Tricyclic derivatives, pharmaceutical compositions and uses

Abstract

The application provides a tri-cyclic derivative, and the structure of the derivative is shown in formula (I). In addition, the application also discloses pharmaceutically acceptable salts of the derivative, stereoisomers of the derivative, a pharmaceutical composition and application of the derivative. The compound of the application has significant CDK12 inhibiting activity and is very practical.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and in particular to a tricyclic derivative, its pharmaceutically acceptable salt, stereoisomer, pharmaceutical composition, and its application. Background Technology

[0002] Cell cycle-dependent kinases (CDKs) belong to the serine / threonine kinase family. They participate in physiological processes such as cell proliferation and transcription. Based on their different functions, CDKs can be divided into two main categories: one category of CDKs participates in cell cycle regulation, mainly including CDK1, CDK2, CDK4, and CDK6; the other category of CDKs participates in transcriptional regulation, mainly including CDK7, CDK8, CDK9, CDK12, and CDK13.

[0003] CDK12 becomes active by binding to cyclin K and transmits signals downstream. The process of copying genetic information from DNA to messenger RNA (mRNA) is called transcription. CDK12 extends the transcriptional response of mRNA by phosphorylating the second serine residue (Ser2) at the C-terminus of RNA polymerase II. CDK12 is thought to be involved in the transcriptional elongation of genes with long gene lengths and affects the expression of genes such as BRCA1, ATM, and FANCD2. All of these genes are a group involved in DNA damage response (DDR) and primarily participate in maintaining the stability of genomic DNA through DNA repair mechanisms. Inhibition of CDK12 reduces the expression of DDR-related genes, thereby inhibiting the DNA repair response. Inhibition of both PARP and CDK12 in DNA completely suppresses the DNA repair response through synthetic lethality and induces cell death in cancer cells. Therefore, inhibiting CDK12 is very useful for the prevention and treatment of cancer.

[0004] In order to better develop new drugs for treating cancer and related diseases, and given the large market demand for CDK-mediated disease drugs, there is an urgent need to develop novel, safe and effective CDK12 inhibitors that can treat a variety of cancers, especially small molecule inhibitors that selectively target CDK12, which may have higher safety. Summary of the Invention

[0005] Based on this, the present invention provides a tricyclic derivative with high activity against CDK12 inhibitors, and its pharmaceutically acceptable salt or stereoisomer.

[0006] The present invention is achieved through the following technical solution.

[0007] A compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0008]

[0009] in:

[0010] R1 and R2 are each independently selected from cyano, -CF3, -OCF3, -Cl, -Br, -F, hydroxyl, nitro, substituted or unsubstituted imidazolyl, substituted or unsubstituted pyrazolyl, straight-chain alkyl having 1 to 20 carbon atoms, alkoxy having 1 to 20 carbon atoms, branched alkyl having 3 to 20 carbon atoms, substituted or unsubstituted cyclic alkyl having 3 to 20 carbon atoms, substituted or unsubstituted cyclic alkoxy having 3 to 20 carbon atoms, substituted or unsubstituted heterocyclic having 3 to 20 ring atoms, -OR, -NH-R, or combinations thereof; each time R appears, it is independently selected from substituted or unsubstituted cyclic alkoxy having 3 to 20 carbon atoms, or substituted or unsubstituted heterocyclic having 3 to 20 ring atoms.

[0011] R3 and R4 are each independently selected from -H, -D, a straight-chain alkyl group having 1 to 20 carbon atoms, a branched or cyclic alkyl group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic group having 6 to 60 cyclic atoms, or a combination of these groups;

[0012] Ar1 is selected from any of the following structures:

[0013]

[0014] Each time X1 appears, it is independently selected from CR5 or N;

[0015] Each time Y1 appears, it is independently selected from NR6, CR6R7, C=O, O, S or S=O;

[0016] R5, R6, and R7 are each independently selected from -H, -D, straight-chain alkyl groups having 1 to 20 carbon atoms, alkoxy groups having 1 to 20 carbon atoms, branched alkyl groups having 3 to 20 carbon atoms, cyclic alkyl groups having 3 to 20 carbon atoms, branched alkoxy groups having 3 to 20 carbon atoms, cyclic alkoxy groups having 3 to 20 carbon atoms, or combinations of these systems.

[0017] In one embodiment, the compound of formula (I) has the structure shown in formula (II):

[0018]

[0019] In one embodiment, R3 and R4 are each independently selected from -H, -D, substituted or unsubstituted aromatic groups having 6 to 30 ring atoms.

[0020] In one embodiment, the compound of formula (I) has the structure shown in formula (III):

[0021]

[0022] In one embodiment, Ar1 is selected from any of the following structures:

[0023]

[0024] In one embodiment, Ar1 is selected from any of the following structures:

[0025]

[0026] In one embodiment, R1 and R2 are each independently selected from cyano, -CF3, -Cl, -Br, -F, or any of the following structures:

[0027]

[0028] In one embodiment, the compound of formula (I) is any one of the following compounds:

[0029]

[0030] The present invention also provides the use of the compound as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of a medicament for treating and / or preventing diseases associated with or mediated by CDK12 activity.

[0031] In one embodiment, the disease associated with or mediated by CDK12 activity is cancer.

[0032] The present invention also provides a pharmaceutical composition comprising the compound as described above, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0033] Compared with the prior art, the compounds of the present invention, or their pharmaceutically acceptable salts, or their stereoisomers, have the following beneficial effects:

[0034] This invention provides a series of novel heterocyclic-substituted tricyclic derivatives that exhibit high inhibitory activity against CDK12, with an IC50 value for CDK12 kinase inhibition. 50 With a value of 42 nM to 367 nM, it can be used as a drug for the treatment and / or prevention of diseases associated with or mediated by CDK12 activity, and has the potential for excellent therapeutic effects on CDK12 activity-mediated diseases. Detailed Implementation

[0035] To facilitate understanding of the present invention, a more complete description will be given below with reference to relevant embodiments. Preferred embodiments of the invention are shown in the embodiments. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0036] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Thus, a feature defined as "first" or "second" may explicitly or implicitly include at least one of that feature. In the description of the invention, "a plurality of" means at least two, such as two, three, etc., unless otherwise explicitly specified. In the description of the present invention, "a number" means at least one, such as one, two, etc., unless otherwise explicitly specified.

[0037] The terms "preferred," "more preferably," etc., used in this invention refer to embodiments of the invention that provide certain beneficial effects under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the description of one or more preferred embodiments does not imply that other embodiments are unavailable, nor is it intended to exclude other embodiments from the scope of this invention.

[0038] When a numerical range is disclosed herein, the range is considered continuous and includes the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0039] Unless otherwise specified, all percentages, fractions, and ratios are calculated based on the total mass of the compositions of the present invention. Unless otherwise specified, all masses of the listed ingredients are given as the content of the active substance, and therefore do not include solvents or byproducts that may be present in commercially available materials. The term "percentage by mass" herein may be expressed by the symbol "%". Unless otherwise specified, all molecular weights herein are weight-average molecular weights expressed in Daltons. Unless otherwise specified, all formulations and tests herein are conducted at 25°C. The terms "comprising," "including," "containing," "containing," "having," or other variations herein are intended to cover non-closed inclusions, and no distinction is made between these terms. The term "comprising" means additional steps and ingredients that may be added without affecting the final result. The compositions and methods / processes of the present invention comprise, consist of, and substantially consist of the essential elements and limitations described herein, as well as any additional or optional ingredients, components, steps, or limitations described herein. No distinction is made between the terms "efficacy," "performance," "effect," and "potency" herein.

