Pharmaceutical combination of KIF18a inhibitor and immune checkpoint inhibitor and use thereof

Abstract

The present invention provides a pharmaceutical combination comprising a kinesin family member 18A (KIF18A) inhibitor and an immune checkpoint inhibitor, a combination product or a pharmaceutical composition comprising the pharmaceutical combination, and use of the pharmaceutical combination, the combination product or the pharmaceutical composition in treating tumors. Formula (K)

Description

Drug combinations and uses of KIF18A inhibitors and immune checkpoint inhibitors

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefits and priority of patent application No. 202410263176.X filed with the State Intellectual Property Office of China on March 7, 2024, the entire contents of which are hereby incorporated by reference into this document in their entirety. Technical Field

[0003] The present disclosure belongs to the field of medicine and relates to a drug combination comprising a kinesin family member 18A (KIF18A) inhibitor and an immune checkpoint inhibitor, a combination product or a pharmaceutical composition comprising the drug combination, and the use of the drug combination, combination product or pharmaceutical composition for treating tumors. Background Art

[0004] 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.

[0005] 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 oscillation during metaphase, and activation of the spindle assembly checkpoint. Therefore, KIF18A may be a viable target for cancer therapy, as KIF18A is overexpressed in multiple types of cancer. Furthermore, in cancer cells, KIF18A gene deletion, knockout, or inhibition affects the mitotic spindle. In particular, KIF18A inhibition has been found to induce mitotic arrest, ultimately leading to mitotic catastrophe or mitotic slippage during interphase in cancer cells, which in turn leads to apoptosis of cancer cells.

[0006] Immunotherapy, represented by immune checkpoint blockade, has achieved significant breakthroughs in many solid tumors, but not all patients benefit. This is because, in addition to PD-1 signaling, other immune checkpoints, immunosuppressive cells or cytokines, and overactive cancer-associated fibroblasts can promote immune tolerance. Therefore, identifying targets that can trigger or enhance tumor immunity remains a key focus. One such target is STING. The cGAS-STING pathway drives the activation of type I interferons and other inflammatory cytokines, recruiting and priming CD8+ T cells that target tumor antigens and mediate tumor cell killing. However, current intratumoral administration of STING agonists has significant limitations, while systemic administration carries the risk of inducing a pathological inflammatory state and may also have off-target effects on other immune cells, such as effector T cells. Therefore, the search for safer combination strategies is urgent. Summary of the Invention

[0007] In a first aspect, the present disclosure provides a drug combination comprising at least one KIF18A inhibitor and at least one immune checkpoint inhibitor, wherein the KIF18A inhibitor is selected from a compound represented by formula (K) or a pharmaceutically acceptable salt thereof:

[0008] in:

[0009] 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);

[0010] 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;

[0011] 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;

[0012] R 2 is H, C1-C6 alkyl, cyano, C1-C6 alkoxy, C(O)R d , halogen, C1-C6 haloalkyl or C1-C6 haloalkoxy;

[0013] R dis selected from OH, C1-C6 alkyl, O(C1-C4 alkyl), NH2, NH(C1-C4 alkyl) or N(C1-C4 alkyl)2; and

[0014] R 3 is selected from C1-C6 alkyl groups optionally substituted by one or more halogen or OH.

[0015] In some embodiments, the ring 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 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).

[0016] In some embodiments, the ring 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 with one or more halogens.

[0017] In some embodiments, the ring A is selected from in Representatives and Rings A common key, and the Optionally substituted with one or more halogens.

[0018] In some embodiments, X1, X2, X3, X4 and X5 are each independently CR a or N, where R a Select from does not exist.

[0019] In some embodiments, X1 and X4 are C, X5 is N, one of X2 and X3 is C, and the other is N.

[0020] In some embodiments, the R 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.

[0021] In some embodiments, the R 1 is H or cyano.

[0022] In some embodiments, the R 2is H, 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.

[0023] In some embodiments, the R 2 For H.

