and MMAF has a synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 31 shows that the combination of compound 6 and docetaxel has no synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 32 shows that the combination of compound 6 and vincristine has a synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 33 shows that the combination of compound 6 and MMAF has a synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 34 shows that the combination of compound 7 and docetaxel has no synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 35 shows that the combination of compound 7 and vincristine has a synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 36 shows that the combination of compound 7 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 37 shows that the combination of compound 9 and docetaxel has no synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 38 shows that the combination of compound 9 and vincristine has no synergistic inhibitory effect on the proliferation of NCI-H292 cells; Figure 39 shows that the combination of compound 9 and MMAE has a synergistic inhibitory effect on the proliferation of NCI-H292 cells.

[0282] Table 3: Summary of synergistic killing experiments on cancer cells using KIF18A inhibitors and microtubule inhibitors * A Bliss score greater than 10 indicates significant synergy between the two molecules.

[0283] Figures 1-39 and Table 3 show that: 1) The KIF18A inhibitor provided herein, when used in combination with a variety of tubulin inhibitors (vincristine, vinblastine, vinorelbine, eribulin, MMAE, MMAF, and maytansine DM1), achieved a synergistic anti-proliferative effect; 2) The combination of the KIF18A inhibitor provided herein with auristatin tubulin inhibitors (MMAE, MMAF, etc.) exhibited significant synergistic anti-proliferative effects in both chromosomal instability-positive (CIN+) and chromosomal instability-negative (CIN-) cancer cell models, and this broad-spectrum synergistic effect was unexpected.

[0284] Test Example 4: Synergistic Killing Test of Cancer Cells by Combining KIF18A Inhibitors and Antibody-Drug Conjugates

[0285] 1. Experimental instruments and materials

[0286] The experimental instruments are as follows

[0287] Cell source: Human ovarian cancer cell line OVCAR3 was purchased from ATCC (catalog number HTB-161 TM ), human colorectal cancer cell line HT29 was purchased from ATCC (catalog number HTB-38 TM ).

[0288] 2. Experimental steps

[0289] Cell Culture: OVCAR3 cells were cultured in RPMI 1640 complete medium (RPMI 1640 medium supplemented with 20% fetal bovine serum, 10 μg / mL bovine insulin, and 1% penicillin-streptomycin); HT29 cells were cultured in McCoy's 5a complete medium (McCoy's 5a medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin). All cells were cultured at 37°C in a 5% CO2 incubator. Cells were used in experiments only when they were in the logarithmic growth phase.

[0290] Compound dilution: A KIF18A inhibitor (e.g., compound 4, prepared in Example 1) was diluted 2-fold starting at 62.5 nM, with six dilution points. The compound and pure DMSO (control) were plated into each well of a 384-well plate using an Echo instrument. The total volume of compound and DMSO was 100 nL. The instrument used different ratios to determine the sample concentrations for the serial dilutions. Ladiratuzumab-MMAE, an antibody-drug conjugate, was serially diluted 2-fold starting at the appropriate concentration (6.25 nM for OVCAR3 and 125 nM for HT29) using complete culture medium from cancer cells (OVCAR3 and HT29).

[0291] Cell proliferation activity assay: Cell-Titer Glo reagent was used to detect the inhibitory effect of KIF18A inhibitor and antibody-drug conjugate on cancer cells (OVCAR3 and HT29). Cancer cells were digested and resuspended in the corresponding complete culture medium, added to a 384-well plate (1000 cells / 40μL / well), and mixed with the KIF18A inhibitor. Then, 10μL of the diluted antibody-drug conjugate Ladiratuzumab-MMAE was added to each well, mixed, and cultured in a 37°C, 5% CO2 incubator for 7 days. 25μL of Cell-Titer Glo reagent was added to each well, shaken to mix, incubated for 10 minutes, and the Cell-Titer Glo reading was detected using Envision.

[0292] The negative control group consisted of the culture medium wells added with 0.2% DMSO, which was defined as 100% proliferation inhibition; the positive control group consisted of the cancer cell (OVCAR3 and HT29) wells added with 0.2% DMSO.