[0040] Unless otherwise stated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0041] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0042] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention, prepared by reacting a compound with a relatively non-toxic acid or base, as discovered in this invention, with a specific substituent. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts, such as formic acid, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; as well as salts of amino acids (such as arginine) and salts of organic acids such as glucuronic acid. Certain compounds of the present invention contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0043] The pharmaceutically acceptable salts of the present invention can be synthesized from parent compounds containing acid radicals or bases by conventional chemical methods. Generally, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of a suitable base or acid in water or an organic solvent or a mixture thereof.

[0044] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0045] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0046] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0047] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being in a non-mirror relationship with each other.

[0048] Unless otherwise stated, "(+)" indicates right-handed rotation, "(-)" indicates left-handed rotation, and "(±)" indicates racemic rotation.

[0049] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key and straight dashed key

[0050] Unless otherwise stated, when a compound contains a double bond structure, such as a carbon-carbon double bond, a carbon-nitrogen double bond, or a nitrogen-nitrogen double bond, and each atom in the double bond is attached to two different substituents (in a double bond containing a nitrogen atom, the lone pair of electrons on the nitrogen atom is considered as one of the substituents it is attached to), if the atoms in the double bond and their substituents in the compound are separated by a wavy line... The ligature indicates that the compound exists as a (Z) type isomer, an (E) type isomer, or a mixture of both isomers. For example, formula (A) indicates that the compound exists as a single isomer of formula (A-1) or formula (A-2) or as a mixture of the two isomers of formula (A-1) and (A-2); formula (B) indicates that the compound exists as a single isomer of formula (B-1) or formula (B-2) or as a mixture of the two isomers of formula (B-1) and (B-2); formula (C) indicates that the compound exists as a single isomer of formula (C-1) or formula (C-2) or as a mixture of the two isomers of formula (C-1) and (C-2).

[0051]

[0052] Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to isomers with different functional groups in dynamic equilibrium at room temperature, capable of rapidly interconverting into each other. Tautomers can be chemically equilibrated if possible (e.g., in solution). For example, proton tautomers (also called prototropic tautomers) involve interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions involving the rearrangement of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between the tautomers pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0053] Unless otherwise stated, the terms "rich in one isomer," "isomer enrichment," "rich in one enantiomer," or "enantiomer enrichment" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0054] Unless otherwise stated, the terms "isomer excess" or "enantiomer excess" refer to the difference between the relative percentages of two isomers or two enantiomers. For example, if one isomer or enantiomer is 90% and the other isomer or enantiomer is 10%, then the isomer or enantiomer excess (ee value) is 80%.

[0055] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of the present invention, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated, and the auxiliary group is cleaved to provide the desired enantiomer in pure form. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase, optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0056] The compounds of this invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium. 3 H), Iodine-125 125 I) or C-14 14 C). For example, deuterium can be used to replace hydrogen to form deuterated drugs. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug biological half-life. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0057] "Optional" or "optionally" means that the event or condition described below may occur but is not required to occur, and the description includes both the scenario in which said event or condition occurs and the scenario in which said event or condition does not occur.

[0058] The term "substitution" refers to the replacement of one or more hydrogen atoms on a specific atom by a substituent. Substituents can include deuterium and hydrogen variants, provided the valence state of the specific atom is normal and the resulting compound is stable. The type and number of substituents can be arbitrary, based on what is chemically possible. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups.

[0059] When any variable (e.g., R1) appears more than once in the composition or structure of a compound, its definition is independent in each case. Therefore, for example, if a group is substituted by 0-2 R1s, the group can optionally be substituted by at most two R1s, and R1 has independent options in each case. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0060] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0061] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0062] When a substituent is vacant, it means that the substituent does not exist. For example, if X is vacant in AX, the structure is actually A. When the listed substituents do not specify which atom they are attached to the substituted group through, such substituents can be bonded to any of their atoms. For example, a pyridinium group as a substituent can be attached to the substituted group through any carbon atom on the pyridine ring.

[0063] When the listed linking groups do not specify their linking direction, the linking direction is arbitrary, for example, The linker group L is -MW-. In this case, -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form a ring. Alternatively, rings A and B can be connected in the opposite direction to the left-to-right reading order to form a ring. The combination of linking groups, substituents, and / or their variants is permitted only if such a combination produces a stable compound.

[0064] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of that group can be connected to other groups by chemical bonds. When the chemical bond connection is non-directional and the connectable site contains H atoms, the number of H atoms at that site will decrease accordingly with the number of chemical bonds connected, resulting in a group with a corresponding valence. The chemical bonds connecting the site to other groups can be straight solid line bonds. Straight dashed key or wavy line For example, a straight solid line bond in -OCH3 indicates that the oxygen atom in that group is connected to other groups; The straight dashed bond in the diagram indicates that the group is connected to other groups through both ends of the nitrogen atom in the group; The wavy lines in the text indicate that the phenyl group is connected to other groups through the carbon atoms at positions 1 and 2 of the phenyl group. This indicates that any connectable site on the piperidinyl group can be linked to other groups via a single chemical bond, including at least... Even if H atoms are drawn on -N- in these four connection methods, Still includes In this type of linkage, when a chemical bond is attached, the number of hydrogen atoms at that site is reduced by one, resulting in a monovalent piperidinyl group.

[0065] Unless otherwise specified, the number of atoms in a ring is usually defined as the elemental number of the ring. For example, a “3-7 elemental ring” refers to a “ring” with 3 to 7 atoms arranged around it.

[0066] Unless otherwise specified, the term "C" 1-6"alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting of 1 to 6 carbon atoms. Preferably, it is C16. 1-4 Alkyl groups can be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). C 1-3 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and butyl (including n-butyl, isobutyl, tert-butyl, and sec-butyl).

[0067] Unless otherwise specified, the term "C" 1-6 "Alkoxy" refers to alkyl groups containing 1 to 6 carbon atoms that are attached to the rest of the molecule by an oxygen atom. Preferably, C16 is used. 1-3 Alkyl group. C 1-3 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), etc.

[0068] Unless otherwise specified, the term "C" 1-6 "Alkylamino" refers to alkyl groups containing 1 to 6 carbon atoms that are attached to the rest of the molecule via an amino group. Preferably, C16 is used. 1-3 Alkylamino. C 1-3 Examples of alkylamino groups include, but are not limited to, -NHCH3, -N(CH3)2, -NHCH2CH3, -N(CH3)CH2CH3, -NHCH2CH2CH3, -NHCH2(CH3)2, etc.

[0069] Unless otherwise specified, the term “halogen” or “halogen” itself or as part of another substituent means a fluorine, chlorine, bromine or iodine atom.

[0070] Unless otherwise specified, the terms "5-membered heteroaryl" and "5-cyclic heteroaryl" are used interchangeably in this invention. The term "5-membered heteroaryl" refers to a monocyclic group consisting of 5 ring atoms with a conjugated π-electron system, wherein 1, 2, 3, or 4 of the ring atoms are heteroatoms independently selected from O, S, and N, and the remainder are carbon atoms. The nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). p(where p is 1 or 2). A 5-membered heteroaryl group can be attached to the rest of the molecule via a heteroatom or a carbon atom. Examples of such 5-membered heteroaryl groups include, but are not limited to, pyrrole (including N-pyrrole, 2-pyrrole, and 3-pyrrole), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl), imidazole (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl, and 5-imidazolyl), oxazolyl (including 2-oxazolyl, 4-oxazolyl, and 5-oxazolyl), and triazolyl (1H-1,2,3-triazolyl, 2H-2H-3H-4H-3H-4H-5 ... H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isooxazolyl, 4-isooxazolyl and 5-isooxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.).