[0024] In some embodiments, the R 3 is selected from C1-C4 alkyl groups optionally substituted by one or more halogen or OH.

[0025] In some embodiments, the R 3 is selected from C1-C4 alkyl groups optionally substituted by one or more OH groups.

[0026] In some embodiments, the R 3 Selected from

[0027] In some embodiments, 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:

[0028] Among them, ring A, R 1 、R 2 As defined above, one of X2 and X3 is C and the other is N.

[0029] 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:

[0030] In a second aspect, the present disclosure further provides a pharmaceutical composition comprising any of the above-mentioned drug combinations and at least one pharmaceutically acceptable excipient.

[0031] 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 compound of the above-mentioned formula (K) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, and the second pharmaceutical composition comprises at least one immune checkpoint inhibitor and a pharmaceutically acceptable excipient.

[0032] 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.

[0033] In another aspect, the present disclosure relates to use of any of the aforementioned drug combinations, pharmaceutical compositions or combination products in anti-tumor treatment.

[0034] In another aspect, the present disclosure relates to any one of the aforementioned pharmaceutical combinations, pharmaceutical compositions or combination products for anti-tumor use.

[0035] In another aspect, the present disclosure relates to an anti-tumor method, which comprises administering a therapeutically effective amount of any of the aforementioned drug combinations, pharmaceutical compositions, or combined products to a patient in need thereof.

[0036] On the other hand, the present disclosure relates to the use of the aforementioned compound of formula (K) or a pharmaceutically acceptable salt thereof in combination with at least one immune checkpoint inhibitor in the preparation of an anti-tumor drug.

[0037] In some embodiments of the present disclosure, the immune checkpoint inhibitor is selected from a PD-1 inhibitor or a PD-L1 inhibitor.

[0038] In some embodiments of the present disclosure, the PD-1 inhibitor or PD-L1 inhibitor is sintilimab, toripalimab, carrelizumab, tislelizumab, penampalimab, slulizumab, cardunilizumab, pembrolizumab, nivolumab, ipalolizumab, QL1706 or IBI363.

[0039] In some embodiments of the present disclosure, the tumor is ovarian cancer, breast cancer, or rectal cancer.

[0040] In some embodiments of the present disclosure, the compound of formula (K) 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.

[0041] In some embodiments of the present disclosure, the immune checkpoint inhibitors in the uses or methods of treatment 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.

[0042] Technical Effects

[0043] Compared with the administration of any drug in the combination alone, the combination of the KIF18A inhibitor disclosed herein and the immune checkpoint inhibitor produces better therapeutic effects in reducing tumor growth or even eliminating tumors, showing excellent anti-tumor synergistic effects.

[0044] Definitions and Explanations of Terms

[0045] 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.

[0046] In this article Indicates the attachment site.

[0047] 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).

[0048] 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.

[0049] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0050] 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.

[0051] 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.

[0052] 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.

[0053] 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.

[0054] The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.

[0055] The term "alkyl" refers to a group of the formula C n H2n+1 The 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".

[0056] 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-".

[0057] 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". 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 include but are not limited to cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0058] The term "C5-C 10 The term "cycloalkenyl" refers to an incompletely saturated non-aromatic carbocyclic group having at least one carbon-carbon double bond and existing in the form of a monocyclic, fused, bridged or spirocyclic ring, and having 5 to 10 ring carbon atoms.

[0059] 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.

[0060] 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. 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.

[0061] 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.

[0062] 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.

[0063] In this paper, the structural unit in Indicates that the ring in which it is located is an aromatic ring.

[0064] The term "halo" or "halogen" refers to fluorine, chlorine, bromine or iodine.

[0065] 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.

[0066] 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.

[0067] 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.

[0068] 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:

[0069] (i) inhibiting a disease or disease state, i.e., arresting its development;

[0070] (ii) ameliorating the disease or condition, i.e., causing regression of the disease or condition.

[0071] 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.

