CN118666851A - Condensed ring KIF18A inhibitor compound, pharmaceutical composition, preparation method and application thereof - Google Patents

Abstract

The present invention provides a fused ring KIF18A inhibitor compound, a pharmaceutical composition, a preparation method and an application thereof. The compound has a good KIF18A inhibitory effect, can regulate KIF18A protein alone or in combination with microtubules to form a binding complex, and is used to treat KIF18A-mediated conditions and/or diseases, such as tumor diseases, and to prepare drugs for such conditions or diseases.

Description

Condensed ring KIF18A inhibitor compounds, pharmaceutical compositions, preparation methods and applications thereof

This application claims priority to the prior application filed with the State Intellectual Property Office of China on March 17, 2023, with patent application number 202310263450.9, entitled “Fused-ring KIF18A inhibitor compounds, pharmaceutical compositions, preparation methods and applications thereof”; and the prior application filed with the State Intellectual Property Office of China on October 11, 2023, with patent application number 202311315881.1, entitled “Fused-ring KIF18A inhibitor compounds, pharmaceutical compositions, preparation methods and applications thereof”. The entire text of the prior application is incorporated herein by reference.

Technical Field

The present invention belongs to the field of medicine, and specifically relates to a condensed-ring KIF18A inhibitor compound, a pharmaceutical composition, and a preparation method and application thereof.

Background Art

Cancer is one of the most serious diseases affecting human health, with mortality and morbidity often ranking among the highest among all diseases. Although the quality of life of some patients has been greatly improved with the continuous development and progress of medical technology and drug research and development, there are still more unmet clinical needs in the search for effective treatment or cure drugs for different cancers, and more new targets will provide new possibilities for future cancer drug research and development.

Cancer cells experience unregulated cell proliferation due to damage or loss of one or more genes that regulate the cell cycle. Various kinases and kinesins have been identified as playing key roles in cell cycle and mitosis regulation and progression in both normally dividing cells and cancer cells.

Kinesin molecules are motor proteins that use intracellular microtubules as tracks, also known as molecular motors, which can convert ATP energy into mechanical energy. In eukaryotic cells, they are closely related to cell mitosis and meiosis, the growth and development of tissues and organs, and the development and signal transduction of neurons. Kinesin members share a relatively conserved motor domain. According to the location of the motor domain in the molecule, the kinesin family is roughly divided into three categories: N-type kinesin, that is, the amino (-NH ₂ ) terminal region of the protein polypeptide chain has a motor domain; M-type kinesin, that is, the middle region has a motor domain; C-type kinesin, that is, the carboxyl (-COOH) terminal region has a motor domain.

Chromosomal instability is a hallmark of cancer and is caused by errors in chromosome segregation during mitosis. Targeting chromosomal instability is an emerging therapeutic strategy in drug development. KIF18A is a member of the N-type Kinesin-8 kinesin family and has been shown to play a role in maintaining the integrity of the bipolar spindle and promoting the viability of cancer cells with chromosomal instability. Mitosis is an effective intervention point, and many anti-mitotic drugs are used in the clinical treatment of human cancers. The most widely used microtubule inhibitor, it can both stabilize microtubules and prevent microtubule assembly. Current anti-mitotic drugs have the limitation of a narrow therapeutic window, and these problems need to be addressed by the development of new targets.

Although tubulin inhibitors are widely used as standard treatments for a variety of human cancer types, these drugs have collateral damage to normal cells, including bone marrow suppression and neurotoxicity. Since KIF18A may not be essential in normal diploid somatic cells (KIF18A knockout mice are viable but have reproductive defects, indicating that KIF18A is not an essential gene for normal somatic cell division), targeting KIF18A may significantly reduce its toxicity, which is conducive to improving the therapeutic safety window of tubulin-targeted drugs in the clinic.

KIF18A protein is highly expressed in a variety of tumors, including colon cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer, and ovarian cancer. KIF18A plays a key role in the occurrence, development, and metastasis of breast cancer, and its high expression indicates a poor prognosis in patients. KIF18A is required for the proliferation of chromosomal instability cells derived from triple-negative breast cancer or colorectal cancer, but KIF18A is not required in diploid cells. Knockout of the KIF18A gene can cause infertility in male mice, but female mice are not affected. KIF18A mRNA expression is significantly associated with higher tumor grade and larger tumors in breast cancer patients, and KIF18A is an independent predictor of lymph node metastasis in breast cancer, with a risk coefficient of 3.2. In addition, inhibition of KIF18A expression not only affects the key function of KIF18A in cell mitosis, but also reduces cancer cell migration by stabilizing leading-edge microtubules, ultimately leading to inactivation of the PI3K-AKT signaling pathway and inducing cell apoptosis.

Therefore, the development of KIF18A protein inhibitors may be a new breakthrough in cancer drugs.

Summary of the invention

In order to improve the above technical problems, the present invention provides a compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound:

in,

R ₁ and R ₂ are the same or different and are independently selected from H, halogen, and C _1-12 alkyl;

A is -NHC(O)- or -C(O)NH-;

Ring E is selected from a C _6-14 aromatic ring, a 5-14 membered heteroaromatic ring;

Ring G is selected from a C _3-12 cycloalkane ring, a C _3-12 cycloalkene ring, a 5-14 membered heterocyclic ring, a C _6-14 aromatic ring, and a 5-14 membered heteroaromatic ring;

Each _Re is the same or different and is independently selected from H, halogen, hydroxy, amino, cyano, C _1-12 alkyl, C _1-12 alkyloxy; m is selected from 0, 1, 2, 3 or 4;

Each _Rg is the same or different and is independently selected from H, halogen, hydroxyl, amino, cyano, the following groups which are unsubstituted or optionally substituted by one, two or more _Rg1 : _C1-12 alkyl, _C1-12 alkyloxy; or, two _Rgs and the atoms to which they are attached form the following groups which are unsubstituted or optionally substituted by one, two or more _Rg2 : _C3-8 cycloalkane ring, _C3-8 cycloalkene ring, 5-14 membered heterocycle, C6-14 aromatic ring or _5-14 membered heteroaromatic ring; each _Rg1 and _Rg2 are the same or different and are independently selected from H, halogen, _C1-12 alkyl, _C1-12 alkyloxy; n is selected from 0, 1, 2, 3 or 4.

According to some embodiments, R ₁ and R ₂ are the same or different and are independently selected from H, F, Cl, and methyl;

According to some embodiments, R ₁ and R ₂ are the same and are F.

According to some embodiments, ring E is selected from a C ₆ aromatic ring, a 6-membered heteroaromatic ring;

According to some embodiments, ring E is selected from a benzene ring, a pyridine ring, a pyrimidine ring, and a pyridazine ring.

According to some embodiments, ring G is selected from a C _5-6 cycloalkane ring, a C _5-6 cycloalkene ring, a 5-6 membered heterocyclic ring, a C ₆ aromatic ring, a 5-6 membered heteroaromatic ring;

According to some embodiments, ring G is selected from a cyclohexane ring, a cyclohexene ring, a benzene ring, a pyridine ring, and an imidazole ring.

According to some embodiments, each _Re is the same or different and is independently selected from H, _C1-6 alkyl, _C1-6 alkyloxy.

According to some embodiments, each _Rg is the same or different and is independently selected from H, halogen, _C1-6 alkyl, _C1-6 alkyloxy; or, two _Rgs and the atoms to which they are attached form the following groups which are unsubstituted or optionally substituted by one, two or more _Rg2 : _C3-8 cycloalkane ring, 5-8 membered heterocycle;

According to some embodiments, each _Rg is the same or different and is independently selected from H, methoxy; or, two _Rgs and their respective atoms to which they are attached form the following groups which are unsubstituted or optionally substituted by one, two or more _Rg2 : (like

According to some embodiments, each R _g2 is the same or different and is independently selected from H, halogen, C _1-6 alkyl, C _1-6 alkyloxy;

According to some embodiments, each R _g2 is the same or different and is independently selected from H, F, and methyl.

According to some embodiments, the compound represented by formula (I) is selected from the following structures:

wherein A, ring E, ring G, R ₁ , R ₂ , _Re , R _g , m, and n have the definitions described herein.

According to some embodiments, the compound represented by formula (I) is selected from the following structures:

wherein A, R ₁ , R ₂ , _Re , R _g , R _g2 , m and n have the meanings described herein.

According to some embodiments, the compound represented by formula (I) is selected from the following structures:

The present invention also provides a method for preparing the compound represented by formula (I), comprising the following steps A:

Step A: reacting compound (a) with 2-hydroxyethanesulfonamide to obtain a compound represented by formula (I):

wherein A, ring E, ring G, R ₁ , R ₂ , _Re , R _g , m, and n have the definitions described herein; and X is selected from halogen, such as F, Cl, Br or I.

