CN116903589A - KIF18A inhibitors and their uses - Google Patents

Abstract

The invention provides a compound represented by formula I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs; the compound has good KIF18A inhibitory effect,

Description

KIF18A inhibitors and uses

Technical Field

The present invention belongs to the field of medicine, and specifically, the present invention relates to a KIF18A inhibitor and its use.

Background Art

KIF18A is a mitotic kinesin that regulates chromosome positioning during cell division and is overexpressed in a portion of human cancers. TP53 mutation-unstable aneuploid cancer cells with chromosomal instability (CIN) characteristics rely on KIF18A motor activity to prevent lethal multipolar cell division. The KIF18A gene belongs to the kinesin-8 subfamily and is a plus-end-directed motor. KIF18A is thought to affect the dynamics of the plus end of the centromere microtubule to control correct chromosome positioning and spindle tension. The depletion of human KIF18A leads to longer spindles in HeLa cervical cancer cells, increased chromosome oscillation in metaphase, and activation of the mitotic spindle assembly checkpoint (MI Mayret al, Current Biology [Contemporary Biology] 17, 488-98, 2007). KIF18A is overexpressed in many types of cancers, including but not limited to colon cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head cancer, neck cancer, cervical cancer, and ovarian cancer. Furthermore, in cancer cell lines, genetic deletion or knockout or inhibition of KIF18A affects the mitotic spindle apparatus. In particular, inhibition of KIF18A has been found to induce mitotic cell arrest, which can promote mitotic cell death by apoptosis, mitotic catastrophe, or multiphase-driven lethality or death after mitotic slippage in interphase.

Currently, there are no drugs on the market that inhibit the KIF18A pathway to treat many diseases, including cancer. Therefore, the development of new compounds that can inhibit the activity of KIF18A has positive significance for the treatment of diseases.

Summary of the invention

The object of the present invention is to provide a novel compound for use as a KIF18A inhibitor.

In the first aspect of the present invention, there is provided a compound of formula I, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof:

in,

X ⁴ is N or -CR ⁸ ;

X ⁵ is N or -CR ⁹ ;

X ⁶ is N or -CR ¹⁰ ; and

When ^X5 is N, ^X4 must be ^CR8 ; when ^X5 is ^CR9 , ^X4 and ^X6 must be N at the same time;

L is -NR ³ -C(=O)- or -C(=O)-NR ³ -;

M is absent or selected from C _1-6 alkylene substituted by R ^c ;

R ¹ is selected from CN or a group -ZR ¹² , wherein Z is independently selected from -C _0-6 alkyl-, -C═N(OH)-, -C _0-6 alkyl-SC _0-6 alkyl-, -C _0-6 alkyl-S(═O)-C _0-6 alkyl-, -C _0-6 alkyl-SO ₂ -C _0-6 alkyl-, -C _0-6 alkyl-NR ¹¹ -C _0-6 alkyl-, -C _0-6 alkyl-NR ¹¹ SO ₂ -C _0-6 alkyl-, -C _0-6 alkyl-SO ₂ NR ¹¹ -C _0-6 alkyl-, -C _0-6 alkyl-NR ¹¹ SO ₂ NR ¹¹ -C _0-6 alkyl-, -C _0-6 alkyl-OC _0-6 alkyl-, -C _0-6 alkyl-C(═O)-C _0-6 alkyl-, -C -C _0-6 alkyl-C(═O)—OC _0-6 alkyl-, -C _0-6 alkyl-(C═O)NR ¹¹ -C _0-6 alkyl-, _{-C 0-6} alkyl-NR ¹¹ (C═O)-C _0-6 alkyl-, -C 0-6 alkyl-S(═O)(═NH)-C _0-6 alkyl-, -C-((C═O)-OR ¹¹ ) ₂ -, -C _0-6 alkyl-NR ¹¹ -S(═O)(═NH)-C _{0-6 alkyl-, -C 0-6 alkyl} -S(═O)(═N ⁺ (CH ₃ ) ₂ )-, -NR ¹¹ SO ₂ NR ¹¹ -C(═O)—O-, -P-, -C _0-6 alkyl-P(═O)(R ¹¹ )-, -C _0-6 alkyl-P(═O) ₂ ; or the group -ZR ¹² is -N=S(=O)-(R ¹² ) ₂ , wherein two R ¹² pairs can be alternatively combined with the sulfur atoms to which they are respectively attached to form a saturated or partially saturated 3-, 4-, 5- or 6-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S;

^R2 is halogen or a group ^-YR13 , wherein Y is _-C0-6alkyl- , _-C0-6alkyl -S(=O)(=NH)-C0-6alkyl-, _{-C0-6alkyl -} SC0-6alkyl-, -C0-6alkyl- _S (=O)-C0-6alkyl- ^, _{-C0-6alkyl - SO2} - _C0-6alkyl- , _-C0-6alkyl - _NR13 -C0-6alkyl-, _-C0-6alkyl -NR13SO2 _{- C0-6alkyl-} ^{, -C0-6alkyl} - _SO2NR13 - ^C0-6alkyl- , _{-C0-6alkyl - NR13SO2NR13} - _C0-6alkyl- , _{-C0-6alkyl - OC0-6alkyl-} ^, _-C0-6alkyl ^- _C (= _O )-C0-6alkyl -YR ¹³ is -N _{= S} (=O)-(R ₁₃ ) 2 , wherein two _{R 13} pairs _can be alternatively combined with the sulfur atoms to which they _are respectively attached to form a saturated or partially saturated ₃ -membered, _4- membered, ^{5 -} membered or 6 _- membered monocyclic ring containing 0, ¹ , 2 or ₃ N atoms and 0, 1 or 2 atoms ^selected from O and S;

^R3 is selected from hydrogen, _C1-6 alkyl, _C1-4 haloalkyl, _C3-8 cycloalkyl, halogenated _C3-8 cycloalkyl;

R ⁴ is selected from hydrogen, halogen, CN, C _1-8 alkyl, -OC _1-8 alkyl, C _1-4 haloalkyl, C _0-6 alkyl-(C=O)-NH-C _0-6 alkyl, -C _0-6 alkyl-SO ₂ NH-C _0-6 alkyl, -C _0-6 alkyl-SO ₂ N(CH ₃ )-C _0-6 alkyl, R ^4a or R ^4b ;

R ⁵ is selected from hydrogen, halogen, C _1-6 alkyl, C _1-4 haloalkyl, C _3-8 cycloalkyl, halogenated C _3-8 cycloalkyl, -OC _1-8 alkyl or -OR ^5a ; wherein R ^5a is a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S; or alternatively, R ² and R ⁵ can be combined with the carbon atoms to which they are each attached to form a saturated or partially saturated 5- or 6-membered monocyclic ring fused to a benzene ring; wherein the 5- or 6-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and further, wherein the 5- or 6-membered monocyclic ring is substituted by 0, 1, 2 or 3 groups selected from halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC 1-8 alkyl or -OR 5a . _1-4 haloalkyl, CN, -NR ^a R ^a , or oxo;

R ⁶ , R ⁸ , and R ⁹ are each independently selected from hydrogen, halogen, CN, C _1-8 alkyl, halogenated C _3-8 cycloalkyl, C _1-4 haloalkyl, or C _3-8 cycloalkyl;

R ⁷ is hydrogen, halogen, CN, C _1-8 alkyl, C _1-4 haloalkyl, C _3-8 cycloalkyl, halogenated C _3-8 cycloalkyl, -OC _1-8 alkyl or R ^7a ; or alternatively, R ² and R ⁷ can be combined with the carbon atoms to which they are each attached to form a saturated or partially saturated 5-membered or 6-membered monocyclic ring fused to a benzene ring; wherein the 5-membered or 6-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and further, wherein the 5-membered or 6-membered monocyclic ring is substituted by 0, 1, 2 or 3 groups selected from halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -NR ^a R ^a or oxo;

R ¹⁰ is hydrogen, halogen, CN, hydroxyl, C _1-8 alkyl, C _3-8 cycloalkyl, halogenated C _3-8 cycloalkyl, C _1-4 haloalkyl, -OR ^10a or -OR ^10b ;

R ¹¹ is hydrogen, R ^11a or R ^11b ;

R ¹² is hydrogen, halogen, OH, CN, R ^12a or R ^12b ;

R ¹³ is hydrogen, halogen, CN, R ^13a or R ^13b ;

R ^4a , R ^7a , R ^10a , R ^11a , R ^12a and R ^13a are each independently selected from the group consisting of a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -C(═O)R ^b , -C(═O)OR ^a , -C(═O)NR ^a R ^a , -C(═NR ^a )NR ^a R ^a , -OC(═O)R ^b , -OC(═O)NR ^a R ^a , -OC _2-6 alkyl NR ^a R ^a , -OC _2-6 alkyl OR ^a , -SR ^a , -S(=O)R ^b , -S(=O) ₂ R ^b , -S(=O) ₂ NR ^a R ^a , -NR ^a R ^a , -N(R ^a )C(=O)OR ^b , -N(R ^a ) ^C (=O)NR ^a R ^a , -N(R ^a )C(=NR ^a )NR ^a R ^a , -N(R ^a )S(= ^O ) ₂ R ^b , -N(R ^a )S(=O) ₂ NR ^a R ^a , -NR ^a C ₂ -C ₆ alkyl NR ^a R ^a , -NR ^a C _2-6 alkyl OR ^a , -C _1-6 alkyl NR ^a R ^a , -C _1-6 alkyl OR ^a , -C _{-C 1-6} alkylN(R ^a )C(═O)R ^b , -C _1-6 alkylOC(═O)R ^b , -C _1-6 alkylC(═O)NR ^a R ^a , -C _1-6 alkylC(═O)OR ^a , R ¹⁴ and oxo;

R ^4b , R ^10b , R ^11b , R ^12b and R ^13b are each independently selected from the group consisting of: C _1-6 alkyl substituted with 0, 1, 2, 3, 4 or 5 groups selected from the group consisting of: halogen, CH ₂ F, CHF ₂ , CF ₃ , -C(═O)OR ^a , -OR ^a , -OC _1-4 haloalkyl, CN, -NH ₂ , NH(CH ₃ ), N(CH ₃ ) ₂ , and a saturated, partially saturated or unsaturated 3-, 4-, 5- or 6-membered monocyclic ring;

R ¹⁴ is independently selected from the group consisting of a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -C(═O)R ^b , -C(═O)OR ^a , -C(═O)NR ^a R ^a , -C(═NR ^a )NR ^a R ^a , -OC(═O)R ^b , -OC(═O)NR ^a R ^a , -OC _2-6 alkylNR ^a R ^a , -OC _2-6 alkylOR ^a , -SR ^a , -S(=O)R ^b , -S(=O) ₂ R ^b , -S(=O) ₂ NR ^a R ^a , -NR ^a R ^a , -N(R ^a )C(=O)R ^b , -N(R ^a )C(=O)OR ^b , -N(R ^a )C(=O)NR ^a R ^a , -N(R ^a )C(=NR ^a )NR ^a R ^a , -N(R ^a )S(=O) ₂ R ^b , -N(R ^a )S(=O) ₂ NR ^a R ^a , -NR ^a C _2-6 alkylNR ^a R ^a , -NR ^a C _2-6 alkylalkyl OR ^a , -C _1-6 alkyl NR ^a R ^a , -C _1-6 alkyl OR ^a , -C _1-6 alkyl N(R ^a )C(=O)R ^b , -C _1-6 alkyl OC(=O)R ^b , -C _1-6 alkyl C(=O)NR ^a R ^a , -C _1-6 alkyl C(=O)OR ^a and oxo;

^Rx is selected from the group consisting of: hydrogen,

or R ^x is C _2-8 alkyl substituted by 0, 1, 2, 3, 4 or 5 groups selected from the group consisting of halogen, -CH ₂ F, CHF ₂ , -CF ₃ , -C(═O)OR ^a , -OR ^a , -OC _1-4 haloalkyl, CN, -NH ₂ , NH(CH ₃ ), N(CH ₃ ) ₂ or R ¹⁵ⁿ ;

or R ^x is phenyl or an unsaturated 5-membered monocyclic ring substituted by 0, 1, 2, 3, 4 or 5 groups selected from the group consisting of halogen, C _1-6 alkyl, C _1-6 haloalkyl, -C(=O)OR ^a , -OR ^a , -OC _1-4 haloalkyl, CN, -NH ₂ , NH(CH ₃ ), N(CH ₃ ) ₂ or R ¹⁵ⁿ ; wherein the 5-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S;

Each of R ^15a , R ^15b , R ^15c , R ^15d , R ^15e , R ^15f , R ^15g , R ^15h , R ¹⁵ⁱ , R ^15j , R ^15k , R ^15l , and R ^15m is hydrogen, halogen, R ^15o or R ^15p ;

or alternatively, each of the pairs of R ^15a and R ^15b , R ^15c and R ^15d , R ^15e and R 15f, R ^15g and R ^15h , R ¹⁵ⁱ and R ^15j , and R ^15k and R ^15l can independently combine with the carbon atoms to which they are respectively attached to form a saturated or partially saturated 3 ^- membered, 4-membered, 5-membered, 6-membered monocyclic ring spiro-attached to the R ^x ring; wherein the 3-membered, 4-membered, 5-membered, 6-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, and further, wherein the 3-membered, 4-membered, 5-membered, 6-membered monocyclic ring is substituted with 0, 1, 2 or 3 groups selected from the following: halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -NR ^a R ^a or oxo;

Or yet alternatively, each of the pairs of R ^15a and R ^15b , R ^15c and R ^15d , R ^15e and R ^15f , R ^15g and R ^15h , R ¹⁵ⁱ and R ^15j , and R ^15k and R ^15l can independently combine to form a double bond;

R ¹⁵ⁿ , R ^15o are each independently selected from the group consisting of a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or an 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, which is substituted by 0, 1, 2, 3 or 4 groups selected from the following: halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -C(═O)R ^b , -C(═O)OR ^a , -C(═O)NR ^a R ^a , -C(═NR ^a )NR ^a R ^a , -OC(═O)R ^b , -OC(═O)NR ^a R ^a , -OC _2-6 alkylNR ^a R ^a , -OC _2-6 alkylOR ^a , -SR ^a , -S(=O)R ^b , -S(=O) ₂ R ^b , -S(=O) ₂ NR ^a R ^a , -NR ^a R ^a , -N(R ^a )C(=O)R ^b , -N(R ^a )C(=O)OR ^b , -N(R ^a )C(=O)NR ^a R ^a , -N(R ^a )C(=NR ^a )NR ^a R ^a , -N(R ^a )S(=O) ₂ R ^b , -N(R ^a )S(=O) ₂ NR ^a R ^a , -NR ^a C ₂ -C ₆ alkylNR ^a R ^a , -NR ^a C _2-6 alkyl OR ^a , -C _1-6 alkyl NR ^a R ^a , -C _1-6 alkyl OR ^a , -C _1-6 alkyl N(R ^a )C(=O)R ^b , -C _1-6 alkyl OC(=O)R ^b , -C _1-6 alkyl C(=O)NR ^a R ^a , -C _1-6 alkyl C(=O)OR ^a and oxo;

R ^15p is C _1-8 alkyl substituted by 0, 1, 2, 3, 4 or 5 groups selected from the group consisting of halogen, CH ₂ F, CHF ₂ , —CF ₃ , —C(═O)OR ^a , —OR ^a , —OC _1-4 haloalkyl, CN, —NH ₂ , NH(CH ₃ ), N(CH ₃ ) ₂ ;

^{Ra is} independently selected from hydrogen or ^Rb ;

R ^b is independently selected from C _1-6 alkyl, phenyl or benzyl, wherein the C _1-6 alkyl is substituted by 0, 1, 2 or 3 substituents selected from halogen, hydroxy, -OC _1-6 alkyl, -NH ₂ , -NHC _1-6 alkyl, -OC(═O)C _1-6 alkyl or -N(C _1-6 alkyl)C _1-6 alkyl; and the phenyl or benzyl is substituted by 0, 1, 2 or 3 substituents selected from halogen, C _1-6 alkyl, C _1-3 haloalkyl, -OH, -OC _1-6 alkyl, -NH ₂ , -NHC _1-6 alkyl, -OC(═O)C _1-6 alkyl or -N(C _1-6 alkyl)C _1-6 alkyl;

R ^c represents absence or is independently selected from halogen, hydroxyl, amino, cyano, carbonyl, oxo, carboxyl, C _2-6 alkenyl, C _1-6 alkyl, C _1-6 deuterated alkyl, C _{2-6 alkynyl, C 1-4 haloalkyl} , hydroxyC _1-6 alkyl, C _1-6 alkylcarbonyl, C _1-6 alkoxy, haloC _1-4 alkoxy, -COO-C _1-6 alkyl, -C(O)NR ^c1 R ^c2 ; wherein, R ^c1 and R ^c2 are each independently hydrogen or C _1-6 alkyl.