[0072] 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.

[0073] The term "administering" means 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 immune checkpoint inhibitors include, but are not limited to, oral, parenteral, intravenous, transdermal, sublingual, intramuscular, and subcutaneous administration. In some specific embodiments, KIF18A inhibitors and immune checkpoint inhibitors are administered orally.

[0074] In the pharmaceutical combination or pharmaceutical composition of the present disclosure, the KIF18A inhibitor and the immune checkpoint inhibitor can be in separate or single formulations. When the KIF18A inhibitor and the immune checkpoint inhibitor are in separate formulations, the KIF18A inhibitor and the immune checkpoint inhibitor can be administered simultaneously, separately or sequentially.

[0075] The term "immune checkpoint" refers to a number of inhibitory signaling pathways present in the immune system. Under normal circumstances, immune checkpoints maintain immune tolerance by regulating the intensity of the body's own immune response. However, when the body is invaded by tumors, activation of immune checkpoints suppresses autoimmunity, favoring the growth and escape of tumor cells. By using immune checkpoint inhibitors, the body's normal anti-tumor immune response can be restored, thereby controlling and eliminating tumors. Immune checkpoints include but are not limited to programmed death receptor 1 (PD1), PD-L1, and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4); they also include some newly discovered immune checkpoints such as lymphocyte activation gene 3 (LAG3), CD160, T cell immunoglobulin and mucin-3 (TIM-3), V-domain immunoglobulin inhibitor of T cell activation (VISTA), and adenosine A2a receptor (A2aR).

[0076] 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.

[0077] 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.”

[0078] The term "ring A is a monocyclic or bicyclic 4-10 membered carbocyclic ring or a 4-10 membered heterocyclic ring" used in the present disclosure 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.

[0079] 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.

[0080] 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.

[0081] 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.

[0082] 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.

[0083] 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.

[0084] 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.

[0085] 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.

[0086] 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.

[0087] 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.

[0088] 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

[0089] Figure 1 shows that KIF18A inhibitor upregulates the expression level of 2',3'-cGAMP in CT26 cell line.

[0090] Figure 2 shows that KIF18A inhibitor upregulates the phosphorylation level of STING.

[0091] Figure 3 shows that KIF18A inhibitor upregulates the expression of interferon-stimulated genes (ISGs). DETAILED DESCRIPTION

[0092] 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 used without further purification.

[0093] Example 1 Preparation of KIF18A Inhibitor

[0094] The following compounds 1, 2, and 3 were prepared according to the preparation methods described in Examples 64, 110, and 86 of PCT patent application PCT / CN2023 / 117694 (filing date: September 8, 2023, the contents of which are incorporated herein by reference in their entirety).

[0095] Compound 4 was prepared according to the preparation methods described in Examples 9 and 10 of PCT patent application PCT / CN2025 / 074827 (filing date: January 24, 2025, the contents of which are incorporated herein by reference in their entirety).

[0096] Test Example 1: KIF18A enzyme activity assay

[0097] Based on ADP-Glo TM Kinase Assay: Detect the amount of ADP generated in the reaction to reflect KIF18A enzyme activity.

[0098] 1. Experimental instruments and materials

[0099] 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).

[0100] Detection kit (ADP-Glo TM The 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.

[0101] Other reagents and consumables required for the experiment are as follows:

[0102] 2. Experimental steps

[0103] 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 50 The values ​​were calculated using the four-parameter logistic model method. In the following formula, x represents the logarithm of the compound concentration; F(x) represents the effect value (the inhibition rate of the enzyme under the conditions of this concentration): F(x) = (A + ((BA) / (1 + ((C / x)^D)))), where A, B, C, and D are four parameters. IC was calculated using Xlfit. 50 The values ​​were further calculated as the compound concentration required for 50% inhibition of enzyme activity in the best fitting curve. The KIF18A inhibitory activities of the disclosed compounds are shown in Table 1.

