WO2024104282A1

WO2024104282A1 - 1h-pyrrolo[2,3-b]pyridine derivative, preparation method therefor, and use thereof in medicine

Info

Publication number: WO2024104282A1
Application number: PCT/CN2023/131185
Authority: WO
Inventors: 邓祖勇; 沈阳; 张蕾
Original assignee: 捷思英达控股有限公司; 邓祖勇
Priority date: 2022-11-14
Filing date: 2023-11-13
Publication date: 2024-05-23

Abstract

A compound as represented by formula (I) and/or a stereoisomer, tautomer or pharmaceutically acceptable salt or solvate of the compound of formula (I), a preparation method therefor, a pharmaceutical composition comprising the compound of formula (I), and a use thereof as an Myt1 kinase inhibitor in medicine.

Description

A 1H-pyrrolo[2,3-b]pyridine derivative, preparation method thereof and application thereof in medicine

Technical Field

The present disclosure relates to the field of medical technology, and in particular to a 1H-pyrrolo[2,3-b]pyridine derivative, a preparation method thereof, and a medical application thereof.

Background technique

Protein kinase, membrane-associated tyrosine and threonine-specific cdc2-inhibitory kinase ("Myt1"), is located at 16p13.3 on human chromosome 16 and encodes an important protein belonging to the WEE kinase family (Qingyi Zhang et al., Cancer Manag Res. 11:7813–7824.2019). Myt1 is responsible for encoding a member of the serine/threonine protein kinase family, and the encoded protein negatively regulates the G2/M transition of the cell cycle by phosphorylating and inactivating cell cycle-dependent kinase 1.

It has been reported that Myt1 kinase phosphorylates Thr14 in the Cdk1-cyclin B complex after Tyr15 is phosphorylated by WEE1 kinase. Since this Tyr14/Tyr15 phosphorylation inactivates the CDK1-cyclin B complex, it prevents the cell cycle from entering mitosis at the G2/M transition (Booher RN et al., J Biol Chem. 272(35):22300–22306.1997). During the cell cycle, when DNA is replicated, a large number of DNA mismatches and DNA defects occur. Normal cells repair DNA at the G1/S checkpoint. However, tumor cells fail to have a G0/G1 and S checkpoints due to mutations in P53, resulting in more DNA damage in the G2/M checkpoint than normal cells (Helen Dixon et al., Cell cycle, 1:362-368.2002). Compared with normal cells, tumor cells repair DNA more during the G2/M checkpoint and avoid immature cells entering mitosis to prevent mitotic catastrophe. Therefore, tumor cells are more dependent on the G2/M checkpoint than normal cells (M Suganuma et al., Cancer Res, 59: 5887-5891. 1999).

A new strategy for cancer treatment is the DNA damage generated after checkpoint failure (Mikhail V Blagosklonn, Cell Cycle, 6(1):70-74.2007). Considering the role of Myt1 in regulating the transition of the G2/M phase and the DNA repair mechanism, more and more scholars have begun to study the effects of Myt1 inhibition on tumor cells. It has been reported that Myt1 inhibitors may effectively reduce the viability of tumor cells (Liu Y et al., Cell Prolif., 53(2):e12741.2020). This suggests that Myt1 inhibitors can be used as potential tumor treatments.

International patent applications WO2021195781A1, WO2001064680A1, WO2000033837A2, etc. disclose some small molecule Myt1 kinase inhibitors, but currently no small molecule Myt1 kinase inhibitors have been approved for marketing. The field still needs to develop new Myt1 kinase inhibitors with good anti-cancer activity and high safety.

Summary of the invention

To solve the above technical problems, the present application discloses a series of 1H-pyrrolo[2,3-b]pyridine compounds that can be used as Myt1 inhibitors. The present application further discloses a pharmaceutical composition comprising at least one such compound, a method for preparing the compound, and a method for using at least one such compound to treat Myt1-related diseases. Methods for treating kinase-related diseases and disorders, such as cancer.

The first aspect of the present application provides a 1H-pyrrolo[2,3-b]pyridine derivative represented by general formula (I), or a pharmaceutically acceptable salt thereof:

in:

R ¹ and R ² are independently selected from H, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl, C _3-8 cycloalkenyl, C _2-9 heterocycloalkyl, C _1-6 alkyl substituted by C _2-9 heterocycloalkyl, C _6-10 aryl, C _1-9 heteroaryl, C _1-6 alkyl substituted by C _1-9 heteroaryl, halogen, cyano, -N(R ^a ) ₂ , -OR ^a , -C(O)N(R ⁹ ) ₂ , -SO ₂ N(R ⁹ ) ₂ , -SO ₂ R ^b and -QR ^c ; or R ¹ and R ² are combined to form an optionally substituted C _3-8 alkylene group, which may have 0, ₁ , 2 or 3 substituents selected from C _1-6 alkyl;

^R3 is selected from halogen, cyano, optionally substituted _C1-6 alkyl, 3-membered, 4-membered, 5-membered or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, the optionally substituted _C1-6 alkyl optionally substituted by 3-membered, 4-membered, 5-membered or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members;

^R4 is selected from cyano, optionally substituted _C1-6 alkyl, 3-, 4-, 5- or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members, the optionally substituted _C1-6 alkyl optionally substituted by 3-, 4-, 5- or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members;

R ⁵ is H or -N(R ^a ) ₂ ;

R ⁶ is -C(O)NH(R ⁹ ), -C(O)R ^b or -SO ₂ R ^b ;

^R7 is selected from H, OH, halogen and _C1-6 alkyl; or, ^R7 and ^R4 or ^R7 and ^R8 are combined to form an optionally substituted _C2-4 alkylene, wherein the optionally substituted _C2-4 alkylene may have 0, 1, 2 or 3 substituents selected from _C1-6 alkyl;

R ⁸ is H or halogen;

Each ^Ra is independently selected from H, _C1-6 alkyl, _C1-6 alkyl substituted by _C6-10 aryl, _C3-8 cycloalkyl, _C6-10 aryl, _C2-9 heterocycloalkyl, _C1-9 heteroaryl, _C1-6 alkyl substituted by _C1-9 heteroaryl, and _-SO2Rb ; or two ^Ra groups and the atoms to which the two ^Ra groups are connected together form a _C2-9 ^{heterocycloalkyl} ;

Each R ^b is independently selected from C _1-6 alkyl, C _3-6 cycloalkyl and C _6-10 aryl;

each R ^c is independently selected from -OH, C _1-6 alkyl, C _6-10 aryl, C _2-9 heterocyclyl, C _1-9 heteroaryl, -N(R ^a ) ₂ , -C(O)N(R ⁹ ) ₂ , -SO ₂ N(R ⁹ ) ₂ , -SO ₂ R ^b and alkoxy;

Each R ⁹ is independently selected from H, C _1-6 alkyl, C _2-6 alkoxyalkyl, C _1-6 substituted by C _6-10 aryl alkyl, C _6-10 aryl, C _3-8 cycloalkyl and C _1-9 heteroaryl; or two R ⁹ groups and the atoms to which the two R ⁹ groups are connected together form a C _2-9 heterocyclic group;

Q is selected from C _1-6 alkylene, C _2-6 alkenylene, C _3-8 cycloalkylene, C _6-10 arylene, C _2-9 heterocycloalkylene and C _1-9 heteroarylene,

When R ³ is halogen or optionally substituted C _1-6 alkyl, said R ⁴ is not optionally substituted C _1-6 alkyl; or

When R ⁴ is an optionally substituted C _1-6 alkyl, said R ³ is not halogen or an optionally substituted C _1-6 alkyl; or

When R ³ is halogen or optionally substituted C _1-6 alkyl and R ⁴ is optionally substituted C _1-6 alkyl, at least one of R ⁷ or R ⁸ is not H.

The compound of the general formula (I) provided in the first aspect of the present invention can be used as a Myt1 inhibitor for treating Myt1-mediated conditions and/or diseases, including cancer or hyperproliferative diseases.

The present application also provides preferred embodiments related to the compounds described by general formula (I).

In one or more embodiments, the compound is enriched in atropisomers of formula (IA):

When ^R7 is -OH, ^R3 and ^R4 are different.

In one or more embodiments, the R ³ is cyano. In one or more embodiments, the R ³ is a 3-, 4-, 5- or 6-membered saturated cycloalkyl or heterocycloalkyl group containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S. In a preferred embodiment, the R ³ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In one or more embodiments, the R ³ is cyclopropyl.

In one or more embodiments, the R ⁴ is cyano. In one or more embodiments, the R ⁴ is a 3-, 4-, 5- or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members. In a preferred embodiment, the R ⁴ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In one or more embodiments, the R ⁴ is cyclopropyl.

In one or more embodiments, R ³ is halogen or C _1-6 alkyl, and R ⁴ is cyano. In one or more embodiments, R ³ is halogen or C _1-6 alkyl, and R ⁴ is a 3-, 4-, 5- or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members. In one or more embodiments, R ⁴ is C _1-6 In one or more embodiments, ^R4 is a _C1-6 alkyl group, and ^R3 is a 3-, 4-, 5- or 6-membered saturated cycloalkyl or heterocycloalkyl group containing 0 ^, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S.

In one or more embodiments, the R ⁷ and R ⁴ or R ⁷ and R ⁸ are combined to form a C _2-4 alkylene group, and the C _2-4 alkylene group may have 0, 1, 2 or 3 substituents selected from C _1-6 alkyl groups. In one or more embodiments, the R ⁴ and R ⁷ groups are combined to form a C ₂ alkylene group, and the C ₂ alkylene group may have 0, 1, 2 or 3 substituents selected from C _1-6 alkyl groups. In an exemplary embodiment, the R ⁴ and R ⁷ groups are combined to form a C ₂ alkylene group. In an exemplary embodiment, the R ⁴ and R ⁷ groups are combined to form a C ₂ alkylene group having a methyl substituent. In one or more embodiments, the R ⁷ and R ⁴ are combined to form a C ₃ alkylene group, and the C ₃ alkylene group may have 0, 1, 2 or 3 substituents selected from C _1-6 alkyl groups. In one or more embodiments, the R ⁷ and R ⁸ are combined to form a C ₂ alkylene group, and the C ₂ alkylene group may have 0, 1, 2 or 3 substituents selected from C _1-6 alkyl groups. In an exemplary embodiment, the R ⁸ and R ⁷ groups are combined to form a C ₂ alkylene group. In an exemplary embodiment, the R ⁸ and R ⁷ groups are combined to form a C ₂ alkylene group having a methyl substituent. In one or more embodiments, the R ⁸ and R ⁷ groups are combined to form a C 3 alkylene group, and the C ₃ alkylene group may have 0, 1, 2 or 3 substituents selected from C _1-6 alkyl groups. In one or more embodiments, the R ⁴ and R ⁷ groups or R ⁷ and R ⁸ groups are combined to form a C ₄ alkylene group, and _{the C 4} _alkylene group may have 0, 1, 2 or 3 substituents selected from C _1-6 alkyl groups.

In one or more preferred embodiments, the R ⁷ is -OH or C _1-6 alkyl. In one or more preferred embodiments, the R ⁷ is -OH. In one or more preferred embodiments, the R ⁷ is C _1-6 alkyl. In a preferred embodiment, the R ⁷ is methyl or isopropyl.

In one or more preferred embodiments, R ² is H, halogen, C _1-6 alkyl, C _1-6 alkyl substituted by C _2-9 heterocycloalkyl, or C _1-6 alkyl substituted by C _1-9 heteroaryl. In one or more preferred embodiments, R ² is H. In one or more preferred embodiments, R ² is C _1-6 alkyl. In a preferred embodiment, R ² is methyl, ethyl, isopropyl or butyl.