[0293] Data Analysis:

[0294] % cell viability = (Cell-Titer Glo reading of compound wells - Cell-Titer Glo reading of negative control wells) / (Cell-Titer Glo reading of positive control wells - Cell-Titer Glo reading of negative control wells) x 100%.

[0295] Combenefit software was used to analyze the cell activity results of the two molecules in combination according to the Bliss model to obtain a Bliss score. A Bliss score greater than 10 indicates significant synergy between the two molecules.

[0296] Experimental results:

[0297] See Figures 40, 41 and Table 4. Figures 40 and 41 demonstrate that the combination of the disclosed KIF18A inhibitor and the antibody-drug conjugate Ladiratuzumab-MMAE achieves a synergistic anti-tumor effect. Figure 40 shows that the combination of Compound 4 and Ladiratuzumab-MMAE has a synergistic inhibitory effect on OVCAR3 cell proliferation; Figure 41 shows that the combination of Compound 4 and Ladiratuzumab-MMAE has a synergistic inhibitory effect on HT29 cell proliferation.

[0298] Table 4: Summary of the synergistic killing test of KIF18A inhibitors and tubulin inhibitors against cancer cells * A Bliss score greater than 10 indicates significant synergy between the two molecules.

[0299] Test Example 5: In vivo efficacy study of the disclosed KIF18A inhibitor combined with an antibody-drug conjugate in the OVCAR3 model

[0300] Experimental reagents:

[0301] OVCAR3 cell source: ATCC, catalog number HTB-161 TM .

[0302] Experimental methods:

[0303] Animal information: BALB / c nude female mice, 5-6 weeks old, weighing approximately 16-20 g, were purchased from Shanghai Lingchang Biotechnology Co., Ltd. The mice were housed in an SPF-grade environment with individual ventilation in each cage. All animals had free access to standard certified commercial laboratory diet and free drinking water.

[0304] Cell Culture: Human ovarian cancer OVCAR3 cells were cultured in RPMI 1640 (cell culture medium) supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin solution in a 37°C, 5% CO2 incubator. Cells were routinely digested and passaged twice weekly using 0.25% trypsin-EDTA. Cells were harvested and counted when cell saturation reached 85%-90% and the required number of cells was reached.

[0305] Cell inoculation: 0.1 mL / (containing 10×10 6 ) OVCAR3 cell suspension was subcutaneously inoculated on the right side of each mouse. On the 20th day after inoculation, the average tumor volume reached 134.5 mm 3 At 4 dpi, randomized stratified groups were assigned based on tumor volume. Four groups (n=6) were administered: vehicle (5% NMP + 15% PEG400 + 80% (20% HP-β-CD), PO, QD), compound 4 (5 mg / kg, PO, QD), ladiratuzumab-MMAE (3 mg / kg, IP, single dose), and ladiratuzumab-MMAE combined with compound 4 (3 mg / kg, IP, single dose + 5 mg / kg, PO, QD).

[0306] Administration: Ladiratuzumab-MMAE was administered at a dose of 3 mg / kg by intraperitoneal injection (IP), a single dose only on the day of grouping; Compound 4 was administered at a dose of 5 mg / kg by PO, once a day (QD) x 21 days.

[0307] Tumor measurements and experimental parameters:

[0308] Tumor diameter was measured twice a week using a vernier caliper. Tumor volume was calculated using the formula: V = 0.5 axb 2 , a and b represent the long diameter and short diameter of the tumor, respectively. The body weight of mice was measured twice a week.

[0309] The tumor inhibition efficacy was evaluated by tumor growth inhibition rate (TGI) (%). TGI (%) = [(1 - (mean tumor volume of a treatment group at the end of drug administration - mean tumor volume of the treatment group at the start of drug administration) / (mean tumor volume of the solvent control group at the end of treatment - mean tumor volume of the solvent control group at the start of treatment)] x 100%.