[0104] Table 1: KIF18A inhibitory activity of the disclosed compounds

[0105] In the above table, “++++” indicates the inhibitory activity of the test compound on the enzyme IC 50 The range is <200nM.

[0106] Test Example 2: Proliferation inhibition test of OVCAR-3 cells by the KIF18A inhibitor disclosed herein

[0107] 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).

[0108] 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.

[0109] 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.

[0110] 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).

[0111] A negative control group (Bottom) was set up. The negative control group consisted of wells containing culture medium added with 0.2% DMSO, which was defined as 100% proliferation inhibition. The positive control group (Top) consisted of wells containing OVCAR3 cells added with 0.2% DMSO.

[0112] Data Analysis:

[0113] 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 .

[0114] Inhibition percentage: %Inhibition=(1-(Signal-Bottom) / (Top-Bottom))×100%.

[0115] Signal: Cell-Titer Glo reading of the wells with added compounds;

[0116] Bottom: Cell-Titer Glo reading of negative control well;

[0117] Top: Cell-Titer Glo readings of positive control wells.

[0118] Experimental results:

[0119] The inhibitory activity of the KIF18A inhibitor disclosed herein on OVCAR3 proliferation is shown in Table 2.

[0120] Table 2: Inhibitory activity of KIF18A inhibitors disclosed herein on OVCAR3 proliferation

[0121] Test Example 3: The KIF18A inhibitor disclosed herein upregulates the level of 2',3'-cGAMP in CT26 cell lines

[0122] Experimental cells and materials:

[0123] CT26 cell line: ATCC CRL-2638; culture conditions: RPMI1640 medium (company: Gibco, catalog number: A10491-01) supplemented with 10% fetal bovine serum (company: Gibco, catalog number: 10091148) and 1% penicillin-streptomycin solution (company: Gibco, catalog number: 15140122), cultured at 37°C, 5% CO2 in a cell culture incubator.

[0124] cGAMP Assay Kit: 2',3'-Cyclic GAMP Enzyme Immunoassay Kit(Cat.No K067-H1,Arbor Assays TM ).

[0125] Preparation of protease inhibitors (50×): cOmplete without EDTA TMTake out one piece of protease inhibitor cocktail (company: Sigma; product number: 04693132001), dissolve it in 1 mL of RIPA lysis buffer, and then divide it into aliquots and store it at -20°C.

[0126] Preparation of phosphatase inhibitor (10ⅹ): PhosSTOP TM (Cat. No.: 4906845001, Company: Sigma) Take out one piece, dissolve it in 1 mL RIPA lysis buffer, divide it into several pieces, and store it at -20℃.

[0127] EDTA solution: 0.5 M, pH 8.0. Company: Invitrogen; Catalog Number: 15575020.

[0128] RIPA lysis buffer: RIPA lysis buffer (Cat. No. 89900, Thermo), add a mixture of protease inhibitors (50ⅹ) and phosphatase inhibitors (10ⅹ) (final concentration is 1ⅹ) and EDTA (final concentration 1 mM) before use, and use it freshly prepared.

[0129] Experimental methods:

[0130] 4×10 6 CT26 cells were seeded in 15 cm cell culture dishes, with 19 mL per dish, and cultured overnight in a 37°C, 5% CO2 incubator. Compound powder was dissolved in DMSO to a 10 mM stock solution and diluted to 20 μM and 200 μM in cell culture medium. 1 mL of cell culture medium containing the compound was added to each dish and incubated in a 37°C, 5% CO2 incubator for 24 h. The culture medium was aspirated, and the cells were digested with trypsin, neutralized with culture medium, and harvested. The cells were then washed twice with ice-cold PBS. Cell counts were performed using Countstar. Cell samples were lysed with RIPA lysis buffer, incubated on ice for 10 min, centrifuged at 4°C, 12,000 rpm, and the supernatant was collected. cGAMP content in the cell lysate was determined using a 2',3'-cGAMP enzyme-linked immunosorbent assay (ELISA) kit, and sample absorbance was measured at OD 450 nm using Envision Biosystems.