[0071] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case with n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 C 11 and C 12 It also includes any range from n to n+m, such as C 1-12 Including C 1-3 C 1-6 C 1-9 C 3-6 C 3-9 C 3-12 C 6-9 C 6-12 and C 9-12 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3-12-membered rings include 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, 10-membered, 11-membered, and 12-membered rings, and also any range from n to n+m. For example, 3-12-membered rings include 3-6-membered, 3-9-membered, 5-6-membered, 5-7-membered, 6-7-membered, 6-8-membered, and 6-10-membered rings, etc.

[0072] Unless otherwise specified, "C 3-7 "Cycloalkyl" refers to a saturated cyclic hydrocarbon group consisting of 3 to 7 carbon atoms, including monocyclic and bicyclic systems, wherein bicyclic systems include spirocyclic, fused, and bridged rings. The C 3-7 Cycloalkyl groups include C 3-6 C 4-6 C 4-5 C 5-7Or C 5-6 Cycloalkyl groups, etc.; they can be monovalent, divalent, or polyvalent. C 3-7 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.

[0073] Unless otherwise specified, the term "3-7 membered heterocyclic alkyl" on its own or in combination with other terms respectively refers to a saturated cyclic group consisting of 3 to 7 ring atoms, wherein 1, 2, 3 or 4 of the ring atoms are heteroatoms independently selected from O, S and N, and the remainder are carbon atoms, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e. NO and S(O)). p (where p is 1 or 2). It includes monocyclic and bicyclic systems, with bicyclic systems including spirocyclic, fused, and bridged rings. Furthermore, regarding the "3-7 membered heterocyclic alkyl", the heteroatom can occupy the connection position between the heterocyclic alkyl group and the rest of the molecule. The 3-7 membered heterocyclic alkyl groups include 5-7, 3, 4, 5, 6, and 7 membered heterocyclic alkyl groups, etc. Examples of 3-7 membered heterocyclic alkyl groups include, but are not limited to, azirrobutyl, oxacyclobutyl, thiocyclobutyl, pyrrolidinyl, pyrazolyl, imidazoalkyl, tetrahydrothiopheneyl (including tetrahydrothiophene-2-yl and tetrahydrothiophene-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxyl, dithiaalkyl, isoxazolyl, isothiazolyl, 1,2-oxazinyl, 1,2-thiaazinyl or hexahydropyridazinyl, etc.

[0074] Unless otherwise specified, the term "3-7 membered nitrogen-containing heterocyclic alkyl" means a 3-7 membered heterocyclic alkyl containing at least one nitrogen atom.

[0075] Alicyclic groups refer to saturated or partially unsaturated all-carbon ring systems. "Partially unsaturated" means a ring moiety comprising at least one double or triple bond. "Partially unsaturated" is intended to encompass rings having multiple unsaturated sites, but does not necessarily include aryl or heteroaryl moieties as defined herein. Non-limiting examples include cyclopropyl rings, cyclobutyl rings, cyclopentyl rings, cyclopentenyl rings, cyclohexyl rings, cyclohexenyl rings, cyclohexadienyl rings, cycloheptyl rings, cycloheptanetrienyl rings, cyclopentanone rings, cyclopentane-1,3-dione rings, etc.

[0076] Alicyclic groups refer to saturated or partially unsaturated alicyclic groups in which one, two, or three ring carbon atoms are selected from nitrogen, oxygen, or S(O). tThe ring is replaced by heteroatoms (where t is an integer from 0 to 2), but does not include the ring portion of -OO-, -OS-, or -SS-, and the remaining ring atoms are carbon. Non-limiting embodiments include propylene oxide rings, azacyclic butane rings, oxacyclic butane rings, tetrahydrofuran rings, tetrahydrothiophene rings, tetrahydropyrrole rings, piperidine rings, pyrrolidine rings, oxazolidine rings, piperazine rings, dioxopentane rings, dioxane-2-one rings, morpholine rings, thiomorpholine rings, thiomorpholine-1,1-dioxide, tetrahydropyran rings, azacyclic butane-2-one rings, oxacyclic butane-2-one rings, pyrrolidine-2-one rings, pyrrolidine-2,5-dione rings, piperidine-2-one rings, dihydrofuran-2(3H)-one rings, dihydrofuran-2,5-dione rings, tetrahydro-2H-pyran-2-one rings, piperazine-2-one rings, and morpholine-3-one rings. Non-limiting examples of partially unsaturated monoheterocyclic rings include 1,2-dihydroaza-butadiene rings, 1,2-dihydrooxo-aza-butadiene rings, 2,5-dihydro-1H-pyrrole rings, 2,5-dihydrofuran rings, 2,3-dihydrofuran rings, 2,3-dihydro-1H-pyrrole rings, 3,4-dihydro-2H-pyrrole rings, 1,2,3,4-tetrahydropyridine rings, 3,6-dihydro-2H-pyrrole rings, 1,2,3,6-tetrahydropyridine rings, 4,5-dihydro-1H-imidazolium rings, 1,4,5,6-tetrahydropyrimidine rings, 3,4,7,8-tetrahydro-2H-1,4,6-oxadiazin rings, 1,6-dihydropyrimidine rings, and 4,5,6,7-tetrahydro-1H-1,3-diaza-aza-butadiene rings. 2,5,6,7-Tetrahydro-1,3,5-oxadiazine Rings, etc.

[0077] "Aryl" and "aromatic ring" are used interchangeably, both referring to an all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group with a conjugated π-electron system. This group can be fused with cycloalkyl, heterocyclic alkyl, cycloalkenyl, heterocyclic alkenyl, or heteroaryl groups. "C" 6-10 "Aryl" refers to a monocyclic or bicyclic aryl group having 6 to 10 carbon atoms. Non-limiting examples of aryl groups include phenyl, naphthyl, etc.

[0078] The terms "heteroaryl" and "heteroaryl ring" are used interchangeably, both referring to a group having a monocyclic, bicyclic, or polycyclic 4n+2 aromatic ring system (e.g., having 6 or 10 shared π electrons arranged in a ring) having a ring carbon atom and a ring heteroatom, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In this invention, the heteroaryl also includes a ring system in which the aforementioned heteroaryl ring is fused with one or more cycloalkyl rings, heterocyclic alkyl rings, cycloalkenyl rings, heterocyclic alkenyl rings, or aromatic rings. The heteroaryl ring may optionally be substituted. "5- to 10-membered heteroaryl" refers to a monocyclic or bicyclic heteroaryl having 5 to 10 ring atoms, wherein 1, 2, 3, or 4 ring atoms are heteroatoms. "5- to 6-membered heteroaryl" refers to a monocyclic heteroaryl group having 5 to 6 ring atoms, of which 1, 2, 3, or 4 ring atoms are heteroatoms. Non-limiting examples include thiophene, furanyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, pyrrole, pyrazolyl, triazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, tetrazolyl, isoxazolyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, thiadiazolyl, pyridinyl, pyridinyl, triazinyl, and tetraazinyl. "8 to 10-membered heteroaryl" refers to a bicyclic heteroaryl group having 8 to 10 ring atoms, of which 1, 2, 3, or 4 ring atoms are heteroatoms. Non-limiting examples include indole, isoindole, indazole, benzotriazolyl, benzothiophene, isobenzothiophene, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzooxazolyl, benzoisooxazolyl, benzooxadiazolyl, benzothiazolyl, and benzoisothiazolyl. Azolyl, benzothiadiazolyl, indazinyl, purine, pyrido[3,2-d]pyrimidinyl, pyrido[2,3-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrido[4,3-d]pyrimidinyl, 1,8-naphthidyl, 1,7-naphthidyl, 1,6-naphthidyl, 1,5-naphthidyl, pteridyl, quinolinyl, isoquinolinyl, zolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. "Heteroatom" refers to nitrogen, oxygen, or sulfur. In heteroaryl groups containing one or more nitrogen atoms, the bonding point can be a carbon or nitrogen atom, provided the valence allows. Heteroaryl bicyclic systems can include one or more heteroatoms in one or both rings.