In one or more preferred embodiments, the R ¹ is H. In one or more preferred embodiments, the R ¹ is C _1-6 alkyl. In a preferred embodiment, the R ¹ is methyl, ethyl, isopropyl or butyl. In one or more preferred embodiments, the R ¹ is halogen. In one or more preferred embodiments, the R ¹ is Cl or Br. In one or more preferred embodiments, the R ¹ is C _1-9 heteroaryl. In one or more preferred embodiments, the R ¹ is 1,3-thiazolyl, 1,2-thiazolyl, 1,3-oxazolyl, benzo-1,3-thiazolyl, benzo-1,3-oxazolyl, indolyl, benzimidazolyl, pyridyl, imidazolyl, pyrimidinyl, pyrazinyl, pyridazinyl or pyrazolyl. In one or more preferred embodiments, the R ¹ is C _3-8 cycloalkyl. In a preferred embodiment, the R ¹ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl. In one or more preferred embodiments, the R ¹ is C _2-9 heterocycloalkyl. In one or more preferred embodiments, R ¹ is a C _6-10 aryl group. In one or more preferred embodiments, R ¹ is a C _2-9 heteroaryl group. In a preferred embodiment, R ¹ is a 1,2,3,6-tetrahydropyridyl group, a piperidyl group, a morpholinyl group, a piperazinyl group, a thiomorpholinyl group, or a thiomorpholinyl group. In one or more preferred embodiments, the R ¹ is a C _3-8 cycloalkyl. For example, R 1 is an optionally substituted cyclohexenyl or an optionally substituted cyclopentenyl. In one or more preferred embodiments, the R ¹ is a C 6-10 aryl. For example, the R ¹ is an optionally substituted phenyl. In one or more preferred embodiments, ^{the R 1} ^is -QR ^c . In one or more preferred embodiments, Q is an optionally substituted C _2-6 alkynylene. In one or more preferred embodiments, Q is an optionally substituted C _1-6 alkylene. In one or more preferred embodiments, Q is an optionally substituted C _6-10 arylene. In one or more preferred embodiments, R ^c is an optionally substituted C _2-9 heterocycloalkyl. In one or more preferred embodiments, R ^c is _{an optionally substituted C 6-10} _aryl . In one or more preferred embodiments, the R ¹ is optionally substituted by 1, 2 or 3 groups selected from methyl, difluoromethyl, trifluoromethyl, fluorine, chlorine, bromine, amino, hydroxyl, cyano, =O, -C(O)NH ₂ , -C(O)NH(Me), -C(O)N(Me) ₂ , -(CH ₂ )nC(O)OH and -(CH ₂ )nC(O)Ot-Bu, wherein n is 0 or 1. In one or more preferred embodiments, the R ¹ is H or halogen. In one or more preferred embodiments, the R ¹ is -N(R ^a ) _2. In a preferred embodiment, the R ¹ is diethylamino.

In one or more preferred embodiments, the R ⁵ is H. In one or more preferred embodiments, the R ⁵ is NH ₂ .

In one or more preferred embodiments, the R ⁶ is -C(O)NH(R ⁹ ). In one or more preferred embodiments, the R ⁶ is -C(O)NH ₂ . In one or more preferred embodiments, the R ⁶ is -C(O)NH(Me). In one or more preferred embodiments, the R ⁶ is -SO ₂ R ^b . In one or more preferred embodiments, the R ⁶ is -SO ₂ Me.

In one or more preferred embodiments, R ⁸ is halogen. For example, R ⁸ is fluorine or chlorine.

Typical compounds of the present invention include, but are not limited to:

or a pharmaceutically acceptable salt thereof.

The second aspect of the present application provides a pharmaceutical composition, which contains an effective dose of a compound of Formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. In some embodiments, the compound is a deuterium-enriched compound.

The third aspect of the present application discloses a method for inhibiting Myt1 in a cell expressing Myt1 protein, comprising contacting the cell with a compound of formula I.

The fourth aspect of the present application discloses a method for treating and/or preventing a disease or condition associated with Myt1 kinase, comprising administering a therapeutically effective amount of the compound or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, to a patient in need thereof.

The diseases or conditions associated with Myt1 kinase include leukemia, solid tumor cancer and metastasis, soft tissue cancer, brain cancer, esophageal cancer, gastric cancer, pancreatic cancer, liver cancer, lung cancer, bladder cancer, bone cancer, prostate cancer, ovarian cancer, breast cancer, colorectal cancer, endometrial cancer, cervical cancer, uterine cancer, testicular cancer, kidney cancer, head and neck cancer, chronic inflammatory proliferative diseases, proliferative cardiovascular diseases, proliferative eye diseases and benign hyperproliferative diseases.

One or more embodiments of the present application provide use of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound of Formula I in the preparation of a medicament for treating a disease or condition associated with Myt1 kinase.

Detailed ways

definition

Unless otherwise stated, some of the terms used in the specification and claims of the present invention are defined as follows:

A dash ("-") that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, -CONR ^a R ^b is attached through a carbon atom.

The term "halogen" herein denotes a halogen selected from bromine, chlorine, iodine and fluorine.

The term "acyl" herein refers to a -C(=O)-R group, wherein R is an alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl or heterocycloalkyl. For each corresponding R group, the acyl group may be substituted with a substituent as described below.

The term "alkanoyl" herein refers to a hydrogen or alkyl group that is connected to a parent molecular group through a carbonyl group, such as formyl (e.g., carboxyaldehyde), acetyl, propionyl, butyryl and isobutyryl. Unsubstituted alkanoyl contains 1 to 7 carbons. Alkanoyl can be unsubstituted or substituted by a substituent as defined below for an alkyl group (e.g., an optionally substituted C _1-7 alkanoyl). "Alkanoyl" can also include "aroyl", "cycloalkanoyl" and "(heterocycloalkyl) acyl". These groups represent a carbonyl group substituted by an aryl, cycloalkyl or heterocycloalkyl group, respectively. Each of "aroyl", "cycloalkanoyl" and "(heterocycloalkyl) acyl" can be optionally substituted by a substituent as defined below for an "aryl", "cycloalkyl" or "heterocyclyl".

The term "alkenyl" herein refers to an acyclic monovalent straight or branched hydrocarbon radical containing one, two or three carbon-carbon double bonds. The limiting examples of alkenyl include vinyl, prop-1-enyl, prop-2-enyl, 1-methylvinyl, but-1-enyl, but-2-enyl, but-3-enyl, 1-methylprop-1-enyl, 2-methylpropyl-1-enyl and 1-methylpropyl-2-enyl. The alkenyl group is optionally replaced by a substituent defined as follows for an alkyl.

The term "alkenylene" herein refers to a divalent alkenyl group. The alkenylene group is optionally substituted with substituents as defined herein for alkenyl groups.

The term "alkylene" as used herein refers to a divalent alkyl group. The alkylene group is optionally substituted with substituents as defined herein for an alkyl group.

The term "alkoxy" herein refers to a chemical substituent of the formula -OR, wherein R is a C _1-6 alkyl, unless otherwise specified. In some embodiments, the alkyl group may be further substituted as defined herein. The term "alkoxy" may be combined with other terms defined herein, such as aryl, cycloalkyl or heterocyclyl, to define "arylalkoxy", "cycloalkylalkoxy" and "(heterocyclyl)alkoxy" groups. These groups represent alkoxy groups substituted by aryl, cycloalkyl or heterocyclyl, respectively. For each separate part, the aryl, cycloalkyl or heterocyclyl moiety in each of "arylalkoxy", "cycloalkylalkoxy" and "(heterocyclyl)alkoxy" may be optionally substituted as defined herein.

The term "alkoxyalkyl" herein refers to a chemical substituent of the formula -LOR, wherein L is C _1-6 alkylene and R is C _1-6 alkyl. The alkyl portion of the alkoxyalkyl may be optionally substituted with substituents as defined below for alkyl.

The term "alkyl" as used herein refers to an acyclic straight or branched saturated hydrocarbon radical having from 1 to 12 carbons when unsubstituted, unless otherwise specified. In certain preferred embodiments, the unsubstituted alkyl radical has from 1 to 6 carbons. Examples of alkyl groups are methyl; ethyl; n-propyl and isopropyl; n-butyl, sec-, iso- and tert-butyl; neopentyl, etc., and may optionally be substituted with one, two, three substituents, or in the case of alkyl radicals containing more than two carbons, four or more substituents, independently selected from the group consisting of the following groups: amino; alkoxy; aryl; aryloxy; azide; cycloalkane alkyl; cycloalkyloxy; cycloalkenyl; cycloalkynyl; halogen; heterocycloalkyl; (heterocycloalkyl)oxy; heteroaryl; hydroxy; nitro; thiol; silyl; cyano; alkylthio; alkylsulfinyl; alkylsulfonyl; =O; =S; -C(O)R or _-SO2R , where R is amino; and =NR', where R' is H, alkyl, aryl or heterocycloalkyl. Each substituent itself may be unsubstituted or, where valence permits, substituted with a substituent as defined herein for the respective group.

The term "alkylamino" as used herein refers to a group having the formula -N( ^RX ) ₂ or ^-NHRX , wherein ^RX is an alkyl group as defined herein. The alkyl portion of the alkylamino group may be optionally substituted as defined herein for an alkyl group.

The term "alkylthio" as used herein refers to a group of the formula -S-(alkyl). The alkyl portion of the alkylsulfinyl group may be optionally substituted as defined herein for an alkyl group.

The term "alkylsulfinyl" as used herein refers to a group of the formula -S(O)-(alkyl). The alkyl portion of the alkylsulfinyl group may be optionally substituted as defined herein for an alkyl group.

The term "alkylsulfonyl" herein denotes a group of formula -S(O) _2- (alkyl). The alkyl part of the alkylsulfonyl group may be optionally substituted as defined herein for an alkyl group.

The term "alkynyl" herein refers to a monovalent straight or branched hydrocarbon group containing two to six carbon atoms and containing at least one carbon-carbon triple bond, such as ethynyl, 1-propynyl, etc. Alkynyl groups may be unsubstituted or may be substituted with substituents as defined herein for alkyl groups.

The term "alkynylene" herein refers to a divalent alkynyl group. The alkynylene group is optionally substituted with substituents as defined herein for an alkynyl group.

The term "amino" as used herein means -N(R ^x ) ₂ , wherein, if the amino group is substituted, both R ^x are H; or, if the amino group is substituted, each R ^x is independently selected from H, -OH, -NO, -N(R ^y ), -SO ₂ OR ^y , -SO ₂ R ^y , -SOR ^y , -C(O)OR ^y or an N-protecting group, alkyl, alkenyl, alkynyl, alkoxy, aryl, aralkyl, aryloxy, cycloalkyl, cycloalkenyl, heteroalkyl and heterocycloalkyl, provided that at least one R ^x is not H and each R ^y is independently selected from H, alkyl or aryl. Each substituent itself may be unsubstituted or substituted with a substituent group as defined herein for each corresponding group. In some embodiments, the amino group is an unsubstituted amino group (i.e., -NH ₂ ) or a substituted amino group (e.g., -NHR ^X ), wherein ^RX is independently selected from -OH, -SO ₂ OR ^y , -SO ₂ R ^y , -SOR ^y , -C(O)OR ^y , an optionally substituted alkyl group, or an optionally substituted aryl group, and each R ^y may be an optionally substituted alkyl group or an optionally substituted aryl group. In some embodiments, the substituted amino group may be an alkylamino group, wherein the alkyl portion may be optionally substituted with a substituent group as defined herein for an alkyl group. In some embodiments, the amino group is -NHR ^X , wherein ^RX is an optionally substituted alkyl group.

The term "aryl" herein means a monocyclic, bicyclic or polycyclic carbocyclic ring system having one or two aromatic rings. The aryl group may include 6 to 10 carbon atoms. All atoms in the unsubstituted carbocyclic aryl group are carbon atoms. Non-limiting examples of carbocyclic aryl groups include phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indanyl, indenyl, etc. The aryl group may be unsubstituted or substituted by one, two, three, four or five groups independently selected from the group consisting of: alkyl; alkenyl; alkynyl; alkoxy; alkylthio; alkylsulfinyl; alkylsulfonyl; amino; aryl; aryloxy; azide; cycloalkyl; cycloalkoxy; cycloalkenyl; Cycloalkynyl; halogen; heteroalkyl; heterocycloalkyl; (heterocycloalkyl)oxy; hydroxy; nitro; thiol; silyl; -( _CH2 ) _n -C(O) ^ORA ; -C(O)R; _-SO2R , wherein R is amino or alkyl, ^RA is H or alkyl, and n is 0 or 1. Each substituent itself may be unsubstituted or substituted by a substituent as defined herein for each corresponding group.

The term "arylene" as used herein refers to a divalent aromatic group. The arylene group is optionally substituted with substituents as defined herein for an aryl group.

The term "aralkyl" herein refers to an alkyl group substituted by an aryl group. Both the aryl and alkyl parts of the aralkyl group may be substituted by substituents defined for the corresponding groups.