[0131] Experimental results: As shown in Figure 1 (1 (1 μM) refers to 1 μM of compound 1 in the cell culture medium, and the same applies to the others), compounds 1, 2, and 3 upregulated the 2',3'-cGAMP content in CT26 cells by approximately 1.5 to 2.0 times at 1 μM. The upregulation of cGAMP content in the cells suggests that the KIF18A inhibitors disclosed herein have the potential to activate the STING pathway.

[0132] Test Example 4: KIF18A inhibitors disclosed herein upregulate p-STING levels in CT26 cell lines

[0133] Experimental cells and materials:

[0134] CT26 cell line: ATCC CRL-2638; culture conditions: RPMI1640 medium (company: Gibco, catalog number: A10491-01) supplemented with 10% fetal bovine serum (company: Gibco, catalog number: 10091148) and 1% penicillin-streptomycin solution (company: Gibco, catalog number: 15140122), cultured at 37°C, 5% CO2 in a cell culture incubator.

[0135] Protease and phosphatase inhibitor cocktail: Company: Beyotime; Product No.: P1046. Contains one bottle of protease inhibitor cocktail (universal type, 50ⅹ, 10mL), 100mM 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride (AEBSF), 15μM aprotinin, 6.5mM bestatin, 0.7mM N-(trans-epoxysuccinyl)-L-leucine-4-guanidinobutylamide (E64) and 0.5mM leupeptin; the solvent is DMSO, and one bottle of phosphatase inhibitor cocktail (50ⅹ, 10mL) (250mM sodium fluoride, 50mM sodium pyrophosphate, 50mM β-glycerophosphate disodium salt pentahydrate and 50mM sodium vanadate). orthovanadate); the solvent is pure water), and 0.05 M ethylenediaminetetraacetic acid (EDTA) (10 mL).

[0136] Loading buffer: Company: CST; Product number: 7722.

[0137] PageRuler TM Prestained protein molecular weight marker: Company: Thermo Scientific; Catalog No.: 26616.

[0138] 4%-20% TGX Stain-Free TM Protein gel: Company: BIO-RAD; Product number: 4568096.

[0139] Trans-Blot Turbo transfer kit: Company: BIO-RAD; Product number: 1704156.

[0140] 10ⅹTris / Glycine / SDS buffer: Company: BIO-RAD; Product number: 1610732.

[0141] EZ-buffer H10×TBST buffer: Company: Sangon; Catalog Number: C520009-0001.

[0142] Bovine serum albumin (BSA): Company: Sigma; Catalog Number: V900933.

[0143] SuperSignal TM West Pico Plus Chemiluminescent Substrate: Company: Thermo Scientific; Product Number: 34577.

[0144] Azure WB Imaging System

[0145] Table 3 List of antibodies used in the test case

[0146] Experimental methods:

[0147] 1.5×10 5 CT26 cells were seeded in 24-well plates and cultured in a 37°C, 5% CO2 incubator overnight. Compound powder was dissolved in DMSO to a 10mM stock solution and diluted to 1mM with cell culture medium. 10μL of cell culture medium containing the compound (final concentration of 10μM) was added to each culture dish and cultured in a 37°C, 5% CO2 incubator for 24h. The culture medium was discarded, and the cells were lysed on ice for 10min with 1× loading buffer (100μL / well) supplemented with a mixture of protease and phosphatase inhibitors (final concentration of 1ⅹ, EDTA concentration of 1mM); the cells were transferred to a 1.5mL EP tube and boiled at 100°C for 10min. 4%-20% TGX Stain-Free TM The protein gel was electrophoresed and the membrane was transferred semi-dry. After the transfer was completed, the membrane was blocked with 1×TBST containing 5% BSA at room temperature for 1 hour. Anti-mouse STING (D1V5L) monoclonal antibody, anti-mouse phosphorylated-STING (Ser365) (D8F4W) monoclonal antibody and anti-mouse β-Actin monoclonal antibody (diluted according to the concentration in the instructions) were incubated overnight at 4°C. The membrane was washed 3 times with 1×TBST, 10 minutes each time. Anti-rabbit IgG (H+L)-horseradish peroxidase conjugate was diluted 1:3000 and the membrane was incubated at room temperature for 1 hour. The membrane was washed 3 times with 1×TBST, 10 minutes each time. SuperSignal TM West Pico Plus Chemiluminescent Substrate Solution A and Solution B were mixed in a 1:1 ratio, incubated with the membrane for 1 minute, and then exposed using the Azure WB imaging system.