[0079] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (such as a nucleophilic substitution reaction). For example, representative leaving groups include trifluoromethanesulfonates; chlorine, bromine, and iodine; sulfonate groups, such as methanesulfonates, toluenesulfonates, p-bromobenzenesulfonates, p-toluenesulfonates, etc.; and acyloxy groups, such as acetoxy and trifluoroacetoxy.

[0080] The term "protecting group" includes, but is not limited to, "amino protecting group," "hydroxy protecting group," or "thiol protecting group." The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the nitrogen position of an amino group. Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenemethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), triphenylmethyl (Tr), 1,1-di-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term "hydroxyl protecting group" refers to a protecting group suitable for preventing hydroxyl side reactions. Representative hydroxyl protecting groups include, but are not limited to: alkyl groups, such as methyl, ethyl, and tert-butyl; acyl groups, such as alkanolyl groups (e.g., acetyl); arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl (diphenylmethyl, DPM); silyl groups, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc.

[0081] Unless otherwise defined, the phrase "substituents selected independently of each other" in this invention means that when one or more hydrogen atoms on a group are replaced by substituents, the types of substituents may be the same or different, and the selected substituents are of independent types.

[0082] Typically, the compounds of this invention, or their pharmaceutically acceptable salts, or stereoisomers, can be formulated with one or more pharmaceutical carriers into suitable dosage forms for administration. These dosage forms are suitable for oral, rectal, topical, intraoral, and other non-gastrointestinal administration (e.g., subcutaneous, intramuscular, intravenous, etc.). For example, suitable dosage forms for oral administration include capsules, tablets, granules, and syrups. The compounds of this invention contained in these formulations can be solid powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; water-in-oil or oil-in-water emulsions, etc. The above dosage forms can be prepared from the active compound and one or more carriers or excipients using common pharmaceutical methods. The carriers mentioned above need to be compatible with the active compound or other excipients. For solid dosage forms, commonly used non-toxic carriers include, but are not limited to, mannitol, lactose, starch, magnesium stearate, cellulose, glucose, and sucrose. Carriers used for liquid dosage forms include water, physiological saline, glucose aqueous solution, ethylene glycol, and polyethylene glycol. The active compound can form solutions or suspensions with the above carriers.

[0083] The compositions of the present invention are formulated, quantified, and administered in accordance with medical practice guidelines. The "therapeutic effective amount" of the compound administered is determined by factors such as the specific condition to be treated, the individual being treated, the cause of the condition, the target of the drug, and the route of administration.

[0084] "Therapeutic effective amount" refers to the amount of the compound of the present invention that will elicit a biological or medical response in an individual, such as reducing or inhibiting enzyme or protein activity or improving symptoms, alleviating symptoms, slowing or delaying disease progression, or preventing disease.

[0085] The therapeutically effective amount of the compound of the present invention or its pharmaceutically acceptable salt or stereoisomer contained in the pharmaceutical composition or the pharmaceutical composition of the present invention is preferably 0.1 mg to 5 g / kg (body weight).

[0086] "Patient" refers to an animal, preferably a mammal, and more preferably a human. The term "mammal" refers to warm-blooded vertebrate mammals, including animals such as cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, rats, pigs, and humans.

[0087] "Treatment" refers to reducing, slowing the progression of, attenuating, preventing, or maintaining an existing disease or condition (such as cancer). Treatment also includes curing, preventing the development of, or reducing to some extent one or more symptoms of a disease or condition.

[0088] The compounds of the present invention can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention.

[0089] The structures of the compounds of this invention can be confirmed using conventional methods well known to those skilled in the art. If this invention relates to the absolute configuration of a compound, the absolute configuration can be confirmed using conventional techniques in the art. For example, single-crystal X-ray diffraction (SXRD) can be used. Diffraction intensity data of the grown single crystals can be collected using a Bruker D8 venture diffractometer with CuKα radiation as the light source and φ-scanning mode. After collecting the relevant data, the crystal structure can be further analyzed using the direct method (Shelxs 97) to confirm the absolute configuration.

[0090] The solvents used in this invention are commercially available. The following abbreviations are used in this invention: Pd / C represents palladium on carbon; H2 represents hydrogen; N2 represents nitrogen; mL represents milliliters; H2O represents water; AcOH represents acetic acid; B2Pin2 represents bis-pinacol borate; Pd(dppf)Cl2 represents 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride; KOAc represents potassium acetate; EtOAc represents ethyl acetate; K2CO3 represents potassium carbonate; MeCN represents acetonitrile; DIEA represents N,N-diisopropylethylamine; MeOH represents methanol; THF represents tetrahydrofuran; DMAP represents 4-dimethylaminopyridine; Pd(Amphos)2Cl2 represents dichloroditert-butyl-(4-dimethylaminophenyl)phosphine palladium(II); SO2Cl2 represents sulfone dichloride; NaHCO3 represents sodium bicarbonate; BrCN represents cyanogen bromide; NaO represents... H represents sodium hydroxide; TEA represents triethylamine; DMF represents N,N-dimethylformamide; H2O represents water; Na2SO4 represents sodium sulfate; FA represents formic acid; Cs2CO3 represents cesium carbonate; NaCl represents sodium chloride; CuI represents cuprous iodide; K3PO4 represents potassium phosphate; NaH represents sodium hydrogen; MeI represents iodomethane; DMSO represents dimethyl sulfoxide; POCl3 represents phosphorus oxychloride; Xphos represents 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl; BH3-Me2S represents borane methyl sulfide; DPPA represents diphenyl azidophosphate; XPhosPdG3 represents methanesulfonic acid (2-dicyclohexylphosphine-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II);

[0091] Compounds are named according to conventional naming principles in the field or using Software naming conventions are used; commercially available compounds use supplier catalog names.

[0092] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0093] This invention provides a compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof:

[0094]

[0095] in:

[0096] R1 and R2 are each independently selected from cyano, -CF3, -OCF3, -Cl, -Br, -F, hydroxyl, nitro, substituted or unsubstituted imidazolyl, substituted or unsubstituted pyrazolyl, straight-chain alkyl having 1 to 20 carbon atoms, alkoxy having 1 to 20 carbon atoms, branched alkyl having 3 to 20 carbon atoms, substituted or unsubstituted cyclic alkyl having 3 to 20 carbon atoms, substituted or unsubstituted cyclic alkoxy having 3 to 20 carbon atoms, substituted or unsubstituted heterocyclic having 3 to 20 ring atoms, -OR, -NH-R, or combinations thereof; each time R appears, it is independently selected from substituted or unsubstituted cyclic alkoxy having 3 to 20 carbon atoms, or substituted or unsubstituted heterocyclic having 3 to 20 ring atoms.

[0097] R3 and R4 are each independently selected from -H, -D, a straight-chain alkyl group having 1 to 20 carbon atoms, a branched or cyclic alkyl group having 3 to 20 carbon atoms, a substituted or unsubstituted aromatic group having 6 to 60 cyclic atoms, or a combination of these groups;

[0098] Ar1 is selected from any of the following structures:

[0099]

[0100] Each time X1 appears, it is independently selected from CR5 or N;

[0101] Each time Y1 appears, it is independently selected from NR6, CR6R7, C=O, O, S or S=O;

[0102] R5, R6, and R7 are each independently selected from -H, -D, straight-chain alkyl groups having 1 to 20 carbon atoms, alkoxy groups having 1 to 20 carbon atoms, branched alkyl groups having 3 to 20 carbon atoms, cyclic alkyl groups having 3 to 20 carbon atoms, branched alkoxy groups having 3 to 20 carbon atoms, cyclic alkoxy groups having 3 to 20 carbon atoms, or combinations of these systems.