In the present invention, the definitions of certain substituents in the heterocyclic compound as shown in Formula I can be as described below, and the definitions of substituents not mentioned are as described in any of the above schemes.

In a preferred embodiment of the present invention, each of R ^15c , R ^15d , R ^15e , R ^15f , R ^15g , R ^15h , R ¹⁵ⁱ and R ^15j is H, halogen, C _1-6 alkyl or C _1-4 haloalkyl; and each of the pair of R ^15a and R ^15b is combined with the carbon atom to which they are respectively attached to form a saturated 3-membered, 4-membered or 5-membered monocyclic ring spiro-attached to the R ^x ring; wherein the ring contains 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S.

In a preferred embodiment of the present invention, each of said R ^15c , R ^15d , R ¹⁵ⁱ and R ^15j is H, methyl or ethyl; and each of the pair of R ^15a and R ^15b combines with the carbon atom to which they are respectively attached to form a cyclopropyl, cyclobutyl or cyclopentyl ring spiro-attached to the R ^x ring.

In a preferred embodiment of the present invention, the group Selected from Preferably, the group Selected from

In a preferred embodiment of the present invention, the group Selected from

Those skilled in the art will appreciate that, according to the conventions used in the art, in the structural formula of the present application, Used to depict chemical bonds, which are the points at which a moiety or substituent is attached to a core or backbone structure.

In a preferred embodiment of the present invention, said R ³ is hydrogen or methyl.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures:

Wherein, ^X4 and ^X6 are defined as described in the first aspect of the present invention; ^R1 , ^R2 , ^R4 , ^R5 , ^R6 , ^R7 and ^Rx are defined as described in the first aspect of the present invention; and M and L are defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: Wherein, ^X4 and ^X5 are defined as described in the first aspect of the present invention; ^R1 , ^R2 , ^R4 , ^R5 , ^R6 , ^R7 and ^Rx are defined as described in the first aspect of the present invention; and M and L are defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: wherein X ⁴ , X ⁵ and X ⁶ are as defined in the first aspect of the present invention; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined in the first aspect of the present invention; R ^15a , R ^15b , R ^15c , R ^15d , R ¹⁵ⁱ and R ^15j are as defined in the first aspect of the present invention; and M and L are as defined in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: Wherein, ^X4 , ^X5 and ^X6 are defined as described in the first aspect of the present invention; ^R1 , ^R2 , ^R4 , ^R5 , ^R6 and ^R7 are defined as described in the first aspect of the present invention; ^R15a , ^R15b , R15c, ^R15d , ^R15i and ^R15j are defined as described in the first aspect of the present invention; and M ^is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: wherein X ⁴ , X ⁵ and X ⁶ are as defined in the first aspect of the present invention; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ are as defined in the first aspect of the present invention; R ^15a , R ^15b , R ^15c , R ^15d , R ¹⁵ⁱ and R ^15j are as defined in the first aspect of the present invention; and M is as defined in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: Wherein, X ⁴ , X ⁵ and X ⁶ are defined as described in the first aspect of the present invention; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ are defined as described in the first aspect of the present invention; and M is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: Wherein, X ⁴ , X ⁵ and X ⁶ are defined as described in the first aspect of the present invention; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ are defined as described in the first aspect of the present invention; and M is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: Wherein, ^X4 and ^X6 are defined as described in the first aspect of the present invention; ^R1 , ^R2 , ^R4 , ^R5 , ^R6 and ^R7 are defined as described in the first aspect of the present invention; and M is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: Wherein, ^X4 and ^X6 are defined as described in the first aspect of the present invention; ^R1 , ^R2 , ^R4 , ^R5 , ^R6 and ^R7 are defined as described in the first aspect of the present invention; and M is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: Wherein, ^X4 is defined as described in the first aspect of the present invention; ^R1 , ^R2 , ^R4 , ^R5 , ^R6 and ^R7 are defined as described in the first aspect of the present invention; and M is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: Wherein, ^X4 is defined as described in the first aspect of the present invention; ^R1 , ^R2 , ^R4 , ^R5 , ^R6 and ^R7 are defined as described in the first aspect of the present invention; and M is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures: Wherein, ^X5 is defined as described in the first aspect of the present invention; ^R1 , ^R2 , ^R4 , ^R5 , ^R6 and ^R7 are defined as described in the first aspect of the present invention; and M is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures:

Wherein, ^X5 is defined as described in the first aspect of the present invention; ^R1 , ^R2 , ^R4 , ^R5 , ^R6 and ^R7 are defined as described in the first aspect of the present invention; and M is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from the following structures:

Wherein, R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ are defined as described in the first aspect of the present invention; and M is defined as described in the first aspect of the present invention.

In a preferred embodiment of the present invention, R ² is selected from: F, Cl, Br or a group -YR ¹³ , wherein Y is a bond, -NH-, -NH-(CH ₂ ) _0-4 -, -O-(CH ₂ ) _0-4 -, -S-, -S＝O-, -S(＝O) _2- , -S(＝O)(＝NH)-, -C(＝O)-; and

and R ¹³ is a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from the following: F, Cl, Br,

C _1-6 alkyl, C _1-4 haloalkyl, -OH, -OC _1-4 haloalkyl, CN, R ¹⁴ and oxo; or R ¹³ is selected from F,

C _1-6 alkyl substituted with 0, 1, 2, 3, 4 or 5 groups selected from Cl, Br, -OH, -OC _1-4 haloalkyl or CN;

Wherein, the definition of R ¹⁴ is as described in the first aspect of the present invention;

Preferably, Y is a bond, R ¹³ is a saturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0 or 1 N atoms and 0 or 1 O atoms; which is substituted by 0, 1, 2 or 3 identical or different groups selected from the following: F, Cl, Br, -C _1-3 alkyl, -OH, CN, -C _1-3 alkyl-OH;

Preferably, it is F, Cl, Br, methyl, -OH, -OCH ₃ , CN, -CH ₂ OH; or,

Y is -NH-, -NH-(CH ₂ ) _0-4 -, -O-(CH ₂ ) _0-4 - or -SO ₂ NH-; R ¹³ is a saturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring containing 0 or 1 N atom and 0 or 1 O atom; which is substituted by 0, 1, 2 or 3 identical or different groups selected from the following: fluorine, methyl, trifluoromethyl, -CN, -OH, CH ₂ OH;

Or Y is a bond, -NH-, -NH-(CH ₂ ) _0-4 -, -O-(CH ₂ ) _0-4 - or -SO ₂ NH-; R ¹³ is a C _1-6 alkyl substituted by 0, 1, 2, 3, 4 or 5 identical or different substituents selected from the following: -OH, methyl, CF ₃ .

In a preferred embodiment of the present invention, said R ² is selected from:

In a preferred embodiment of the present invention, the M is absent or selected from -NH-, _-CH2- ,

In a preferred embodiment of the present invention, Z is selected from a bond, -NH-, -NHSO ₂ -, -SO ₂ NH-, -S(=O)(=

NH)-, -S-, -S(=O)-, -SO ₂ -, -O-, -P-, -P(=O)CH ₃ -, -P(=O) ₂ , -(C =O)-, -(C=O)NH- or -NH(C=O)-;

Preferably, Z is selected from -SO ₂ NH-, -S(=O)(=NH)-, and -SO ₂ -.

In a preferred embodiment of the present invention, R ¹² is selected from (a) hydrogen; (b) C _1-6 alkyl substituted by 0, 1, 2 or 3 groups selected from F, Cl, Br, -OH, -OCH ₃ or cyclopropyl; or (c) a saturated, partially saturated or unsaturated 3-membered, 4-membered, 5-membered, 6-membered or 7-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from the following: F, Cl, Br, C _1-6 alkyl, C _1-4 haloalkyl, -C _1-6 alkylOH, -OH, -OCH ₃ , -NH ₂ or oxo.

In a preferred embodiment of the present invention, the R ¹² is selected from (a) hydrogen; (b) -OH

or (c) cyclopropyl _{, cyclobutyl, cyclopentyl, oxetanyl, azetidinyl, tetrahydrofuranyl, 1,3,4-oxathiazinyl, or C 3-8 cycloalkyl} substituted with -OH.

In a preferred embodiment of the present invention, the R ¹ is selected from:

In a preferred embodiment of the present invention, said R ⁴ is selected from (a) hydrogen, F, Cl, Br, CN, -OC _1-6 alkyl; (b)

C _1-6 alkyl substituted by 0, 1, 2 or 3 OH groups; (c) cyclopropyl or (d) C _1-3 haloalkyl.

In a preferred embodiment of the present invention, R ⁴ is selected from hydrogen, methyl, F, Cl, Br, CN, cyclopropyl, methoxy, -(C=O)NH ₂ , CF ₃ , CHF ₂ , furanyl, pyridyl, morpholinyl or azetidine.

In a preferred embodiment of the present invention, said R ⁵ is selected from hydrogen, F or methyl.

In a preferred embodiment of the present invention, said R ⁶ is selected from hydrogen.

In a preferred embodiment of the present invention, said R ⁷ is selected from hydrogen, F, Cl, Br, CN, methyl, methoxy, cyclopropyl; or R ² and R ⁷ are combined with the carbon atoms to which they are attached to form In a preferred embodiment of the present invention, said R ⁸ is selected from hydrogen, F, Cl, Br, cyclopropyl, -(C=O)CH ₃ or CF ₃ ;

Preferably, R ⁸ is selected from hydrogen.

In a preferred embodiment of the present invention, said R ⁹ is selected from hydrogen or F.

In a preferred embodiment of the present invention, said R ¹⁰ is selected from hydrogen, F, methylmethoxy or cyclopropyl.

In a preferred embodiment of the present invention, the heterocyclic compound as shown in Formula I is selected from any of the following compounds:

The second aspect of the present invention provides a pharmaceutical composition, which comprises: a compound of formula I as described in the first aspect of the present invention, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a pharmaceutically acceptable carrier.

In the third aspect of the present invention, there is provided the use of the compound of formula I as described in the first aspect of the present invention, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or the use of the pharmaceutical composition as described in the fifth aspect of the present invention, the uses comprising: inhibiting KIF18A; and/or, preventing and/or treating diseases related to KIF18A; and/or, preparing drugs, pharmaceutical compositions or preparations for inhibiting KIF18A, and/or preventing and/or treating diseases related to KIF18A.

Preferably, the KIF18A-related diseases include: cancer, psoriasis, atopic dermatitis, autoimmune diseases or proliferative disorders of inflammatory bowel disease.

Preferably, the cancer is selected from mesothelioma, neuroblastoma, rectal cancer, colon cancer, familiar adenomatous polyposis and hereditary non-polyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine body cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer, breast cancer, urinary cancer, Melanoma, brain tumor, lymphoma, head and neck cancer, acute lymphoid leukemia, chronic lymphoid leukemia, acute myeloid leukemia, chronic myeloid leukemia, hepatocellular carcinoma, gallbladder cancer, bronchial carcinoma, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal sarcoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myoma, liposarcoma, fibrosarcoma, Ewing sarcoma, and plasmacytoma.

Preferably, the autoimmune disease is selected from rheumatoid arthritis, systemic lupus erythematosus, Sjögren's syndrome, scleroderma, mixed connective tissue disease, dermatomyositis, polymyositis, Reiter's syndrome, autoimmune lymphoproliferative syndrome, multiple sclerosis, myasthenia gravis and encephalomyelitis.

Preferably, the inflammatory bowel disease is selected from ulcerative colitis or Crohn's disease.

In a fourth aspect of the present invention, a method for inhibiting KIF18A, or preventing and/or treating a disease related to KIF18A is provided, comprising the steps of administering to a subject in need thereof the compound of formula I described in the first aspect of the present invention, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof.

Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention.

Terms and Definitions

Unless otherwise specified, the definitions of groups and terms recorded in the specification and claims of this application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be arbitrarily combined and combined with each other. The group definitions and compound structures after such combinations and combinations shall fall within the scope of the description of this application.

Unless otherwise defined, all technical terms used herein have the same meaning as those commonly understood by those skilled in the art to which the subject matter of the claims pertains. Unless otherwise indicated, all patents, patent applications, and publications cited herein are incorporated herein by reference in their entirety. If there are multiple definitions of a term herein, the definition in this chapter shall prevail.

It should be understood that the above brief description and the detailed description below are exemplary and are only used for explanation, and do not impose any restrictions on the subject matter of the present invention. In this application, unless otherwise specifically stated, the use of the singular also includes the plural. It must be noted that unless otherwise clearly stated in the text, the singular form used in this specification and claims includes the plural form of the referred thing. It should also be noted that unless otherwise stated, the "or" and "or" used mean "and/or". In addition, the term "including" and other forms, such as "including", "containing" and "containing" are not restrictive.

Definitions of standard chemical terms can be found in references (including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4 THE D." Vols. A (2000) and B (2001), Plenum Press, New York). Unless otherwise stated, conventional methods within the technical scope of the art, such as mass spectrometry, NMR, IR and UV/VIS spectroscopy and pharmacological methods, are used. Unless specifically defined, the terms used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and drugs and medicinal chemistry are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug preparation, formulation and delivery, and in the treatment of patients. For example, the manufacturer's instructions for use of the kit can be used, or the reaction and purification can be carried out in a manner known in the art or the description of the present invention. The above-mentioned techniques and methods can usually be implemented according to the descriptions in a plurality of summary and more specific documents cited and discussed in this specification, according to conventional methods well known in the art. In this specification, groups and substituents thereof can be selected by those skilled in the art to provide stable structural parts and compounds.