According to an embodiment of the present invention, the method for preparing the compound represented by formula (I) may further comprise the following steps B1 or B2:

Step B1: for a compound represented by formula (I) wherein A is -C(O)NH-, reacting compound (a2) with compound (a3) to obtain a corresponding compound represented by formula (a); or

Step B2: For the compound represented by formula (I) wherein A is -NHC(O)-, reacting compound (a4) with compound (a5) to obtain the corresponding compound represented by formula (a);

or

wherein A, ring E, ring G, R ₁ , R ₂ , _Re , R _g , m, and n have the definitions described herein; and X is selected from halogen, such as F, Cl, Br or I.

According to an embodiment of the present invention, the method for preparing the compound represented by formula (I) may further comprise the following steps C1 or C2:

Step C1:

Compound (b1) and compound (b2) are coupled to obtain compound (a1), and compound (a1) is subjected to reduction reaction to obtain compound (a2):

Step C2:

Compound (b1) and compound (b3) are reacted to obtain compound (a4):

wherein R ₁ and R ₂ have the definitions described herein, and X and Y are independently selected from halogen, such as F, Cl, Br or I.

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof.

According to an embodiment of the present invention, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

According to an embodiment of the present invention, the pharmaceutical composition may further contain one or more additional therapeutic agents.

The present invention also provides a method for treating tumor diseases, comprising administering to a patient a preventive or therapeutically effective amount of at least one of the compounds represented by formula (I), its racemates, stereoisomers, tautomers, isotope-labeled substances, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof.

The present invention also provides a method for treating tumor diseases, comprising administering to a patient an effective amount of the above-mentioned pharmaceutical composition for prevention or treatment.

The tumor diseases include colorectal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer and ovarian cancer.

In some embodiments, the patient comprises a mammal, preferably a human.

The present invention also provides a compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or at least one of its prodrug compounds for treating tumor diseases, or a pharmaceutical composition thereof.

The present invention also provides the use of at least one of the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope-labeled substance, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof in the preparation of a drug.

According to an embodiment of the present invention, the use may be use in preparing a medicament for treating KIF18A-mediated disorders and/or diseases, such as use in preparing a KIF18A inhibitor drug.

According to an embodiment of the present invention, the disease is, for example, cancer, including colon cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer or ovarian cancer.

Beneficial Effects

The compound of the present invention has a good KIF18A inhibitory effect. The compound can regulate KIF18A protein alone or by forming a binding complex with microtubules, so as to treat KIF18A-mediated disorders and/or diseases, such as tumor diseases, and to prepare drugs for such disorders or diseases.

Definition and explanation of terms

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

Unless otherwise specified, the numerical ranges recorded in this specification and claims are equivalent to recording at least each specific integer value therein. For example, the numerical range "1-12" is equivalent to recording each integer value in the numerical range "1-12", that is, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

The term "C _1-12 alkyl" is understood to mean straight-chain and branched alkyl groups having 1 to 12 carbon atoms, "C _1-8 alkyl" means straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms, and "C _1-6 alkyl" means straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, or the like or isomers thereof.

The term "C _3-12 cycloalkyl/cycloalkane ring" is understood to mean a saturated monovalent monocyclic, bicyclic (such as condensed, bridged, spiro) hydrocarbon ring or tricyclic alkane having 3 to 12 carbon atoms, preferably a "C _3-10 cycloalkyl", more preferably a "C _3-8 cycloalkyl". The term "C _3-12 cycloalkyl" is understood to mean a saturated monovalent monocyclic, bicyclic (such as bridged, spiro) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. The _C3-12 cycloalkyl group may be a monocyclic hydrocarbon group, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, or a bicyclic hydrocarbon group, such as borneol, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonanyl, 2,6-diazaspiro[3,4]octanyl, or a tricyclic hydrocarbon group, such as adamantyl.

The term "C _3-12 cycloalkenyl/cycloalkene ring" is understood to mean a monovalent monocyclic, bicyclic (such as fused, bridged, spiro) hydrocarbon ring or tricyclic alkene containing a carbon-carbon double bond, which has 3 to 12 carbon atoms, preferably "C _3-10 cycloalkenyl", and more preferably "C _3-8 cycloalkenyl". The term "C _3-12 cycloalkenyl" is understood to mean a monovalent monocyclic, bicyclic (such as bridged, spiro) hydrocarbon ring or tricyclic alkene containing a carbon-carbon double bond, which has 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms. The C _3-12 cycloalkenyl group may be a monocyclic hydrocarbon group, such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclononenyl or cyclodecenyl, or a bicyclic hydrocarbon group such as spiro[2.5]oct-5-enyl, spiro[3.5]non-6-enyl, spiro[4.5]dec-7-enyl.

The term "C _6-14 aromatic ring" should be understood to preferably mean a monovalent aromatic or partially aromatic monocyclic, bicyclic (such as fused, bridged, spiro) or tricyclic hydrocarbon ring having 6 to 14 carbon atoms, which may be a single aromatic ring or a polyaromatic ring fused together, preferably a "C _6-10 aromatic group". The term "C _6-14 aryl" is to be understood as preferably meaning a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring ("C _6-14 aryl") having 6, 7, 8, 9, 10, 11, 12, 13 or 14 carbon atoms, in particular a ring having 6 carbon atoms ("C ₆ aryl"), such as phenyl; or biphenyl, or a ring having 9 carbon atoms ("C ₉ aryl"), such as indanyl or indenyl, or a ring having 10 carbon atoms ("C ₁₀ aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, or a ring having 13 carbon atoms ("C ₁₃ aryl"), such as fluorenyl, or a ring having 14 carbon atoms ("C ₁₄ aryl"), such as anthracenyl. When the C _6-20 aryl is substituted, it may be mono- or polysubstituted. Furthermore, there is no limitation on the substitution site, and for example, substitution may be at the ortho, para or meta position.

The term "5-14 membered heteroaromatic ring" is understood to include monocyclic, bicyclic (e.g. fused, bridged, spiro) or tricyclic aromatic ring systems having 5 to 14 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S, for example "5-10 membered heteroaryl". The term "5-14 membered heteroaryl" is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3 heteroatoms each independently selected from N, O and S and, in each case, furthermore, may be benzo-fused. "Heteroaryl" also refers to a radical in which a heteroaromatic ring is fused to one or more aryl, alicyclic or heterocyclyl rings, wherein the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indolizinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4-, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinolizinyl, 2-, 3- , 4-, 5-, 6-, 7- or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolinyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-, 4-, 6- or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-carbazolylcarbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8- or 9-carbolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-phenanthridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-phenanthridinyl, -acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-oxadiazole, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthroline, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenothiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenazinyl , 2-, 3-, 4-, 5-, 6- or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-benzoisoquinolyl, 2-, 3-, 4- or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-7H-pyrazino[2,3-c]carbazolyl, 2-, 3-, 5-, 6- or 7-2H-furo[3,2-b]pyranyl , 2-, 3-, 4-, 5-, 7-, or 8-5H-pyrido[2,3-d]-o-oxazinyl, 1-, 3-, or 5-1H-pyrazolo[4,3-d]-oxazolyl, 2-, 4-, or 54H-imidazo[4,5-d]thiazolyl, 3-, 5-, or 8-pyrazino[2,3-d]pyridazinyl, 2-, 3-, 5-, or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8-, or 9 -furo[3,4-c]cinnolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10 or 11-4H-pyrido[2,3-c]carbazolyl, 2-, 3-, 6- or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thienyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 2-, 4-, 5-, 6- or 7-benzimidazolyl, 2-, 4- , 4-, 5-, 6-, or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-4H-pyrrolo[1,2-b][2]benzazepinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzoxazolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. When the 5-14 membered heteroaryl group is connected to other groups to form the compound of the present invention, it can be a carbon atom on the 5-14 membered heteroaryl ring connected to other groups, or it can be a heteroatom on the 5-14 membered heteroaryl ring connected to other groups. When the 5-14 membered heteroaryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there is no limitation on the substitution site, for example, a hydrogen atom connected to a carbon atom on a heteroaryl ring may be substituted, or a hydrogen atom connected to a heteroatom on a heteroaryl ring may be substituted.