[0103] In a specific example, the compound of formula (I) has the structure shown in formula (II):

[0104]

[0105] In a specific example, R3 and R4 are independently selected from -H, -D, substituted or unsubstituted aromatic groups having 6 to 30 ring atoms, respectively.

[0106] In a specific example, R3 and R4 are independently selected from -H, -D, substituted or unsubstituted aromatic groups having 6 to 15 ring atoms, respectively.

[0107] In a specific example, R3 and R4 are independently selected from -H, -D, substituted or unsubstituted phenyl groups, respectively.

[0108] In a specific example, the compound of formula (I) has the structure shown in formula (III):

[0109]

[0110] In a specific example, Ar1 is selected from any of the following structures:

[0111]

[0112] In a specific example, R5, R6, and R7 each appear independently selected from -H, -D, straight-chain alkyl with 1 to 10 carbon atoms, alkoxy with 1 to 10 carbon atoms, branched alkyl with 3 to 10 carbon atoms, cyclic alkyl with 3 to 10 carbon atoms, branched alkoxy with 3 to 10 carbon atoms, cyclic alkoxy with 3 to 10 carbon atoms, or combinations of these systems.

[0113] In a specific example, R5, R6, and R7 each time they appear, are independently selected from -H, -D, straight-chain alkyl with 1 to 5 carbon atoms, alkoxy with 1 to 5 carbon atoms, branched alkyl with 3 to 5 carbon atoms, cyclic alkyl with 3 to 5 carbon atoms, branched alkoxy with 3 to 5 carbon atoms, cyclic alkoxy with 3 to 5 carbon atoms, or combinations of these systems.

[0114] In a specific example, Ar1 is selected from any of the following structures:

[0115]

[0116] In a specific example, R1 and R2 are independently selected from cyano, -CF3, -OCF3, -Cl, -Br, -F, hydroxyl, nitro, unsubstituted imidazolyl, imidazolyl substituted with alkyl, Cl or F, unsubstituted pyrazolyl, pyrazolyl substituted with alkyl, Cl or F, unsubstituted heterocyclic group having 3 to 20 ring atoms, heterocyclic group having 3 to 20 ring atoms substituted with alkyl, hydroxyl, Cl, F or a combination thereof, -OR, -NH-R.

[0117] In a specific example, R1 and R2 are independently selected from cyano, -CF3, -OCF3, -Cl, -Br, -F, hydroxyl, nitro, unsubstituted imidazolyl, imidazolyl substituted with alkyl, Cl or F, unsubstituted pyrazolyl, pyrazolyl substituted with alkyl, Cl or F, unsubstituted heterocyclic group having 3 to 10 ring atoms, heterocyclic group having 3 to 10 ring atoms substituted with alkyl, hydroxyl, Cl, F or a combination thereof, -OR, and -NH-R.

[0118] In a specific example, R1 and R2 are independently selected from cyano, -CF3, -OCF3, -Cl, -Br, -F, hydroxyl, nitro, unsubstituted imidazolyl, imidazolyl substituted with alkyl, Cl or F, unsubstituted pyrazolyl, pyrazolyl substituted with alkyl, Cl or F, unsubstituted heterocyclic group having 3 to 5 ring atoms, heterocyclic group having 3 to 5 ring atoms substituted with alkyl, hydroxyl, Cl, F or a combination thereof, -OR, -NH-R.

[0119] More specifically, the heterocyclic group is a nitrogen-containing cycloalkyl group.

[0120] In a specific example, each time R appears, it is independently selected from substituted or unsubstituted cyclic alkoxy groups having 3 to 10 C atoms, or substituted or unsubstituted heterocyclic groups having 3 to 10 ring atoms.

[0121] In a specific example, each time R appears, it is independently selected from substituted or unsubstituted cyclic alkoxy groups having 3 to 5 C atoms, or substituted or unsubstituted heterocyclic groups having 3 to 5 ring atoms.

[0122] More specifically, the heterocyclic group is a nitrogen-containing cycloalkyl group.

[0123] In a specific example, R1 and R2 are independently selected from cyano, -CF3, -Cl, -Br, -F or any of the following structures:

[0124]

[0125] In a specific example, the compound of formula (I) is any one of the following compounds:

[0126]

[0127] In a specific example, a pharmaceutically acceptable salt is an alkyl salt. Further, a pharmaceutically acceptable salt is a formate.

[0128] The present invention also provides the use of the above-mentioned compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of a medicament for treating and / or preventing diseases associated with or mediated by CDK12 activity.

[0129] In a specific example, the disease associated with or mediated by CDK12 activity is cancer.

[0130] The present invention also provides a pharmaceutical composition comprising the above-described compound, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier.

[0131] It is understood that, within the scope of this invention, the above-described technical features of this invention and the technical features specifically described below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be described in detail here.

[0132] The tricyclic derivatives of the present invention and their preparation methods are further described in detail below with reference to specific embodiments. Unless otherwise specified, all raw materials used in the following embodiments are commercially available products.

[0133] Example 1

[0134] This embodiment provides compound 1, whose structural formula is as follows:

[0135]

[0136] The reaction route is shown below:

[0137]

[0138] The preparation process is as follows:

[0139] Step 1:

[0140] In a 25 mL three-necked flask, compound 1-1 (5.00 g, 26.7 mmol, 1.0 eq) and BrCN (3.87 g, 36.5 mmol, 2.69 mL, 1.37 eq) were added to a mixed solution of ethanol (25.0 mL) and water (25.0 mL), and stirred at 70 °C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove ethanol (25.0 mL). The product was then dissolved in NaOH (2 mol / L) solution and adjusted to pH 9.0, and filtered to remove soluble substances. The crude product, compound 1-2, was given. LCMS:MS(ESI) m / z = 213.9 [M+H]+. 1H NMR: DMSO-d6, 400MHz δ = 7.22 (d, J = 2.0Hz, 1H), 7.05-7.00 (m, 1H), 6.99-6.94 (m, 1H), 6.36 (s, 2H).

[0141] Step 2:

[0142] At room temperature, compounds 1-2 (3.00 g, 14.1 mmol, 1.00 eq), 1-3 (1.54 g, 14.1 mmol, 1.00 eq), and TEA (14.1 g, 14.1 mmol, 1.8 mL, 1.00 eq) were added to toluene (10.0 mL) and stirred at 100 °C for 2 hours. After the reaction was complete, 30.0 mL of water was added and stirred for 30 min. After the solids had completely precipitated, the mixture was filtered to obtain insoluble crude products 1-4 and 1-4a. LCMS:MS(ESI) m / z = 365.8 [M+H]+.

[0143] Step 3:

[0144] Compounds 1-4 and 1-4a (5.70 g, 15.6 mmol, 1.00 eq) were degassed with NaOH (1.25 g, 31.2 mmol, 2.00 eq) in MeCN (10.0 mL) at room temperature, with three displacements under a N2 atmosphere, followed by stirring at 70 °C under a N2 atmosphere for 1 hour. After the reaction was complete, 30.0 mL of water was added and stirred for 30 min. After the solids had completely precipitated, the mixture was filtered to obtain insoluble crude products, compounds 1-5 and 1-5a. LCMS:MS(ESI) m / z = 366.8 [M+H]+. 1 H NMR: DMSO-d6, 400MHz δ = 8.77 (t, J = 8.0Hz, 1H), 8.23 ​​(d, J = 1.6Hz, 1H), 7.88 (d, J = 8.0Hz, 1H), 7.72 (br d, J = 1.6Hz, 1H), 7.64 (br d, J=2.0Hz, 1H), 7.41 (brdd, J=1.6, 8.4Hz, 1H), 6.17-6.11 (m, 1H).