When substituents are described by conventional chemical formulas written from left to right, the substituents also include chemically equivalent substituents that would result if the formula were written from right to left. For example, CH ₂ O is equivalent to OCH ₂ . As used herein, Indicates the attachment site of a group. As used herein, "R ₁ ", "R1" and "R ¹ " have the same meaning and can be replaced with each other. For other symbols such as R ₂ , similar definitions have the same meaning.

The section headings used herein are only for the purpose of organizing the article and should not be interpreted as limitations on the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, operating manuals and papers, are incorporated herein by reference in their entirety.

In addition to the foregoing, when used in the specification and claims of the present application, the following terms have the meanings indicated below unless otherwise specifically stated.

The numerical ranges recorded in the specification and claims of this application, when the numerical range is understood as an "integer", should be understood as recording the two endpoints of the range and each integer in the range. For example, "an integer from 1 to 6" should be understood as recording each integer of 0, 1, 2, 3, 4, 5 and 6. When the numerical range is understood as a "number", it should be understood as recording the two endpoints of the range and each integer in the range and each decimal in the range. For example, "a number from 1 to 10" should be understood as recording not only each integer of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, but also at least recording the sum of each integer therein and 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, respectively.

In the present application, "saturated, partially saturated or unsaturated" includes substituents saturated with hydrogen, substituents completely unsaturated with hydrogen and substituents partially saturated with hydrogen.

As used herein, the term "halogen" alone or as part of another substituent refers to fluorine, chlorine, bromine, iodine; preferably fluorine or chlorine.

As used herein, the term "cyano" by itself or as part of another substituent means -CN.

As used herein, the term "amino" by itself or as part of another substituent means _-NH2 .

The term "alkyl" when used alone or as part of other substituents means a straight or branched hydrocarbon chain radical consisting only of carbon and hydrogen atoms, having, for example, 1 to 6 carbon atoms and connected to the rest of the molecule by a single bond. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, pentyl, isopentyl, neopentyl and hexyl. Alkyl groups may be unsubstituted or substituted with one or more suitable substituents. Alkyl groups may also be isotopomeric isomers of natural abundance alkyl groups that are rich in isotopes of carbon and/or hydrogen (i.e., deuterium or tritium).

The term "C _α-β alkyl" refers to an alkyl group containing a minimum of α and a maximum of β carbon atoms in a branched or linear relationship or any combination of the three, where α and β represent integers, and can also be represented in the form of "C _α- C _β alkyl". The alkyl groups described in this section may also contain one or two double bonds or triple bonds. The designation of C ₀ alkyl represents a direct bond. Examples of _{C 1-6} alkyl groups include, but are not limited to, the following:

"Benzo group" alone or in combination refers to a divalent _{group C4H4} =, one of which is represented by -CH=CH-CH=CH-, which forms a benzene-like ring when attached to another ring in the ortho position, for example, tetralin, indole, and the like.

The term "C _α-β haloalkyl" by itself or as part of another substituent refers to an alkyl group as described above wherein any number (at least one) of the hydrogen atoms attached to the alkyl chain are replaced by fluorine, chlorine, bromine or iodine.

When alone or as part of other substituents, the term "cycloalkyl" refers to a cyclic alkyl group. The term "mn-membered cycloalkyl" or "C _m -C _n cycloalkyl" should be understood to mean a saturated, unsaturated or partially saturated carbocyclic ring having m to n atoms. For example, "3-15-membered cycloalkyl" or "C ₃ -C ₁₅ cycloalkyl" refers to a cyclic alkyl group containing 3 to 15, 3 to 9, 3 to 6 or 3 to 5 carbon atoms, which may contain 1 to 4 rings. "3-10-membered cycloalkyl" contains 3-10 carbon atoms. Including monocyclic, bicyclic, tricyclic, spirocyclic or bridged rings. Examples of unsubstituted cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and adamantyl, or a bicyclic hydrocarbon group such as a decalin ring. Cycloalkyl can be substituted with one or more substituents. In some embodiments, the cycloalkyl can be a cycloalkyl fused to an aryl or heteroaryl ring group. The term "cycloalkyl" can be used interchangeably with the term "carbocyclic group".

The term "heterocycloalkyl" when used alone or as part of another substituent refers to a cycloalkyl group in which one or more (in some embodiments, 1 to 3) carbon atoms are replaced by heteroatoms, such as, but not limited to, N, O, S, and P. The term "mn-membered heterocycloalkyl" or "C _m -C _n heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having m to n atoms, wherein the heteroatoms are selected from N, O, S, P, preferably N, O or S. For example, the term "4-8 membered heterocycloalkyl" or "C ₄ -C ₈ heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having 4 to 8 atoms, wherein 1, 2, 3, or 4 ring atoms are selected from N, O, S, P, preferably N, O or S. "4-10 membered heterocyclyl" means a saturated, unsaturated or partially saturated ring having 4 to 10 atoms. In some embodiments, the heterocycloalkyl group may be a heterocycloalkyl group fused to an aromatic ring group or a heteroaromatic ring group. When prefixes such as 4-8 yuan or 4-10 yuan are used to represent heterocycloalkyl, the number of carbons also means to include heteroatoms. Include monocyclic, bicyclic, tricyclic, spirocyclic or bridged rings. Examples of heterocycloalkyl are: pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridyl, tetrahydropyrrolyl, azetidinyl, thiazolidinyl, oxazolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, azepanyl, diazepanyl, oxazepanyl, etc. The term "heterocycloalkyl" can be used interchangeably with the term "heteroalkyl ring".

The term "alkoxy" by itself or as part of another substituent refers to the group ^-ORQ , wherein ^RQ is an "alkyl" as defined above.

The term "oxo" when used alone or as part of another substituent refers to the replacement of two hydrogen atoms on a methylene group by oxygen atoms, ie, the methylene group is replaced by a carbonyl group, representing =0.

The term "thio" by itself or as part of another substituent refers to the replacement of 2 hydrogens on a methylene group with sulfur, representing =S.

The term "aryl" when used alone or as part of another substituent refers to a monocyclic or polycyclic carbon ring having from 6 to 20 carbon atoms, at least one of which is aromatic. When one of the rings is non-aromatic, the group may be attached through either the aromatic ring or the non-aromatic ring. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl, biphenyl, phenanthrenyl, anthracenyl, and acenaphthenyl.

When alone or as part of other substituents, the term "heteroaromatic ring" refers to a monocyclic or polycyclic carbocyclic ring in which at least one ring atom is a heteroatom independently selected from oxygen, sulfur and nitrogen, and the remaining ring atoms are C, wherein at least one ring is an aromatic ring. The group may be a carbon group or a heteroatom group (i.e., it may be C-connected or N-connected, as long as it is possible). When one of the rings is a non-aromatic ring, the group may be connected through an aromatic ring or through a non-aromatic ring. Examples of heteroaryl include, but are not limited to, imidazolyl, acridinyl, carbazolyl, cinnolinyl, quinoxalinyl, pyrazolyl, indolyl, benzotriazolyl, furanyl, thienyl, benzothienyl, benzofuranyl, quinolyl, isoquinolyl, oxazolyl, isoxazolyl, indolyl, pyrazinyl, pyridazinyl, pyridinyl, pyrimidinyl, pyrrolyl, N-methylpyrrolyl and tetrahydroquinoline. The term "heteroaromatic ring" may be used interchangeably with the terms "heteroaromatic ring", "heteroaryl" or "heteroaromatic ring group".

The term "bicyclic" when used alone or as part of another substituent refers to a group having two connected rings. The bicyclic ring can be a carbocyclic ring (all ring atoms are carbon atoms) or a heterocyclic ring (in addition to carbon atoms, the ring atoms include, for example, 1, 2 or 3 heteroatoms, such as N, O or S). Both rings can be aliphatic (e.g., decalin and norbornane), or can be aromatic (e.g., naphthalene), or a combination of aliphatic and aromatic (e.g., tetralin).

Bicyclic rings include (a) spiro compounds, in which the two rings share only one single atom (the spiro atom, which is usually a quaternary carbon). Examples of spiro compounds include, but are not limited to:

It also includes spirocycloalkyl groups that share a spiro atom with a heterocycloalkyl group. Non-limiting examples include:

(b) Fused bicyclic compounds, where the two rings share two adjacent atoms. In other words, the rings share one covalent bond, i.e., the bridgehead atoms are directly connected (e.g., α-thujene and decalin). Examples of fused bicyclic rings include, but are not limited to:

and (c) bridged bicyclic compounds, wherein the two rings share three or more atoms and the two bridgehead atoms are separated by a bridge comprising at least one atom. For example, norbornane, also known as bicyclo[2.2.1]heptane, can be considered as a pair of cyclopentane rings, each ring sharing three of their five carbon atoms. Examples of bridged bicyclic rings include, but are not limited to:

When alone or as part of another substituent, the NR ^a R ^a group can be In the form of, or may include two ^Ra groups together to form a ring, the ring optionally contains N, O or S atoms, and may also include the following groups, for example:

The group N(Cα _-βalkyl )Cα _-βalkyl (wherein α and β are as defined above) when alone or as part of other substituents includes substituents wherein two Cα _-βalkyl groups are taken together to form a ring (optionally containing N, O or S atoms), and includes groups such as:

Compounds provided herein include intermediates that can be used to prepare compounds provided herein, which contain reactive functional groups (such as but not limited to carboxyl, hydroxyl and amino moieties), and also include protected derivatives thereof. "Protected derivatives" are those compounds in which one or more reactive sites are blocked by one or more protecting groups (also referred to as protecting groups). Suitable carboxyl moiety protecting groups include benzyl, tert-butyl, etc., and isotopes, etc. Suitable amino and amido protecting groups include acetyl, trifluoroacetyl, tert-butyloxycarbonyl, benzyloxycarbonyl, etc. Suitable hydroxyl protecting groups include benzyl, etc. Other suitable protecting groups are well known to those of ordinary skill in the art.

In the present application, "optional" or "optionally" means that the event or situation described later may or may not occur, and the description includes both the occurrence and non-occurrence of the event or situation. For example, "optionally substituted aryl" means that aryl is substituted or unsubstituted, and the description includes both substituted aryl and unsubstituted aryl.

In the present application, the term "salt" or "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms that are suitable for use in contact with human and animal tissues within the scope of sound medical judgment without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.

"Pharmaceutically acceptable acid addition salts" refers to salts formed with inorganic or organic acids that retain the biological effectiveness of the free base without other side effects. "Pharmaceutically acceptable base addition salts" refers to salts formed with inorganic or organic bases that retain the biological effectiveness of the free acid without other side effects. In addition to pharmaceutically acceptable salts, other salts are contemplated by the present invention. They can serve as intermediates in the purification of compounds or in the preparation of other pharmaceutically acceptable salts or can be used for the identification, characterization or purification of the compounds of the present invention.

The term "amine salt" refers to the product obtained by neutralizing an alkyl primary amine, secondary amine or tertiary amine with an acid. The acid includes the inorganic acid or organic acid described in the present application.

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

Depending on the choice of starting materials and methods, the compounds of the invention may exist in the form of one of the possible isomers or a mixture thereof, for example as a pure optical isomer, or as a mixture of isomers, such as a racemic and diastereomeric mixture, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral center (or multiple chiral centers) in the molecule. The prefixes D and L or (+) and (–) are the symbols used to specify the rotation of plane polarized light caused by the compound, where (–) or L indicates that the compound is levorotatory. Compounds prefixed with (+) or D are dextrorotatory.

When the bonds to the chiral carbon in the formula of the present invention are depicted as straight lines, it should be understood that both the (R) and (S) configurations of the chiral carbon and the enantiomerically pure compounds and mixtures thereof produced therefrom are included within the scope of the general formula. The graphic representation of racemates or enantiomerically pure compounds herein is from Maehr, J. Chem. Ed. 1985, 62: 114-120. The absolute configuration of a stereocenter is indicated by a wedge-shaped bond and a dashed bond.

The term "tautomer" refers to functional group isomers resulting from the rapid movement of an atom in a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Prototropic tautomers arise from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to separate a single tautomer usually produce a mixture whose physicochemical properties are consistent with a mixture of compounds. The position of equilibrium depends on the chemical characteristics 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 invention encompasses all tautomeric forms of the compounds.

The term "solvate" refers to a compound of the present invention or a salt thereof including a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent forces between molecules. When the solvent is water, it is a hydrate.

The term "prodrug" refers to a compound of the present invention that can be converted into a biologically active compound under physiological conditions or by solvolysis. The prodrug of the present invention is prepared by modifying the functional groups in the compound, and the modification can be removed by conventional operations or in vivo to obtain the parent compound. The prodrug includes a compound formed by connecting a hydroxyl or amino group in the compound of the present invention to any group. When the prodrug of the compound of the present invention is administered to a mammalian subject, the prodrug is cleaved to form a free hydroxyl group and a free amino group, respectively.

In this application, "pharmaceutical composition" refers to a preparation of the compound of the present invention and a medium generally accepted in the art for delivering the biologically active compound to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of the pharmaceutical composition is to promote administration of the organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

In this application, "pharmaceutically acceptable carrier" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye/colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the relevant governmental regulatory authorities as acceptable for human or livestock use.

The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of the term "excipient" include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, and diluents. Excipients can enhance the handling characteristics of a pharmaceutical formulation, i.e., make the formulation more suitable for direct compression by increasing fluidity and/or adhesion.

The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) ameliorating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade leading to or causing the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

The term "prevent" refers to the reduction of the risk of acquiring or developing a disease or disorder.

The term "patient" refers to any animal, preferably a mammal, that is about to or has been administered the compound or composition according to embodiments of the present invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., preferably humans.

The term "therapeutically effective amount" refers to an amount of a compound that is effective in treating a disease or condition described herein when administered to a patient."Therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, and can be adjusted as needed by those skilled in the art.

The reaction temperature of each step can be appropriately selected according to the solvent, starting materials, reagents, etc., and the reaction time can also be appropriately selected according to the reaction temperature, solvent, starting materials, reagents, etc. After the reaction of each step is completed, the target compound can be separated and purified from the reaction system by common methods, such as filtration, extraction, recrystallization, washing, silica gel column chromatography, etc. In the case of not affecting the next step reaction, the target compound can also be directly entered into the next step reaction without separation and purification.

Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain the preferred embodiments of the present invention.

Beneficial Effects

The present inventors have unexpectedly developed a compound or a pharmaceutically acceptable salt thereof, a preparation method and use thereof through extensive and in-depth research.

The present invention provides a compound represented by formula I, and its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs. The compound represented by formula I has a significant inhibitory effect on KIF18A and has high safety and drug-forming properties.