Unless otherwise defined, the term "3-14 membered heterocyclyl" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5-, 6- or 7-membered monocyclic ring, a 7-, 8-, 9-, 10-, 11- or 12-membered bicyclic ring (such as a fused ring, a bridged ring, a spirocyclic ring) or a 10-, 11-, 12-, 13- or 14-membered tricyclic ring system, and contains at least one, for example 1, 2, 3, 4, 5 or more heteroatoms selected from O, S and N, wherein N and S may also be optionally oxidized to various oxidation states to form nitrogen oxides, -S(O)- or -S(O) ₂ -. Preferably, the heterocyclyl may be selected from "3-10 membered heterocyclyl". The term "3-10 membered heterocyclyl" means a saturated or unsaturated non-aromatic ring or ring system, and contains at least one heteroatom selected from O, S and N. The heterocyclic group can be connected to the rest of the molecule by any one of the carbon atoms or the nitrogen atom (if present). The heterocyclic group can include fused or bridged rings and spirocyclic rings. In particular, the heterocyclic group can include but is not limited to: 4-membered rings, such as azetidinyl, oxetanyl; 5-membered rings, such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or 7-membered rings, such as diazepanyl. Optionally, the heterocyclic group can be benzo-fused. The heterocyclic group may be bicyclic, for example but not limited to a 5,5-membered ring, such as a hexahydrocyclopenta[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, such as a hexahydropyrrolo[1,2-a]pyrazine-2(1H)-yl ring. The heterocyclic group may be partially unsaturated, i.e., it may contain one or more double bonds, such as but not limited to dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl or 4H-[1,4]thiazinyl, or it may be benzo-fused, such as but not limited to dihydroisoquinolinyl. When the 3-14-membered heterocyclic group is connected to other groups to form the compound of the present invention, the carbon atoms on the 3-14-membered heterocyclic group may be connected to other groups, or the heterocyclic atoms on the 3-14-membered heterocyclic group may be connected to other groups. For example, when the 3-14 membered heterocyclic group is selected from piperazinyl, the nitrogen atom on the piperazinyl group may be connected to other groups. Or when the 3-14 membered heterocyclic group is selected from piperidinyl, the nitrogen atom on the piperidinyl ring and the carbon atom at the para position thereof may be connected to other groups.

The term "spirocyclic" refers to a ring system in which two rings share one ring-forming atom.

The term "fused ring" refers to a ring system in which two rings share two ring atoms.

The term "bridged ring" refers to a ring system in which two rings share three or more ring atoms.

The term "halogen" refers to fluorine, chlorine, bromine and iodine.

"Halo" means substituted with one or more halogens.

It will be appreciated by those skilled in the art that the compounds of formula (I) may exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they may form acid addition salts; if these compounds have an acidic center, they may form a base addition salt; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they may also form an inner salt.

The compounds of the present invention may exist in the form of solvates (e.g., hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of the polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

According to its molecular structure, the compounds of the present invention may be chiral, and therefore various enantiomeric forms may exist. Thus, these compounds may exist in racemic form or optically active form. The compounds of the present invention encompass isomers or mixtures, racemates in which each chiral carbon is in R or S configuration. The compounds of the present invention or their intermediates can be separated into enantiomeric compounds by chemical or physical methods known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are prepared from the mixture by reaction with an optically active resolution agent. Examples of suitable resolution agents are optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-phenylsulfonylproline) or various optically active camphorsulfonic acids in R and S forms. Chromatographic enantiomer resolution can also be advantageously carried out with the aid of optically active resolving agents such as dinitrobenzoylphenylglycine, cellulose triacetate or other carbohydrate derivatives or chiral derivatized methacrylate polymers immobilized on silica gel. Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example, hexane/isopropanol/acetonitrile.

The corresponding stable isomers can be separated according to known methods, for example by extraction, filtration or column chromatography.

The term "patient" refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses or primates, and most preferably humans.

The term "therapeutically effective amount" refers to the amount of an active compound or drug that elicits the biological or medical response that a researcher, veterinarian, physician or other clinician is seeking in a tissue, system, animal, individual or human, and includes one or more of the following: (1) Preventing disease: e.g., preventing a disease, disorder or condition in an individual who is susceptible to the disease, disorder or condition but does not yet experience or develop the pathology or symptoms of the disease. (2) Inhibiting disease: e.g., inhibiting a disease, disorder or condition (i.e., preventing further development of the pathology and/or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition. (3) Alleviating disease: e.g., alleviating a disease, disorder or condition (i.e., reversing the pathology and/or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder or condition.

DETAILED DESCRIPTION

The technical scheme of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only exemplary descriptions and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope that the present invention is intended to protect.

Unless otherwise specified, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods.

Example 1

2-(4-(Difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)-N-((1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl)benzamide (Compound 1)

The first step is the synthesis of (1S, 4R)-2-(3-bromo-2-nitrophenyl)-2-azabicyclo[2.2.1]heptan-3-one (compound 1-3):

Under nitrogen protection, palladium acetate (159.85 mg, 0.712 mmol, 0.1 eq) and 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthene (823.96 mg, 1.424 mmol, 0.2 eq) were added to a solution of 1,3-dibromo-2-nitrobenzene (compound 1-1, 2.00 g, 7.120 mmol, 1 eq), cesium carbonate (6.96 g, 21.360 mmol, 3 eq) and (1S,4R)-2-azabicyclo[2.2.1]heptan-3-one (compound 1-2, 791.33 mg, 7.120 mmol, 1 eq) in 1,4-dioxane (1 mL) at room temperature, and the reaction solution was heated to 80°C and stirred for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3×80 mL). The organic phases were combined, backwashed with saturated brine (2×30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (1:1) to obtain the product (1S, 4R)-2-(3-bromo-2-nitrophenyl)-2-azabicyclo[2.2.1]heptan-3-one (compound 1-3, 700 mg, 31.60%).

MS: (ESI,m/z):311.25[M+H] ⁺ ,RT(min):1.026.

Step 2 Synthesis of (1S, 4R)-6-bromo-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine (Compound 1-4):

Under nitrogen protection, acetic acid (5 mL) was added dropwise to a solution of (1S, 4R)-2-(3-bromo-2-nitrophenyl)-2-azabicyclo[2.2.1]heptan-3-one (compound 1-3, 700 mg, 2.250 mmol, 1 eq) and reduced iron powder (592.31 mg, 10.606 mmol, 5.00 eq) in ethanol (10 mL) at room temperature. The reaction solution was heated to 110°C and stirred for 2 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The crude product was diluted with water (50 mL) and extracted with ethyl acetate (3×50 mL). The organic phases were combined, backwashed with saturated brine (2×60 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with dichloromethane/methanol (10:1) to give (1S,4R)-6-bromo-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine (Compound 1-4, 560 mg, 94.59%).

MS: (ESI,m/z):263.30[M+H] ⁺ ,RT(min):0.779.

Step 3 Synthesis of tert-butyl ((1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl)carbamate (Compound 1-5):

To a solution of (1S,4R)-6-bromo-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine (compound 1-4, 500 mg, 1.900 mmol, 1 eq), cesium carbonate (1857.31 mg, 5.700 mmol, 3 eq) and tert-butyl carbamate (445.20 mg, 3.800 mmol, 2 eq) in N,N-dimethylformamide (5 mL) was added (methanesulfonic acid {biscyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphine}(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II)4 (174.54 mg, 0.190 mmol, 0.1 eq), bicyclohexyl[3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphine}(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II)4 (174.54 mg, 0.190 mmol, 0.1 eq), dicyclopent ... Hexyl (3-isopropoxy-2', 4', 6'-triisopropyl-[1,1'-biphenyl]-2-yl) phosphane (203.24 mg, 0.380 mmol, 0.2 eq), the reaction solution was heated to 100 ° C and stirred for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3×50 mL). The organic phases were combined, backwashed with saturated brine (1×50 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (1:1) to obtain ((1S, 4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl)carbamic acid tert-butyl ester (Compound 1-5, 560 mg, 98.44%).

MS: (ESI,m/z):300.35[M+H] ⁺ ,RT(min):1.053.

Step 4: Synthesis of (1S, 4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-amine (Compound 1-6):

To a solution of tert-butyl ((1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl)carbamate (compound 1-5, 560 mg, 1.871 mmol, 1 eq) in dichloromethane (5 mL) was added HCl in 1,4-dioxane (5 mL, 4 M) at room temperature. After the addition was complete, the system was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure to obtain (1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-amine (compound 1-6, 500 mg). The crude product was not further purified and was directly used in the next step.

MS:(ESI,m/z):200.40[M+H] ⁺ ,RT(min):0.629.