[0145] Step 4:

[0146] At room temperature, compounds 1-5 and 1-5a (4.00 g, 15.1 mmol, 1.00 eq), CH3I (2.58 g, 18.1 mmol, 1.13 mL, 1.20 eq), and K2CO3 (4.19 g, 30.2 mmol, 2.00 eq) were added to DMF (20.0 mL) and stirred at 95 °C for 12 hours. After dilution with 50.0 mL of H2O, the mixture was extracted three times with 45.0 mL of EtOAc (15.0 mL × 3), washed three times with NaCl (15.0 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. Purification was performed using preparative-high performance liquid chromatography (FA conditions) to obtain compounds 1-6 and 1-6a. LCMS:MS(ESI) m / z = 279.9 [M+H]+. 1 H NMR: DMSO-d6, 400MHz δ = 8.84 (t, J = 8.0Hz, 1H), 8.28 (d, J = 1.6Hz, 1H), 7.92 (d, J = 8.0Hz, 1H),7.82(d,J=1.6Hz,1H),7.57(d,J=8.0Hz,1H),7.50-7.41(m,1H),6.31-6.25(m,1H).

[0147] Step 5:

[0148] At room temperature, compounds 1-6 (100 mg, 359 μmol, 1.00 eq), 1-7 (154 mg, 719 μmol, 2.0 eq), Xphos (32.6 mg, 35.9 μmol, 0.10 eq), BrettPhosPd G3 (34.2 mg, 71.9 μmol, 0.20 eq), and Cs2CO3 (351 mg, 1.08 mmol, 3.00 eq) were added to DMSO (1.00 mL), and the mixture was stirred in a microwave at 90 °C for 12 hours. After the reaction was complete, 10.0 mL of water was added to the reaction mixture, and the mixture was extracted three times with 15.0 mL of EtOAc (5.00 mL × 3). The mixture was then washed three times with NaCl (5.00 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain crude compounds 1-8. LCMS: MS (ESI) m / z=412.1 [M+H]+. 1H NMR: DMSO-d6, 400MHz δ = 8.77 (d, J = 8.0Hz, 1H), 8.44-8.40 (m, 1H), 7.59 (d, J = 8.4H z,1H),7.29(d,J=8.4Hz,1H),7.08-7.05(m,1H),6.71(d,J=2.0Hz,1H),6.66-6.53 (m,1H),6.14-6.08(m,1H),5.42-5.31(m,1H),3.53-3.50(m,4H),3.19-3.11(m,2H ),2.36-2.31(m,1H),2.07-1.98(m,2H),1.87-1.75(m,3H),1.39(d,J=1.6Hz,9H).

[0149] Step 6:

[0150] Compounds 1-8 (400 mg, 969 μmol, 1.00 eq), benzyl isocyanate (258 mg, 1.94 mmol, 236 μL, 2.00 eq), and TEA (294 mg, 2.91 mmol, 404 μL, 3.00 eq) were added to THF (4.00 mL) at room temperature. The mixture was stirred at 25 °C for 2 hours. 10.0 mL of water was added to the reaction mixture, and the mixture was extracted three times with EtOAc (5.00 mL × 3), washed three times with NaCl (5.00 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product is compounds 1-9. LCMS: MS (ESI) m / z = 545.0 [M+H]+.

[0151] Step 7:

[0152] At room temperature, compounds 1-9 (500 mg, 918.03 μmol, 1.00 eq) and HCl / ethyl acetate (4.0 M, 229 μL, 1.00 eq) were added to ethyl acetate (5.00 mL), and the mixture was stirred at 25 °C for 1 hr. The crude product, the hydrochloride salt of compound 1-10, was concentrated under reduced pressure. LCMS:MS(ESI) m / z = 411.1 [M+H]+.

[0153] Step 8:

[0154] Compounds 1-10 (100 mg, 224 μmol, 1.00 eq), 1-11 (68.7 mg, 269 μmol, 1.20 eq), and DIEA (87.2 mg, 674 μmol, 117 μL, 3.00 eq) were added to DMF (0.50 mL) at room temperature and stirred at 25 °C for 10 hrs. The mixture was then filtered to remove insoluble substances. The formate salt of compound 1 was obtained by preparative high-performance liquid chromatography (pLC-HPLC) (column: Phenomenexluna C18 150 × 25.0 mm × 10.0 μm; mobile phase: [water(FA)-ACN]; B%: 39.0%-69.0%, 9.0 min). LC-MS (ESI) m / z = 663.0 [M+H]+. 1 H NMR: DMSO-d6,400MHzδ=8.93-8.87(m,1H),8.32-8.26(m,1H),8.05-7.92(m ,2H),7.47-7.39(m,1H),7.33-7.22(m,3H),7.20-7.08(m,4H),6.30-6.26(m ,1H),5.72-5.50(m,2H),4.90-4.83(m,1H),4.72-4.65(m,1H),4.56-4.50(m ,2H),4.37-4.26(m,1H),4.20-4.13(m,2H),3.57(s,3H),1.58-0.96(m,8H).

[0155] Example 2

[0156] This embodiment provides compound 2, whose structural formula is as follows:

[0157]

[0158] The reaction route is shown below:

[0159]

[0160] The preparation process is as follows:

[0161] Step 1:

[0162] In a 25 mL three-necked flask, compound 2-1 (10.0 g, 53.1 mmol, 1.00 eq) and BrCN (7.79 g, 73.5 mmol, 5.41 mL, 1.38 eq) were added to a mixed solution of ethanol (25.0 mL) and water (25.0 mL), and stirred at 70 °C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove ethanol. The product was then dissolved in NaOH (2.0 mol / L) solution and adjusted to pH 9.0, and filtered to remove soluble substances. The crude product was compound 2-2. LCMS:MS(ESI) m / z = 215.0 [M+H]+. 1 HNMR: DMSO-d6, 400MHz δ = 7.29 (d, J = 8.0Hz, 1H), 6.97-6.90 (m, 1H), 6.86 (br s, 2H), 3.30 (s, 1H).

[0163] Step 2:

[0164] Compounds 2-2 (8.00 g, 37.5 mmol, 1.00 eq), 1-3 (8.17 g, 37.5 mmol, 1.00 eq), and TEA (3.80 g, 37.5 mmol, 5.23 mL, 1.00 eq) were added to 80.0 mL of DMSO and stirred at 120 °C for 4 hours at room temperature. After the reaction was complete, 30.0 mL of water was added and the mixture was stirred for 30.0 min. After the solids had completely precipitated, the mixture was filtered to obtain insoluble crude products 2-4 and 2-4a. LCMS:MS(ESI) m / z = 267.0 [M+H]+.

[0165] Step 3:

[0166] At room temperature, compounds 2-4 and 2-4a (19.0 g, 71.6 mmol, 1.00 eq), CH3I (10.1 g, 71.6 mmol, 4.46 mL, 1.00 eq), and K2CO3 (9.91 g, 71.6 mmol, 1.00 eq) were added to 190 mL of DMF and stirred at 95 °C for 12 hours. After dilution with 200 mL of H2O, the mixture was extracted three times with EtOAc (200 mL × 3), washed three times with NaCl (200 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. Purification was performed using preparative-high performance liquid chromatography (FA conditions) to obtain compounds 2-5 and 2-5a. LCMS:MS(ESI) m / z = 280.9 [M+H]+.