[0310] The efficacy of compound combinations was evaluated using Kim Jong-kyun's formula. CI = E(A+B) / (EA+EB-EA*EB), where E(A+B) represents the tumor growth inhibition rate (TGI) of compound A and B combined, and EA and EB represent the tumor growth inhibition rates of compound A and B alone. A CI < 0.85 indicates antagonism; a CI between 0.85 and 1.15 indicates simple additive effects; and a CI > 1.15 indicates potentiation.

[0311] Experimental results:

[0312] On day 21 of group-based dosing, once-daily oral administration of 5 mg / kg Compound 4 combined with a single intraperitoneal injection of 3 mg / kg Ladiratuzumab-MMAE (on the day mice were grouped) to tumor-bearing mice showed a significant tumor inhibition effect compared to the vehicle control group (P<0.0001). The efficacy was significantly improved compared to the treatment with Compound 4 (5 mg / kg) or Ladiratuzumab-MMAE (3 mg / kg) alone (P<0.0001). See Table 7 and Figure 42.

[0313] Table 7. Tumor volume and significance analysis of each experimental group of OVCAR3 tumor-bearing mice a Statistical analysis was performed using the two-way anova method with GraphPad Prism 9.0. P < 0.05 was considered to indicate a significant difference (*: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001).

[0314] Experimental conclusion:

[0315] Compared to the vehicle control group, once-daily oral administration of compound 4 at 5 mg / kg, combined with a single intraperitoneal injection of ladiratuzumab-MMAE at 3 mg / kg, significantly reduced tumor volume in OVCAR3 human ovarian cancer model mice compared to the vehicle control group, and also significantly reduced tumor volume compared to the two single-agent groups. The combination index (CI) calculated by Kim Jung-kyun's formula was 1.43, indicating a synergistic effect.

Claims

1. A drug combination comprising at least one KIF18A inhibitor and at least one tubulin inhibitor.

2. The drug combination according to claim 1, wherein the tubulin inhibitor is selected from a small molecule cytotoxic drug and an antibody-cytotoxic drug conjugate (ADC), and the payload of the small molecule cytotoxic drug and the antibody-cytotoxic drug conjugate (ADC) are each independently selected from the group consisting of dolastatins, auristatins, maytansines, tubulysins, and cryptomycins; Preferably, the payload of the small molecule cytotoxic drug and the antibody-cytotoxic drug conjugate (ADC) are each independently selected from vincristine, vinblastine, vinorelbine, eribulin, MMAE, MMAF and maytansine DM1; Preferably, the payload of the small molecule cytotoxic drug and the antibody-cytotoxic drug conjugate (ADC) are each independently selected from eribulin, MMAE, MMAF and maytansine DM1; Preferably, the small molecule cytotoxic drug and the payload of the antibody-cytotoxic drug conjugate (ADC) are each independently selected from MMAE and MMAF.