Step 5 Synthesis of tert-butyl 4-(difluoromethylene)piperidine-1-carboxylate (Compound 1-9):

Under nitrogen protection, add tert-butyl 4-oxopiperidin-1-carboxylate (compound 1-7, 5g, 25.09mmol, 1eq) and 2-difluoromethanesulfonylpyridine (compound 1-8, 4.07g, 21.08mmol, 0.84eq) in N', N'-dimethylformamide (20mL) to a solution of potassium tert-butoxide (4.22g, 37.64mmol, 1.5eq) in N', N'-dimethylformamide (50mL) at -50°C. Then continue stirring at this temperature for 30 minutes, add saturated sodium bicarbonate aqueous solution (12.5mL) and 3M hydrochloric acid (32.5mL) in sequence at -50°C, and then naturally warm the reaction solution to room temperature. The reaction mixture is extracted with ethyl acetate (3×80mL). Combine the organic phases, backwash with saturated brine (2×100mL), and dry over anhydrous sodium sulfate. The obtained mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain tert-butyl 4-(difluoromethylene)piperidine-1-carboxylate (Compound 1-9, 5 g, crude product).

Step 6 Synthesis of 4-(difluoromethylene)piperidine hydrochloride (Compound 1-10):

To a solution of tert-butyl 4-(difluoromethylene)piperidine-1-carboxylate (compound 1-9, 5 g, 21.44 mmol, 1 eq) in dichloromethane (50 mL) was added a solution of HCl in 1,4-dioxane (4 mL) at room temperature, and the reaction solution was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure to obtain 4-(difluoromethylene)piperidine hydrochloride (compound 1-10, 4.2 g, crude product).

¹ H NMR (400MHz, DMSO-d ₆ ) δ9.16 (s, 2H), 3.14–3.00 (m, 4H), 2.44–2.29 (m, 4H).

Step 7 Synthesis of 2-[4-(difluoromethylene)piperidin-1-yl]-4-iodobenzoic acid (Compound 1-12):

Under nitrogen protection, potassium carbonate (3.11 g, 22.53 mmol, 3 eq) and 2-fluoro-4-iodobenzoic acid (compound 2-11, 2.60 g, 9.76 mmol, 1.3 eq) were added to a solution of 4-(difluoromethylene)piperidine hydrochloride (compound 1-10, 1 g, 7.51 mmol, 1 eq) in dimethyl sulfoxide (10 mL, 140.79 mmol, 18.75 eq) at room temperature. The reaction solution was heated to 140°C and stirred for 18 hours. The reaction solution was cooled to room temperature, diluted with water (60 mL), and the reaction mixture was extracted with ethyl acetate (3×20 mL). The aqueous phase was collected, the pH value was adjusted to 4 with 1 M hydrochloric acid solution, the mixture was extracted with ethyl acetate (3×40 mL), the organic phases were combined, backwashed with saturated brine (2×60 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure to obtain 2-[4-(difluoromethylene)piperidin-1-yl]-4-iodobenzoic acid (Compound 1-12, 1 g, 31.60%).

MS: (ESI,m/z):379.85[M+H] ⁺ ,RT(min):0.878.

Step 8 Synthesis of 2-(4-(difluoromethylene)piperidin-1-yl)-4-iodo-N-((1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl)benzamide (Compound 1-13):

Under nitrogen protection, N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (888.04 mg, 3.16 mmol, 4 eq) and 1-methyl-1H-imidazole (649.67 mg, 7.91 mmol, 10 eq) were added to a dichloromethane solution of 2-[4-(difluoromethylene)piperidin-1-yl]-4-iodobenzoic acid (compound 1-12, 300 mg, 0.79 mmol, 1 eq) and (1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-amine (compound 1-6, 315.33 mg, 1.58 mmol, 2 eq) at room temperature. The reaction solution was heated to 60°C and stirred for 1 hour. The reaction solution was cooled to room temperature and quenched with water (20 mL). The reaction mixture was extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (1×30 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (5:1) to obtain 2-(4-(difluoromethylene)piperidin-1-yl)-4-iodo-N-((1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl)benzamide (Compound 1-13, 500 mg, 78.93%).

MS: (ESI,m/z):561.05[M+H] ⁺ ,RT(min):1.013.

Step 9 Synthesis of 2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)-N-((1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl)benzamide (Compound 1):

Under nitrogen protection, 2-(methylamino)acetic acid (9.54 mg, 0.11 mmol, 0.2 eq), potassium carbonate (221.96 mg, 1.61 mmol, 3 eq), cuprous iodide (10.20 mg, 0.054 mmol, 0.1 eq) and 2-hydroxyethanesulfonamide (100.49 mg, 0.80 mmol, 1.5 eq) were added to a solution of 2-(4-(difluoromethylene)piperidin-1-yl)-4-iodo-N-((1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl)benzamide (compound 1-13, 300 mg, 0.53 mmol, 1 eq) in N',N'-dimethylformamide (3 mL) at room temperature. The reaction solution was heated to 100°C and reacted for 16 hours. The reaction solution was cooled to room temperature and quenched with water. The reaction mixture was extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated brine (3×30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions: chromatography column specifications: XBridge BEH Shield RP18 5μm, 30mm*150mm; mobile phase A: water (10mmol/L sodium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL/min; elution gradient: 38%B to 56%B in10min; detection wavelength: 254nm/220nm; retention time (min): 8.3, to obtain 2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)-N-((1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl)benzamide (compound 1, 51.29mg, 17.2%).

MS:(ESI,m/z):557.90[M+H] ⁺ ,RT(min):1.570.

¹ H-NMR: (400MHz, DMSO-d ₆ )δ12.28(s,1H),10.20(s,1H),8.32–8.27(m,1H),8.03(d,1H),7.27–7.22(m,1H),7.19(d,1H),7.15(d,1H),7.12–7.07(m,1H),5.14(s,1H),4.94( s,1H),3.76(t,2H),3.51(d,1H),3.36(d,2H),2.99(t,4H),2.71–2.56(m,4H),2.23(d,1H),2.14–2.08(m,1H),2.02–1.92(m,2H),1.13–1.00(m,2 H).

Example 2

N-(3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)benzamide (Compound 2)

Compound 2

The first step is the synthesis of (2Z)-N-(2,6-dibromophenyl)-3,3-difluoropyrrolidine-2-imine (compound 2-3):

Under nitrogen protection, POCl ₃ (837.11 mg, 5.460 mmol, 1 eq) was added to a solution of 3,3-difluoropyrrolidin-2-one (compound 2-2, 0.99 g, 8.176 mmol, 1.50 eq) in ultra-dry toluene (15 mL) at 0°C, and the reaction solution was stirred at room temperature for 2 hours. A solution of 2,6-dibromoaniline (compound 2-1, 1.37 g, 5.460 mmol, 1 eq) in toluene (5 mL) was added to the above solution, and the reaction solution was heated to 110°C and stirred for 16 hours. The reaction solution was cooled to room temperature and quenched with water. The reaction mixture was extracted with dichloromethane (3×60 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (1×80 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was recrystallized from petroleum ether/dichloromethane (100:1) to obtain (2Z)-N-(2,6-dibromophenyl)-3,3-difluoropyrrolidine-2-imine (Compound 2-3, 1.2 g, 62.09%).

MS(ESI,m/z):352.85[M+H] ⁺ ,RT(min):0.674

Step 2 Synthesis of 5-bromo-3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole (Compound 2-4):

Under nitrogen protection, N,N-dimethylethylenediamine (12.45 mg, 0.141 mmol, 0.1 eq), anhydrous potassium carbonate (195.21 mg, 1.412 mmol, 1 eq) and cuprous iodide (13.45 mg, 0.071 mmol, 0.05 eq) were added to (2Z)-N-(2,6-dibromophenyl)-3,3-difluoropyrrolidine-2-imine (compound 2-3, 500 mg, 1.412 mmol, 1 eq) in N,N-dimethylformamide (10.00 mL) at room temperature. The reaction solution was heated to 100°C and stirred for 1 h. The reaction solution was cooled to room temperature, quenched with water, and the reaction mixture was extracted with dichloromethane (3×50 mL). The organic phases were combined, backwashed with saturated saline solution (1×50 mL), and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (2:1) to give 5-bromo-3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole (Compound 2-4, 240 mg, 62.22%).