[0167] Step 4:

[0168] At room temperature, compounds 2-5 (100 mg, 358 μmol, 1.00 eq), compounds 1-7 (153 mg, 716 μmol, 2.00 eq), CuI (68.2 mg, 358 μmol, 1.00 eq), and K3PO4 (228 mg, 1.07 mmol, 3.00 eq) were added to DMSO (1.00 mL), and the mixture was stirred at 90 °C for 3 hours. After the reaction was complete, 10.0 mL of water was added to the reaction mixture, and the mixture was extracted three times with EtOAc1 (5.00 mL × 3), washed three times with NaCl (5.00 mL × 3), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. Compounds 2-6 were purified by preparative high-performance liquid chromatography (FA conditions: column: Phenomenexluna C18 150×25.0mm×10.0μm; mobile phase: [water (FA)-ACN]; B%: 17.0%-47.0%, 9.0min). LCMS: MS (ESI) m / z = 413.0 [M+H]+. 1 H NMR: DMSO-d6, 400MHz δ = 8.68-8.63 (m, 1H), 7.89 (d, J = 8.8Hz, 1H), 6.82-6.75 (m, 1H), 6.47-6.40 (m, 1H), 6.36-6.31 (m, 1H), 6. 21-6.14(m,1H),3.73-3.63(m,1H),3.54(s,3H),3.24-3.14(m,1H),2.08-1.95(m,2H),1.88-1.73(m,4H),1.42-1.32(m,15H).

[0169] Step 5:

[0170] At room temperature, a solution of compound 2-6 (100 mg, 242 μmol, 1.00 eq) and DIEA (94.0 mg, 727 μmol, 126 μL, 3.00 eq) was added to THF (1.00 mL), followed by bis(trichloromethyl) carbonate (359 mg, 1.21 mmol, 5.00 eq). The mixture was stirred at 60 °C for 1 hour. Then, compound 2-7 (51.9 mg, 484 μmol, 52.8 μL, 2.00 eq) and DMAP (29.6 mg, 242 μmol, 1.00 eq) were added. After the reaction was complete, 10.0 mL of water was added to the reaction mixture, and the mixture was extracted three times with EtOAc (5.00 mL × 3), washed three times with NaCl (5.00 mL × 3), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure to obtain the crude product, compound 2-8. LCMS: MS (ESI) m / z=546.2[M+H]+.

[0171] Step 6:

[0172] Compound 2-8 (30.0 mg, 55.0 μmol, 1.00 eq) and HCl / ethyl acetate (4.00 M, 12.5 μL, 1.00 eq) were added to ethyl acetate (5.00 mL) at room temperature and stirred at 25 °C for 0.5 hr. The crude product, compound 2-9 hydrochloride, was obtained by concentration under reduced pressure. LCMS:MS(ESI) m / z = 446.1 [M+H]+.

[0173] Step 7:

[0174] Compounds 2-9 (30.0 mg, 67.4 μmol, 1.00 eq), 1-11 (20.6 mg, 80.9 μmol, 1.20 eq), and DIEA (26.1 mg, 202 μmol, 35.2 μL, 3.00 eq) were added to DMF (0.50 mL) at room temperature and stirred at 25 °C for 10 hrs. The mixture was then filtered to remove insoluble substances. Compound 2 was purified by preparative high performance liquid chromatography (p-HPLC) (column: Phenomenexluna C18 150 × 25.0 mm × 10 μm; mobile phase: [water (FA)-ACN]; B%: 36.0%-66.0%, 9.0 min). LCMS: MS (ESI) m / z = 663.9 [M+H]+. 1 H NMR: DMSO-d6, 400MHzδ=9.23-8.76(m,2H),8.42-8.36(m,1H),8.33-8.26(m,1H),7.34-7.19(m,7H),6.37(d, J=8.0Hz,1H),5.85-5.52(m,2H),4.89-4.84(m,1H),4.76-4.70(m,1H),4.63(dd,J=4.8,8.0Hz,2H),4.55(br d,J=6.4Hz,1H),4.18(br d,J=4.8Hz,2H),3.60(s,3H),1.92-1.83(m,4H),1.43-1.36(m,4H).

[0175] Example 3

[0176] This embodiment provides compound 3, whose structural formula is as follows:

[0177]

[0178] The reaction route is shown below:

[0179]

[0180] The preparation process is as follows:

[0181] Step 1:

[0182] Compound 3-1 (6.00 g, 23.9 mmol, 1.00 eq) was dissolved in tetrahydrofuran (50.0 mL) at 0 °C. Methanol (4.75 g, 148 mmol, 6.00 mL, 6.19 eq) and potassium tert-butoxide (4.03 g, 35.9 mmol, 1.5 eq) were added, and the mixture was heated to 20 °C and reacted for 3 hours. The reaction was quenched by adding ammonium chloride (50.0 mL) at 25 °C, followed by extraction three times with ethyl acetate (100 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (150 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 3-2. LCMS:MS(ESI) m / z = 246.0 [M+H]+. 1 HNMR: (400MHz, CDCl3) δ7.75 (d, J = 8.8 Hz, 1H), 6.73 (d, J = 8.8 Hz, 1H), 3.97 (s, 3H), 3.94 (s, 3H).

[0183] Step 2:

[0184] Compound 3-2 (4.00 g, 16.2 mmol, 1.00 eq), Pin2B2 (8.26 g, 32.5 mmol, 2.00 eq), KOAc (3.19 g, 32.5 mmol, 2.00 eq), and Pd(dppf)Cl2 (356 mg, 487 μmol, 0.03 eq) were dissolved in dioxane (40.0 mL) at room temperature, and the mixture was purged with nitrogen three times. The reaction was carried out at 90 °C under a nitrogen atmosphere for 3 hours. The mixture was extracted three times with ethyl acetate (100 mL). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 3-3. LCMS:MS(ESI) m / z = 293.9 [M+H]+.

[0185] Step 3:

[0186] Compounds 3-3 (4.77 g, 16.2 mmol, 1.00 eq), 3-4 (7.34 g, 24.4 mmol, 1.50 eq), Cs₂CO₃ (10.6 g, 32.5 mmol, 2.00 eq), and Pd(dppf)Cl₂ (398 mg, 488 μmol, 0.0300 eq) were dissolved in dioxane (40.0 mL) and water (4.00 mL) at room temperature, purged three times with nitrogen. The reaction was carried out at 90 °C under a nitrogen atmosphere for 3 hours. The mixture was extracted three times with ethyl acetate (100 mL). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 1 / 0 to 5 / 1) to obtain compound 3-5. LCMS: MS (ESI) m / z=339.9[M+H]+. 1 HNMR: (400MHz, CDCl3) δ7.56 (d, J = 8.4Hz, 1H), 7.36-7.33 (m, 1H), 7.30 (dd, J = 1.6 ,9.2Hz,1H),7.13(t,J=8.0Hz,1H),6.97-6.92(m,1H),4.03(s,3H),3.78(s,3H).

[0187] Step 4:

[0188] Compounds 3-5 (2.70 g, 7.94 mmol, 1.00 eq) and potassium hydroxide (2.23 g, 39.7 mmol, 5.00 eq) were added to water (25.0 mL) at 25 °C and stirred at 100 °C for 3 hours. The aqueous layer was then acidified to pH 1.0 with 1 M hydrochloric acid solution. The reaction mixture was extracted three times with ethyl acetate (20.0 mL). The combined organic phases were washed three times with saturated sodium chloride aqueous solution (50.0 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compounds 3-6. LCMS: m / z = 325.8(M+H)+. 1 HNMR: (400MHz, CDCl3) δ7.63 (d, J = 8.4Hz, 1H), 7.35 (dd, J = 1.6, 8.4Hz, 1H), 7.31 (dd, J = 1.6, 9.2Hz, 1H), 7.12-7.08 (m, 2H), 4.05 (s, 3H).