3. The pharmaceutical combination according to claim 1 or 2, wherein the KIF18A inhibitor is selected from a compound represented by formula (K) or a pharmaceutically acceptable salt thereof, a compound represented by formula (L) or a pharmaceutically acceptable salt thereof, a compound represented by formula (M) or a pharmaceutically acceptable salt thereof, a compound represented by formula (N) or a pharmaceutically acceptable salt thereof, a compound represented by formula (P) or a pharmaceutically acceptable salt thereof, wherein: The formula (K) is selected from the following structures: A is a monocyclic or bicyclic ring selected from a 4-10 membered carbocyclic ring or a 4-10 membered heterocyclic ring, wherein the 4-10 membered carbocyclic ring or the 4-10 membered heterocyclic ring is optionally substituted by one or more R b Replace, each R b Independently selected from halogen, OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2, the C1-C6 alkyl is optionally substituted by one or more groups independently selected from halogen, cyano, OH, NH2 or O(C1-C4 alkyl); X1, X2, X3, X4 and X5 are each independently CR a or nitrogen, where R a are each independently selected from absent, H or halogen; R 1 is absent, H, C1-C6 alkyl, cyano, C1-C6 alkoxy, (CH2) r -C(O)R d 、Halogen、OH、NH2、(CH2) r -C3-C 10 Cycloalkyl, (CH2) r -C5-C 10 Cycloalkenyl, (CH2) r -4-10 membered heterocyclic group, (CH2) r -C6-C 10 Aryl or (CH2) r -5-10 membered heteroaryl, wherein r is 0, 1, 2 or 3, the C1-C6 alkyl, C1-C6 alkoxy, NH2, C3-C 10 Cycloalkyl, C5-C 10 Cycloalkenyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted with one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; R 2 is H, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy, C(O)R d , halogen, C1-C6 haloalkyl, C1-C6 haloalkoxy, NH2, C3-C6 cycloalkyl, 4-7 heterocyclyl, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; R d is selected from OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; Ring E is absent, or is selected from C5-C6 cycloalkenyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl, said C5-C6 cycloalkenyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl being optionally substituted with one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl), or N(C1-C4 alkyl); and R 3 Selected from C1-C6 alkyl optionally substituted by one or more halogen or OH; The formula (L) is selected from the following structures: R 4 、R 5 Each is independently selected from H, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclyl; R 6 Selected from C1-C6 alkyl optionally substituted by one or more halogen or OH; Ring B is selected from C5-C7 cycloalkyl or 4-7 membered heterocyclic group; Each R 7 The groups are independently selected from halogen, oxo, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy, C1-C6 haloalkyl or C1-C6 haloalkoxy, or two adjacent R 7 The group together with the carbon atom to which it is attached forms a C3-C6 cycloalkyl group, or two geminal R 7 The group together with the carbon atom to which it is attached forms a C3-C6 cycloalkyl group; m is 0, 1, 2, 3 or 4; The formula (M) is selected from the following structures: Each R 8 The groups are independently selected from halogen, oxo, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy, C1-C6 haloalkyl or C1-C6 haloalkoxy, n is 0, 1, 2, 3 or 4; Y1, Y2, Y3 are each independently selected from C, CH or N; Indicates a single bond or a double bond; Ring C is selected from 5-6 membered heteroaryl, phenyl or 5-6 membered heterocyclyl, wherein the 5-6 membered heteroaryl, phenyl or 5-6 membered heterocyclyl is optionally substituted with one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl); R 9 is selected from 4-10 membered heterocyclyl, wherein the 4-10 membered heterocyclyl is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl); R 10 Selected from C1-C6 alkyl optionally substituted by one or more halogen or OH; The formula (N) is selected from the following structures: R 11 Selected from C3-C 10 Cycloalkyl or 4-10 membered heterocyclic group, the C3-C 10 Cycloalkyl or 4-10 membered heterocyclyl is optionally substituted with one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl); R 12 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; Z1 and Z2 are each independently selected from CR 14 or N, the R 14 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; Ring D is absent, or is selected from C5-C6 cycloalkenyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl, wherein the C5-C6 cycloalkenyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl is optionally substituted with one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl), or N(C1-C4 alkyl); R 13 Selected from C1-C6 alkyl optionally substituted by one or more halogen or OH; The formula (P) is selected from the following structures: R 18 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; Z4 and Z5 are each independently selected from CR 22 or N, the R 22 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; R 19 Selected from H, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl or C1-C6 haloalkyl; R 20 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2, or R 19 、R 20 and the atoms to which they are attached together form a 4-7 membered heterocyclyl, which is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl); L1 is selected from (L2) a -(L3) b -(L4) c -(L5) d , L2 and L4 are each independently selected from CH2 or L3 and L5 are each independently selected from O, NH or NH(C1-C4 alkyl), a is selected from 0, 1, 2 or 3, c is selected from 1, 2, 3 or 4, b and d are each independently selected from 0 or 1, said (L2) a -(L3) b -(L4) c -(L5) d Optionally substituted with one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl); R 21 is selected from H, halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; R 23 is selected from C1-C6 alkyl groups optionally substituted by one or more halogen or OH.