MS(ESI,m/z):272.90[M+H] ⁺ ,RT(min):0.837

Step 3 Synthesis of tert-butyl (3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)carbamate (Compound 2-5):

To a solution of 5-bromo-3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole (compound 2-4, 230 mg, 0.842 mmol, 1 eq) in N,N-dimethylformamide (5.00 mL) was added tert-butyl carbamate (197.33 mg, 1.684 mmol, 2 eq), (methanesulfonic acid {dicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl)phosphine}(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) (154.73 mg, 0.168 mmol, 0.20 eq), dicyclohexyl(3-isopropyl-2',4',6'-triisopropyl)phosphine}(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) (154.73 mg, 0.168 mmol, 0.20 eq), -[1,1'-biphenyl]-2-yl)phosphine (45.04 mg, 0.084 mmol, 0.10 eq), cesium carbonate (823.25 mg, 2.526 mmol, 3.00 eq), the reaction solution was heated to 100 ° C and stirred for 1 h. The reaction mixture was extracted with ethyl acetate (3×50 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (1×50 mL), and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (2:1) to obtain (3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)carbamic acid tert-butyl ester (Compound 2-5, 189 mg, 72.55%).

MS(ESI,m/z):310.05[M+H] ⁺ ,RT(min):1.004

Step 4 Synthesis of 3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-amine (Compound 2-6):

Under nitrogen protection, a solution of hydrochloric acid in 1,4-dioxane (2 mL) was added to a dichloromethane solution (2 mL) of (3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)carbamic acid tert-butyl ester (compound 2-5, 189 mg, 0.611 mmol, 1 eq) at room temperature, and the reaction solution was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain 3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-amine (compound 2-6, 127 mg, 99.35%).

MS(ESI,m/z):210.10[M+H] ⁺ ,RT(min):0.193

Step 5 Synthesis of N-(3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-2-(4-(difluoromethylene)piperidin-1-yl)-4-iodobenzamide (Compound 2-8):

Under nitrogen protection, N-methylimidazole (313.98 mg, 3.820 mmol, 10 eq) was added to a solution of 2-[4-(difluoromethylene)piperidin-1-yl]-4-iodobenzoic acid (compound 1-12, 144.99 mg, 0.382 mmol, 1.00 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (429.19 mg, 1.528 mmol, 4 eq) in dichloromethane (4 mL) at room temperature, and stirred for 5 minutes. Then, 3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-amine (compound 2-6, 144.99 mg, 0.382 mmol, 1.00 eq) was added in batches at room temperature. The temperature was raised to 60°C and stirred for 1.5 hours. The reaction solution was cooled to room temperature and extracted with dichloromethane (3×10 mL). The organic phases were combined, backwashed with saturated brine (1×10 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (1:1) to obtain N-(3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-2-(4-(difluoromethylene)piperidin-1-yl)-4-iodobenzamide (160 mg, 73.36%).

MS:(ESI,m/z):571.30[M+H] ⁺ ,RT(min):1.447

Step 6 Synthesis of N-(3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)benzamide (Compound 2):

Under nitrogen protection, potassium carbonate (58.16 mg, 0.420 mmol, 3 eq), sarcosine (2.5 mg, 0.028 mmol, 0.2 eq) and cuprous iodide (2.67 mg, 0.014 mmol, 0.1 eq) were added in batches to a solution of N-(3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-2-(4-(difluoromethylene)piperidin-1-yl)-4-iodobenzamide (compound 2-8, 80 mg, 0.140 mmol, 1 eq) and 2-hydroxyethanesulfonamide (35.11 mg, 0.280 mmol, 2 eq) in N,N-dimethylformamide (3 mL) at room temperature. The temperature was raised to 120°C and stirred for 6 hours. The reaction solution was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (3×10 mL). The organic phases were combined, backwashed with saturated brine (1×10 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions: chromatographic column specifications: Ultimate μXB-C18; mobile phase A: water (10 mM ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 100 ml/min; elution gradient: 50% B to 90% B in 23 minutes; detection wavelength: UV 254 nm/220 nm; retention time (min): 20, and N-(3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)benzamide (compound 2, 35.57 mg, 44.46%) was obtained.

MS:(ESI,m/z):567.85[M+H] ⁺ ,RT(min):1.446

¹ H NMR (400MHz, DMSO-d ₆ ) δ12.58(s,1H),10.23(s,1H),8.52–8.38(m,1H),8.07(d,1H),7.43–7.28(m,2H ),7.22(d,1H),7.16–7.08(m,1H),4.94(s,1H),4.44(t,2H),3.76(t,2H),3.36(t,2H),3.31–3.22( m,2H),3.00(t,4H),2.64(t,4H).

Example 3

2-(4-(Difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)-N-(7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepin-4-yl)benzamide (Compound 3)

Compound 3

The first step is the synthesis of (2Z)-N-(2,6-dibromoaniline)azepine-2-imine (compound 3-2):

Under nitrogen protection, phosphorus oxychloride (6.1 g, 39.85 mmol, 5 eq) was added dropwise to a toluene (10 mL) solution of caprolactam (1.36 g, 15.94 mmol, 2 eq) at 0°C, and the reaction solution was reacted at 0°C for 2 hours. Subsequently, a toluene solution (5 mL) of 2,6-dibromoaniline (compound 2-1, 2 g, 7.97 mmol, 1 eq) was added, and the reaction solution was heated to 110°C and stirred for 16 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, quenched with ice water (50 mL), adjusted to pH 10 with saturated sodium bicarbonate solution, and the mixture was extracted with ethyl acetate (3×60 mL). The organic phases were combined, backwashed with saturated brine (1×50 mL), and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (10:1) to obtain (2Z)-N-(2,6-dibromoanilino)azepine-2-imine (compound 3-2, 2.7 g, 27.65%).

MS:(ESI,m/z):345.10[M+H] ⁺ ,RT(min):0.595.

Step 2 Synthesis of 4-bromo-7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepine (Compound 3-3):

Under nitrogen protection, N,N-dimethylethylenediamine (25.47 mg, 0.289 mmol, 0.10 eq), cuprous iodide (27.52 mg, 0.145 mmol, 0.05 eq), and potassium carbonate (599.04 mg, 4.335 mmol, 3 eq) were added to a solution of (2Z)-N-(2,6-dibromoaniline)azepine-2-imine (compound 3-2, 500 mg, 1.445 mmol, 1 eq) in acetonitrile (3 mL) at room temperature. The reaction solution was heated to 100°C and stirred for 1 hour. The reaction solution was cooled to room temperature, diluted with water (40 mL), and extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated brine (1×40 mL), and dried over anhydrous sodium sulfate. Filtered, and the filtrate was concentrated under reduced pressure. 4-Bromo-7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepine (Compound 3-3, 250 mg, 65.26%) was obtained.

MS:(ESI,m/z):264.90[M+H] ⁺ ,RT(min):1.151.

Step 3 Synthesis of tert-butyl (7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepine-4-yl)carbamate (Compound 3-4):

Under nitrogen protection, tert-butyl carbamate (220.91 mg, 1.886 mmol, 2 eq), dicyclohexyl (3-isopropyl-2',4',6'-triisopropyl-[1,1 '-biphenyl]-2-yl)phosphine (50.42mg, 0.094mmol, 0.1eq), (methanesulfonic acid {bicycloethyl (3-isopropyl-2,4,6-triisopropyl-(1,1-biphenyl)-2-yl)phosphine})(2-methylamino-1,1-biphenyl-2-yl)palladium (II) (173.21mg, 0.189mmol, 0.2eq) and cesium carbonate (921.59mg, 2.829mmol, 3eq). The reaction solution was heated to 100°C and stirred for 1 hour. The reaction was cooled to room temperature and extracted with ethyl acetate (3×50mL). The organic phases were combined, backwashed with saturated brine (1×50mL), and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (5:1) to obtain tert-butyl (7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepin-4-yl)carbamate (Compound 3-4, 150 mg, 52.79%).

MS:(ESI,m/z):302.35[M+H] ⁺ ,RT(min):1.106.

Step 4: Synthesis of 7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepine-4-amine (Compound 3-5):

To a solution of (7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepin-4-yl)carbamic acid tert-butyl ester (compound 3-4, 150 mg, 0.498 mmol, 1 eq) in dichloromethane (2 mL) was added a solution of hydrochloric acid in 1,4-dioxane (4M, 2 mL) at room temperature. The reaction solution was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain 7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepin-4-amine (compound 3-5, 150 mg, 66.67%). It was directly used for the next step without purification.

MS:(ESI,m/z):202.40[M+H] ⁺ ,RT(min):0.544.