The term "saturated" as used herein refers to a moiety that does not contain any carbon-carbon double bonds or carbon-carbon triple bonds, for example, only carbon-carbon single bonds.

The term "aryloxy" as used herein refers to a chemical substituent of the formula -OR, wherein R is aryl, unless otherwise specified. The aryl portion of the aryloxy group is optionally substituted with substituents as described herein for aryl groups.

The term "azido" as used herein refers to a _-N3 group.

The term "carbonyl" as used herein refers to a -C(O)- group.

The term "cyano" as used herein refers to a -CN group.

The term "cycloalkenyl" herein refers to a non-aromatic carbocyclic group (for example, cycloalkenyl) having at least one double bond and three to ten carbons in a ring, unless otherwise indicated. The limiting examples of cycloalkenyl include cycloprop-1-alkenyl, cycloprop-2-alkenyl, cyclobut-1-alkenyl, cyclobut-1-alkenyl, cyclobut-2-alkenyl, cyclopent-1-alkenyl, cyclopent-2-alkenyl, cyclopentyl-3-alkenyl, norbornene-1-base, norbornene-2-base, norbornene-5-base and norbornene-7-base. Cycloalkenyl can be unsubstituted or substituted by a substituent defined for cycloalkyl.

The term "cycloalkenylalkyl" herein refers to an alkyl group substituted by a cycloalkenyl group, wherein the cycloalkenyl group and the alkyl group are as defined herein. Both the cycloalkenyl group and the alkyl group are optionally substituted by substituents as defined for the cycloalkenyl group and the alkyl group.

The term "cycloalkenylene" herein refers to a divalent cycloalkenyl group. The cycloalkenylene group is optionally substituted with substituents as defined for the cycloalkyl group.

The term "cycloalkyl" herein refers to a saturated or partially unsaturated cycloalkyl group.

The term "cycloalkyl" herein refers to a cyclic alkyl group (e.g., cycloalkyl) having three to eight carbon atoms, unless otherwise specified. Cycloalkyl groups may be monocyclic or bicyclic. Bicyclic cycloalkyl groups may be bicyclic [pq0] alkyl groups, wherein p and q are each independently 1, 2, 3, 4, 5, 6, or 7, and the sum of p and q is 2, 3, 4, 5, 6, 7, or 8. Alternatively, bicyclic cycloalkyl groups may include bridged cycloalkyl structures, such as bicyclic [pqr] alkyl groups, wherein r is 1, 2, or 3, and p and q are each independently 1, 2, 3, 4, 5, or 6, and the sum of p, q, and r is 3, 4, 5, 6, 7, or 8. Cycloalkyl groups may be spirocyclic groups, such as spiro [pq] alkyl groups, wherein each of p and q is independently 2, 3, 4, 5, 6, or 7, and the sum of p and q is 4, 5, 6, 7, 8, or 9. Cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 1-bicyclo[2.2.1.]heptyl, 2-bicyclo[2.2.1.]heptyl, 5-bicyclo[2.2.1.]heptyl, 7-bicyclo[2.2.1.]heptyl and decahydronaphthyl. Cycloalkyl groups may be unsubstituted or substituted with one, two, three, four or more cycloalkyl radicals. The invention is substituted with five substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, amino, aryl, aryloxy, azide, cycloalkyl, cycloalkyloxy, cycloalkenyl, cycloalkynyl, halogen, heteroalkyl, heterocycloalkyl, (heterocycloalkyl)oxy, heteroaryl, hydroxy, nitro, thiol, silyl, cyano, =O, =S, _-SO2R , wherein R is an optionally substituted amino, =NR', wherein R' is H, alkyl, aryl or heterocycloalkyl, and -CON( ^Rz ) ₂ , wherein each ^Rz is independently H or alkyl, or two ^Rz, together with the atoms to which they are attached, are combined to form heterocycloalkyl. Each substituent itself may be unsubstituted or substituted with a substituent as defined herein for each corresponding group.

The term "cycloalkylalkyl" as used herein refers to an alkyl group substituted by a cycloalkyl group, wherein each group is as defined herein. Both the cycloalkyl and alkyl portions of the cycloalkylalkyl group are optionally substituted by substituents as defined herein for each group.

The term "cycloalkylene" herein refers to a divalent cycloalkyl group. The cycloalkylene group is optionally substituted with substituents as defined herein for cycloalkyl groups.

The term "cycloalkynyl" herein refers to a monovalent carbocyclic group having one or two carbon-carbon triple bonds and eight to twelve carbons. The cycloalkynyl group may include a trans-ring bond or bridge bond. Non-limiting examples of cycloalkynyl groups include cyclooctynyl, cyclononynyl, cyclodecynyl and cyclodecadiynyl. The cycloalkynyl group may be unsubstituted or substituted with a substituent defined for cycloalkyl.

The term "heteroalkyl" herein refers to an alkyl, alkenyl, or alkynyl group interrupted once by one or two heteroatoms; an alkyl, alkenyl, or alkynyl group interrupted twice, three times, or four times by one or two heteroatoms each time. Each heteroatom is independently O, N, or S. In some embodiments, the heteroatom is O or N. Two consecutive oxygen or sulfur atoms are not included in the heteroalkyl group. The heteroalkyl group may be unsubstituted or substituted (e.g., an optionally substituted heteroalkyl group). When the heteroalkyl group is substituted and a substituent is attached to a heteroatom, the substituent is selected according to the nature and valence of the heteroatom. Thus, the substituents bonded to the heteroatom are selected, where valence permits, from =O, -N(R ^y ), -SO ₂ OR ^w , -SO ₂ R ^y , -SOR ^w , -COOR ^w , N protecting groups, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocycloalkyl or cyano, wherein each R ^y is independently selected from H, alkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and heterocycloalkyl, and each R ^{w is} independently selected from alkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl and heterocycloalkyl. Each of these substituents may itself be unsubstituted or substituted with substituents defined for each group. When the heteroalkyl group is substituted and the substituent is attached to carbon, the substituent is selected from the substituent groups defined for the alkyl group, as long as the substituent on the carbon atom attached to the heteroatom is not Cl, Br or I. It should be understood that the carbon atom is at the end of the heteroalkyl group.

The term "heteroarylalkyl" herein refers to an alkyl group substituted by a heteroaryl group, wherein heteroaryl and alkyl are as defined herein. The heteroaryl and alkyl portions of the heteroarylalkyl group may be substituted by substituents as defined herein for each corresponding group.

The term "heteroarylene" herein refers to a divalent heteroaryl group. An optionally substituted heteroarylene group refers to a heteroarylene group substituted with a substituent group as defined herein for a heteroaryl group.

The term "heteroaryloxy" as used herein refers to the structure -OR, wherein R is heteroaryl. The heteroaryl portion of the heteroaryloxy group is optionally substituted with substituents as described herein for aryl.

The term "heterocyclyl" herein refers to a fused, bridged and/or spiro 3-, 4-, 5-, 6-, 7- or 8-membered monocyclic, bicyclic, tricyclic or tetracyclic ring system having one, two, three or four heteroatoms independently selected from nitrogen, oxygen and sulfur, unless otherwise specified. In some embodiments, "heterocyclyl" refers to a fused or bridged 5-, 6-, 7- or 8-membered monocyclic, bicyclic, tricyclic or tetracyclic ring system having one, two, three or four heteroatoms independently selected from nitrogen, oxygen and sulfur, unless otherwise specified. Heterocyclyl can be aromatic or non-aromatic. Aromatic heterocyclyls are referred to as heteroaryls. Non-limiting examples of heteroaryl groups include benzimidazolyl, benzofuranyl, benzothiazolyl, benzothiophenyl, benzoxazolyl, furanyl, imidazolyl, indolyl, isoindazolyl, isoquinolyl, isothiazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, purinyl, pyrrolyl, pyridinyl, pyrazinyl, pyrazinyl, quinolyl, thiadiazolyl (e.g., 1,3,4-thiadiazole), thiazolyl, thienyl, triazolyl, tetrazolyl, etc. The term "heterocyclyl" also refers to heterocyclic compounds having a bridged polycyclic structure in which one or more carbon and/or heteroatoms bridge two non-adjacent ring atoms of a single ring, such as quinuclidine, tropane, or diazabicyclo[2.2.2]octane. The term "heterocyclic group" includes bicyclic, tricyclic and tetracyclic groups in which any of the above heterocyclic rings is fused to one, two or three carbon rings, such as an aromatic ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a cyclopentene ring or another monocyclic heterocycle. Examples of fused heterocyclic groups include 1,2,3,5,8,8a-hexahydroindolizine; 2,3-dihydrobenzofuran; 2,3-dihydroindole; and 2,3-dihydrobenzothiophene. The heterocyclyl group may be unsubstituted or substituted by one, two, three, four or five substituents independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylsulfinyl, alkylsulfonyl, amino, aryl, aryloxy, azide, cycloalkyl, cycloalkyloxy, cycloalkenyl, cycloalkynyl, halocycle, heteroalkyl, heterocyclyl, (heterocyclyl)oxy, hydroxy, nitro, thiol, silyl, cyano, -C(O)R or _-SO2R , wherein R is amino or alkyl, =O, =S, =NR', wherein R' is H, alkyl, aryl or heterocyclyl. Each substituent itself may be unsubstituted or substituted by a substituent as defined herein for each corresponding group.

The term "heterocycloalkyl" herein refers to a non-aromatic heterocyclic radical, for example, a non-aromatic 5-membered heterocyclic radical having 0 or 1 double bonds, a non-aromatic 6-membered and 7-membered heterocyclic radical having 0 to 2 double bonds, a non-aromatic 8-membered heterocyclic radical having 0 to 2 double bonds and/or 0 or 1 carbon-carbon triple bonds. Unless otherwise indicated, heterocycloalkyl includes 1 to 16 carbon atoms. Some heterocycloalkyls may include up to 9 carbon atoms. Non-limiting embodiments of heterocycloalkyl include pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, isopiperidinyl, piperazinyl, pyridazinyl. Oxazolinyl, isoxazolinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, isothiazolidinyl, thiazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, dihydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyranyl, dihydropyranyl, dithiazolyl and the like.

The term "heterocyclylalkyl" herein refers to an alkyl group substituted by a heterocyclyl group, wherein each group is as defined herein. The heterocyclyl and alkyl portions are optionally substituted by substituents as defined herein for each group.

The term "heterocyclylene" herein refers to a divalent heterocyclyl group. An optionally substituted heterocyclylene group is optionally substituted with substituents as defined herein for a heterocyclyl group.

The term "(heterocyclyl)oxy" as used herein refers to a chemical substituent of the formula -OR, wherein R is a heterocyclic The heterocyclyl part of the (heterocyclyl)oxy group is optionally substituted with substituents as defined for the heterocyclyl group.

[0026] As used herein, the terms "hydroxy" and "hydroxyl" are used interchangeably to refer to an -OH group.

As used herein, the term "isotopically enriched" means that the pharmaceutically active agent has an isotopic content of an isotope at a predetermined position within the molecule that is at least 100 times greater than the natural abundance of the isotope. For example, a deuterium isotopically enriched composition includes an active agent having an abundance of deuterium at at least one hydrogen atom position that is at least 100 times the natural abundance of deuterium. Preferably, the isotopic enrichment of deuterium is at least 1000 times the natural abundance of deuterium. More preferably, the isotopic enrichment of deuterium is at least 4000 times the natural abundance of deuterium.

As used herein, the term "Myt1" refers to the membrane-associated tyrosine and threonine-specific cdc2-inhibitory kinase (Myt1) (gene name PKMYT1).

The term "Myt1 inhibitor" herein refers to a compound that, when in contact with the enzyme Myt1, whether in vitro, in cell culture or in an animal, reduces the activity of Myt1 such that the measured IC ₅₀ for Myt1 is 10 μM or less (e.g., 5 μM or less or 1 μM or less). For certain Myt1 inhibitors, the IC ₅₀ for Myt1 may be 100 nM or less (e.g., 10 nM or less, or 3 nM or less). Preferably, the IC ₅₀ for Myt1 is 1 nM to 1 μM (e.g., 1 nM to 750 nM, 1 nM to 500 nM, or 1 nM to 250 nM). Even more preferably, the IC ₅₀ for Myt1 is less than 20 nM (e.g., 1 nM to 20 nM).

The term "nitro" as used herein refers to a _-NO2 group.