4. The pharmaceutical combination according to claim 3, wherein the ring A in the formula (K) is a monocyclic or bicyclic ring selected from a 4-10 membered carbocyclic ring or a 4-10 membered heterocyclic ring, wherein the 4-10 membered carbocyclic ring or the 4-10 membered heterocyclic ring is optionally substituted by one or more R b Replace, each R b independently selected from F or C1-C4 alkyl, said C1-C4 alkyl being optionally substituted with one or more groups independently selected from F or O(C1-C4 alkyl); or Ring A in the formula (K) is a monocyclic or bicyclic ring selected from a 4-10 membered carbocyclic ring or a 4-10 membered heterocyclic ring, wherein the 4-10 membered carbocyclic ring or the 4-10 membered heterocyclic ring is optionally substituted by one or more halogens.

5. The pharmaceutical combination according to claim 3 or 4, wherein X1, X2, X3, X4 and X5 in the formula (K) are each independently CR a or N, where R a Select from does not exist; or In the formula (K), X1 and X4 are C, X5 is N, and one of X2 and X3 is C, and the other is N.

6. The pharmaceutical combination according to any one of claims 3 to 5, wherein R in the formula (K) 1 is absent, H, C1-C6 alkyl, cyano, C1-C6 alkoxy, (CH2) r -C(O)R d , halogen, OH, (CH2) r -C3-C 10 Cycloalkyl, (CH2) r -C5-C 10 Cycloalkenyl, (CH2) r -4-10 membered heterocyclic group, (CH2) r -C6-C 10 Aryl or (CH2) r -5-10 membered heteroaryl, wherein r is 0, 1, 2 or 3, the C1-C6 alkyl, C1-C6 alkoxy, C3-C 10 Cycloalkyl, C5-C 10 Cycloalkenyl, 4-10 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally substituted with one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl); or R in the formula (K) 1 is absent, H, OH, C1-C6 alkyl, halogen, cyano, C1-C6 alkoxy or C(O)R d , the C1-C6 alkyl group is optionally substituted by one or more halogens, R d is selected from OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; or R in the formula (K) 1 is H or cyano.

7. The pharmaceutical combination according to any one of claims 3 to 6, wherein R in formula (K) 2 is H, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy, C(O)R d , halogen, C1-C6 haloalkyl or C1-C6 haloalkoxy; or R in the formula (K) 2 is H, OH, C1-C6 alkyl, cyano, C1-C6 alkoxy or C(O)R d , R d is selected from OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; or R in the formula (K) 2 For H.

8. The pharmaceutical combination according to any one of claims 3 to 7, wherein R in formula (K) 3 is selected from C1-C4 alkyl optionally substituted by one or more halogen or OH; or R in the formula (K) 3 is selected from C1-C4 alkyl groups optionally substituted by one or more OH groups.

9. The pharmaceutical combination according to any one of claims 3 to 8, wherein R in formula (K) 3 Selected from 10. The pharmaceutical combination according to any one of claims 3 to 9, wherein Ring E in Formula (K) is absent, or Ring E is a C5-C6 cycloalkenyl group, which is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

11. The pharmaceutical combination according to any one of claims 3 to 10, wherein the compound represented by formula (K) or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (K-1) or a pharmaceutically acceptable salt thereof: Among them, ring E, ring A, R 1 、R 2 、R 3 As defined in any one of claims 3 to 10, one of X2 and X3 is C, and the other is N.

12. The pharmaceutical combination according to claim 3, wherein the compound of formula (K) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

13. The pharmaceutical combination according to claim 3, wherein R in the formula (L) 4 、R 5 Each is independently selected from H or C1-C6 alkyl.

14. The pharmaceutical combination according to claim 3 or 13, wherein the structural unit in the formula (L) Selected from wherein * indicates the point of attachment to the rest of formula (L).

15. The pharmaceutical combination according to any one of claims 3, 13, and 14, wherein R in the formula (L) 6 Selected from C1-C6 alkyl.

16. The pharmaceutical combination according to claim 3, wherein the compound of formula (L) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

17. The pharmaceutical combination according to claim 3, wherein each R in the formula (M) 8 The groups are independently selected from halogen; preferably, the halogen is F.