Step 5 Synthesis of 2-(4-(difluoromethylene)piperidin-1-yl)-4-iodo-N-(7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepin-4-yl)benzamide (Compound 3-6)

Under nitrogen protection, tetramethyl chlorouronium hexafluorophosphate (278.81 mg, 0.992 mmol, 4 eq) and N-methylimidazole (203.97 mg, 2.480 mmol, 10 eq) were added to a solution of 2-[4-(difluoromethylene)piperidin-1-yl]-4-iodobenzoic acid (compound 1-12, 113.02 mg, 0.298 mmol, 1.2 eq) and 7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepine-4-amine (compound 3-5, 50 mg, 0.248 mmol, 1 eq) in dichloromethane (2.0 mL) at room temperature, and the reaction solution was stirred at room temperature for 1.5 hours. Water (10 mL) was added to the reaction mixture at room temperature to quench it. The reaction mixture was extracted with dichloromethane (3×20 mL). The organic phases were combined, backwashed with saturated brine (1×20 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (3:1) to obtain 2-(4-(difluoromethylene)piperidin-1-yl)-4-iodo-N-(7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepin-4-yl)benzamide (compound 3-6, 50 mg, 35.79%).

MS:(ESI,m/z):563.00[M+H] ⁺ ,RT(min):1.643

Step 6 Synthesis of 2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)-N-(7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepin-4-yl)benzamide (Compound 3)

Under nitrogen protection, sarcosine (58 mg, 0.018 mmol, 0.2 eq), potassium carbonate (6.86 mg, 0.267 mmol, 3 eq) and cuprous iodide (08 mg, 0.027 mmol, 0.3 eq) were added to a solution of 2-(4-(difluoromethylene)piperidin-1-yl)-4-iodo-N-(7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepin-4-yl)benzamide (compound 3-6, 50 mg, 0.089 mmol, 1 eq) and 2-hydroxyethanesulfonamide (22.25 mg, 0.178 mmol, 2 eq) in N,N-dimethylformamide (2 mL) at room temperature. The reaction solution was heated to 120°C and stirred for 3 hours. The reaction solution was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (3×20 mL). Combine the organic phases, backwash with saturated brine (2×20 mL), and dry over anhydrous sodium sulfate. After the obtained mixture is filtered, the filtrate is concentrated under reduced pressure. The crude product is purified by high-performance liquid chromatography under the following conditions: Chromatographic column specifications: Kinetex 5μmEVO C18, 30mm*150mm; Mobile phase A: water (10mM ammonium bicarbonate), mobile phase B: acetonitrile; Flow rate: 60ml/min; Elution gradient: 30% B to 65% B in 8 minutes; Detection wavelength: UV 254nm/221nm; Retention time (min): 7.5. 2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)-N-(7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepin-4-yl)benzamide (Compound 3, 28.98 mg, 57.96%) was obtained.

MS:(ESI,m/z):560.30[M+H] ⁺ ,RT(min):1.723

¹ H NMR (400MHz, DMSO-d ₆ ) δ12.39(s,1H),10.21(s,1H),8.35–8.28(m,1H),8.06(d,1H),7.28–7.22(m,1H ),7.21–7.14(m,2H),7.13–7.08(m,1H),4.94(s,1H),4.30–4.23(m,2H),3.76(t,2H),3.36(t,2H), 3.00(t,6H),2.69(d,4H),1.87(s,2H),1.73(s,2H),1.65(s,2H).

Example 4

2-(4-(Difluoromethylene)piperidin-1-yl)-N-(2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-4-2-hydroxyethanesulfonylamino)benzamide (Compound 4)

Compound 4

The first step is the synthesis of (2Z)-N-(2,6-dibromophenyl)pyrrolidine-2-imine (compound 4-2):

Under nitrogen protection, phosphorus oxychloride (1.22 g, 85.106 mmol, 2 eq) was added to a toluene (10 mL) solution of pyrrolidone (compound 4-1, 1.36 g, 15.942 mmol, 2 eq) at 0°C, and the reaction solution was reacted at 0°C for 2 hours, followed by the addition of 2,6-dibromoaniline (compound 2-1, 2 g, 7.971 mmol, 1 eq), and the reaction solution was heated to 110°C and stirred for 16 hours. The reaction solution was cooled to room temperature, concentrated under reduced pressure, quenched with ice water (50 mL), adjusted to pH 10 with saturated sodium bicarbonate solution, extracted with ethyl acetate (3×50 mL), combined the organic phases, backwashed with saturated brine (1×50 mL), and dried over anhydrous sodium sulfate. Filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative chromatography using ethyl acetate/petroleum ether (1:10) to give (2Z)-N-(2,6-dibromophenyl)pyrrolidine-2-imine (compound 4-2, 2.7 g, 27.65%).

MS(ESI,m/z):317.10[M+H] ⁺ ,RT(min):0.919

Step 2 Synthesis of 5-bromo-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole (Compound 4-3):

Under nitrogen protection, N,N'-dimethyl-1,2-ethylenediamine (74.84 mg, 0.849 mmol, 0.10 eq), cuprous iodide (80.85 mg, 0.425 mmol, 0.05 eq), potassium carbonate (1.17 g, 8.490 mmol, 1 eq) were added to a solution of (2Z)-N-(2,6-dibromophenyl)pyrrolidine-2-imine (compound 4-2, 2.7 g, 8.490 mmol, 1 eq) in acetonitrile (3 mL) at room temperature. The reaction solution was heated to 100 ° C and stirred for 2 hours. The reaction solution was cooled to room temperature, quenched with water (80 mL), and extracted with ethyl acetate (3×60 mL). The organic phases were combined, backwashed with saturated brine (1×80 mL), and dried over anhydrous sodium sulfate. The mixture was filtered and the filtrate was concentrated under reduced pressure to obtain 5-bromo-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole (compound 4-3, 2 g, 74.07%).

MS(ESI,m/z):237.15[M+H] ⁺ ,RT(min):0.657

Step 3 Synthesis of tert-butyl (2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)carbamate (Compound 4-4):

Under nitrogen protection, dicyclohexyl (3-isopropyl-2',4',6'-triisopropyl-[1,1'-biphenyl]) was added to a solution of 5-bromo-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole (compound 4-3, 2 g, 8.436 mmol, 1 eq) and tert-butyl carbamate (1.99 g, 16.87 mmol, 2 eq) in N,N-dimethylformamide (10 mL) at room temperature. -2-yl)phosphine (451.12mg, 0.844mmol, 0.1eq), (methanesulfonic acid {bicycloethyl (3-isopropyl-2,4,6-triisopropyl-(1,1-biphenyl)-2-yl)phosphine}) (2-methylamino-1,1-biphenyl-2-yl) palladium (II) (1.549g, 1.688mmol, 0.2eq) and cesium carbonate (8.245g, 25.308mmol, 3eq). The reaction solution was heated to 100°C and stirred for 1 hour. The reaction solution was cooled to room temperature and quenched with water (80mL). It was extracted with ethyl acetate (3×60mL). The organic phases were combined, backwashed with saturated brine (1×80mL), and dried over anhydrous sodium sulfate. Filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative chromatography using ethyl acetate/petroleum ether (1:5) to give tert-butyl (2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)carbamate (compound 4-4, 700 mg, 35.77%).

MS(ESI,m/z):274.35[M+H] ⁺ ,RT(min):0.592

Step 4 Synthesis of 2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-amine (Compound 4-5):

The reaction mixture of tert-butyl (2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)carbamate (compound 4-4, 300 mg, 1.098 mmol, 1 eq), hydrochloric acid in 1,4-dioxane (2 mL), and dichloromethanol (2 mL) was stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure to obtain 2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-amine (compound 4-5, 300 mg, 57.87%), which was directly used in the next step.

MS(ESI,m/z):174.35[M+H] ⁺ ,RT(min):0.602

Step 5 Synthesis of 2-(4-(difluoromethylene)piperidin-1-yl)-N-(2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-4-iodobenzamide (Compound 4-6)

Under nitrogen protection, tetramethyl chlorouronium hexafluorophosphate (647.92 mg, 2.308 mmol, 4 eq) and N-methylimidazole (474.0 mg, 5.770 mmol, 10 eq) were added to a solution of 2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-amine (compound 4-5, 100 mg, 0.577 mmol, 1 eq) in DCM (1 mL, 0.006 mmol, 0.01 eq) at room temperature. After stirring for 2 minutes, 2-[4-(difluoromethylene)piperidin-1-yl]-4-iodobenzoic acid (compound 1-12, 262.66 mg, 0.692 mmol, 1.2 eq) was added at room temperature and reacted for one hour. Water (3 mL) was added to the reaction mixture at room temperature to quench it. The reaction mixture was extracted with dichloromethane (3×2 mL). The organic phases were combined, backwashed with saturated brine (1×3 mL), and dried over sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (5:1) to obtain 2-(4-(difluoromethylene)piperidin-1-yl)-N-(2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-4-iodobenzamide (Compound 4-6, 60 mg, 19.45%).