The term "oxo" as used herein refers to a divalent oxygen atom (eg, the structure of oxo can be shown as =0).

The term "Ph" herein refers to a phenyl group.

The term "pharmaceutical composition" herein refers to a composition containing a compound described herein, formulated with a pharmaceutically acceptable excipient and approved by a governmental regulatory agency for manufacture or sale as part of a therapeutic regimen for treating a disease in a mammal. For example, the pharmaceutical composition can be formulated into a unit dosage form for oral administration (e.g., tablets, capsules, caplets, gel capsules, or syrups); a dosage form for topical administration (e.g., as a cream, gel, lotion, or ointment); a dosage form for intravenous administration (e.g., as a sterile solution without particulate plugs and a solvent system suitable for intravenous use); or any other dosage form described herein.

"Pharmaceutically acceptable salts" refer to certain salts of the above compounds that can retain the original biological activity and are suitable for medical use. Pharmaceutically acceptable salts of the compound represented by formula (I) include, but are not limited to, acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrobromide, hydrochloride, hydroiodide, -hydroxy-ethanesulfonate, lactobionate, lactic acid, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate, and the like. Representative alkali metal or alkaline earth metal salts include but are not limited to Limited to sodium, lithium, potassium, calcium, magnesium, etc., and non-toxic ammonium, quaternary ammonium and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, etc.

The term "protecting group" in the text means a group intended to protect a hydroxyl, amino or carbonyl group from participating in one or more undesirable reactions during a chemical synthesis. As used herein, the term "O-protecting group" means a group intended to protect a hydroxyl or carbonyl group from participating in one or more undesirable reactions during a chemical synthesis. As used herein, the term "N-protecting group" means a group intended to protect a nitrogen-containing group (e.g., an amino, amide, heterocyclic NH or hydrazine) from participating in one or more undesirable reactions during a chemical synthesis. Exemplary O- and N-protecting groups include alkanoyl, aroyl or carbamoyl, such as formyl, acetyl, propionyl, pivaloyl, tert-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthaloyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, tert-butyldimethylsilyl I, triisopropylsilyloxymethyl, 4,4'-dimethoxytrityl, isobutyryl, phenoxyacetyl, 4-isopropylphenoxyacetyl, dimethylformamidinyl and 4-nitrobenzoyl. Exemplary O-protecting groups for protecting carbonyl-containing groups include, but are not limited to, acetal, acyl acetal, 1,3-dithiolane, 1,3-dioxane, 1,3-dioxolane and 1,3-dithiolane.

The term "tautomer" herein refers to structural isomers that are generally susceptible to interconversion by the relocation of protons. Tautomers are different chemical species that can be identified by different spectral features, but generally cannot be separated individually. Non-limiting examples of tautomers include keto-enols, enamines-imines, amides-imidic acids, nitroso-oximes, enones-alkynols, and amino acids-ammonium carboxylates.

[00136] The term "disease" or "disorder" as used herein refers to a state of being or health condition in a patient or subject that can be treated with the compounds or methods provided herein.

"Carrier" refers to a material that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

"Excipient" refers to an inert substance added to a pharmaceutical composition to facilitate administration of a compound. Non-limiting examples include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, and disintegrants.

"Prodrug" refers to a compound of the present invention that can be converted into a biologically active compound through in vivo metabolism. The prodrug of the present invention is prepared by modifying the amino or carboxyl group in the compound of the present invention, and the modification can be removed by conventional operations or in vivo to obtain the parent compound. When the prodrug of the present invention is administered to a mammalian subject, the prodrug is cleaved to form a free amino or carboxyl group.

"Stereoisomers" refer to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and conformational isomers.

"Optional" or "optionally" or "selective" or "optionally" means that the subsequently described event or situation may but need not occur, and the description includes instances where the event or situation occurs and instances where it does not occur. For example, "heterocyclyl optionally substituted with alkyl" means that the alkyl group may but need not be present, and the description includes instances where the heterocyclyl group is substituted with alkyl group and instances where the heterocyclyl group is not substituted with alkyl group.

“Treating,” “treat,” “treatment,” or “alleviation” refers to the need for The invention relates to administering at least one compound disclosed herein, and/or at least one stereoisomer thereof (if any), at least one stable isotope thereof, or at least one pharmaceutically acceptable salt thereof, to a subject suffering from, for example, cancer.

The term "subject" refers to a human or animal (eg, mammal) suffering from or at risk of a disease. Preferably, the subject is a human. Non-limiting examples of diseases and disorders include diseases with symptoms of cell hyperproliferation, such as cancer.

The term "effective amount" refers to an amount of at least one compound disclosed herein, and/or at least one stereoisomer thereof (if any), at least one stable isotope thereof, or at least one pharmaceutically acceptable salt thereof, that is effective for "treating" (as defined above) a disease or disorder in a subject.

The carbon, hydrogen, oxygen, sulfur, nitrogen or F, Cl, Br, I involved in the groups and compounds described in the present invention all include their isotopes, and the carbon, hydrogen, oxygen, sulfur or nitrogen involved in the groups and compounds described in the present invention are optionally further replaced by one or more of their corresponding isotopes, wherein carbon isotopes include ¹² C, ¹³ C and ¹⁴ C, hydrogen isotopes include protium (H), deuterium (D, also called heavy hydrogen), tritium (T, also called super tritium), oxygen isotopes include ¹⁶ O, ¹⁷ O and ¹⁸ O, sulfur isotopes include ³² S, ³³ S, ³⁴ S and ³⁶ S, nitrogen isotopes include ¹⁴ N and ¹⁵ N, fluorine isotopes include ¹⁷ F and ¹⁹ F, chlorine isotopes include ³⁵ Cl and ³⁷ Cl, and bromine isotopes include ⁷⁹ Br and ⁸¹ Br.

Any asymmetric atom (e.g., carbon, etc.) of one or more compounds of the present disclosure may exist in a racemic or enantiomerically enriched configuration (e.g., (R)-, (S)-, or (R,S)-configuration). In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess of (R)- or (S)-configuration; that is, for optically active compounds, for example, one enantiomer is typically used to substantially exclude the other enantiomer. Where possible, substituents at atoms having carbon-carbon double bonds may exist in cis-(Z)- or trans-(E)-form, and unless otherwise indicated, both are included in the present disclosure.

Thus, as used herein, the compounds of the present disclosure may be in the form of one of the possible isomers, rotational isomers, atropisomers, or as a mixture thereof (e.g., as substantially pure geometric (cis or trans) isomers, diastereomers, optical isomers (enantiomers), racemates, or mixtures thereof). As used herein, "substantially pure" or "substantially free of other isomers" means that the product contains less than 5% by weight (e.g., less than 2% by weight) of other isomers, calculated by weight relative to the amount of the preferred isomer.

Various embodiments are disclosed herein. It will be appreciated that the features specified in each embodiment can be combined with other specified features to provide additional embodiments of the present disclosure. The following non-limiting listed embodiments are representative of the present disclosure.

Scheme 1 shows a general method for preparing the compounds and intermediates of the present disclosure. Detailed descriptions and syntheses are disclosed in the following examples. Those skilled in the art will be able to find other synthetic methods or modify the following methods using conventional chemistry to prepare suitable compounds covered by Formula I. These methods are also applicable to the preparation of compounds of other embodiments. Although specific starting materials and reagents are depicted in the schemes and discussed below, other starting materials and reagents can be easily substituted to provide a variety of compounds and/or reaction conditions.

The following examples provide the preparation of representative compounds represented by formula (I) and related structural identification data. It must be noted that the following examples are used to illustrate the present invention rather than to limit the present invention. ¹ H NMR spectra were obtained using a Bruker instrument (400 MHz) and chemical shifts are expressed in ppm. Tetramethylsilane internal standard (0.00 ppm) was used. ¹ H NMR representation method: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublets, dt = doublet of triplet. If coupling constants are provided, the unit is Hz.

The mass spectrum is obtained by LC/MS, and the ionization method can be ESI or APCI.

Thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate, thin layer chromatography (TLC)

The silica gel plate used has a specification of 0.15 mm to 0.2 mm, and the thin layer chromatography separation and purification product has a specification of 0.4 mm to 0.5 mm.

Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

In the following examples, unless otherwise specified, all temperatures are in degrees Celsius. Unless otherwise specified, various starting materials and reagents are commercially available or synthesized according to known methods. Commercially available raw materials and reagents are used directly without further purification. Unless otherwise specified, commercial manufacturers include but are not limited to Aldrich Chemical Company, ABCR GmbH & Co. KG, Acros Organics, Guangzan Chemical Technology Co., Ltd. and Jingyan Chemical Technology Co., Ltd.

CD ₃ OD: deuterated methanol.

CDCl ₃ : deuterated chloroform.

DMSO-d ₆ : deuterated dimethyl sulfoxide.

Argon atmosphere means that the reaction bottle is connected to an argon balloon with a capacity of about 1L.

Unless otherwise specified in the examples, the solution in the reaction refers to an aqueous solution.

The compound is purified using a silica gel column chromatography eluent system and thin layer chromatography, wherein the eluent system is selected from: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: dichloromethane: ethyl acetate; wherein the volume ratio of the solvent varies according to the polarity of the compound, and a small amount of acidic or alkaline reagents, such as acetic acid or triethylamine, may also be added for adjustment.

The following examples illustrate certain embodiments of the present disclosure and how to make and use them. Therefore, the following examples are not intended to limit the scope of the present invention. Those skilled in the art will readily recognize that various non-critical parameters and conditions can be changed or modified to produce substantially the same results. According to one or more of the assays described herein, the following example compounds were found to be inhibitors of the Myt1 protein.

The present invention is further described below with reference to the embodiments, but these embodiments are not intended to limit the scope of the present invention.

In the following examples, the following abbreviations are used:

PE: Petroleum ether

EA: Ethyl acetate

DMF: Dimethylformamide

Synthesis of intermediate A1

Preparation of 4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-trien-2-amine

Step 1: 3-Methoxy-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1b)

3-Bromo-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1a) (10 g, 54.63 mmol), sodium methoxide (29.5 g, 163.39 mmol, 30 w% methanol solution) and cuprous bromide (783.67 mg, 5.46 mmol) were dissolved in DMF (60 mL). The mixture was stirred at 100 °C for 6 h. After cooling to room temperature, the reaction solution was slowly poured into water (300 mL) and extracted with dichloromethane (3×50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation to obtain a crude product. The title compound 3-methoxy-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1b) was obtained by column chromatography purification (PE:EA=100:1 to 10:1).

Step 2: 3-Bromo-4-methoxy-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1c)

Dissolve 3-methoxy-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1b) (5 g, 37.26 mmol) and N-bromosuccinimide (77.30 g, 40.99 mmol) in trifluoroethanol (80 mL). Stir the reaction at room temperature for 4 h. The solvent was dried by rotary evaporation, water (200 mL) was added, and the mixture was extracted with dichloromethane (3×50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation to obtain a crude product. The title compound 3-bromo-4-methoxy-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1c) was obtained by column chromatography purification (PE:EA=100:1 to 10:1).

Step 3: 4-Bromo-2-chloro-3-methoxy-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1d)

Dissolve 3-bromo-4-methoxy-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1c) (3 g, 14.08 mmol) and N-bromosuccinimide (2.07 g, 15.49 mmol) in trifluoroethanol (30 mL). Stir the reaction at room temperature for 4 h. Dry the solvent, add water (100 mL), and extract with dichloromethane (3×30 mL). Dry the organic phase with anhydrous sodium sulfate, filter, and concentrate by rotary evaporation to obtain the crude product. Purify by column chromatography (PE:EA=100:1 to 10:1) to give the title compound 4-bromo-2-chloro-3-methoxy-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1d).

Step 4: 2-Chloro-3-methoxy-4-methyl-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1e)

Dissolve 4-bromo-2-chloro-3-methoxy-bicyclo[4.2.0]octa-1,3,5-triene (compound 1d) (1g, 4.04mmol), methylboronic acid (365mg, 6.10mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (293mg, 0.40mmol) in dioxane (30mL) and water (5mL). Slowly add nitric acid (280mg, 5.86mmol). Heat the reaction solution to 100℃ and stir for 4h. Cool to room temperature, add water (100mL), and extract with dichloromethane (3×30mL). Dry the organic phase with anhydrous sodium sulfate, filter, and concentrate by rotary evaporation to obtain a crude product. Purification by column chromatography (PE:EA=100:1 to 10:1) gave the title compound 2-chloro-3-methoxy-4-methyl-bicyclo[4.2.0]octa-1,3,5-triene (compound 1e).