18. The pharmaceutical combination according to claim 3 or 17, wherein the ring C in the formula (M) is selected from 5-6 membered heteroaryl, phenyl or 5-6 membered heterocyclic group, and the 5-6 membered heteroaryl, phenyl or 5-6 membered heterocyclic group is optionally substituted with one or more halogens; preferably, the halogen is F; or Ring C in the formula (M) is selected from a 5-6 membered heteroaryl or a phenyl group, and the 5-6 membered heteroaryl or phenyl group is optionally substituted by one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C4 alkyl), NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

19. The pharmaceutical combination according to any one of claims 3, 17, and 18, wherein R in the formula (M) 9 Selected from 20. The pharmaceutical combination according to any one of claims 3, 17-19, wherein R in the formula (M) 10 is selected from C1-C6 alkyl groups optionally substituted with one or more OH groups.

21. The pharmaceutical combination according to claim 3, wherein the compound of formula (M) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

22. The pharmaceutical combination according to claim 3, wherein R in the formula (N) 11 C5-C7 cycloalkyl or 4-7 membered heterocyclyl, wherein the C5-C7 cycloalkyl or 4-7 membered heterocyclyl is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl); or R in the formula (N) 11 is selected from 5-6 membered heterocyclyl, which is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl); or R in the formula (N) 11 is selected from 5-6 membered heterocyclyl, wherein the 5-6 membered heterocyclyl is optionally substituted by one or more halogens.

23. The pharmaceutical combination according to claim 3 or 22, wherein R in the formula (N) 12 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

24. The pharmaceutical combination according to any one of claims 3, 22, and 23, wherein Z1 and Z2 in the formula (N) are each independently selected from CR 14 or N, the R 14 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

25. The pharmaceutical combination according to any one of claims 3, 22-24, wherein ring D in the formula (N) does not exist, or ring D is a C5-C6 cycloalkenyl group, which is optionally substituted by one or more groups independently selected from halogen, oxo, CN, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

26. The pharmaceutical combination according to claim 3, wherein the compound of formula (N) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

27. The pharmaceutical combination according to claim 3, wherein R in the formula (P) 18 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

28. The pharmaceutical combination according to claim 3 or 27, wherein Z4 and Z5 in the formula (P) are each independently selected from CR 22 or N, the R 22 Selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl.

29. The pharmaceutical combination according to any one of claims 3, 27, and 28, wherein R in the formula (P) 19 is selected from H, C1-C6 alkyl or C1-C6 haloalkyl, R in the formula (P) 20 is selected from H, halogen, CN, C1-C6 alkyl or C1-C6 haloalkyl, or R 19 、R 20 and the atoms to which they are attached together form a 5-6 membered heterocyclic group, which is optionally substituted by one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

30. The pharmaceutical combination according to any one of claims 3, 27-29, wherein L1 in the formula (P) is selected from (CH2) a -O-(CH2) c , a is selected from 1 or 2, c is selected from 1 or 2, the (CH2) a -O-(CH2) c Optionally substituted by one or more groups independently selected from halogen, CN, OH, NH2, C1-C6 alkyl, O(C1-C6 alkyl), C1-C6 haloalkyl, C1-C6 haloalkoxy, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2.

31. The pharmaceutical combination according to any one of claims 3, 27-30, wherein R in the formula (P) 21 Selected from H, C1-C6 alkyl or C1-C6 haloalkyl.

32. The pharmaceutical combination according to claim 3, wherein the compound of formula (P) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

33. The pharmaceutical combination according to claim 3, wherein the KIF18A inhibitor is selected from the following compounds or pharmaceutically acceptable salts thereof:

34. A combination product comprising a first pharmaceutical composition and a second pharmaceutical composition, wherein the first pharmaceutical composition comprises at least one KIF18A inhibitor according to any one of claims 3 to 33 and a pharmaceutically acceptable excipient, and the second pharmaceutical composition comprises at least one tubulin inhibitor according to claim 2 and a pharmaceutically acceptable excipient.

35. A pharmaceutical composition comprising the pharmaceutical combination according to any one of claims 1 to 33, and at least one pharmaceutically acceptable excipient.

36. Use of the pharmaceutical combination according to any one of claims 1 to 33, the combination product according to claim 34, and the pharmaceutical composition according to claim 35 in the preparation of an anti-tumor drug.

Images