[0148] Experimental results: As shown in FIG2 , compounds 1, 2, and 3 significantly activated the phosphorylation of STING, suggesting that the KIF18A inhibitor disclosed herein has the potential to activate the STING pathway.

[0149] Test Example 5: KIF18A inhibitors disclosed herein upregulate the expression of interferon-stimulated genes (ISGs) in CT26 cell lines

[0150] Experimental cells and materials:

[0151] CT26 cell line: ATCC CRL-2638; culture conditions: RPMI1640 medium (company: Gibco, catalog number: A10491-01) supplemented with 10% fetal bovine serum (company: Gibco, catalog number: 10091148) and 1% penicillin-streptomycin solution (company: Gibco, catalog number: 15140122), cultured at 37°C, 5% CO2 in a cell culture incubator.

[0152] QIAwave RNA Kit: Qiagen, Cat. No. 74536.

[0153] PrimeScript TM II 1st Strand cDNA Synthesis Kit: TAKARA, Cat. No.: 6210A.

[0154] TB Fast qPCR Mix: TAKARA, Catalog Number: RR430A.

[0155] Nanodrop8000 Ultra-Micro Spectrophotometer: Company: Thermo Scientific

[0156] Table 4 List of qPCR primers used in Test Example 5

[0157] Experimental methods:

[0158] 2×10 5 CT26 cells were seeded in 12-well plates and cultured overnight in a 37°C, 5% CO2 incubator. Cell culture medium containing the compound (final concentration of 10 μM) was used to continue the culture for 24 hours. RNA samples were extracted using the QIAwave RNA kit and RNA concentration was measured using the Nanodrop 8000 ultra-micro spectrophotometer. PrimeScript was used. TM II cDNA synthesis kit was used to reverse transcribe the sample RNA, and TB Green kit was used for PCR reaction.

[0159] Experimental results: As shown in Figure 3, compounds 1, 2 and 3 significantly upregulated the expression of IFNβ, TNFα, CCL5 and CXCL10 in CT26 cells.

[0160] The results of Test Examples 3-5 showed that the KIF18A inhibitor disclosed herein can activate the cGAS-TING pathway of CT26 cells, revealing the great potential of the KIF18A inhibitor disclosed herein in combination with immune checkpoint inhibitors in the treatment of tumors.

[0161] Test Example 6: In vivo efficacy experiment in CT26 mice

[0162] Experimental reagents:

[0163] CT26 cells: ATCC CRL-2638

[0164] Experimental methods:

[0165] Animal information: BALB / c female mice, 5-6 weeks old, weighing approximately 16-20 g, were purchased from Beijing Weitonglihua 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.

[0166] Anti-mouse PD-1 antibody: purchased from Bioxcell, clone number: RMP1-14, product number: BE0146.

[0167] Cell Culture: Mouse colorectal cancer CT-26 cells were cultured in RPMI 1640 medium (Gibco, Catalog No. A10491-01) supplemented with 10% fetal bovine serum (Gibco, Catalog No. 10091148) and 1% penicillin-streptomycin solution (Gibco, Catalog No. 15140122) at 37°C in a 5% CO2 incubator. Cells were routinely digested and passaged twice weekly using 0.25% trypsin-EDTA solution (Gibco, Catalog No. 25200-072). Cells were harvested and counted when cell saturation reached 85%-90% and the required number of cells was reached.