MS:(ESI,m/z):534.95[M+H] ⁺ ,RT(min):1.094

Step 6 Synthesis of 2-(4-(difluoromethylene)piperidin-1-yl)-N-(2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-4-(2-hydroxyethylsulfonylamino)benzamide (Compound 4)

Under nitrogen protection, sarcosine (1.67 mg, 0.019 mmol, 0.2 eq), potassium carbonate (38.8 mg, 0.282 mmol, 3 eq) and cuprous iodide (5.35 mg, 0.028 mmol, 0.3 eq) were added in batches to 2-(4-(difluoromethylene)piperidin-1-yl)-N-(2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-4-iodobenzamide (compound 4-6, 50 mg, 0.094 mmol, 1 eq) and 2-hydroxyethanesulfonamide (23.42 mg, 0.188 mmol, 2 eq) in N,N-dimethylformamide (1 mL) at room temperature. The temperature was raised to 120°C, stirred for reaction for 1.5 hours, and the reaction solution was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (3×10 mL). The organic phases were combined, backwashed with saturated brine (1×10 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid phase to obtain 2-(4-(difluoromethylene)piperidin-1-yl)-N-(2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-4-(2-hydroxyethanesulfonylamino)benzamide (compound 4, 19.7 mg, 39.6%), under the following conditions: Column specifications: XBridge BEH Shield RP18 5μm, 30mm*150mm; Mobile phase A: water (10mM ammonium bicarbonate), mobile phase B: acetonitrile; Flow rate: 60mL/min; Elution gradient: 35%B to 58%B in 8 minutes; Detection wavelength: UV 254nm/220nm; Retention time (min): 7.2.

MS:(ESI,m/z):532.30[M+H] ⁺ ,RT(min):1.564

¹ H NMR: (400MHz, DMSO-d ₆ ) δ12.30(s,1H),10.16(s,1H),8.32(dd,1H),8.03(d,1H),7.18(d,1H),7.17 –7.13(m,2H),7.09(dd,1H),5.00(s,1H),4.13(t,2H),3.76(t,2H),3.36(s,2H),3.00(t,4H), 2.93–2.88(m,2H),2.71–2.60(m,6H).

Example 5

1S,4R-N-(2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)phenyl)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carboxamide (Compound 22)

Compound 22

The first step is the synthesis of 1-(5-bromo-2-nitrophenyl)-4-(difluoromethylene)piperidine (compound 22-2):

Under nitrogen protection, potassium carbonate (28.02 g, 202.791 mmol, 3 eq) and 4-bromo-2-fluoro-1-nitrobenzene (compound 22-1, 16.36 g, 74.357 mmol, 1.1 eq) were added to a solution of 4-(difluoromethylene)piperidine hydrochloride (compound 1-10, 9 g, 67.597 mmol, 1 eq) in dimethyl sulfoxide (90 mL) at room temperature. The reaction system was heated to 90°C and stirred for 16 hours. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3×150 mL). The organic phases were combined, backwashed with saturated brine (2×150 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (5:1) to obtain 1-(5-bromo-2-nitrophenyl)-4-(difluoromethylene)piperidine (compound 22-2, 4.5 g, 19.98%).

¹ H NMR (400MHz, DMSO-d ₆ ) δ7.80(d,1H),7.48(d,1H),7.33–7.25(m,1H),3.06(t,4H),2.27(d,4H).

Step 2 Synthesis of 4-bromo-2-[4-(difluoromethylene)piperidin-1-yl]aniline (Compound 22-3):

Iron powder (41.91 mg, 0.750 mmol, 5 eq) and ammonium chloride (441.56 mg, 8.255 mmol, 5 eq) were added to a mixed solution of 1-(5-bromo-2-nitrophenyl)-4-(difluoromethylene)piperidine (compound 22-2, 550 mg, 1.651 mmol, 1 eq) in ethanol (11.0 mL) and water (2.2 mL) at room temperature, and the reaction solution was heated to 80°C and stirred for 2 hours. The reaction solution was filtered, the filter cake was washed with ethyl acetate (3×20 mL), and the filtrate was concentrated under reduced pressure. The reaction mixture was extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated brine (3×20 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. 4-bromo-2-[4-(difluoromethylene)piperidin-1-yl]aniline (compound 22-3, 330 mg, 65.93%) was obtained. The crude product was not further purified and was directly used in the next step.

MS:(ESI,m/z):303.05[M+H]+,RT(min):1.406

Step 3 Synthesis of (1S, 4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carbonitrile (Compound 22-4):

Under nitrogen protection, tetrakis(triphenylphosphine)palladium (439.16 mg, 0.380 mmol, 0.2 eq) was added to a solution of (1S,4R)-6-bromo-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine (compound 1-4, 500 mg, 1.900 mmol, 1 eq) and zinc cyanide (267.74 mg, 2.280 mmol, 1.2 eq) in N,N-dimethylformamide (5 mL) at room temperature. The reaction solution was heated to 100°C and stirred for 3 hours. The reaction solution was cooled to room temperature, diluted with water, and extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated brine (2×30 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography with petroleum ether/ethyl acetate (5:1) to give (1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carbonitrile (compound 22-4, 300 mg, 75.45%).

MS:(ESI,m/z):210.10[M+H]+,RT(min):0.901

Step 4 Synthesis of (1S, 4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carboxylic acid (Compound 22-5):

At room temperature, sulfuric acid (0.5 mL, 9.381 mmol, 6.54 eq) and acetic acid (1 mL, 17.452 mmol, 12.17 eq) were added dropwise to a solution of (1S, 4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carbonitrile (compound 22-4, 300 mg, 1.434 mmol, 1 eq) in water (4 mL). The reaction solution was heated to 110°C and stirred for 2 days. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography under the following conditions: C18 chromatography column, mobile phase, water and acetonitrile, 10% to 50% gradient in 10 minutes, UV254 nanometer detector to obtain (1S, 4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carboxylic acid (compound 22-5, 200 mg, 61.12%).

MS:(ESI,m/z):229.40[M+H] ⁺ ,RT(min):0.480

Step 5: Synthesis of (1S, 4R)-N-(4-bromo-2-(4-(difluoromethylene)piperidin-1-yl)phenyl)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carboxamide (Compound 22-6):

Under nitrogen protection, N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (860.48 mg, 3.070 mmol, 10 eq) and N-methylimidazole (100.72 mg, 1.228 mmol, 4 eq) were added to a solution of 4-bromo-2-[4-(difluoromethylene)piperidin-1-yl]aniline (compound 22-3, 185.94 mg, 0.614 mmol, 2 eq) and (1S,4R)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carboxylic acid (compound 22-5, 70 mg, 0.307 mmol, 1.00 eq) in dichloromethane (4 mL) at room temperature. After the addition was complete, the system was stirred at 60°C for 1 hour. The reaction solution was cooled to room temperature and extracted with ethyl acetate (3×30 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (3×20 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (2:01) to obtain (1S, 4R)-N-(4-bromo-2-(4-(difluoromethylene)piperidin-1-yl)phenyl)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carboxamide (compound 22-6, 120 mg, 57.16%).

MS:(ESI,m/z):513.40[M+H] ⁺ ,RT(min):1.496

Step 6: Synthesis of 1S,4R-N-(2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)phenyl)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carboxamide (Compound 22):

Under nitrogen protection, (1S,4R)-N-(4-bromo-2-(4-(difluoromethylene)piperidin-1-yl)phenyl)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carboxamide (compound 22-6, 50 mg, 0.097 mmol, 1 eq) and 2-hydroxyethanesulfonamide (24.38 mg, 0.19 To a solution of 1,4-dioxane (2 mL) containing 2-(4 mmol, 2 eq) of 1,4-dioxane, dicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl) phosphane (10.42 mg, 0.019 mmol, 0.2 eq) and (methanesulfonic acid {dicyclohexyl (3-isopropoxy-2',4',6'-triisopropyl-[1,1'-biphenyl]-2-yl) phosphane (10.42 mg, 0.019 mmol, 0.2 eq) The reaction mixture was heated to 100 °C and stirred for 1 hour. The reaction mixture was cooled to room temperature and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (3 × 20 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography under the following conditions (chromatographic column specifications: Ultimate μXB-C18; mobile phase A: water (10 mM ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 100 mL/min; elution gradient: 30% B to 65% B in 20 minutes; detection wavelength: UV 254nm/221nm). 1S,4R-N-(2-(4-(difluoromethylene)piperidin-1-yl)-4-(2-hydroxyethanesulfonylamino)phenyl)-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine-6-carboxamide (Compound 22, 25.8mg, 47.17%) was obtained.