Step 5: 2-Chloro-3-methoxy-4-methyl-5-nitro-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1f)

Under ice bath, 2-chloro-3-methoxy-4-methyl-bicyclo[4.2.0]octa-1,3,5-triene (compound 1e) (1g, 5.48mmol) was dissolved in concentrated sulfuric acid (8mL). Nitric acid (385mg, 8.20mmol) was slowly added. Stir the reaction for 2h under ice bath. The reaction solution was slowly poured into ice water (100mL) and extracted with dichloromethane (3×50mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation to obtain a crude product. The title compound 2-chloro-3-methoxy-4-methyl-5-nitro-bicyclo[4.2.0]octa-1,3,5-triene (compound 1f) was obtained by column chromatography purification (PE:EA=100:1 to 5:1).

Step 6: 4-methoxy-3-methyl-bicyclo[4.2.0]oct-1,3,5-triene-2-amine (Intermediate A1)

2-Chloro-3-methoxy-4-methyl-5-nitro-bicyclo[4.2.0]octa-1,3,5-triene (Compound 1f) (1 g, 4.39 mmol) and palladium on carbon (100 mg) were dissolved in methanol (20 mL). The mixture was reacted for 16 h under a hydrogen atmosphere. The solid was filtered off and the organic phase was concentrated to give the title compound 4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-triene-2-amine (Intermediate A1).

The intermediates listed in Table 1 were prepared by referring to the preparation method of Intermediate A1.

Table 1

Synthesis of intermediate A3

Preparation of 5-chloro-4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-trien-2-amine

Step 1: 5-Chloro-4-methoxy-3-methyl-bicyclo[4.2.0]oct-1,3,5-trien-2-amine (Intermediate A3)

2-Chloro-3-methoxy-4-methyl-5-nitro-bicyclo[4.2.0]octa-1,3,5-triene (compound 1f) (1g, 4.39mmol) and iron powder (980mg, 17.57mmol) were dissolved in ethanol (10mL) and saturated aqueous ammonium chloride solution (5mL). The reaction solution was heated to 80°C and stirred for 2h. After cooling to room temperature, the ethanol was dried by rotary evaporation, water (100mL) was added, and the mixture was extracted with dichloromethane (3×30mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation to obtain a crude product. The title compound 5-chloro-4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-triene-2-amine (intermediate A3) was obtained by column chromatography purification (PE:EA=100:1 to 5:1).

Synthesis of intermediate A4

Preparation of 6-cyclopropyl-3-methoxy-2-methyl-aniline

Step 1: 1-cyclopropyl-3-methyl-2-nitrobenzene (compound 4b)

1-Bromo-3-methyl-2-nitrobenzene (Compound 4a) (5 g, 23.14 mmol) and cyclopropylboronic acid (2.98 g, 34.72 mmol) were dissolved in toluene (54 mL) and water (6 mL). Potassium phosphate (114.74 g, 69.43 mmol), Pd(OAc) ₂ (780 mg, 3.47 mmol) and tricyclohexylphosphine (1.3 g, 4.63 mmol) were added to the reaction. The reaction solution was heated to 30 °C for 5 h. After cooling to room temperature, water (100 mL) was added and extracted with dichloromethane (3×50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation to obtain a crude product. The title compound 1-cyclopropyl-3-methyl-2-nitrobenzene (Compound 4b) was obtained by column chromatography purification (PE:EA=100:1 to 10:1).

Step 2: 1-Bromo-4-cyclopropyl-2-methyl-3-nitrobenzene (Compound 4c)

Dissolve 1-cyclopropyl-3-methyl-2-nitrobenzene (compound 4b) (5g, 28.22mmol) in concentrated dichloromethane (25mL). Add iron powder (398.66g, 7.14mmol) and ferrous bromide (36.51g, 169.30mmol) in sequence. Slowly add liquid bromine (4.96g, 31.04mmol). The reaction temperature does not exceed 30℃ and stir for 2h. Slowly pour the reaction solution into ice water (100mL) and extract with dichloromethane (3×50mL). The organic phase is dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation to obtain a crude product. Purify by column chromatography (PE:EA=100:1 to 8:1) to obtain the title compound 1-bromo-4-cyclopropyl-2-methyl-3-nitrobenzene (compound 4c).

Step 3: 1-cyclopropyl-4-methoxy-3-methyl-2-nitrobenzene (Compound 4d)

1-Bromo-4-cyclopropyl-2-methyl-3-nitrobenzene (compound 4c) (5g, 19.52mmol), sodium methoxide (10.55g, 58.57mmol, 30w% methanol solution) and cuprous bromide (280mg, 1.95mmol) were dissolved in DMF (30mL). The mixture was heated to 100°C and stirred for 5h. After cooling to room temperature, the reaction solution was slowly poured into water (200mL) and extracted with dichloromethane (3×50mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation to obtain a crude product. The title compound 1-cyclopropyl-4-methoxy-3-methyl-2-nitrobenzene (compound 4d) was obtained by column chromatography purification (PE:EA=100:1 to 10:1).

Step 4: 6-Cyclopropyl-3-methoxy-2-methyl-aniline (Intermediate A4)

1-Cyclopropyl-4-methoxy-3-methyl-2-nitrobenzene (Compound 4d) (1g, 4.83mmol) and iron powder (1.08g, 19.30mmol) were dissolved in ethanol (10mL) and saturated aqueous ammonium chloride solution (5mL). The reaction solution was heated to 80°C and stirred for 2h. After cooling to room temperature, the ethanol was dried by rotary evaporation, water (100mL) was added, and the mixture was extracted with dichloromethane (3×30mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated by rotary evaporation to obtain a crude product. The title compound 6-cyclopropyl-3-methoxy-2-methyl-aniline (Intermediate A4) was obtained by column chromatography purification (PE:EA=100:1 to 5:1).

The intermediates listed in Table 2 were prepared by referring to the preparation method of Intermediate A4.

Table 2

Example 1

2-amino-1-(4-hydroxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-1H-pyrrolidone Preparation of pyrrolidine-3-carboxamide

Method A:

Step 1: 3-Bromo-N-(4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-pyridin-2-amine (Compound 1h)

4-Methoxy-3-methyl-bicyclo[4.2.0]oct-1,3,5-triene-2-amine (intermediate A1) (1.78 g, 10.91 mmol), 5,6-dibromo-2,3-dimethylpyridine (compound 1g) (2.75 g, 10.38 mmol), tris(dibenzylideneacetone)dipalladium (428 mg, 0.47 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethylxanthene (540 mg, 0.94 mmol) and cesium carbonate (8.46 g, 25.95 mmol) were dissolved in ethylene glycol dimethyl ether (40 mL). The mixture was heated to 110° C. and stirred for 6 h under nitrogen protection, cooled to room temperature and concentrated by rotary evaporation. Purification by column chromatography (PE:EA=10:1 to 1:1) gave the title compound 3-bromo-N-(4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-pyridin-2-amine (Compound 1h).

Step 2: 2-amino-1-(4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (Compound 1i)

3-Bromo-N-(4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-pyridin-2-amine (Compound 1h) (2.07 g, 5.97 mmol), malononitrile (833 mg, 12.59 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (373 mg, 0.46 mmol) and sodium tert-butoxide (1.16 g, 12.05 mmol) was dissolved in ethylene glycol dimethyl ether (25 mL). Under nitrogen protection, the reaction was heated to 110°C and stirred for 6 h, cooled to room temperature and concentrated by rotary evaporation. The title compound 2-amino-1-(4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-triene-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (Compound 1i) was obtained by column chromatography purification (PE:EA=10:1 to 1:2).

Step 3: 2-amino-1-(4-hydroxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-triene-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 1)

A mixture of concentrated sulfuric acid (1.2 mL) and water (0.8 mL) was slowly added to methanesulfonic acid (7.5 mL), and then 2-amino-1-(4-methoxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-triene-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (Compound 1i) (1 g, 3 mmol) was slowly added. The reaction solution was stirred at room temperature for 1.5 h. After adding methionine (1.87 g, 12.49 mmol), the reaction solution was heated to 40 ° C and stirred for 6 h. After cooling to room temperature, the reaction solution was added to a solution of dimethyl hydrogen phosphate (1.25 g) and sodium hydroxide (6.8 g) in water (63 mL). Then extracted with ethyl acetate (3×100 mL). The organic phases were combined, dried over sodium sulfate, filtered, concentrated, and purified by reverse phase column to obtain the title compound 2-amino-1-(4-hydroxy-3-methyl-bicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 1).

Method B:

Step 1: (3,5-dibromophenoxy)triisopropylsilane (Compound 1B)

Dissolve 3,5-dibromophenol (Compound 1A) (18 g, 71.46 mmol), imidazole (9.72 g, 142.92 mmol) and triisopropylsilyl chloride (14.48 g, 75.03 mmol) in DMF (150 mL) and stir at room temperature over the weekend. Add water (500 mL) and extract with ethyl acetate (300 mL×3). Combine the organic phases and wash with saturated brine (200 mL). Dry over anhydrous sodium sulfate, filter and concentrate. The residue is purified twice by column chromatography (petroleum ether) to obtain the target compound (3,5-dibromophenoxy)triisopropylsilane (Compound 1B) (26.4 g, yield: 90%).

Step 2: 2-(2,6-dibromo-4-((triisopropylsilyl)oxy)phenyl)1-ethanol (Compound 1C)

Under nitrogen, (3,5-dibromophenoxy)triisopropylsilane (Compound 1B) (23.4 g, 57.32 mmol) was dissolved in ultra-dry THF (200 mL), the reaction solution was cooled to -80 °C, and LDA (6.75 g, 63.05 mmol). After 10 minutes of dripping, solids precipitated in the reaction solution. The temperature of the reaction system was always controlled at about -80°C. After about 40 minutes, ethylene oxide (3.03 g, 68.78 mmol) was slowly added dropwise for a total of 15 minutes. The temperature of the reaction solution was always maintained at -80°C to -77°C. The reaction solution was slowly restored to room temperature and stirred for a total of 3.5 hours. Saturated NH ₄ Cl aqueous solution (200 mL) was added to quench the reaction and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether) to obtain the target compound 2-(2,6-dibromo-4-((triisopropylsilyl)oxy)phenyl)1-ethanol (Compound 1C) (15 g, yield: 58%).

Step 3: (3,5-dibromo-4-(2-bromoethyl)phenoxy)triisopropylsilane (Compound 1D)

2-(2,6-dibromo-4-((triisopropylsilyl)oxy)phenyl)1-ethanol (Compound 1C) (16 g, 35.37 mmol) and triphenylphosphine (18.55 g, 70.75 mmol) were dissolved in DCM (300 mL), and carbon tetrabromide (17.60 g, 53.06 mmol) was slowly added. Stir at room temperature overnight. TLC monitoring showed that the raw material had been completely converted to the target compound. Concentrate. The residue was purified by column chromatography (petroleum ether) to obtain the target compound (3,5-dibromo-4-(2-bromoethyl)phenoxy)triisopropylsilane (Compound 1D) (17 g, yield: 93%).

Step 4: ((5-bromobicyclo[4.2.0]oct-1,3,5-trien-3-yl)oxy)triisopropylsilane (Compound 1E)

In an anhydrous and oxygen-free environment under nitrogen, (3,5-dibromo-4-(2-bromoethyl)phenoxy)triisopropylsilane (Compound 1D) (16g, 31.06mmol) was dissolved in THF (200mL), the reaction solution was cooled to -80°C, and n-BuLi (31ml, 31mmol, 1M) was slowly added dropwise for about 20 minutes. After the addition was completed, the temperature of the reaction solution system was maintained between -80°C and -70°C for a total of 3 hours. After 3 hours, the reaction solution was slowly warmed up and stirred at room temperature overnight. The reaction solution was slowly poured into an ice-cold saturated aqueous ammonium chloride solution (500mL), and ethyl acetate (100mL×3) was added for extraction. The organic phases were combined, washed with saturated brine (300mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate=500/1) to obtain the target compound ((5-bromobicyclo[4.2.0]octa-1,3,5-trien-3-yl)oxy)triisopropylsilane (Compound 1E) (10 g, yield: 90%).