The present invention provides a method for preparing the compound shown in I, its tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs and intermediates. The method is simple to operate, high in yield and purity, and can be used for industrial production of medicines.

DETAILED DESCRIPTION

The present invention is further described below in conjunction with specific examples. It should be understood that the following description is only the most preferred embodiment of the present invention and should not be considered as limiting the scope of protection of the present invention. On the basis of a full understanding of the present invention, the experimental methods in the following examples that do not specify specific conditions are usually carried out under conventional conditions or under conditions recommended by the manufacturer. Those skilled in the art may make non-essential changes to the technical solution of the present invention, and such changes should be deemed to be included in the scope of protection of the present invention.

This application has the following definitions:

Symbol or unit:

IC ₅₀ : Half inhibitory concentration, which refers to the concentration at which half of the maximum inhibitory effect is achieved

M: mol/L, for example, n-butyllithium (14.56 mL, 29.1 mmol, 2.5 M n-hexane solution) means a n-butyllithium n-hexane solution with a molar concentration of 2.5 mol/L

N: equivalent concentration, for example, 2N hydrochloric acid means 2 mol/L hydrochloric acid solution

Reagents:

DIPEA: N,N-diisopropylethylamine

Boc: tert-butyloxycarbonyl

-HATU: 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate Intermediate A1: Preparation of 3-(4,4-difluoropiperidin-1-yl)aniline

The synthetic route of intermediate A1 is as follows:

Step 1: Synthesis of 4,4-difluoro-1-(3-nitrophenyl)piperidine (A1-3)

Under nitrogen protection, palladium acetate (1.07 g, 4.75 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthene (2.75 g, 4.75 mmol) and cesium carbonate (38.7 g, 119 mmol) were added to a solution of 1-bromo-3-nitrobenzene (A1-3) (8.0 g, 39.6 mmol) and 4,4-difluoropiperidine hydrochloride (8.74 g, 55.4 mmol) in 1,4-dioxane (80 mL), and the reaction was carried out at 80 ° C for 16 h under nitrogen protection. After the reaction was completed, the mixture was diluted with water (100 mL) at room temperature, and then extracted with ethyl acetate (200 mL*3). The organic layers were combined, dried over sodium sulfate, and concentrated to obtain a crude product, which was then separated and purified using a silica gel column (petroleum ether: ethyl acetate (V/V) = 15:1 to 5:1) to obtain compound 4,4-difluoro-1-(3-nitrophenyl)piperidine (A1-3) (yellow solid, 9.0 g, yield 93.8%).

LC-MS, M/Z(ESI):243.0[M+H] ⁺ .

Step 2: Synthesis of 3-(4,4-difluoropiperidin-1-yl)aniline (A1)

Under nitrogen protection, iron powder (18.4 g, 330 mmol) and ammonium chloride (17.7 g, 330 mmol) were added to a solution of 4,4-difluoro-1-(3-nitrophenyl)piperidine (A1-3) (8.0 g, 33.0 mmol) in ethanol (80 mL) and water (20 mL), and the reaction was carried out at 75 ° C for 16 h under nitrogen protection. After the reaction was completed, it was diluted with water (300 mL) and ethyl acetate (500 mL) at room temperature, then filtered, and the filtrate was extracted with ethyl acetate (300 mL*2), the organic phases were combined, dried over sodium sulfate, and concentrated to obtain a crude product, which was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 10:1 to 2:1) to obtain compound 3-(4,4-difluoropiperidin-1-yl)aniline (intermediate A1) (brown solid, 5.7 g, yield 81.3%).

¹ H NMR (400MHz, CDCl ₃ ) δ7.08(t,1H),6.38-6.40(m,1H),6.25-6.29(m,2H),3.65(s,2H),3.35(t,4H), 2.06-2.15(m,4H).

LC-MS, M/Z(ESI):213.2[M+H] ⁺ .

Intermediate A2: Preparation of 5-bromo-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylic acid

The synthetic route of intermediate A2 is as follows:

5-Bromo-3-fluoropyridine-2-carboxylic acid (10 g, 35.47 mmol) was added to N-methylpyrrolidone (100 mL), and then 6-azaspiro[2.5]octane hydrochloride (7.83 g, 53.2 mmol) and DIPEA (22.88 g, 177.35 mmol) were added. The mixture was heated to 140°C for 3 h, extracted with ethyl acetate (100 mL*2), and the pH of the aqueous phase was adjusted to 1 with 1 M dilute hydrochloric acid, and then extracted with dichloromethane (100 mL*2). The organic phase was dried and concentrated to give compound 5-bromo-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylic acid (intermediate A2) (light yellow solid, 10 g, yield 91.8%).

LC-MS, M/Z(ESI): 311.0[M+H] ⁺ .

Intermediate A3: Preparation of 3-(4,4-difluoropiperidin-1-yl)-5-methylaniline

The synthetic route of intermediate A3 is as follows:

Step 1: Synthesis of 4,4-difluoro-1-(3-methyl-5-nitrophenyl)piperidine (A3-2)

1-Bromo-3-methyl-5-nitrobenzene (A3-1) (1 g, 4.63 mmol) was dissolved in dry 1,4-dioxane (10 mL), and 4,4-difluoropiperidine hydrochloride (1.1 g, 6.98 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (535 mg, 0.93 mmol), tris(dibenzylideneacetone)dipalladium (424 mg, 0.46 mmol) and cesium carbonate (4.53 g, 13.9 mmol) were added in sequence, and the argon gas was replaced three times, and the temperature was raised to 100°C for 18 h. The mixture was cooled to room temperature, the solvent was concentrated, and the mixture was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 20:1) to obtain 4,4-difluoro-1-(3-methyl-5-nitrophenyl)piperidine (A3-2) (850 mg, yield 71.7%).

Step 2: Synthesis of 3-(4,4-difluoropiperidin-1-yl)-5-methylaniline (A3)

4,4-difluoro-1-(3-methyl-5-nitrophenyl)piperidine (A3-2) (850 mg, 3.32 mmol) and ammonium chloride (531 mg, 9.93 mmol) were dissolved in ethanol (20 mL) and water (5 mL), iron powder (1.12 g, 20 mmol) was added, the temperature was raised to 50°C for reaction for 2 h, diatomaceous earth was filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, and the residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 3:1) to obtain 3-(4,4-difluoropiperidin-1-yl)-5-methylaniline (A3) (720 mg, yield 96.0%)

LC-MS, M/Z(ESI): 227.2[M+H] ⁺ .

Intermediate A4: Preparation of 3-(4,4-difluoropiperidin-1-yl)-5-fluoroaniline

The synthetic route of intermediate A4 is as follows:

Step 1: Synthesis of 4,4-difluoro-1-(3-fluoro-5-nitrophenyl)piperidine (A4-2)

1-Bromo-3-fluoro-5-nitrobenzene (A4-1) (1 g, 4.5 mmol) was dissolved in dry 1,4-dioxane (10 mL), and 4,4-difluoropiperidine hydrochloride (1.06 g, 6.7 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (521 mg, 0.9 mmol), tris(dibenzylideneacetone)dipalladium (366 mg, 0.4 mmol) and cesium carbonate (4.40 g, 13.5 mmol) were added in sequence, and the argon gas was replaced three times, and the temperature was raised to 100°C for reaction for 18 h. The mixture was cooled to room temperature, the solvent was concentrated, and the mixture was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 20:1) to obtain 4,4-difluoro-1-(3-fluoro-5-nitrophenyl)piperidine (A4-2) (750 mg, yield 63.4%).

Step 2: Synthesis of 3-(4,4-difluoropiperidin-1-yl)-5-fluoroaniline (A4)

4,4-difluoro-1-(3-fluoro-5-nitrophenyl)piperidine (A4-2) (750 mg, 2.9 mmol) and ammonium chloride (465 mg, 8.7 mmol) were dissolved in ethanol (20 mL) and water (5 mL), iron powder (972 mg, 17.4 mmol) was added, the temperature was raised to 50°C for reaction for 2 h, diatomaceous earth was filtered, the filter cake was rinsed with dichloromethane, the filtrate was concentrated, and the residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 3:1) to obtain 3-(4,4-difluoropiperidin-1-yl)-5-fluoroaniline (A4) (560 mg, yield 84.4%)

LC-MS, M/Z(ESI): 231.2[M+H] ⁺ .

Intermediate A5: Preparation of 3-amino-5-(4,4-difluoropiperidin-1-yl)benzonitrile

The synthetic route of intermediate A5 is as follows:

Step 1: Synthesis of 3-(4,4-difluoropiperidin-1-yl)-5-nitrobenzonitrile (A5-2)

3-Fluoro-5-nitrobenzonitrile (2 g, 0.01 mol), 4,4-difluoropiperidine hydrochloride (2.2 g, 0.02 mol) and potassium carbonate (3.3 g, 0.03 mol) were added to N,N-dimethylformamide (15 mL), stirred at 80 ° C for 2 h, diluted with water (100 mL), extracted with ethyl acetate (30 mL×2), and the organic phase was concentrated to dryness. The residue was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 50:1) to obtain 3-(4,4-difluoropiperidin-1-yl)-5-nitrobenzonitrile (A5-2) (1.8 g, yield 50%).

Step 2: Synthesis of 3-amino-5-(4,4-difluoropiperidin-1-yl)benzonitrile (A5)

3-(4,4-difluoropiperidin-1-yl)-5-nitrobenzonitrile (A5-2) (1.5 g, 5.62 mmol), iron powder (1 g, 16.2 mmol), and saturated ammonium chloride solution (5 mL) were added to ethanol (15 mL) and reacted at 80° C. for 2 h. Water (60 mL) was added for dilution, and the mixture was extracted with ethyl acetate (30 mL×2). The organic phase was concentrated to dryness, and the residue was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 10:1) to obtain 3-amino-5-(4,4-difluoropiperidin-1-yl)benzonitrile (800 mg, yield 50%).

LC-MS, M/Z(ESI):238.1[M+H] ⁺ .

Intermediate A6: Preparation of 3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylaniline

The synthetic route of intermediate A6 is as follows:

Step 1: Synthesis of 1-bromo-2-fluoro-3-methyl-5-nitrobenzene (A6-2)

1-Fluoro-2-methyl-4-nitrobenzene (A6-1) (2 g, 12.9 mmol) was dissolved in dilute sulfuric acid (2N, 20 mL), cooled to about 0°C, and N-bromosuccinimide (2.76 g, 15.5 mmol) was added in batches. After addition, the temperature was restored to room temperature and the reaction was continued for 20 h. Water (30 mL) was added to dilute the mixture, and dichloromethane was used for extraction (30 mL*3). The solvent was concentrated, and the residue was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 50:1) to obtain 1-bromo-2-fluoro-3-methyl-5-nitrobenzene (A6-2) (2.0 g, yield 66.4%).

Step 2: Synthesis of 4,4-difluoro-1-(2-fluoro-3-methyl-5-nitrophenyl)piperidine (A6-3)

1-Bromo-2-fluoro-3-methyl-5-nitrobenzene (A6-2) (2 g, 8.5 mmol) was dissolved in dry 1,4-dioxane (30 mL), and 4,4-difluoropiperidine hydrochloride (2.02 g, 12.8 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (983 mg, 1.7 mmol), tris(dibenzylideneacetone)dipalladium (824 mg, 0.9 mmol) and cesium carbonate (8.31 g, 25.5 mmol) were added in sequence, and the argon gas was replaced three times, and the temperature was raised to 100°C for reaction for 18 h. The mixture was cooled to room temperature, the solvent was concentrated, and the mixture was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 20:1) to obtain 4,4-difluoro-1-(2-fluoro-3-methyl-5-nitrophenyl)piperidine (A6-3) (1.8 g, yield 85.3%).

Step 3: Synthesis of 3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylaniline (A6)

4,4-difluoro-1-(2-fluoro-3-methyl-5-nitrophenyl)piperidine (A6-3) (1.8 g, 6.6 mmol) and ammonium chloride (1.06 g, 19.8 mmol) were dissolved in ethanol (60 mL) and water (15 mL), iron powder (2.21 g, 39.6 mmol) was added, the temperature was raised to 50°C for reaction for 2 h, diatomaceous earth was filtered, the filter cake was rinsed with methanol, the filtrate was concentrated, and the residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 3:1) to obtain 3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylaniline (A6) (500 mg, yield 31.2%)

LC-MS, M/Z(ESI): 245.2[M+H] ⁺ .

Intermediate A7: Preparation of 3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylaniline

The synthetic route of intermediate A7 is as follows:

Step 1: Synthesis of 3-bromo-2-fluoro-5-methylbenzamide (A7-2)

3-Bromo-2-fluoro-5-methylbenzoic acid (A7-1) (1 g, 4.3 mmol) was dissolved in dry N,N-dimethylformamide (10 mL), and ammonium chloride (690 mg, 12.9 mmol), N,N-diisopropylethylamine (1.67 g, 12.9 mmol) and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (2.47 g, 6.5 mmol) were added in sequence. The reaction was stirred at room temperature for 20 h, and water (30 mL) was added to dilute, and ethyl acetate was extracted (30 mL*3). The solvent was concentrated, and the residue was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 10:1) to obtain 3-bromo-2-fluoro-5-methylbenzamide (A7-2) (930 mg, yield 93.4%).

Step 2: Synthesis of 3-bromo-2-fluoro-5-methylaniline (A7-3)

Sodium hydroxide (1.6 g, 40 mmol) was dissolved in water (20 mL), cooled to 0-5°C, bromine (1.09 g, 6.8 mmol) was added dropwise to form a yellow clear liquid, 3-bromo-2-fluoro-5-methylbenzamide (A7-2) (930 mg, 4.0 mmol) was added, and the temperature was raised to 85°C for reaction for 2 h, cooled to room temperature, extracted with dichloromethane (20 ml*3), the organic phases were combined, the solvent was concentrated, and the mixture was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 20:1) to obtain 3-bromo-2-fluoro-5-methylaniline (A7-3) (600 mg, yield 73.4%).

Step 3: Synthesis of tert-butyl (3-bromo-2-fluoro-5-methylphenyl)carbamate (A7-4)

3-Bromo-2-fluoro-5-methylaniline (A7-3) (600 mg, 2.94 mmol) was dissolved in a sodium hydroxide aqueous solution (2N, 10 mL, 20 mmol), di-tert-butyl dicarbonate (1.02 g, 4.7 mmol) was added, the temperature was raised to 100 ° C and reacted for 20 h, cooled to room temperature, ethyl acetate was added for extraction (10 mL*3), the organic phases were combined and concentrated, and the residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 20:1) to obtain tert-butyl (3-bromo-2-fluoro-5-methylphenyl)carbamate (A7-4) (440 mg, yield 49.2%)

Step 4: Synthesis of tert-butyl (3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylphenyl)carbamate (A7-5)

Tert-butyl (3-bromo-2-fluoro-5-methylphenyl)carbamate (A7-4) (300 mg, 0.99 mmol) was dissolved in dry 1,4-dioxane (5 mL), and 4,4-difluoropiperidine hydrochloride (233 mg, 1.48 mmol), methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl) palladium (II) (82.6 mg, 0.1 mmol), 2-dicyclohexylphosphino-2',6'-diisopropoxybiphenyl (88.6 mg, 0.19 mmol) and cesium carbonate (965 mg, 2.96 mmol) were added in sequence. The mixture was replaced with argon three times, and the temperature was raised to 100°C under argon protection for 18 h. The reaction solution was quenched by adding water (50 mL), the layers were separated, the aqueous phase was extracted with ethyl acetate (30 mL*3), the organic phases were combined and concentrated, and the residue was separated and purified by silica gel column (petroleum ether: ethyl acetate (V/V) = 20:1) to obtain tert-butyl (3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylphenyl)carbamate (A7-5) (180 mg, yield 53.1%).