MS:(ESI,m/z):558.25[M+H] ⁺ ,RT(min):1.612

¹ H NMR (400 MHz, DMSO-d ₆ )δ12.19(s,1H),9.46(s,1H),8.51(d,1H),8.01–7.95(m,1H),7.87–7.80(m,1H),7.36(t,1H),7.10(d,1H),7.03–6.96(m,1H),5.29(s,1H),4.95( s,1H),3.75(t,2H),3.58(d,1H),3.22(t,2H),2.99–2.77(m,4H),2.60–2.50(m,4H),2.31(d,1H),2.23–2.12(m,1H),2.11–2.00(m,2H),1.28–1.1 4(m,2H).

Biological evaluation

Test Example 1

Test Name: Imaging-based Nuclear Count Analysis (NCA) in OVCAR-3 cells

Day 0 Compound dilution and treatment

a) The final tested concentrations of AM-5308 were: 10000, 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, and 0.5 nM.

b) Test compounds, final test concentrations: 10000, 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, 0.5 nM.

c) The cells were cultured in a 37°C, 5% CO ₂ incubator for 4 days.

d) The DMSO concentration was 0.1%.

On day 1, cells were seeded into 384-well cell culture plates.

a) When the cell confluence reached 80%-90%, cells were processed.

b) Resuspend the cells in culture medium, then count and dilute the cells at the desired density.

c) Add 30 μL/well of the cell suspension containing appropriate cells to a 384-well plate: 600 cells/well.

Day 4 Testing

a) Add 30 μL of 8% fixative (final concentration 4%) and incubate the plate at room temperature for 30 minutes.

b) Centrifuge the plate at 1000 RPM for 30 seconds.

c) Wash twice with 60 μl/well PBS.

d) After fixation, cells were permeabilized and stained in 60 μL of wash buffer (1% BSA, 0.2% Triton X-100, IX PBS) containing 2 μg/mL Hoechst 33342 DNA dye.

e) Seal the plate and incubate at room temperature for 1 hour in the dark.

f) Wash 3 times with PBS.

g) Add 50 μL PBS/well and scan the plate using HCS.

h) Data collection and detection

Data analysis

Inhibition rate (%) = 100-(compound well reading value-low reading control well reading value)/(high reading control well reading value-low reading control well reading value)*100

High reading control wells: cells were added with 30 nL DMSO; low reading control wells: 10 μM AM-5308 wells.

GraphPad Prism 8 software was used to calculate IC ₅₀ (nM) and plot the effect-dose curve of the compound.

Table 1 Biological activity data of the compounds of the present application

The above is an exemplary description of the implementation of the technical solution of the present invention. It should be understood that the protection scope of the present invention is not limited to the above implementation. Any modification, equivalent substitution, improvement, etc. made by those skilled in the art within the spirit and principle of the present invention should be included in the protection scope of the claims of this application.

Claims (10)

1. A compound of formula (I), racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof:

Wherein,

R ₁、R₂, identical or different, are independently of one another selected from H, halogen, C _1-12 alkyl;

a is-NHC (O) -or-C (O) NH-;

Ring E is selected from C _6-14 aromatic ring, 5-14 membered heteroaromatic ring;

Ring G is selected from the group consisting of C _3-12 cycloalkane ring, C _3-12 cycloalkene ring, 5-14 membered heterocycle, C _6-14 aryl ring, 5-14 membered heteroaryl ring;

Each R _e, which are the same or different, are independently selected from H, halogen, hydroxy, amino, cyano, C _1-12 alkyl, C _1-12 alkyloxy; m is selected from 0, 1, 2,3 or 4;

Each R _g, which is the same or different, is independently selected from H, halogen, hydroxy, amino, cyano, the following groups, unsubstituted or optionally substituted with one, two or more R _g1: c _1-12 alkyl, C _1-12 alkyloxy; or two R _g form with the atom to which they are each attached the following group, unsubstituted or optionally substituted with one, two or more R _g2: a C _3-8 cycloalkyl ring, a C _3-8 cycloalkyl ring, a 5-14 membered heterocyclic ring, a C _6-14 aromatic ring, or a 5-14 membered heteroaromatic ring; each R _g1、R_g2, which are identical or different, are independently of one another selected from H, halogen, C _1-12 alkyl, C _1-12 alkyloxy; n is selected from 0, 1,2, 3 or 4.

2. The compound, racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof according to claim 1, wherein R ₁、R₂ is the same or different and is independently selected from H, F, cl, methyl;

Preferably, R ₁、R₂ is the same and is F.

3. The compound, racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof according to claim 1 or 2, wherein ring E is selected from a C ₆ aromatic ring, a 6 membered heteroaromatic ring;

Preferably, ring E is selected from benzene ring, pyridine ring, pyrimidine ring, pyridazine ring.

4. A compound according to any one of claims 1 to 3, racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof, wherein ring G is selected from C _5-6 cycloalkyl ring, C _5-6 cycloalkyl ring, 5-6 membered heterocycle, C ₆ aryl ring, 5-6 membered heteroaryl ring;

Preferably, ring G is selected from cyclohexane ring, cyclohexene ring, benzene ring, pyridine ring, imidazole ring;

preferably, each R _e, equal to or different from each other, is selected independently from H, C _1-6 alkyl, C _1-6 alkyloxy;

Preferably, each R _g, equal to or different from each other, is selected independently from H, halogen, C _1-6 alkyl, C _1-6 alkyloxy; or two R _g form with the atom to which they are each attached the following group, unsubstituted or optionally substituted with one, two or more R _g2: a C _3-8 cycloalkane ring, a 5-8 membered heterocycle;

Preferably, each R _g, which are the same or different, are independently selected from H, methoxy; or two R _g form with the atom to which they are each attached the following group, unsubstituted or optionally substituted with one, two or more R _g2: (e.g )、

Preferably, each R _g2, equal to or different from each other, is selected independently from H, halogen, C _1-6 alkyl, C _1-6 alkyloxy;

Preferably, each R _g2, identical or different, is independently selected from H, F, methyl.

5. The compound, racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof of any one of claims 1 to 4, wherein the compound of formula (I) is selected from the group consisting of:

Wherein A, ring E, ring G, R ₁、R₂、R_e、R_g, m, n have the definition as defined in any one of claims 1 to 4.

6. The compound, racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof of any one of claims 1 to 5, wherein the compound of formula (I) is selected from the group consisting of:

Wherein A, R ₁、R₂、R_e、R_g、R_g2, m, n have the definition as defined in any one of claims 1 to 5.

7. The compound according to any one of claims 1-6, racemate, stereoisomer, tautomer, isotopic label, solvate, polymorph, pharmaceutically acceptable salt or prodrug compound thereof, wherein the compound of formula (I) is selected from the group consisting of:

8. a process for the preparation of a compound according to any one of claims 1 to 7, comprising the following step a:

step A: reacting the compound (a) with 2-hydroxyethanesulfonamide to obtain a compound represented by the formula (I):

Wherein a, ring E, ring G, R ₁、R₂、R_e、R_g, m, n have the definition of any one of claims 1 to 7; x is selected from halogen, such as F, cl, br or I;

preferably, the preparation method of the compound shown in the formula (I) can further comprise the following steps B1 or B2:

Step B1: reacting a compound (a 2) with a compound (a 3) to obtain a corresponding compound represented by formula (a) for a compound represented by formula (I) wherein a is-C (O) NH-; or alternatively

Step B2: for a compound represented by formula (I) wherein A is-NHC (O) -, reacting compound (a 4) with compound (a 5) to obtain the corresponding compound represented by formula (a);

Wherein a, ring E, ring G, R ₁、R₂、R_e、R_g, m, n have the definition of any one of claims 1 to 7; x is selected from halogen, such as F, cl, br or I;

preferably, the preparation method of the compound shown in the formula (I) can further comprise the following steps C1 or C2:

Step C1:

Coupling the compound (b 1) and the compound (b 2) to obtain a compound (a 1), and carrying out reduction reaction on the compound (a 1) to obtain a compound (a 2):

Step C2:

Reacting the compound (b 1) with the compound (b 3) to obtain a compound (a 4):

Wherein R ₁ and R ₂ have the definition as defined in any one of claims 1 to 7, X and Y are each independently selected from halogen, such as F, cl, br or I, respectively.

9. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the compounds of any one of claims 1-7, racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof.

10. Use of at least one of the compounds of any one of claims 1-7, racemates, stereoisomers, tautomers, isotopic labels, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof for the preparation of a medicament;

Preferably, the use may be in the manufacture of a medicament for the treatment of a KIF18A mediated disorder and/or disease, such as in the manufacture of a KIF18A inhibitor medicament;

Preferably, the disease is, for example, cancer, including bowel cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer or ovarian cancer.