Step 5: tert-Butyl (4-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-2-yl)carbamate (Compound 1F)

Dissolve ((5-bromobicyclo[4.2.0]oct-1,3,5-trien-3-yl)oxy)triisopropylsilane (Compound 1E) (6 g, 16.88 mmol), NH ₂ Boc (3.95 g, 33.77 mmol), Pd ₂ (dba) ₃ (775 mg, 0.85 mmol), JohnPhos (504 mg, 1.69 mmol) and t-BuONa (3.25 g, 33.77 mmol) in 1,4-dioxane (250 mL). Stir and heat to 70°C under nitrogen protection for 15 min. TLC monitors that the raw material has reacted completely. Cool the reaction solution to room temperature, add water (100 mL), and extract with ethyl acetate (200 mL×3). Combine the organic phases, wash with saturated brine (200 mL), dry over anhydrous sodium sulfate, filter, and concentrate. The residue was purified by column chromatography (petroleum ether/ethyl acetate=100/1) to obtain Target compound tert-butyl (4-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-2-yl)carbamate (Compound 1F) (5 g, yield: 76%).

Step 6: 4-((Triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-trien-2-amine (Compound 1G)

Tert-butyl (4-((triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-trien-2-yl)carbamate (Compound 1F) (1 g, 2.55 mmol) was dissolved in HFIP (5 mL) and heated to 150°C in a microwave reactor for 15 min. TLC monitoring showed that the raw material had been completely converted. The reaction was carried out in parallel for 5 times and combined. Concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate = 100/1) to obtain the target compound 4-((triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-trien-2-amine (Compound 1G) (3 g, yield: 81%).

Step 7: 3-Bromo-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-trien-2-amine (Compound 1H)

4-((Triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-triene-2-amine (Compound 1G) (2.5 g, 8.51 mmol) and AcOH (0.51 g, 8.51 mmol) were dissolved in toluene (100 mL). NBS (1.53 g, 8.51 mmol) was added in batches under an ice-salt bath and stirred for 30 minutes. The reaction solution was returned to room temperature, water (100 mL) was added, and extracted with ethyl acetate (100 mL×3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse phase column separation (mobile phase: methanol) to obtain the target compound 3-bromo-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-triene-2-amine (Compound 1H) (yellow solid, 1.1 g, yield: 35%).

Step 8: 3-methyl-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-trien-2-amine (Compound 1I)

3-Bromo-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-2-amine (Compound I) (2.64 g, 7.13 mmol), trimethylcyclotriboroxane (4.48 g, 35.65 mmol), Pd(dppf) ₂ Cl ₂ (0.42 g, 0.57 mmol) and K ₂ CO ₃ (2.46 g, 17.82 mmol) were dissolved in 1,4-Dioxane (32 mL) and H ₂ O (8 mL), and heated to 130°C in a microwave reactor under nitrogen protection for 30 minutes. The reaction solution was cooled to room temperature, water (50 mL) was added, and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate = 10/1) to give the target compound 3-methyl-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-trien-2-amine (Compound 1I) (yellow oil, 1.56 g, yield: 72%).

Step 9: 3-Bromo-5,6-dimethyl-N-(3-methyl-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-trien-2-yl)pyridin-2-amine (Compound 1J)

3-Methyl-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-trien-2-amine (Compound 1I) (1.88 g, 6.15 mmol), 5,6-dibromo-2,3-dimethylpyridine (1.79 g, 6.77 mmol), Pd ₂ (dba) ₃ (281.36 mg, 0.31 mmol), XantPhos (356.09 mg, 0.62 mmol) and Cs ₂ CO ₃ (4.01 g, 12.3 mmol) were dissolved in tert-amyl alcohol (20 mL), and stirred and heated to 120°C under nitrogen protection for 3 h. The reaction solution was cooled to room temperature, water (50 mL) was added, and extracted with ethyl acetate (30 mL×3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether/ethyl acetate = 10/1) to give the target compound 3-bromo-5,6-dimethyl-N-(3-methyl-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]oct-1,3,5-trien-2-yl)pyridine-2-amine (Compound 1J) (yellow solid, 2.57 g, yield: 85%).

Step 10: 2-amino-5,6-dimethyl-1-(3-methyl-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (Compound 1K)

Under nitrogen, dissolve malononitrile (805 mg, 12.18 mmol) in DME (30 mL), cool to 0°C, add NaH (464 mg, 11.6 mmol), stir for 20 min, continue to add 3-bromo-5,6-dimethyl-N-(3-methyl-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-2-yl)pyridin-2-amine (Compound 1J) (2.84 g, 5.80 mmol) and Pd(dppf)Cl ₂ (340 mg, 0.46 mmol), replace nitrogen protection, heat to 105°C for 5 h. Cool the reaction solution to room temperature, pour into ice water (30 mL), and extract with ethyl acetate (50 mL×3). Combine the organic phases, wash with saturated brine (50 mL), dry over anhydrous sodium sulfate, filter, and concentrate. The residue was purified by column chromatography (petroleum ether/ethyl acetate=10/1) to give the target compound 2-amino-5,6-dimethyl-1-(3-methyl-4-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-2-yl)-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (Compound 1K) (white solid, 985 mg, yield: 36%).

Step 11: 2-amino-1-(4-((tert-butyldimethylsilyl)oxy)-3-methylbicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 1L)

2-Amino-1-(4-((tert-butyldimethylsilyl)oxy)-3-methylbicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 1L) (985 mg, 2.07 mmol) and LiOH (261 mg, 6.21 mmol) were dissolved in H ₂ O ₂ (4 mL) and EtOH (60 mL), stirred and heated to 50°C, and reacted for 3 h. LCMS monitoring showed that the raw material had been completely converted. The reaction solution was cooled to room temperature, water (100 mL) was added, and extracted with DCM/MeOH=10/1 (50 mL×3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product of 2-amino-1-(4-((tert-butyldimethylsilyl)oxy)-3-methylbicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 1L) (1 g) was used directly in the next step.

Step 12: 2-amino-1-(4-hydroxy-3-methylbicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 1)

2-Amino-1-(4-((tert-butyldimethylsilyl)oxy)-3-methylbicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 1L) (1 g) was dissolved in THF (10 mL), the reaction solution was cooled to 0°C, TBAF (0.60 g, 2.28 mmol) was added dropwise, and stirred for 10 min. LCMS monitoring showed that the residue had been completely converted into the target product. The reaction solution was warmed to room temperature, water (50 mL) was added, and extracted with DCM/MeOH=10/1 (50 mL×3). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by reverse phase column to obtain the target product 2-amino-1-(4-hydroxy-3-methylbicyclo[4.2.0]octa-1,3,5-trien-2-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 1) (70 mg, yield: 10%).

MS (ESI) m/z: 337.2 [M+H] +

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.56 (s, 1H), 7.83 (s, 1H), 6.85 (s, 2H), 6.79 (s, 1H), 6.66 (s, 2H), 3.12-3.07 (m, 2H), 2.96-2.89 (m, 1H), 2.84-2.76 (m, 1H), 2.30 (s, 3H), 2.29 (s, 3H), 1.74 (s, 3H).

The compounds in Examples 2-12 were prepared by using different intermediates and referring to the method of Example 1. For the specific compound structures, please refer to Table 3-4 below.

table 3

Table 4

Example 13

2-Amino-1-(5-hydroxy-2,4-dimethyl-bicyclo[4.2.0]octa-1,3,5-trien-3-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide

Step 1: 5-allyloxy-1,3-dibromo-2-methylbenzene (Compound 13b)

Potassium carbonate (52.49 g, 379.8 mmol) and allyl bromide (27.57 g, 227.88 mmol) were added to a solution of 3,5-dibromo-4-methylphenol (50.5 g, 189.90 mmol) in N,N-dimethylformamide (400 mL). The mixture was stirred at room temperature for 16 hours, diluted with ethyl acetate (2000 mL), washed with water (400 mL×5), dried over anhydrous sodium sulfate, and concentrated to obtain a crude brown oily title compound (58.1 g, yield: 100%), which was directly used in the next step.

Step 2: 2-allyl-3,5-dibromo-4-methylphenol (Compound 13c)

A solution of 5-allyloxy-1,3-dibromo-2-methylbenzene (58.1 g, 189.88 mmol) in 1,2-dichlorobenzene (120 mL) was heated to 250° C. in a microwave reactor, stirred for 20 minutes, cooled to room temperature and concentrated under reduced pressure to give a crude brown oily title compound (58 g, yield: 100%), which was used directly in the next reaction.

Step 3: (2-allyl-3,5-dibromo-4-methylphenoxy)triisopropylsilane (Compound 13d)

Triisopropylsilyl chloride (38.37 g, 199.03 mmol) was added to a solution of 2-allyl-3,5-dibromo-4-methylphenol (58 g, 189.55 mmol) and imidazole (25.78 g, 379.1 mmol) in N,N-dimethylformamide (400 mL). The reaction mixture was stirred at room temperature under nitrogen atmosphere for 16 hours, diluted with ethyl acetate (2000 mL), and washed with water (500 mL × 4) and saturated brine (200 mL). The residue was washed, dried over anhydrous sodium sulfate and concentrated. The residue was purified by column chromatography (petroleum ether) to obtain the compound (85 g, yield: 97%) as a yellow solid.

Step 4: 2-(2,4-dibromo-3-methyl-6-((triisopropylsilyl)oxy)phenyl)acetaldehyde (Compound 13e)

To a solution of (2-allyl-3,5-dibromo-4-methylphenoxy)triisopropylsilane (85 g, 183.85 mmol) in tetrahydrofuran (600 mL) at room temperature, potassium osmate aqueous solution (677 mL, 3.68 mmol, w/w: 0.2%) and sodium periodate (78.65 g, 367.7 mmol) were added successively. The reactants were stirred at room temperature overnight, diluted with ethyl acetate (2000 mL), washed successively with sodium sulfite (400 mL), water (300 mL) and saturated brine (300 mL), dried over anhydrous sodium sulfate and concentrated to give the title compound as a solid (85.4 g, yield: 100%).

MS (ESI) m/z: 465.1 [M+H]+

Step 5: 2-(2,4-dibromo-3-methyl-6-((triisopropylsilyl)oxy)phenyl)ethane-1-ol (Compound 13f)

Sodium borohydride (8.35 g, 220.72 mmol) was added in portions to a solution of 2-(2,4-dibromo-3-methyl-6-((triisopropylsilyl)oxy)phenyl)acetaldehyde (85.4 g, 183.93 mmol) in ethanol (600 mL) at 0°C. The mixture was stirred at room temperature for 1 hour, saturated aqueous ammonium chloride solution (200 mL) was added, diluted with ethyl acetate (2000 mL), washed with water (200 mL) and saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by flash column chromatography on silica gel (petroleum ether: ethyl acetate = 40: 1 to 10: 1) to give the title compound (55.5 g, yield: 65%) as a colorless oil.

MS (ESI) m/z: 467.1 [M+H]+

Step 6: (3,5-dibromo-4-methyl-2-(2-(tetrahydro-2H-pyran-2-yl)oxy)ethyl)phenoxy)triisopropylsilane (Compound 13g)

To a solution of 2-(2,4-dibromo-3-methyl-6-[(tri(propan-2-yl)silyl)oxy]phenyl)ethane-1-ol (55.5 g, 119.02 mmol) in tetrahydrofuran (500 mL) was added dihydropyran (30.04 g, 357.06 mmol) and 4-methylbenzenesulfonic acid pyridinium-1-ium salt (2.99 g, 11.90 mmol) at room temperature. The mixture was stirred at 80 ° C for 3 hours and concentrated. The residue was dissolved in ethyl acetate (600 mL), washed with water (200 mL) and saturated brine (200 ml), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel flash column chromatography (petroleum ether: ethyl acetate = 80: 1 to 40: 1) to give the title compound (48.7 g, yield: 74%) as a yellow oil.