Step 5: Synthesis of 3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylaniline (A7)

To tert-butyl (3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylphenyl)carbamate (A7-5) (180 mg, 0.52 mmol) was added a solution of hydrochloric acid in 1,4-dioxane (4N, 2 mL, 8 mmol), the reaction was stirred at room temperature for 1 h, and the solvent was concentrated to give 3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylaniline (A7) (130 mg, yield 100%).

Intermediate A8: Preparation of 3-chloro-5-(4,4-difluoropiperidin-1-yl)aniline

The synthetic route of intermediate A8 is as follows:

Step 1: Synthesis of 1-(3-chloro-5-nitrophenyl)-4,4-difluoropiperidine (A8-2)

1-Chloro-3-fluoro-5-nitrobenzene (A8-1) (1 g, 5.7 mmol) was dissolved in N, N-dimethylformamide (10 mL), and then 4,4-difluoropiperidine hydrochloride (1.38 g, 11.4 mmol) and potassium carbonate (3.15 g, 22.8 mmol) were added and reacted at 120 ° C for 2 h under microwave conditions. The reaction solution was poured into water (100 mL), extracted with ethyl acetate, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V) = 1/0-1/1) to obtain 1-(3-chloro-5-nitrophenyl)-4,4-difluoropiperidine (A8-2) (0.5 g, yield 31.7%).

Step 2: Synthesis of 3-chloro-5-(4,4-difluoropiperidin-1-yl)aniline (A8)

Iron powder (0.5 g, 9 mmol) and ammonium chloride (0.96 g, 18 mmol) were added to a solution of 1-(3-chloro-5-nitrophenyl)-4,4-difluoropiperidine (A8-2) (0.5 g, 1.8 mmol) in ethanol (4 mL) and water (1 mL), and the mixture was reacted at 85°C for 2 h. The reaction solution was poured into water (50 mL) and filtered, and the filtrate was extracted with ethyl acetate (20 mL*3). The organic phase was washed with saturated brine (20 mL*3), dried over anhydrous sodium sulfate, filtered and concentrated to give 3-chloro-5-(4,4-difluoropiperidin-1-yl)aniline (A8) (0.42 g, yield 94.2%).

LC-MS, M/Z(ESI):247.4[M+H] ⁺

Example 1: Preparation of N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-5-(2-hydroxyethylsulfonamide)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 1) The synthetic route of the target compound 1 is as follows:

Step 1: Synthesis of 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)phenyl-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B1-2)

5-Bromo-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylic acid (400 mg, 1.285 mmol) was dissolved in dichloromethane (5 mL) and N,N-dimethylformamide (0.1 mL), and thionyl chloride (0.469 mL, 6.43 mmol) was slowly added dropwise at 0°C, and then stirred at 25°C for 0.5 h. The reaction solution was spin-dried, and dichloromethane (5 mL) was added to dissolve the residue, and then N,N-diisopropylethylamine (1.123 mL, 6.43 mmol), potassium phosphate (1364 mg, 6.43 mmol) and 3-(4,4-difluoropiperidin-1-yl)aniline (273 mg, 1.285 mmol) were added to the reaction solution, and stirred at 25°C for 3 h. The reaction solution was poured into water (30 mL) and extracted with dichloromethane (20 mL*3). The organic phase was washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, filtered, and spin-dried to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V) = 1/0-0/1) to obtain compound 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)phenyl-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B1-2) (yellow solid, 450 mg, yield 69.3%).

LC-MS, M/Z(ESI):505.1[M+H] ⁺ .

Step 2: Synthesis of N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-5-(2-hydroxyethylsulfonamide)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 1)

To a solution of 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)phenyl-3(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (350 mg, 0.693 mmol) in N,N-dimethylformamide (5 mL) were added 2-hydroxyethanesulfonamide (130 mg, 1.039 mmol), N,N-dimethylglycine (14.28 mg, 0.139 mmol), cuprous iodide (13.19 mg, 0.069 mmol) and potassium carbonate (191 mg, 1.385 mmol). The reaction solution was reacted at 130°C for 18 h. The reaction solution was added with water (50 mL) and extracted with ethyl acetate (20 mL*3). The organic phase was washed with saturated brine (10 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was subjected to reverse phase preparation (column: Phenomenex Synergi C ₁₈ 100*25mm*4μm; solvent: A=water+0.1 volume % formic acid (99%), B=acetonitrile; gradient: 5%-95%, 7 minutes) to obtain compound N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-5-(2-hydroxyethylsulfonamide)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 1) (174.8 mg, yield 44.4%).

1H NMR (400MHz, DMSO-d6) δ10.52(s,1H),7.91(s,1H),7.40(s,1H),7.31-7.22(m,2H),7.18(t,J＝8.1Hz, 1H),6.75-6.67(m,1H),3.75(t,J＝6.5Hz,2H),3.32(s,4H),3.19(t,J＝6.5Hz,2H),3.03-2.95(m,4H ),2.13-1.97(m,4H),1.46(s,4H),0.31(s,4H).

LC-MS, M/Z(ESI):550.3[M+H] ⁺ .

Example 2: Preparation of N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-6-(2-hydroxyethylsulfonamide)-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxamide (Compound 2) The synthetic route of target compound 2 is as follows:

Step 1: Synthesis of ethyl 6-chloro-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxylate (B2-2)

6-Azaspiro [2.5] octane hydrochloride (302 mg, 2.71 mmol) and N, N-diisopropylethylamine (585 mg, 4.52 mmol) were added to a solution of 4,6-dichloropyridazine-3-carboxylic acid ethyl ester (500 mg, 2.262 mmol) in acetonitrile (5 mL), and stirred at 25 ° C for 18 h. Water (50 mL) was added to the mixture and extracted with ethyl acetate (20 mL * 3). The organic layer was washed with saturated brine (20 mL * 2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate (V / V) = 1 / 0-1 / 1) to give compound 6-chloro-4- (6-azaspiro [2.5] octan-6-yl) pyridazine-3-carboxylic acid ethyl ester (B2-2) (colorless oil, 430 mg, yield 64.3%).

LC-MS, M/Z(ESI): 296.2[M+H] ⁺ .

Step 2: Synthesis of 6-chloro-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxylic acid (B2-3)

Ethyl 6-chloro-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxylate (430 mg, 1.454 mmol) was dissolved in tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), and then lithium hydroxide (174 mg, 7.27 mmol) was added. The reaction solution was stirred at 25 ° C for 18 h. Water (50 mL) was added to the reaction solution, and the pH was adjusted to 4-5 with 1N hydrochloric acid aqueous solution, and then lyophilized. The solid was dissolved in ethyl acetate (50 mL), filtered, and the filtrate was dried to obtain compound 6-chloro-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxylic acid (B2-3) (white solid, 380 mg, yield 98%).

LC-MS, M/Z(ESI):268.2[M+H] ⁺ .

Step 3: Synthesis of 6-chloro-N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-4-(6-azaspiro[2.5]oct-6-yl)pyridazine-3-carboxamide (B2-4)

To a solution of 6-chloro-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxylic acid (370 mg, 1.382 mmol) in dichloromethane (5 mL) was added O-(7-nitrobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1577 mg, 4.15 mmol) and N,N-diisopropylethylamine (536 mg, 4.15 mmol). The reaction solution was stirred at 25 ° C for 0.5 h and 3-(4,4-difluoropiperidin-1-yl)aniline (352 mg, 1.659 mmol) was added. The reaction solution was then stirred at 25 ° C for 18 h. Water (50 mL) was added to the reaction solution and extracted with dichloromethane (10 mL*3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V) = 1/0-1/1) to give compound 6-chloro-N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxamide (B2-4) (white solid, 400 mg, yield 62.7%).

LC-MS, M/Z(ESI): 462.2[M+H] ⁺ .

Step 4: Synthesis of N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-6-(2-hydroxyethylsulfonamide)-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxamide (Compound 2)

2-Hydroxyethanesulfonamide (171 mg, 1.364 mmol), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (88 mg, 0.152 mmol), potassium carbonate (262 mg, 1.894 mmol) and bis(dibenzylideneacetone)palladium (43.6 mg, 0.076 mmol) were added to a solution of 6-chloro-N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxamide (350 mg, 0.758 mmol) in 1,4-dioxane (3 mL). The reaction solution was replaced with N ₂ three times and reacted at 90° C. for 18 h under a nitrogen atmosphere. The reaction solution was concentrated to obtain a crude product. The crude product was subjected to reverse phase preparation (column: Phenomenex Synergi C ₁₈ 100*25mm*4μm; solvent: A=water+0.1 volume% formic acid (99%), B=acetonitrile; gradient: 5%-95%, 7 minutes) to obtain compound N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-6-(2-hydroxyethylsulfonamide)-4-(6-azaspiro[2.5]octan-6-yl)pyridazine-3-carboxamide (compound 2) (28.1 mg, yield 6.7%).

1H NMR (400MHz, DMSO-d6) δ10.38(s,1H),7.40(s,1H),7.25(d,J＝8.0Hz,1H),7.18(t,J＝8.1Hz,1H),6.75 (d,J＝8.2Hz,1H),6.49(s,1H),3.72(t,J＝6.8Hz,2H),3.30(s,5H),3.20(dd,J＝16.4,9.2Hz,5H) ,2.05(dt,J＝19.8,7.2Hz,4H),1.42(d,J＝4.6Hz,4H),0.31(s,4H).

LC-MS, M/Z(ESI):551.2[M+H] ⁺ .

Example 3: Preparation of N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-2-(2-hydroxyethylsulfonamide)-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxamide (Compound 3) The synthetic route of target compound 3 is as follows:

Step 1: Synthesis of methyl 2-chloro-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxylate (B3-2)

6-Azaspiro [2.5] octane hydrochloride (322 mg, 2.90 mmol) and N, N-diisopropylethylamine (624 mg, 4.83 mmol) were added to a solution of 2,4-dichloropyrimidine-5-carboxylic acid methyl ester (500 mg, 2.415 mmol) in acetonitrile (5 mL), and stirred at 25 ° C for 18 h. Water (50 mL) was added to the mixture and extracted with ethyl acetate (20 mL * 3). The organic layer was washed with saturated brine (20 mL * 2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product. The crude product was purified by column chromatography (petroleum ether / ethyl acetate (V / V) = 1 / 0-1 / 1) to give the compound 2-chloro-4- (6-azaspiro [2.5] octan-6-yl) pyrimidine-3-carboxylic acid methyl ester (colorless oil, 450 mg, yield 66.1%).

LC-MS, M/Z(ESI):282.0[M+H] ⁺ .

Step 2: Synthesis of 2-chloro-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxylic acid (B3-3)

2-Chloro-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-3-carboxylic acid methyl ester (420 mg, 1.491 mmol) was dissolved in tetrahydrofuran (3 mL), methanol (1 mL) and water (1 mL), and then lithium hydroxide (179 mg, 7.45 mmol) was added. The reaction solution was stirred at 25 ° C for 18 h. Water (10 mL) was added to the reaction solution, and the pH was adjusted to 4-5 with 1N hydrochloric acid aqueous solution, and then lyophilized. The solid was dissolved in ethyl acetate (50 mL), filtered, and the filtrate was dried to obtain compound 2-chloro-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxylic acid (white solid, 390 mg, yield 98%).

LC-MS, M/Z(ESI):268.2[M+H] ⁺ .

Step 3: Synthesis of 2-chloro-N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxamide (B3-4)

To a solution of 2-chloro-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxylic acid (390 mg, 1.457 mmol) in dichloromethane (5 mL) was added O-(7-nitrobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1662 mg, 4.37 mmol) and N,N-diisopropylethylamine (565 mg, 4.37 mmol). The reaction solution was stirred at 25 ° C for 0.5 h and 3-(4,4-difluoropiperidin-1-yl)aniline (464 mg, 2.185 mmol) was added. The reaction solution was then stirred at 25 ° C for 18 h. Water (50 mL) was added to the reaction solution and extracted with dichloromethane (10 mL*3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V) = 1/0-1/1) to give compound 2-chloro-N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxamide (white solid, 350 mg, yield 52%).

LC-MS, M/Z(ESI): 462.2[M+H] ⁺ .

Step 4: Synthesis of N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-2-(2-hydroxyethylsulfonamide)-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxamide (Compound 3)

2-Hydroxyethanesulfonamide (171 mg, 1.364 mmol), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (88 mg, 0.152 mmol), potassium carbonate (262 mg, 1.894 mmol) and bis(dibenzylideneacetone)palladium (43.6 mg, 0.076 mmol) were added to a solution of 2-chloro-N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxamide (350 mg, 0.758 mmol) in 1,4-dioxane (3 mL), the reaction solution was replaced with N ₂ three times, and then reacted at 90° C. for 18 h. The reaction solution was concentrated to obtain a crude product. The crude product was subjected to reverse phase preparation (column: Phenomenex Synergi C ₁₈ 100*25mm*4μm; solvent: A=water+0.1 volume% formic acid (99%), B=acetonitrile; gradient: 5%-95%, 7 minutes) to obtain compound N-(3-(4,4-difluoropiperidin-1-yl)phenyl)-2-(2-hydroxyethylsulfonamide)-4-(6-azaspiro[2.5]octan-6-yl)pyrimidine-5-carboxamide (compound 3) (165.7 mg, yield 38.7%).

¹ H NMR (400MHz, DMSO-d6) δ10.13(s,1H),8.07(s,1H),7.38(s,1H),7.19-7.07(m,2H),6.72(d,J＝7.0Hz ,1H),3.71(t,J＝7.0Hz,2H),3.62-3.47(m,4H),3.42(t,J＝7.0Hz,2H),3.36-3.23(m,4H),2.05(ddd, J＝19.7,13.9,5.6Hz,4H),1.44-1.27(m,4H),0.31(s,4H).

LC-MS, M/Z(ESI):551.4[M+H] ⁺ .