[0168] Cell inoculation: 0.1 mL (containing 1×10 6 )CT26 cell suspension was subcutaneously inoculated on the right side of each mouse. On the 8th day after inoculation, the average tumor volume was measured to be 50-80 mm 3At 4 hr, the patients were randomly divided into stratified groups according to tumor volume and the drug was administered. There were 8 mice in each group, for a total of 6 groups: vehicle (aqueous solution containing 0.5% methylcellulose and 0.25% Tween-80, PO, QD), anti-mouse PD-1 antibody (10 mg / kg, IP, Q2D), compound 1 (30 mg / kg, PO, QD), compound 1 (100 mg / kg, PO, QD), anti-mouse PD-1 antibody combined with compound 1 (10 mg / kg + 30 mg / kg, IP + PO, Q2D + QD), and anti-mouse PD-1 antibody combined with compound 1 (10 mg / kg + 100 mg / kg, IP + PO, Q2D + QD).

[0169] Dosing: The anti-mouse PD-1 antibody was administered at a dose of 10 mg / kg via intraperitoneal injection (IP) every two days (Q2D) for 21 days; Compound 1 was administered at a dose of 30 mg / kg or 100 mg / kg PO once daily (QD) for 21 days. Each group consisted of 8 mice.

[0170] Tumor measurements and experimental parameters:

[0171] 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.

[0172] 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)] × 100%.

[0173] Experimental results: Compared with the single-drug group, compound 1 combined with PD-1 antibody achieved a synergistic inhibitory effect in the mouse tumor model.

Claims

1. A drug combination comprising at least one KIF18A inhibitor and at least one immune checkpoint inhibitor, wherein the KIF18A inhibitor is selected from a compound represented by formula (K) or a pharmaceutically acceptable salt thereof: in: 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, (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; R 2 is H, C1-C6 alkyl, cyano, C1-C6 alkoxy, C(O)R d , halogen, C1-C6 haloalkyl or C1-C6 haloalkoxy; 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 R 3 is selected from C1-C6 alkyl groups optionally substituted by one or more halogen or OH.

2. The pharmaceutical combination according to claim 1, 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.

3. The pharmaceutical combination according to claim 1, 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.

4. The pharmaceutical combination according to claim 1, wherein 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.

5. The pharmaceutical combination according to claim 1, wherein R in the formula (K) 2 is H, 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.

6. The pharmaceutical combination according to claim 1, 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 optionally substituted with one or more OH groups; or R in the formula (K) 3 Selected from 7. The pharmaceutical combination according to any one of claims 1 to 6, 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: Wherein one of X2 and X3 is C and the other is N, and the ring A, R 1 、R 2 As defined in any one of claims 1 to 6.

8. The pharmaceutical combination according to claim 1, wherein the compound of formula (K) or a pharmaceutically acceptable salt thereof is selected from the following compounds or pharmaceutically acceptable salts thereof:

9. A combination product comprising a first pharmaceutical composition and a second pharmaceutical composition, wherein the first pharmaceutical composition comprises at least one compound of formula (K) according to any one of claims 1 to 8 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient, and the second pharmaceutical composition comprises at least one immune checkpoint inhibitor and a pharmaceutically acceptable excipient.

10. The pharmaceutical combination according to any one of claims 1 to 8 or the combination product according to claim 9, wherein the immune checkpoint inhibitor is selected from a PD-1 inhibitor or a PD-L1 inhibitor.

11. A pharmaceutical composition comprising the pharmaceutical combination according to any one of claims 1 to 8, and at least one pharmaceutically acceptable excipient.

12. Use of the pharmaceutical combination according to any one of claims 1 to 8, the combination product according to claim 9 or the pharmaceutical composition according to claim 11 in the preparation of an anti-tumor drug.

Images