MS (ESI) m/z: 573.1 [M+23] +

Step 7: N-benzyl-3-bromo-2-methyl-4-(2-(tetrahydro-2H-pyran-2-yl)oxy)ethyl)-5-(triisopropylsilyl)oxy)aniline (Compound 13h)

(3,5-dibromo-4-methyl-2-(2-(tetrahydro-2H-pyran-2-yl)oxy)ethyl)phenoxy)triisopropylsilane (42 g, 76.30 mmol) was dissolved in 1,4-dioxane (100 mL), and benzylamine (8.18 g, 76.3 mmol), Pd ₂ (dba) ₃ (3.49 g, 3.81 mmol), JohnPhos (3.42 g, 11.44 mmol) and sodium tert-butoxide (11.00 g, 114.45 mmol) were added. The reaction system was replaced with nitrogen and stirred at 80°C for 6 hours. The mixture was cooled to room temperature and concentrated, diluted with ethyl acetate (2000 mL), washed with water (400 mL) and saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered and concentrated under pressure. The residue was purified by flash chromatography on silica gel (petroleum ether/ethyl acetate=120/1 to 40/1) to give the title compound (12 g, yield: 27%) as a colorless liquid.

MS (ESI) m/z: 576.2 [M+H] +

Step 8: N,N-dibenzyl-3-bromo-2-methyl-4-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-5-(triisopropylsilyl)oxyaniline (Compound 13i)

Potassium carbonate (5.74 g, 41.62 mmol) and benzyl bromide (5.34 g, 31.21 mmol) were added to a solution of N-benzyl-3-bromo-2-methyl-4-(2-(tetrahydro-2H-pyran-2-yl)oxy)ethyl)-5-(triisopropylsilyl)oxy)aniline (12 g, 20.81 mmol) in acetonitrile (50 mL) at room temperature. The reaction solution was stirred at 100 ° C for 16 hours, cooled to room temperature and concentrated. The residue was dissolved in ethyl acetate (400 mL), washed with water (100 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel flash column chromatography (petroleum ether: ethyl acetate = 120: 1 to 80: 1) to give the title compound (10.7 g, yield: 77%) as a colorless oil.

MS (ESI) m/z: 666.2 [M+H]+

Step 9: 2-(2-bromo-4-(dibenzylamino)-3-methyl-6-((triisopropylsilyl)oxy)phenyl)ethan-1-ol (Compound 13j)

At room temperature, 4-methylbenzenesulfonic acid pyridinium-1-ium salt (1.21 g, 4.82 mmol) was added to a solution of N,N-dibenzyl-3-bromo-2-methyl-4-((tetrahydro-2H-pyran-2-yl)oxy)ethyl)-5-(triisopropylsilyl)oxyaniline (10.7 g, 16.05 mmol) in methanol (120 mL). The mixture was stirred at 80 ° C for 4 hours, cooled to room temperature and concentrated. The residue was dissolved in ethyl acetate (400 mL), washed with water (50 mL) and saturated brine (50 ml), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by flash column chromatography on silica gel (petroleum ether: ethyl acetate = 80: 1 to 20: 1) to give the title compound (7.6 g, yield: 81%) as a colorless oil.

MS (ESI) m/z: 584.1 [M+H]+

Step 10: N,N-dibenzyl-3-bromo-4-(2-bromoethyl)-2-methyl-5-((triisopropylsilyl)oxy)aniline (Compound 13k)

Carbon tetrabromide (8.80 g, 26.91 mmol) was dissolved in dichloromethane (120 mL), and triphenylphosphine (7.94 g, 30.28 mmol) was added under ice-cooling. After stirring for 10 minutes, 2-(2-bromo-4-(dibenzylamino)-3-methyl-6-((triisopropylsilyl)oxy)phenyl)ethane-1-ol (9.8 g, The reaction solution was stirred at room temperature for 16 hours, concentrated under reduced pressure, and the residue was purified by silica gel flash column chromatography (petroleum ether) to obtain the title compound (8.9 g, yield: 82%) as a white solid.

MS (ESI) m/z: 646.2 [M+H] +

Step 11: N,N-dibenzyl-2-methyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-3-amine (Compound 131)

Dissolve N,N-dibenzyl-3-bromo-4-(2-bromoethyl)-2-methyl-5-((triisopropylsilyl)oxy)aniline (8.9 g, 13.79 mmol) in dry tetrahydrofuran (100 mL), cool to -78 ° C, and slowly drop n-butyl lithium solution (5.52 mL, 13.79 mmol, 2.5 M n-hexane solution). After the addition is complete, continue to stir at -78 ° C for 2 hours, slowly warm the reaction solution to room temperature and stir overnight. Add water (50 mL) to quench the reaction, extract with ethyl acetate (200 mL), wash with saturated brine (30 mL), dry with anhydrous sodium sulfate, filter and concentrate to obtain the target compound (6.7 g, yield: 100%) as a yellow oil.

MS (ESI) m/z: 486.3 [M+H] +

Step 12: 2-methyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-triene-3-amine (Compound 13m)

At room temperature, formic acid (4.17 g, 90.60 mmol) and potassium hydroxide (2.54 g, 45.30 mmol) and palladium carbon (800 mg, containing 10% palladium) were added to a solution of N, N-dibenzyl-2-methyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-triene-3-amine (4.4 g, 9.06 mmol) in ethanol (60 mL) successively. The reaction mixture was stirred at 70 ° C for 1 hour, cooled, filtered and concentrated. The residue was purified by flash column chromatography on silica gel (petroleum ether: ethyl acetate = 80: 1 to 40: 1) to give the title compound (2.51 g, yield: 91%) as a colorless oil.

MS (ESI) m/z: 306.2 [M+H]+

Step 13: 4-Bromo-2-methyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-triene-3-amine (Compound 13n)

To a solution of 2-methyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-triene-3-amine (2.51 g, 8.22 mmol) in acetonitrile (50 mL) was added NBS (1.32 g, 7.40 mmol) at -30°C. The mixture was stirred at room temperature for 2 hours, concentrated under reduced pressure, and the residue was purified by silica gel flash column chromatography (petroleum ether:ethyl acetate=120:1) to give the title compound (3.1 g, yield: 67%) as a yellow solid.

MS (ESI) m/z: 384.2 [M+H] +

Step 14: 2,4-dimethyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-triene-3-amine (Compound 13o)

Methylmagnesium bromide (13.43 mL, 40.30 mmol) was added to a 250 mL three-necked flask at 0°C under nitrogen atmosphere, and zinc dichloride (80.60 mL, 40.30 mmol, 0.5 M tetrahydrofuran solution) was slowly added dropwise. The reaction solution was stirred at 0°C for 5 minutes, and a solution of 4-bromo-2-methyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-triene-3-amine (3.1 g, 8.06 mmol) and bis(tri-tert-butylphosphine)palladium(0) (411.87 mg, 0.81 mmol) in N,N-dimethylformamide (45 mL) was slowly added dropwise to the bottle. After the addition was complete, the reaction solution was warmed to room temperature and stirred for 6 hours. The reaction solution was cooled in an ice bath again, saturated aqueous ammonium chloride solution (80 mL) was added dropwise to quench the reaction, diluted with water, extracted with ethyl acetate (400 mL), washed with water (100 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by silica gel flash chromatography (petroleum ether/ethyl acetate = 60/1) to obtain the title compound (2.51 g, yield 97%).

MS (ESI) m/z: 320.2 [M+H]+

Step 15: 3-Bromo-N-(2,4-dimethyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-3-yl)-5,6-dimethylpyridin-2-amine (Compound 13p)

2,3-Dibromo-5,6-dimethylpyridine (2.29 g, 8.63 mmol), tris(dibenzylideneacetone)dipalladium(0) (359.42 mg, 0.39 mmol), 4,5-bis(diphenylphosphino-9,9-dimethylxanthene) (454.22 mg, 0.79 mmol) and cesium carbonate (7.67 g, 23.55 mmol) were added to a solution of 2,4-dimethyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-triene-3-amine (2.51 g, 7.85 mmol) in ethylene glycol dimethyl ether (40 mL) in succession. The reaction solution was stirred at 100° C. under nitrogen atmosphere for 6 hours, cooled to room temperature and concentrated. The residue was dissolved in ethyl acetate (60 mL), washed with water (10 mL) and brine (10 ml), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate=40:1) to give the title compound (3.9 g, yield: 99%) as a white solid.

MS (ESI) m/z: 503.2 [M+H] +

Step 16: 2-amino-1-(2,4-dimethyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-3-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (Compound 13q)

To a solution of malononitrile (0.83 g, 12.52 mmol) in ethylene glycol dimethyl ether (50 mL) was added sodium tert-butoxide (1.15 g, 11.92 mmol) at room temperature. The mixture was stirred under a nitrogen atmosphere for 10 minutes, and 3-bromo-N-(2,4-dimethyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-3-yl)-5,6-dimethylpyridin-2-amine (3 g, 5.96 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (486.72 mg, 0.60 mmol) were added. The reaction solution was stirred at 110°C under a nitrogen atmosphere for 12 hours and concentrated. The residue was dissolved in ethyl acetate (200 mL), washed with water (60 mL) and brine (60 ml), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by flash column chromatography on silica gel (petroleum ether:ethyl acetate=40:1 to 10:1) to give the title compound (1.37 g, yield: 47%) as a white solid.

MS (ESI) m/z: 489.3 [M+H] +

Step 17: 2-amino-1-(2,4-dimethyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0] Octa-1,3,5-trien-3-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 13r)

To a mixed solution of 2-amino-1-(2,4-dimethyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-trien-3-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (1.37 g, 2.80 mmol) in ethanol (20 mL) and water (10 mL) was added (dimethylphosphonic acid) platinum (II) hydrogen complex (11.96 mg, 0.028 mmol). The reaction solution was stirred at 80°C for 8 hours, cooled and concentrated. The residue was dissolved in ethyl acetate (200 mL), washed with water (50 mL) and brine (50 ml), dried over anhydrous sodium sulfate, filtered and concentrated to give the title compound as a white solid (1.4 g, yield: 99%).

MS (ESI) m/z: 507.2 [M+H]+

Step 18: 2-amino-1-(5-hydroxy-2,4-dimethylbicyclo[4.2.0]octa-1,3,5-trien-3-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (Compound 13)

CsF (1.26 g, 8.28 mmol) was added to a solution of 2-amino-1-(2,4-dimethyl-5-((triisopropylsilyl)oxy)bicyclo[4.2.0]octa-1,3,5-triene-3-yl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carboxamide (1400 mg, 2.76 mmol) in N,N-dimethylformamide (15 mL). The mixture was stirred at room temperature for 2 hours. The mixture was purified by preparative HPLC (methanol/10 mM aqueous ammonium bicarbonate solution, 10%-100%) and lyophilized to give the title compound (770 mg, yield: 80%) as a white solid.

MS (ESI) m/z: 351.1 [M+H]+

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 9.41 (s, 1H), 7.82 (s, 1H), 6.68 (s, 2H), 6.62 (s, 2H), 3.11–3.06 (m, 2H), 3.06–2.98 (m, 2H), 2.28 (s, 3H), 2.26 (s, 3H), 1.63 (s, 3H), 1.60 (s, 3H).

Biological evaluation

The following assays were used to test exemplary compounds of the invention. The data for those examples tested according to the following procedure are shown in Table 5 below.

Test Example 1: In vitro enzyme activity test

The Myt1 kinase activity was detected using a recombinant human Myt1 kinase assay and the ADP-Glo fluorescence luminescence assay was used to measure ATP hydrolysis.

Human recombinant Myt1 (Thermo Fisher #A33387) was diluted with enzyme reaction buffer (70mM Hepes, 3mM MgCl2, 3mM MnCl2, 50ug/mL PEG20000, 3μM sodium orthovanadate, 1.2mM dithiothreitol) to obtain the enzyme reaction buffer of human recombinant Myt1. 5μL of human recombinant Myt1 enzyme reaction buffer was added to a 384-well plate. The test compound was diluted 5 times in a 9-point concentration range in DMSO. A series of diluted DMSO solutions of the test compound or pure DMSO control solution was added to a 384-well plate. Then, incubate at room temperature for 15 minutes. Finally, 5μL of adenosine triphosphate (ATP) was added to the 384-well plate to start the enzymatic reaction. At the beginning of the enzymatic reaction, the concentration of ATP was 10μM and the concentration of Myt1 enzyme was 82nM. Then, the reactant was incubated at 37°C for 1 hour. After the incubation, 15 μL ADP-GLO reagent was added and the 384-well plate was incubated at room temperature for 40 minutes. Finally, 30 μL kinase detection reagent was added and incubated at room temperature for 30 minutes before measuring the chemiluminescence value using a microplate reader. Each experiment was repeated 3 times.