Example 4: Preparation of N-(3-(4,4-difluoropiperidin-1-yl)-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]-6-yl)pyridine-2-carboxamide (Compound 4)

The synthetic route of target compound 4 is as follows:

Step 1: 5-Bromo-N-(3-(4,4-difluoropiperidin-1-yl)-5-methylphenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylate

Synthesis of amide (B4-1)

5-Bromo-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylic acid (500 mg, 1.61 mmol) was dissolved in thionyl chloride (5 mL), the temperature was raised to 70 °C and the reaction was carried out for 2 h. The solvent was concentrated, dry dichloromethane (10 mL) was added to the residue, and 3-(4,4-difluoropiperidin-1-yl)-5-methylaniline (A3) (545 mg, 2.41 mmol) and N,N-diisopropylethylamine (1.03 g, 7.98 mmol) were added in sequence, and the reaction was stirred at room temperature for 3 h. The reaction solution was quenched by adding water (50 mL), the liquids were separated, the aqueous phase was extracted with dichloromethane (30 mL*2), the organic phases were combined and concentrated, and the residue was separated and purified on a silica gel plate (petroleum ether: ethyl acetate (V/V) = 3:1) to obtain 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)-5-methylphenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B4-1) (430 mg, yield 51.5%).

LC-MS, M/Z(ESI): 519.2[M+H] ⁺ .

Step 2: Synthesis of N-(3-(4,4-difluoropiperidin-1-yl)-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 4)

5-Bromo-N-(3-(4,4-difluoropiperidin-1-yl)-5-methylphenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B4-1) (130 mg, 0.25 mmol) was dissolved in dry N,N-dimethylformamide (5 mL), and 2-hydroxyethane-1-sulfonamide (63 mg, 0.50 mmol), cuprous iodide (24 mg, 0.13 mmol), sarcosine (22 mg, 0.25 mmol) and potassium carbonate (104 mg, 0.75 mmol) were added in sequence. The mixture was replaced with argon three times and the temperature was raised to 130 °C under argon protection for 3 h. The solvent of the reaction solution was concentrated, and the residue was separated by high pressure preparative chromatography and freeze-dried to give N-(3-(4,4-difluoropiperidin-1-yl)-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 4) (40.2 mg, yield 28.5%).

¹ H NMR (400MHz, DMSO-d6): δ10.18(s,1H),10.14(s,1H),8.03(brs,1H),7.34(s,1H),7.20-7.07(m,2H), 6.57(s,1H),3.77(t,2H),3.34-3.16(m,10H),2.24(s,3H),2.07-1.98(m,4H),1.44-1.42(m,4H),0.30( s,4H).

LC-MS, M/Z(ESI): 564.4[M+H] ⁺ .

Example 5: Preparation of N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 40)

The synthetic route of target compound 40 is as follows:

Step 1: Synthesis of 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)-5-fluorophenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B40-1)

5-Bromo-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylic acid (342 mg, 1.1 mmol) was dissolved in thionyl chloride (3 mL), the temperature was raised to 70 °C and the reaction was carried out for 2 h. The solvent was concentrated, dry dichloromethane (10 mL) was added to the residue, and 3-(4,4-difluoropiperidin-1-yl)-5-fluoroaniline (A4) (385 mg, 1.7 mmol) and N,N-diisopropylethylamine (737 mg, 5.7 mmol) were added in sequence, and the reaction was stirred at room temperature for 18 h. The reaction solution was quenched by adding water (50 mL), the liquids were separated, the aqueous phase was extracted with dichloromethane (30 mL*2), the organic phases were combined and concentrated, and the residue was separated and purified on a silica gel plate (petroleum ether: ethyl acetate (V/V) = 5:1) to obtain 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)-5-fluorophenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B40-1) (280 mg, yield 32.0%).

LC-MS, M/Z(ESI): 523.2[M+H] ⁺ .

Step 2: Synthesis of N-(3-(4,4-difluoropiperidin-1-yl)-5-fluorophenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 40)

5-Bromo-N-(3-(4,4-difluoropiperidin-1-yl)-5-fluorophenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B40-1) (280 mg, 0.5 mmol) was dissolved in dry N,N-dimethylformamide (10 mL), and 2-hydroxyethane-1-sulfonamide (188 mg, 1.5 mmol), cuprous iodide (57.1 mg, 0.3 mmol), N,N-dimethylglycine (51.6 mg, 0.5 mmol) and potassium carbonate (207 mg, 1.5 mmol) were added in sequence. The mixture was replaced with argon three times and the temperature was raised to 130 °C under argon protection for 3 h. The solvent of the reaction solution was concentrated, and the residue was separated by high pressure preparative chromatography and freeze-dried to give N-(3-(4,4-difluoropiperidin-1-yl)-5-fluorophenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 40) (136 mg, yield 45.9%).

¹ H NMR (400MHz, DMSO-d6) δ10.39(s,1H),10.16(s,1H),8.03(s,1H),7.33(s,1H),7.21(d,1H),7.12(s ,1H),6.58(d,1H),4.97(s,1H),3.77(t,2H),3.34(d,6H),3.05(s,4H),2.12-1.93(m,4H),1.43( s,4H),0.30(s,4H).

LC-MS, M/Z(ESI): 568.4[M+H] ⁺ .

Example 6: Preparation of N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 41) The synthetic route of the target compound 41 is as follows:

Step 1: Synthesis of 5-bromo-N-(3-cyano-5-(4,4-difluoropiperidin-1-yl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B41-1)

5-Bromo-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylic acid (A2) (285 mg, 0.8 mmol) was added to dichloromethane (10 mL), and dichlorothionyl (2.85 mL) and two drops of N,N-dimethylformamide were added dropwise. The mixture was stirred at room temperature for 2 h, and then concentrated to dryness to obtain a crude product. The crude product was dissolved in dichloromethane (10 mL), and 3-amino-5-(4,4-difluoropiperidin-1-yl)benzonitrile (200 mg, 0.8 mmol) and N,N-diisopropylethylamine (2 mL) were added, and the reaction was carried out at room temperature for 14 h. Water (30 mL) was added to dilute, and the mixture was extracted with dichloromethane (10 mL×2). The organic phase was concentrated to dryness, and the residue was separated and purified by silica gel column (petroleum ether: ethyl acetate = 10:1) to obtain 5-bromo-N-(3-cyano-5-(4,4-difluoropiperidin-1-yl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B41-1) (150 mg, yield 51%).

LC-MS, M/Z(ESI):578.4[M+H] ⁺ .

Step 2: Synthesis of N-(3-cyano-5-(4,4-difluoropiperidin-1-yl)phenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 41)

5-Bromo-N-(3-cyano-5-(4,4-difluoropiperidin-1-yl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B41-1) (150 mg, 0.26 mmol) was added to 2 mL of N,N-dimethylformamide, and then 2-hydroxyethane-1-sulfonamide (72 mg, 0.58 mmol), N-methylglycine (10 mg, 0.10 mmol), cuprous iodide (11 mg, 0.05 mmol), and potassium carbonate (160 mg, 1.16 mmol) were added, and the mixture was heated to 120°C for 1 h. Water (10 mL) was added to dilute, and the mixture was extracted with ethyl acetate (10 mL×2). The organic phase was concentrated to dryness, and the residue was separated and purified by silica gel column (petroleum ether: ethyl acetate = 1:1) to obtain N-(3-cyano-5-(4,4-difluoropiperidin-1-yl)phenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)picolinamide (Compound 41) (50 mg, yield 30%).

¹ H NMR (400MHz, DMSO-d6) δ10.55(s,1H),10.21(s,1H),8.05(s,1H),7.74(s,1H),7.61(s,1H),7.36(s ,1H),7.20(s,1H),3.88(m,2H),3.44(m,7H),3.07(m,4H),2.06(m,4H),1.49(m,4H),0.31(s, 4H).

LC-MS, M/Z(ESI):575.2[M+H] ⁺ .

Example 7: Preparation of N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 50) The synthetic route of the target compound 50 is as follows:

Step 1: Synthesis of 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B50-1)

5-Bromo-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylic acid (300 mg, 1.0 mmol) was dissolved in thionyl chloride (3 mL), the temperature was raised to 70 °C and the reaction was carried out for 2 h. The solvent was concentrated, dry dichloromethane (10 mL) was added to the residue, and 3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylaniline (A6) (366 mg, 1.5 mmol) and N,N-diisopropylethylamine (646 mg, 5 mmol) were added in sequence, and the reaction was stirred at room temperature for 18 h. The reaction solution was quenched by adding water (50 mL), the liquids were separated, the aqueous phase was extracted with dichloromethane (30 mL*2), the organic phases were combined and concentrated, and the residue was separated and purified on a silica gel plate (petroleum ether: ethyl acetate (V/V) = 3:1) to obtain 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B50-1) (230 mg, yield 44.4%).

LC-MS, M/Z(ESI): 537.2[M+H] ⁺ .

Step 2: Synthesis of N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 50)

5-Bromo-N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B50-1) (230 mg, 0.4 mmol) was dissolved in dry N,N-dimethylformamide (6 mL), and 2-hydroxyethane-1-sulfonamide (150 mg, 1.2 mmol), cuprous iodide (38.1 mg, 0.2 mmol), N,N-dimethylglycine (41.2 mg, 0.4 mmol) and potassium carbonate (166 mg, 1.2 mmol) were added in sequence. The mixture was replaced with argon three times and the temperature was raised to 130 °C under argon protection for 3 h. The solvent of the reaction solution was concentrated, and the residue was separated by high pressure preparative chromatography and freeze-dried to give N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 50) (26.6 mg, yield: 10.7%).

¹ H NMR (400MHz, DMSO-d6) δ10.29(s,1H),7.99(s,1H),7.37(d,1H),7.29(d,2H),3.76(t,2H),3.29-3.20 (m,2H),3.11(s,4H),3.04(s,4H),2.20(s,3H),2.11(t,4H),1.43(s,4H),0.30(s,4H).

LC-MS, M/Z(ESI): 582.4[M+H] ⁺ .

Example 8: Preparation of N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 51) The synthetic route of the target compound 51 is as follows:

Step 1: Synthesis of 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylphenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B51-1)

5-Bromo-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylic acid (110 mg, 0.353 mmol) was dissolved in thionyl chloride (2 mL), the temperature was raised to 70 °C and the reaction was carried out for 2 h. The solvent was concentrated, dry dichloromethane (10 mL) was added to the residue, and 3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylaniline (A5) (130 mg, 0.53 mmol) and N,N-diisopropylethylamine (291 mg, 2.26 mmol) were added in sequence, and the reaction was stirred at room temperature for 18 h. The reaction solution was quenched by adding water (50 mL), the liquids were separated, the aqueous phase was extracted with dichloromethane (30 mL*2), the organic phases were combined and concentrated, and the residue was separated and purified on a silica gel plate (petroleum ether: ethyl acetate (V/V) = 5:1) to give 5-bromo-N-(3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylphenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B51-1) (90 mg, yield 47.4%).

LC-MS, M/Z(ESI): 537.2[M+H] ⁺ .

Step 2: Synthesis of N-(3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 51)

5-Bromo-N-(3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylphenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B51-1) (90 mg, 0.17 mmol) was dissolved in dry N,N-dimethylformamide (3 mL), and 2-hydroxyethane-1-sulfonamide (63 mg, 0.5 mmol), cuprous iodide (15.9 mg, 0.08 mmol), N,N-dimethylglycine (17.3 mg, 0.17 mmol) and potassium carbonate (69.3 mg, 0.5 mmol) were added in sequence. The mixture was replaced with argon three times and the temperature was raised to 130 °C under argon protection for 3 h. The reaction solution was concentrated to dryness and the residue was separated by high pressure preparative chromatography and freeze-dried to give N-(3-(4,4-difluoropiperidin-1-yl)-2-fluoro-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 51) (13 mg, yield 13.3%).

¹ H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.04(s,1H),7.58(s,1H),7.37(s,1H),6.70(d,1H),3.76(t ,2H),3.35(t,2H),3.12(s,4H),3.04(s,4H),2.26(s,3H),2.10(m,4H),1.47(s,4H),0.32(s, 4H).

LC-MS, M/Z(ESI): 582.4[M+H] ⁺ .

Example 9: Preparation of N-(3-(4,4-difluoropiperidin-1-yl)-4-fluoro-5-methylphenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 52) The synthetic route of the target compound 52 is as follows:

Step 1: Synthesis of 5-bromo-N-(3-chloro-5-(4,4-difluoropiperidin-1-yl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B52-1)

5-Bromo-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxylic acid (A3) (249 mg, 0.8 mmol) was added to thionyl chloride (1 mL) and stirred at 70°C for 2 h. The reaction solution was concentrated and dissolved in dichloromethane (4 mL), and then N,N-diisopropylethylamine (620 mg, 4.8 mmol) and 3-chloro-5-(4,4-difluoropiperidin-1-yl)aniline (A8) (400 mg, 1.6 mmol) were added and stirred at room temperature for 18 h. The reaction solution was concentrated to obtain a crude product. The crude product was purified by column chromatography (petroleum ether/ethyl acetate (V/V) = 1/0-1/1) to give 5-bromo-N-(3-chloro-5-(4,4-difluoropiperidin-1-yl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B52-1) (260 mg, yield 29.7%).

LC-MS, M/Z(ESI): 539.3[M+H] ⁺ .

Step 2: Synthesis of N-(3-chloro-5-(4,4-difluoropiperidin-1-yl)phenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 52)

2-(Dimethylamino)acetic acid (10 mg, 0.1 mmol), cuprous iodide (10 mg, 0.05 mmol) and potassium carbonate (138 mg, 1 mmol) were added to a solution of 5-bromo-N-(3-chloro-5-(4,4-difluoropiperidin-1-yl)phenyl)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (B52-1) (260 mg, 0.5 mmol) and 2-hydroxyethanesulfonamide (100 mg, 0.8 mmol) in N,N-dimethylformamide (1 mL), and the mixture was reacted at 90°C for 3 h. The reaction solution was poured into water (20 mL) and extracted with ethyl acetate (10 mL*3). The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was subjected to reverse phase preparation (column: Phenomenex Synergi C18 100*25mm*4μm; solvent: A=water+0.1 volume% formic acid (99%), B=acetonitrile; gradient: 5%-95%, 7 minutes) to give N-(3-chloro-5-(4,4-difluoropiperidin-1-yl)phenyl)-5-((2-hydroxyethyl)sulfonylamino)-3-(6-azaspiro[2.5]octan-6-yl)pyridine-2-carboxamide (Compound 52) (55.4 mg, yield 19.7%).

¹ H NMR (400MHz, DMSO-d6) δ10.39(s,1H),10.18(s,1H),8.04(s,1H),7.46(s,1H),7.34(s,1H),7.26(s ,1H),6.79(s,1H),3.78(t,2H),3.36(dd,7H),3.15-2.95(m,4H),2.11-1.96(m,4H),1.56-1.35(m,4H ),0.31(s,4H).

LC-MS, M/Z(ESI):584.1[M+H] ⁺ .

The following target compounds were prepared similarly to the synthesis method of compound 1.

Test Example 1. Compound inhibition of tumor cell OVCAR3 cell proliferation test

NIH:OVCAR-3 (ATCC, Cat No.HTB-161, purchased from Co-Bier) cells were seeded in T75 culture flasks and cultured in RPMI 1640 medium containing 20% FBS for 3 days in preparation for subsequent culture and seeding in 96-well plates for cell proliferation assays.