Molecular Devices ID5 operating software was used to perform 4-parameter nonlinear regression equation (variable slope) curve fitting and determine _IC50 values, which are shown in Table 5. _IC50 refers to the concentration of the compound at which the activity of Myt1 is inhibited by 50%.

Test Example 2: Cell Activity Test

The following method was used to determine the effects of preferred compounds of the present invention on tumor cell proliferation.

The experimental method is briefly described as follows: 90 μL of tumor cell OVCAR-3 cell suspension (2500 cells per well) was inoculated into a 96-well plate. The culture plate was then placed in a CO2 incubator for 24 hours to allow the cells to fully adhere. On the second day, the compound to be tested was dissolved in DMSO and then prepared into a 10 mM stock solution. The compound was continuously diluted 5-fold with DMSO to 9 different concentrations; 5 μL of the above gradient diluted compound solution was transferred to a 96-well plate containing 45 μL of culture medium and mixed thoroughly to obtain a 100× compound stock solution; then 10 μL of the above 100× compound was transferred to a 96-well plate inoculated with 90 μL of cells to obtain a 10× concentration; finally, 10 μL of the above 10× compound was transferred to a 96-well plate inoculated with 90 μL of cells to obtain a final 1× concentration compound, in which the final concentration of DMSO was 0.1%. The plate was gently shaken to fully mix the drug, and the cell culture plate was placed in a _CO2 incubator for further culture for 4 days. On the 4th day, CellTiter-Glo reagent (25 μL) was added to each test well, mixed on a shaker for 2 minutes, and then incubated at room temperature for 25 minutes. The luminescence intensity was measured using an ID5 multifunctional microplate reader. The Molecular Devices ID5 operating software was used to perform a 4-parameter nonlinear regression equation (variable slope) curve fitting and determine the IC ₅₀ value, which is shown in Table 5. Among them, IC ₅₀ refers to the concentration of the compound when cell viability is inhibited by 50%.

In the following Table 5, A represents less than 0.2 μM; B represents 0.2 to 1 μM; C represents 1 to 10 μM; and D represents >10 μM.

Table 5 IC ₅₀ values of the preferred compounds of the present invention for inhibition of Myt1 kinase activity

Claims

A compound represented by general formula (I) or a pharmaceutically acceptable salt thereof:

in:

R 1 and R 2 are independently selected from H, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-8 cycloalkyl, C 3-8 cycloalkenyl, C 2-9 heterocycloalkyl, C 1-6 alkyl substituted by C 2-9 heterocycloalkyl, C 6-10 aryl, C 1-9 heteroaryl, C 1-6 alkyl substituted by C 1-9 heteroaryl, halogen, cyano, -N(R a ) 2 , -OR a , -C(O)N(R 9 ) 2 , -SO 2 N(R 9 ) 2 , -SO 2 R b and -QR c ; or R 1 and R 2 form an optionally substituted C 3-8 alkylene group, which may be substituted by 0 , 1, 2 or 3 substituents selected from C 1-6 alkyl;

R3 is selected from halogen, cyano, optionally substituted C1-6 alkyl, 3-membered, 4-membered, 5-membered or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S, the optionally substituted C1-6 alkyl optionally substituted by 3-membered, 4-membered, 5-membered or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members;

R4 is selected from cyano, optionally substituted C1-6 alkyl, 3-, 4-, 5- or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members, the optionally substituted C1-6 alkyl optionally substituted by 3-, 4-, 5- or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members;

R 5 is H or -N(R a ) 2 ;

R 6 is -C(O)NH(R 9 ), -C(O)R b or -SO 2 R b ;

R7 is selected from H, OH, halogen and C1-6 alkyl; or, R7 and R4 or R7 and R8 are combined to form an optionally substituted C2-4 alkylene, wherein the optionally substituted C2-4 alkylene is optionally substituted by 0, 1, 2 or 3 substituents selected from C1-6 alkyl;

R 8 is H or halogen;

Each Ra is independently selected from H, C1-6 alkyl, C1-6 alkyl substituted by C6-10 aryl, C3-8 cycloalkyl, C6-10 aryl, C2-9 heterocycloalkyl, C1-9 heteroaryl, C1-6 alkyl substituted by C1-9 heteroaryl, and -SO2Rb ; or two Ra groups and the atoms to which the two Ra groups are connected together form a C2-9 heterocycloalkyl ;

Each R b is independently selected from C 1-6 alkyl, C 3-6 cycloalkyl and C 6-10 aryl;

each R c is independently selected from -OH, C 1-6 alkyl, C 6-10 aryl, C 2-9 heterocycloalkyl, C 1-9 heteroaryl, -N(R a ) 2 , -C(O)N(R 9 ) 2 , -SO 2 N(R 9 ) 2 , -SO 2 R b , and alkoxy;

Each R 9 is independently selected from H, C 1-6 alkyl, C 2-6 alkoxyalkyl, C 1-6 substituted by C 6-10 aryl alkyl, C 6-10 aryl, C 3-8 cycloalkyl and C 1-9 heteroaryl; or two R 9 groups and the atoms to which the two R 9 groups are connected together form a C 2-9 heterocyclic group;

Q is selected from C 1-6 alkylene, C 2-6 alkenylene, C 3-8 cycloalkylene, C 6-10 arylene, C 2-9 heterocycloalkylene and C 1-9 heteroarylene,

When R 3 is halogen or optionally substituted C 1-6 alkyl, said R 4 is not optionally substituted C 1-6 alkyl; or

When R 4 is an optionally substituted C 1-6 alkyl, said R 3 is not halogen or an optionally substituted C 1-6 alkyl; or

When R 3 is halogen or optionally substituted C 1-6 alkyl and R 4 is optionally substituted C 1-6 alkyl, at least one of R 7 or R 8 is not H.
The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is enriched in atropisomers of formula (IA):

Wherein, when R7 is -OH, the R3 and R4 groups are different.
The compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 3 is cyano.
The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein R3 is a 3-membered, 4-membered, 5-membered or 6-membered saturated cycloalkyl or heterocycloalkyl group containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S.
The compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 3 is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
The compound or pharmaceutically acceptable salt thereof according to claim 5, wherein R 3 is cyclopropyl.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein R 4 is cyano.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein R 4 is a 3-, 4-, 5- or 6-membered saturated cycloalkyl or heterocycloalkyl group containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members.
The compound or pharmaceutically acceptable salt thereof according to claim 8, wherein R 4 is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
The compound or pharmaceutically acceptable salt thereof according to claim 9, wherein R 4 is cyclopropyl.
The compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 3 is halogen or C 1-6 alkyl, and R 4 is cyano.
The compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 3 is halogen or C 1-6 alkyl, and R 4 is a 3-membered, 4-membered, 5-membered or 6-membered saturated cycloalkyl or heterocycloalkyl containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S as ring members.
The compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 4 is C 1-6 alkyl and R 3 is cyano.
The compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein R 4 is a C 1-6 alkyl group, and R 3 is a 3-membered, 4-membered, 5-membered or 6-membered saturated cycloalkyl or heterocycloalkyl group containing 0, 1, 2 or 3 N atoms and 0, 1 or 2 atoms selected from O and S.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein R 7 and R 4 or R 7 and R 8 combine to form a C 2-4 alkylene group, and the C 2-4 alkylene group may have 0, 1, 2 or 3 substituents selected from C 1-6 alkyl groups.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein R 7 and R 4 or R 7 and R 8 combine to form a C 2 alkylene group, and the C 2 alkylene group may have 0, 1, 2 or 3 substituents selected from C 1-6 alkyl groups.
The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein R 7 and R 4 or R 7 and R 8 combine to form a C 3 alkylene group, and the C 3 alkylene group may have 0, 1, 2 or 3 substituents selected from C 1-6 alkyl groups.
The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein R 7 and R 4 or R 7 and R 8 combine to form a C 4 alkylene group, and the C 4 alkylene group may have 0, 1, 2 or 3 substituents selected from C 1-6 alkyl groups.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, wherein R 7 is -OH or C 1-6 alkyl.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 14, wherein R 2 is H, halogen, C 1-6 alkyl, C 1-6 alkyl substituted by C 2-9 heterocycloalkyl, or C 1-6 alkyl substituted by C 1-9 heteroaryl.
The compound according to claim 20 or a pharmaceutically acceptable salt thereof, wherein R2 is H.
The compound or pharmaceutically acceptable salt thereof according to claim 20, wherein R 2 is C 1-6 alkyl.
The compound or pharmaceutically acceptable salt thereof according to claim 20, wherein R2 is methyl, ethyl, isopropyl or butyl.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein R 1 is C 1-6 alkyl.
The compound according to claim 24 or a pharmaceutically acceptable salt thereof, wherein R 1 is methyl, ethyl, isopropyl or butyl.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein R 1 is C 1-9 heteroaryl.
The compound according to claim 26 or a pharmaceutically acceptable salt thereof, wherein R 1 is 1,3-thiazolyl, 1,2-thiazolyl, 1,3-oxazolyl, benzo-1,3-thiazolyl, benzo-1,3-oxazolyl, indolyl, benzimidazolyl, pyridyl, imidazolyl, pyrimidinyl, pyrazinyl, pyridazinyl or pyrazolyl.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein R 1 is C 3-8 cycloalkyl.
The compound or pharmaceutically acceptable salt thereof according to claim 28, wherein R 1 is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein R 1 is C 2-9 heterocycloalkyl.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein R 1 is C 6-10 aryl.
The compound according to any one of claims 1 to 31, or a pharmaceutically acceptable salt thereof, wherein R 1 may be substituted by 1, 2 or 3 groups selected from methyl, difluoromethyl, trifluoromethyl, fluorine, chlorine, bromine, amino, hydroxyl, cyano, =O, -C(O)NH 2 , -C(O)NH(Me), -C(O)N(Me) 2 , -(CH 2 )nC(O)OH and -(CH 2 )nC(O)Ot-Bu, wherein n is 0 or 1.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein R 1 is H or halogen.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 23, wherein R 1 is -N(R a ) 2 .
The compound or pharmaceutically acceptable salt thereof according to claim 34, wherein R 1 is diethylamino.
The compound according to any one of claims 1 to 35, or a pharmaceutically acceptable salt thereof, wherein R 5 is H.
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 35, wherein R 5 is NH 2 .
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 37, wherein R 6 is -C(O)NH(R 9 ).
The compound or pharmaceutically acceptable salt thereof according to claim 38, wherein R 6 is -C(O)NH 2 .
The compound or pharmaceutically acceptable salt thereof according to claim 38, wherein R 6 is -C(O)NH(Me).
The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 37, wherein R 6 is -SO 2 R b .
The compound according to claim 41 or a pharmaceutically acceptable salt thereof, wherein R 6 is -SO 2 Me.
The compound according to any one of claims 1 to 42, or a pharmaceutically acceptable salt thereof, wherein said R 8 is halogen.
A compound according to any one of claims 1 to 43 or a pharmaceutically acceptable salt thereof, wherein the compound comprises:
A pharmaceutical composition comprising an effective dose of the compound according to any one of claims 1 to 44 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
A method for treating and/or preventing a disease or disorder associated with Myt1 kinase, comprising administering to a patient in need thereof a therapeutically effective amount of a compound according to any one of claims 1 to 44 or a pharmaceutically acceptable salt thereof.
A method for treating and/or preventing a disease or disorder associated with Myt1 kinase, comprising administering a therapeutically effective amount of the composition of claim 45 to a patient in need thereof.
The method of claim 46 or 47, wherein the disease or condition associated with Myt1 kinase comprises leukemia, solid tumor cancer and metastasis, soft tissue cancer, brain cancer, esophageal cancer, gastric cancer, pancreatic cancer, liver cancer, lung cancer, bladder cancer, bone cancer, prostate cancer, ovarian cancer, breast cancer, colorectal cancer, endometrial cancer, cervical cancer, uterine cancer, testicular cancer, kidney cancer, head and neck cancer, chronic inflammatory proliferative diseases, proliferative cardiovascular diseases, proliferative eye diseases and benign hyperproliferative diseases.
Use of a compound according to any one of claims 1 to 44 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a disease or disorder associated with Myt1 kinase.
Use of the pharmaceutical composition according to claim 45 in the preparation of a medicament for treating a disease or condition associated with Myt1 kinase.