OVCAR3 cells were inoculated at a density of 3000 cells/100 μL/well in a 96-well cell plate, and the cell plate was placed in an incubator for 18 hours (37°C, 5% CO ₂ ). On the second day, drug treatment was started. 100 μL/well gradient dilutions of the compound to be tested (the starting concentration of each drug was 10 μM, the diluent was DMSO, the dilution ratio was 1:3, and each drug was diluted to nine gradient points) or DMSO (negative control) were added to the culture medium of the culture plate. A blank group was set up in which no cell line was inoculated but only the culture medium was added. After adding the drug, the culture plate was placed in an incubator for another 3 days (37°C, 5% CO ₂ ). On the fourth day, 100 μl of CellTiter-Glo (Promega G9243) reagent was added to each well, shaken for 5 minutes, and then allowed to stand at room temperature for 5 minutes. The plate was analyzed using an ELISA reader (PerkinElmer 2104) The chemiluminescent signal value of each well was measured. Cell proliferation inhibition rate = (1-(compound group-blank group)/(DMSO group-blank group)) × 100%, and the IC ₅₀ value of each compound was calculated using GraphPad Prism8 software.

Table 1. Inhibitory effects of compounds on tumor cell proliferation

Compound No. IC ₅₀ (nM) Compound 41 650 Compound 50 256 Compound 51 150 Compound 52 230

The experimental results show that the OVCAR3 cell proliferation test results show that the compounds of the present invention can significantly inhibit the proliferation of OVCAR3 cells.

Test Example 2. Compound Inhibition Test on KIF18A Enzyme Activity

The recombinant protein KIF18A (kinesin domain: aa 1-467) used in this test was expressed by Invitrogen's Bac-to-Bac baculovirus expression system, and the resulting protein was purified and used in this test (Seki, M. et al. 2003 Nucleic Acids Res.). The quantitative detection of KIF18A enzyme activity was completed using the ADP-Glo ^TM Kinase Assay Kit (Promega Inc). The relevant operations strictly followed the product instructions and are briefly described as follows:

The reaction system consists of the test compound, recombinant protein KIF18A (aa 1-467), ATP (Promega Inc) and experimental buffer. The test compound was prepared into a 0.5mM stock solution with DMSO (Sigma Inc) and gradient dilution was performed using DMSO. The experimental buffer consists of 15mM Tris, 10mM MgCl ₂ (Sigma Inc), 0.01% Pluronic F-68 (Life Technologies Inc), 1μM paclitaxel (Cytoskeleton Inc), 30μg/mL porcine tubulin (Cytoskeleton Inc) and 2% DMSO in water. KIF18A protein (final concentration of 80nM) and different concentrations of compounds (1μL) were added to the prepared experimental buffer (50μL) and incubated at room temperature for 15min; ATP (final concentration of 80μΜ) was then added to the reaction mixture and incubated at room temperature for 3hr. After the reaction, 5 μL of ADP-Glo ^TM reagent and 2.5 μL of reaction mixture were added to a 384-well plate (Grenier Inc), mixed well, sealed with aluminum foil sealing film and incubated at room temperature in the dark for 40 min; Finally, 10 μL of ADP-Glo ^TM detection reagent was added to each reaction well and incubated at room temperature in the dark for 40 min. After all reactions were completed, the luminescence value of each well was read using an ELISA reader (Molecular Device_SpectraMax Id5), and the inhibition rate was calculated. The inhibition rate calculation formula is as follows: inhibition rate = (1-(compound group-blank group)/(negative control group-blank group)) × 100%, and the IC50 value of the compound to be tested was calculated in GraphPad Prism8 software according to the inhibition rate of compounds at different concentrations.

Table 2. Inhibitory effects of compounds on KIF18A enzyme activity

Test compound IC ₅₀ (μM) Compound 41 0.002 Compound 50 0.012 Compound 51 0.018 Compound 52 0.003

The experimental results show that: the KIF18A enzyme activity inhibition experiment shows that the compound of the present invention has a good inhibitory activity on the KIF18A enzyme.

Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims (36)

1. A compound as shown in formula I, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug: in, X ⁴ is N or -CR ⁸ ; X ⁵ is N or -CR ⁹ ; X ⁶ is N or -CR ¹⁰ ; and When ^X5 is N, ^X4 must be ^CR8 ; when ^X5 is ^CR9 , ^X4 and ^X6 must be N at the same time; L is -NR ³ -C(=O)- or -C(=O)-NR ³ -; M is absent or selected from C _1-6 alkylene substituted by R ^c ; R ¹ is selected from CN or a group -ZR ¹² , wherein Z is independently selected from -C _0-6 alkyl-, -C═N(OH)-, -C _0-6 alkyl-SC _0-6 alkyl-, -C _0-6 alkyl-S(═O)-C _0-6 alkyl-, -C _0-6 alkyl-SO ₂ -C _0-6 alkyl-, -C _0-6 alkyl-NR ¹¹ -C _0-6 alkyl-, -C _0-6 alkyl-NR ¹¹ SO ₂ -C _0-6 alkyl-, -C _0-6 alkyl-SO ₂ NR ¹¹ -C _0-6 alkyl-, -C _0-6 alkyl-NR ¹¹ SO ₂ NR ¹¹ -C _0-6 alkyl-, -C _0-6 alkyl-OC _0-6 alkyl-, -C _0-6 alkyl-C(═O)-C _0-6 alkyl-, -C -C _0-6 alkyl-C(═O)—OC _0-6 alkyl-, -C _0-6 alkyl-(C═O)NR ¹¹ -C _0-6 alkyl-, _{-C 0-6} alkyl-NR ¹¹ (C═O)-C _0-6 alkyl-, -C 0-6 alkyl-S(═O)(═NH)-C _0-6 alkyl-, -C-((C═O)-OR ¹¹ ) ₂ -, -C _0-6 alkyl-NR ¹¹ -S(═O)(═NH)-C _{0-6 alkyl-, -C 0-6 alkyl} -S(═O)(═N ⁺ (CH ₃ ) ₂ )-, -NR ¹¹ SO ₂ NR ¹¹ -C(═O)—O-, -P-, -C _0-6 alkyl-P(═O)(R ¹¹ )-, -C _0-6 alkyl-P(═O) ₂ ; or the group -ZR ¹² is -N=S(=O)-(R ¹² ) ₂ , wherein two R ¹² pairs can be alternatively combined with the sulfur atoms to which they are respectively attached to form a saturated or partially saturated 3-, 4-, 5- or 6-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S; ^R2 is halogen or a group ^-YR13 , wherein Y is _-C0-6alkyl- , _-C0-6alkyl -S(=O)(=NH)-C0-6alkyl-, _{-C0-6alkyl -} SC0-6alkyl-, -C0-6alkyl- _S (=O)-C0-6alkyl- ^, _{-C0-6alkyl - SO2} - _C0-6alkyl- , _-C0-6alkyl - _NR13 -C0-6alkyl-, _-C0-6alkyl -NR13SO2 _{- C0-6alkyl-} ^{, -C0-6alkyl} - _SO2NR13 - ^C0-6alkyl- , _{-C0-6alkyl - NR13SO2NR13} - _C0-6alkyl- , _{-C0-6alkyl - OC0-6alkyl-} ^, _-C0-6alkyl ^- _C (= _O )-C0-6alkyl -YR ¹³ is -N _{= S} (=O)-(R ₁₃ ) 2 , wherein two _{R 13} pairs _can be alternatively combined with the sulfur atoms to which they _are respectively attached to form a saturated or partially saturated ₃ -membered, _4- membered, ^{5 -} membered or 6 _- membered monocyclic ring containing 0, ¹ , 2 or ₃ N atoms and 0, 1 or 2 atoms ^selected from O and S; ^R3 is selected from hydrogen, _C1-6 alkyl, _C1-4 haloalkyl, _C3-8 cycloalkyl, halogenated _C3-8 cycloalkyl; R ⁴ is selected from hydrogen, halogen, CN, C _1-8 alkyl, -OC _1-8 alkyl, C _1-4 haloalkyl, C _0-6 alkyl-(C=O)-NH-C _0-6 alkyl, -C _0-6 alkyl-SO ₂ NH-C _0-6 alkyl, -C _0-6 alkyl-SO ₂ N(CH ₃ )-C _0-6 alkyl, R ^4a or R ^4b ; R ⁵ is selected from hydrogen, halogen, C _1-6 alkyl, C _1-4 haloalkyl, C _3-8 cycloalkyl, halogenated C _3-8 cycloalkyl, -OC _1-8 alkyl or -OR ^5a ; wherein R ^5a is a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S; or alternatively, R ² and R ⁵ can be combined with the carbon atoms to which they are each attached to form a saturated or partially saturated 5- or 6-membered monocyclic ring fused to a benzene ring; wherein the 5- or 6-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and further, wherein the 5- or 6-membered monocyclic ring is substituted by 0, 1, 2 or 3 groups selected from halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC 1-8 alkyl or -OR 5a . _1-4 haloalkyl, CN, -NR ^a R ^a , or oxo; R ⁶ , R ⁸ , and R ⁹ are each independently selected from hydrogen, halogen, CN, C _1-8 alkyl, halogenated C _3-8 cycloalkyl, C _1-4 haloalkyl, or C _3-8 cycloalkyl; R ⁷ is hydrogen, halogen, CN, C _1-8 alkyl, C _1-4 haloalkyl, C _3-8 cycloalkyl, halogenated C _3-8 cycloalkyl, -OC _1-8 alkyl or R ^7a ; or alternatively, R ² and R ⁷ can be combined with the carbon atoms to which they are each attached to form a saturated or partially saturated 5-membered or 6-membered monocyclic ring fused to a benzene ring; wherein the 5-membered or 6-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, and further, wherein the 5-membered or 6-membered monocyclic ring is substituted by 0, 1, 2 or 3 groups selected from halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -NR ^a R ^a or oxo; R ¹⁰ is hydrogen, halogen, CN, hydroxyl, C _1-8 alkyl, C _3-8 cycloalkyl, halogenated C _3-8 cycloalkyl, C _1-4 haloalkyl, -OR ^10a or -OR ^10b ; R ¹¹ is hydrogen, R ^11a or R ^11b ; R ¹² is hydrogen, halogen, OH, CN, R ^12a or R ^12b ; R ¹³ is hydrogen, halogen, CN, R ^13a or R ^13b ; R ^4a , R ^7a , R ^10a , R ^11a , R ^12a and R ^13a are each independently selected from the group consisting of a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -C(═O)R ^b , -C(═O)OR ^a , -C(═O)NR ^a R ^a , -C(═NR ^a )NR ^a R ^a , -OC(═O)R ^b , -OC(═O)NR ^a R ^a , -OC _2-6 alkyl NR ^a R ^a , -OC _2-6 alkyl OR ^a , -SR ^a , -S(=O)R ^b , -S(=O) ₂ R ^b , -S(=O) ₂ NR ^a R ^a , -NR ^a R ^a , -N(R ^a )C(=O)OR ^b , -N(R ^a ) ^C (=O)NR ^a R ^a , -N(R ^a )C(=NR ^a )NR ^a R ^a , -N(R ^a )S(=O) ₂ R ^b , -N(R ^{a )} S(=O) ₂ NR ^a R ^a , -NR ^a C ₂ -C ₆ alkyl NR ^a R ^a , -NR ^a C _2-6 alkyl OR ^a , -C _1-6 alkyl NR ^a R ^a , -C _1-6 alkyl OR ^a , -C _{-C 1-6} alkylN(R ^a )C(═O)R ^b , -C _1-6 alkylOC(═O)R ^b , -C _1-6 alkylC(═O)NR ^a R ^a , -C _1-6 alkylC(═O)OR ^a , R ¹⁴ and oxo; R ^4b , R ^10b , R ^11b , R ^12b and R ^13b are each independently selected from the group consisting of: C _1-6 alkyl substituted with 0, 1, 2, 3, 4 or 5 groups selected from the group consisting of: halogen, CH ₂ F, CHF ₂ , CF ₃ , -C(═O)OR ^a , -OR ^a , -OC _1-4 haloalkyl, CN, -NH ₂ , NH(CH ₃ ), N(CH ₃ ) ₂ , and a saturated, partially saturated or unsaturated 3-, 4-, 5- or 6-membered monocyclic ring; R ¹⁴ is independently selected from the group consisting of a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from halogen, Ci _-6 alkyl, Ci _-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -C(═O)R ^b , -C(═O)OR ^a , -C(═O)NR ^a R ^a , -C(═NR ^a )NR ^a R ^a , -OC(═O)R ^b , -OC(═O)NR ^a R ^a , -OC _2-6 alkylNR ^a R ^a , -OC _2-6 alkylOR ^a , -SR ^a , -S(=O)R ^b , -S(=O) ₂ R ^b , -S(=O) ₂ NR ^a R ^a , -NR ^a R ^a , -N(R ^a )C(=O)R ^b , -N(R ^a )C(=O)OR ^b , -N(R ^a )C(=O)NR ^a R ^a , -N(R ^a )C(=NR ^a )NR ^a R ^a , -N(R ^a )S(=O) ₂ R ^b , -N(R ^a )S(=O) ₂ NR ^a R ^a , -NR ^a C _2-6 alkylNR ^a R ^a , -NR ^a C _2-6 alkylalkyl OR ^a , -C _1-6 alkyl NR ^a R ^a , -C _1-6 alkyl OR ^a , -C _1-6 alkyl N(R ^a )C(=O)R ^b , -C _1-6 alkyl OC(=O)R ^b , -C _1-6 alkyl C(=O)NR ^a R ^a , -C _1-6 alkyl C(=O)OR ^a and oxo; ^Rx is selected from the group consisting of: hydrogen, or R ^x is C _2-8 alkyl substituted by 0, 1, 2, 3, 4 or 5 groups selected from the group consisting of halogen, CH ₂ F, CHF ₂ , CF ₃ , —C(═O)OR ^a , —OR ^a , —OC _1-4 haloalkyl, CN, NH ₂ , NH(CH ₃ ), N(CH ₃ ) ₂ or R ¹⁵ⁿ ; or R ^x is phenyl or an unsaturated 5-membered monocyclic ring substituted by 0, 1, 2, 3, 4 or 5 groups selected from the group consisting of halogen, C _1-6 alkyl, C _1-6 haloalkyl, -C(=O)OR ^a , -OR ^a , -OC _1-4 haloalkyl, CN, NH ₂ , NH(CH ₃ ), N(CH ₃ ) ₂ or R ¹⁵ⁿ ; wherein the 5-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S; Each of R ^15a , R ^15b , R ^15c , R ^15d , R ^15e , R ^15f , R ^15g , R ^15h , R ¹⁵ⁱ , R ^15j , R ^15k , R ^15l , and R ^15m is hydrogen, halogen, R ^15o or R ^15p ; or alternatively, each of the pairs of R ^15a and R ^15b , R ^15c and R ^15d , R ^15e and R 15f, R ^15g and R ^15h , R ¹⁵ⁱ and R ^15j , and R ^15k and R ^15l can independently combine with the carbon atoms to which they are respectively attached to form a saturated or partially saturated 3 ^- membered, 4-membered, 5-membered, 6-membered monocyclic ring spiro-attached to the R ^x ring; wherein the 3-membered, 4-membered, 5-membered, 6-membered monocyclic ring contains 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, and further, wherein the 3-membered, 4-membered, 5-membered, 6-membered monocyclic ring is substituted with 0, 1, 2 or 3 groups selected from the following: halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -NR ^a R ^a or oxo; Or yet alternatively, each of the pairs of R ^15a and R ^15b , R ^15c and R ^15d , R ^15e and R ^15f , R ^15g and R ^15h , R ¹⁵ⁱ and R ^15j , and R ^15k and R ^15l can independently combine to form a double bond; R ¹⁵ⁿ , R ^15o are each independently selected from the group consisting of a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or an 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, which is substituted by 0, 1, 2, 3 or 4 groups selected from the following: halogen, C _1-6 alkyl, C _1-4 haloalkyl, -OR ^a , -OC _1-4 haloalkyl, CN, -C(═O)R ^b , -C(═O)OR ^a , -C(═O)NR ^a R ^a , -C(═NR ^a )NR ^a R ^a , -OC(═O)R ^b , -OC(═O)NR ^a R ^a , -OC _2-6 alkylNR ^a R ^a , -OC _2-6 alkylOR ^a , -SR ^a , -S(=O)R ^b , -S(=O) ₂ R ^b , -S(=O) ₂ NR ^a R ^a , -NR ^a R ^a , -N(R ^a )C(=O)R ^b , -N(R ^a )C(=O)OR ^b , -N(R ^a )C(=O)NR ^a R ^a , -N(R ^a )C(=NR ^a )NR ^a R ^a , -N(R ^a )S(=O) ₂ R ^b , -N(R ^a )S(=O) ₂ NR ^a R ^a , -NR ^a C ₂ -C ₆ alkylNR ^a R ^a , -NR ^a C _2-6 alkyl OR ^a , -C _1-6 alkyl NR ^a R ^a , -C _1-6 alkyl OR ^a , -C _1-6 alkyl N(R ^a )C(=O)R ^b , -C _1-6 alkyl OC(=O)R ^b , -C _1-6 alkyl C(=O)NR ^a R ^a , -C _1-6 alkyl C(=O)OR ^a and oxo; R ^15p is C _1-8 alkyl substituted by 0, 1, 2, 3, 4 or 5 groups selected from the group consisting of halogen, CH ₂ F, CHF ₂ , CF ₃ , —C(═O)OR ^a , —OR ^a , —OC _1-4 haloalkyl, CN, NH ₂ , NH(CH ₃ ), N(CH ₃ ) ₂ ; ^{Ra is} independently selected from hydrogen or ^Rb ; R ^b is independently selected from C _1-6 alkyl, phenyl or benzyl, wherein the C _1-6 alkyl is substituted by 0, 1, 2 or 3 substituents selected from halogen, hydroxy, -OC _1-6 alkyl, NH ₂ , -NHC _1-6 alkyl, -OC(═O)C _1-6 alkyl or -N(C _1-6 alkyl)C _1-6 alkyl; and the phenyl or benzyl is substituted by 0, 1, 2 or 3 substituents selected from halogen, C _1-6 alkyl, C _1-3 haloalkyl, -OH, -OC _1-6 alkyl, NH ₂ , -NHC _1-6 alkyl, -OC(═O)C _1-6 alkyl or -N(C _1-6 alkyl)C _1-6 alkyl; R ^c represents absence or is independently selected from halogen, hydroxyl, amino, cyano, carbonyl, oxo, carboxyl, C _2-6 alkenyl, C _1-6 alkyl, C _1-6 deuterated alkyl, C _{2-6 alkynyl, C 1-4 haloalkyl} , hydroxyC _1-6 alkyl, C _1-6 alkylcarbonyl, C _1-6 alkoxy, haloC _1-4 alkoxy, -COO-C _1-6 alkyl, -C(O)NR ^c1 R ^c2 ; wherein, R ^c1 and R ^c2 are each independently hydrogen or C _1-6 alkyl. 2. The compound of formula (I) as claimed in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that each of R ^15c , R ^15d , R ^15e , R 15f , R ^15g , R ^15h , R ¹⁵ⁱ and R ^15j is H, halogen, C _1-6 alkyl or C _1-4 haloalkyl; and each of ^the R ^15a and R ^15b pairs is combined with the carbon atom to which they are attached to form a saturated 3-, 4- or 5-membered monocyclic ring spiro-connected to the R ^x ring; wherein the ring contains 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S. 3. The compound shown in formula I as described in any one of claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that each of the R ^15c , R ^15d , R ¹⁵ⁱ and R ^15j is H, methyl or ethyl; and each of the R ^15a and R ^15b pairs is combined with the carbon atom to which they are attached to form a cyclopropyl, cyclobutyl or cyclopentyl ring that is spiro-connected to the R ^x ring. 4. The compound of formula (I) as described in any one of claims 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the group Selected from Preferably, the group Selected from 5. The compound of formula I as described in any one of claims 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the group Selected from 6. The compound of formula (I) as claimed in any one of claims 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that R ³ is hydrogen or methyl. 7. The compound of formula (I) according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the compound is selected from the following structures: Wherein, ^X4 and ^X6 have the definitions as described in claim 1; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ , R ⁷ and R ^x have the same meaning as defined in claim 1; M and L have the same meanings as those defined in claim 1. 8. The compound of formula (I) according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the compound is selected from the following structures: Wherein, ^X4 and ^X5 have the definitions as described in claim 1; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ , R ⁷ and R ^x have the same meaning as defined in claim 1; M and L have the same meanings as those defined in claim 1. 9. The compound of formula (I) according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the compound is selected from the following structures: Wherein, X ⁴ , X ⁵ and X ⁶ have the definitions as described in claim 1; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as those defined in claim 1; R ^15a , R ^15b , R ^15c , R ^15d , R ¹⁵ⁱ and R ^15j have the meanings as defined in claim 1; M and L have the same meanings as those defined in claim 1. 10. The compound of formula (I) according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the compound is selected from the following structures: Wherein, X ⁴ , X ⁵ and X ⁶ have the definitions as described in claim 1; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as those defined in claim 1; R ^15a , R ^15b , R ^15c , R ^15d , R ¹⁵ⁱ and R ^15j have the meanings as defined in claim 1; M has the definition as described in claim 1. 11. The compound of formula (I) according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the compound is selected from the following structures: Wherein, X ⁴ , X ⁵ and X ⁶ have the definitions as described in claim 1; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as those defined in claim 1; M has the definition as described in claim 1. 12. The compound of formula (I) according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the compound is selected from the following structures: Wherein, ^X4 and ^X6 have the definitions as described in claim 1; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as those defined in claim 1; M has the definition as described in claim 1. 13. The compound of formula (I) according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the compound is selected from the following structures: Wherein, ^X4 has the definition as described in claim 1; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as those defined in claim 1; M has the definition as described in claim 1. 14. The compound of formula (I) according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the compound is selected from the following structures: Wherein, ^X5 has the definition as described in claim 1; R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ and R ⁷ have the same meanings as those defined in claim 1; M has the definition as described in claim 1. 15. The compound of formula I according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that R ² is selected from: F, Cl, Br or a group -YR ¹³ , wherein Y is a bond, -NH-, -NH-(CH ₂ ) _0-4 -, -O-(CH ₂ ) _0-4 -, -S-, -S＝O-, -S(＝O) _2- , -S(＝O)(＝NH)-, -C(＝O)-; and R ^{R 13} is a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from the following: F, Cl, Br, C _1-6 alkyl, C _1-4 haloalkyl, -OH, -OC _1-4 haloalkyl, CN, R ¹⁴ and oxo; or R ¹³ is a C _1-6 alkyl substituted by 0, 1, 2, 3, 4 or 5 groups selected from F, Cl, Br, -OH, -OC _1-4 haloalkyl or CN; wherein R ¹⁴ has the definition as described in claim 1; Preferably, Y is a bond, R ¹³ is a saturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring or a 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring containing 0 or 1 N atoms and 0 or 1 O atoms; which is substituted by 0, 1, 2 or 3 identical or different groups selected from the following: F, Cl, Br, -C _1-3 alkyl, -OH, CN, -C _1-3 alkyl-OH; Preferably, it is F, Cl, Br, methyl, -OH, -OCH ₃ , CN, -CH ₂ OH; or, Y is -NH-, -NH-(CH ₂ ) _0-4 -, -O-(CH ₂ ) _0-4 - or -SO ₂ NH-; R ¹³ is a saturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring containing 0 or 1 N atom and 0 or 1 O atom; which is substituted by 0, 1, 2 or 3 identical or different groups selected from the following: fluorine, methyl, trifluoromethyl, -CN, -OH, CH ₂ OH; or Y is a bond, -NH-, -NH-(CH ₂ ) _0-4 -, -O-(CH ₂ ) _0-4 - or -SO ₂ NH-; R ¹³ is selected from the following 0, 1, C _1-6 alkyl substituted with 2, 3, 4 or 5 identical or different substituents, wherein the substituent is -OH, methyl or CF ₃ . 16. The compound of formula I according to any one of claims 1 or 7 to 15, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that R ² is selected from: 17. The compound of formula I as claimed in any one of claims 7 to 14, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that M is absent or selected from -NH-, 18. The compound of formula I according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that Z is selected from a bond, -NH-, -NHSO ₂ -, -SO ₂ NH-, -S(=O)(=NH)-, -S-, -S(=O)-, -SO ₂ -, -O-, -P-, -P(=O)CH ₃ -, -P(=O) ₂ , -(C=O)-, -(C=O)NH- or -NH(C=O)-; Preferably, Z is selected from -SO ₂ NH-, -S(=O)(=NH)-, and -SO ₂ -. 19. The compound as claimed in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the R ¹² is selected from (a) hydrogen; (b) C 1-6 alkyl substituted by 0, 1, 2 or 3 groups selected from F, Cl, Br, -OH, -OCH ₃ or _cyclopropyl ; or (c) a saturated, partially saturated or unsaturated 3-, 4-, 5-, 6- or 7-membered monocyclic ring containing 0, 1, 2 or 3 N atoms and 0 or 1 atom selected from O and S, which is substituted by 0, 1, 2 or 3 groups selected from the following: F, Cl, Br, C _1-6 alkyl, C _1-4 haloalkyl, -C _1-6 alkylOH, -OH, -OCH ₃ , -NH ₂ or oxo. 20. The compound of formula (I) as claimed in claim 19, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that R ¹² is selected from (a) hydrogen; (b) C _1-6 alkyl substituted with -OH; or (c) cyclopropyl, cyclobutyl, cyclopentyl, oxetanyl, azetidinyl, tetrahydrofuranyl, 1,3,4-oxathiazinyl, C _3-8 cycloalkyl substituted with -OH. 21. The compound of formula I according to any one of claims 7 to 14, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that R ¹ is selected from: 22. A compound of formula I as claimed in claim 1 or any one of claims 7 to 14, a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug thereof, characterized in that R ⁴ is selected from (a) hydrogen, F, Cl, Br, CN, -OC _1-6 alkyl; (b) C _1-6 alkyl substituted with 0, 1, 2 or 3 OH groups; (c) cyclopropyl or (d) C _1-3 haloalkyl. 23. The compound of formula I according to claim 22, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that ^R4 is selected from hydrogen, methyl, F, Cl, Br, CN, cyclopropyl, methoxy, -(C=O) _NH2 , _CF3 , _CHF2 , furanyl, pyridyl, morpholinyl or Azetidine. 24. The compound of formula I according to claim 1 or any one of claims 7 to 14, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that R ⁵ is selected from hydrogen, F or methyl. 25. The compound of formula I according to claim 1 or any one of claims 7 to 14, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that ^R6 is selected from hydrogen. 26. The compound of formula I according to any one of claims 1 or 7 to 14, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that R ⁷ is selected from hydrogen, F, Cl, Br, CN, methyl, methoxy, cyclopropyl; or R ² and R ⁷ are combined with the carbon atoms to which they are attached to form 27. The compound of formula I according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that R ⁸ is selected from hydrogen, F, Cl, Br, cyclopropyl, -(C=O)CH ₃ or CF ₃ ; Preferably, R ⁸ is selected from hydrogen. 28. The compound of formula (I) as claimed in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that R ⁹ is selected from hydrogen or F. 29. The compound of formula (I) according to claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that ^R10 is selected from hydrogen, F, methyl, methoxy or cyclopropyl. 30. The compound of formula (I) as claimed in claim 1, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that the compound comprises: 31. A pharmaceutical composition, characterized in that the pharmaceutical composition comprises: a compound of formula I as described in any one of claims 1 to 30, its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug; and a pharmaceutically acceptable carrier. 32. A use of a compound of formula I as claimed in any one of claims 1 to 30, a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof, or a use of a pharmaceutical composition as claimed in claim 31, comprising: Inhibition of KIF18A; and/or, preventing and/or treating KIF18A-related diseases; And/or, preparing a drug, pharmaceutical composition or preparation for inhibiting KIF18A, and/or preventing and/or treating a KIF18A-related disease. 33. The use according to claim 32, characterized in that the KIF18A-related disease comprises: cancer, psoriasis, atopic dermatitis, autoimmune diseases or proliferative disorders of inflammatory bowel disease. 34. The method of claim 33, wherein the cancer is selected from the group consisting of mesothelioma, neuroblastoma, rectal cancer, colon cancer, familiar adenomatous polyposis and hereditary non-polyposis colorectal cancer, esophageal cancer, lip cancer, laryngeal cancer, hypopharyngeal cancer, tongue cancer, salivary gland cancer, gastric cancer, adenocarcinoma, medullary thyroid cancer, papillary thyroid cancer, kidney cancer, renal parenchymal cancer, ovarian cancer, cervical cancer, uterine corpus cancer, endometrial cancer, choriocarcinoma, pancreatic cancer, prostate cancer, bladder cancer, testicular cancer , breast cancer, urological cancer, melanoma, brain tumor, lymphoma, head and neck cancer, acute lymphatic leukemia, chronic lymphatic leukemia, acute myeloid leukemia, chronic myeloid leukemia, hepatocellular carcinoma, gallbladder cancer, bronchial cancer, small cell lung cancer, non-small cell lung cancer, multiple myeloma, basal sarcoma, teratoma, retinoblastoma, choroidal melanoma, seminoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma, myoma, liposarcoma, fibrosarcoma, Ewing sarcoma, and plasmacytoma. 35. purposes as claimed in claim 33, it is characterized in that, described autoimmune disease is selected from rheumatoid arthritis, systemic lupus erythematosus, Sjögren's syndrome, scleroderma, mixed connective tissue disease, dermatomyositis, polymyositis, Reiter's syndrome, autoimmune lymphoproliferative syndrome, multiple sclerosis, myasthenia gravis and encephalomyelitis. 36. The use according to claim 33, wherein the inflammatory bowel disease is selected from ulcerative colitis or Crohn's disease.