[0189] Step 5:

[0190] Compound 3-6 (1.57 g, 4.81 mmol, 1.00 eq) was dissolved in THF (15.0 mL) at 0 °C, and BH3-Me2S (10 M, 1.20 mL, 2.50 eq) was slowly added dropwise. The mixture was stirred at 50 °C for 6 hours. The reaction mixture was then slowly added to water (30 mL × 3), and extracted three times with ethyl acetate (30.0 mL). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (50.0 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 3-7. LCMS: m / z = 311.9 (M + H) +.

[0191] Step 6:

[0192] Compound 3-7 (1.28 g, 4.10 mmol, 1.00 eq) was dissolved in THF (10.0 mL) at 0 °C, and NaH (410 mg, 10.2 mmol, 60% purity, 2.50 eq) was slowly added. The mixture was stirred at 50 °C for 12 hours. At 0 °C, the reaction mixture was slowly added to ammonium chloride (5.00 mL), and extracted three times with ethyl acetate (20.0 mL). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (30.0 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by prep-HPLC (FA condition) to obtain compound 3-8. LCMS: m / z = 291.9 (M+H)+. 1 HNMR: (400MHz, CDCl3) δ7.83 (d, J=8.4Hz, 1H), 7.42-7.38 (m, 1H), 7.18-7.15 (m, 2H), 6.75 (d, J=8.4Hz, 1H), 5.15 (s, 2H), 3.95 (s, 3H).

[0193] Step 7:

[0194] Compounds 3-8 (100 mg × 2, 342 μmol, 1.00 eq), compounds 1-7 (110 mg, 513 μmol, 1.50 eq), RuPhosPdG3 (28.6 mg, 34.2 μmol, 0.100 eq), and Cs2CO3 (334 mg, 1.03 mmol, 3.00 eq) were dissolved in DMSO (1.00 mL) at room temperature, substituted with N2 three times, and reacted at 100 °C under a nitrogen atmosphere for 12 hours. The reaction mixture was diluted with water (5.00 mL) and extracted three times with ethyl acetate (10.0 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (15.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by prep-HPLC (FA condition) to obtain compounds 3-9. LCMS: m / z = 426.1 (M+H)+. 1 HNMR: (400MHz, CDCl3) δ7.72(d,J=8.4Hz,1H),7.34(d,J=8.4Hz,1H),6.69(d,J=8.4Hz,1H),6.44-6.11(m,2H),5.09(s,2H),4.40(br s,1H),3.93(s,3H),3.48(br s,1H),3.23(br s,1H),2.23-2.13(m,2H),2.13-2.04(m,2H),1.46(s,9H),1.32-1.21(m,4H).

[0195] Step 8:

[0196] Compound 3-9 (110 mg, 258 μmol, 1.00 eq), BnNCO (68.8 mg, 517 μmol, 63.1 μL, 2.00 eq), and DMAP (3.16 mg, 25.8 μmol, 0.100 eq) were dissolved in tetrahydrofuran (0.50 mL) at room temperature, substituted with N2 three times, and reacted at 50 °C under a nitrogen atmosphere for 12 hours. The reaction system was concentrated to remove the solvent, yielding compound 3-10. LCMS: m / z = 559.0(M+H)+.

[0197] Step 9:

[0198] Compound 3-10 (114 mg, 204 μmol, 1.00 eq) was dissolved in HCl / EtOAc (2.00 mL) at room temperature and reacted at 20 °C for 2 hours. The reaction mixture was concentrated to remove the solvent, yielding compound 3-11. LCMS: m / z = 459.0 (M+H)+.

[0199] Step 10:

[0200] Compound 3-11 (93.0 mg, 202 μmol, 1.00 eq) and compound 1-11 (51.6 mg, 202 μmol, 1.00 eq) were dissolved in DMF (1.00 mL) at room temperature, followed by the addition of DIEA (78.6 mg, 608 μmol, 105 μL, 3.00 eq). The reaction mixture was then reacted at 20 °C for 12 hours. The reaction mixture was diluted with water (3.00 mL) and extracted three times with ethyl acetate (5.00 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (8.00 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by prep-HPLC (FA condition) to obtain the formate of compound 3. LCMS: m / z = 676.9 [M+H]+. 1 HNMR: (400MHz, DMSO-d6) δ8.30 (d, J=6.0Hz, 1H), 8.23 ​​(dd, J=6.0, 8.8Hz, 1H), 7.9 8-7.72(m,2H),7.31-7.25(m,2H),7.21-7.15(m,3H),6.90(s,1H),6.79(dd,J=2.0 ,11.2Hz,1H),6.03-5.95(m,1H),5.57(quin,J=5.7Hz,1H),5.17(s,2H),4.85(t,J =7.2Hz,1H),4.74(t,J=7.2Hz,1H),4.60-4.50(m,2H),4.30-4.20(m,1H),4.16(br d,J=5.6Hz,2H),3.89(s,3H),1.94-1.79(m,4H),1.43-1.33(m,2H),1.27-1.16(m,2H).

[0201] Example 4

[0202] This embodiment provides compound 4, whose structural formula is as follows:

[0203]

[0204] The reaction route is shown below:

[0205]

[0206] The preparation process is as follows:

[0207] Step 1:

[0208] Compound 4-1 (6.00 g, 23.9 mmol, 1.00 eq) and potassium hydroxide (6.21 g, 110 mmol, 5.00 eq) were dissolved in H₂O (60.0 mL) at room temperature and reacted at 100 °C for 3 hours. The reaction system was adjusted to pH 1 with hydrochloric acid and then extracted three times with dichloromethane (100 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (150 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 4-2. LCMS:MS(ESI) m / z = 256.9 [M+H]+. 1 HNMR: (400MHz, CDCl3) δ8.11 (s, 1H), 8.06 (d, J = 0.8Hz, 1H), 7.78 (d, J = 8.4Hz, 1H), 7.55 (dd, J = 1.6, 8.4Hz, 1H).

[0209] Step 2:

[0210] Compound 4-2 (5.60 g, 21.7 mmol, 1.00 eq), TEA (2.42 g, 23.9 mmol, 3.33 mL, 1.10 eq), and DPPA (6.59 g, 23.9 mmol, 5.19 mL, 1.10 eq) were dissolved in tert-butanol (60.0 mL) at room temperature. The mixture was reacted at 20 °C for 0.5 hours, then heated to 80 °C and reacted for 8 hours. The mixture was diluted with NaHCO3 (aq, 40.0 mL) and extracted three times with ethyl acetate (80.0 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1) to give compound 4-3. LCMS:MS(ESI) m / z = 271.9 [M+H]+.

[0211] Step 3:

[0212] DMF (1.34 g, 18.2 mmol, 1.41 mL, 2.00 eq) was dissolved in tetrahydrofuran (15.0 mL) at 0 °C, and POCl3 (4.20 g, 27.4 mmol, 2.55 mL, 3.00 eq) was added dropwise. The reaction was carried out at 0 °C for 0.5 h. Then, compound 4-3 (3.00 g, 9.14 mmol, 1.00 eq) was dissolved in tetrahydrofuran (15.0 mL) and added dropwise to the reaction system. The reaction was carried out at 20 °C for 1.5 h. The reaction system was quenched with sodium hydroxide (2.0 M, 10.0 mL) and extracted three times with ethyl acetate (30.0 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (40.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 4-4. LCMS:MS(ESI) m / z = 299.8 [M+H]+.1 HNMR: (400MHz, CDCl3) δ11.11-10.99(m,1H),10.11(s,1H),7.78(d,J=1.6Hz,1H),7.68(d,J=8.8Hz,1H),7.45(dd,J=1.6,8.4Hz,1H),1.51(s,9H).

[0213] Step 4:

[0214] Compound 4-5 (7.55 g, 33.7 mmol, 6.68 mL, 4.00 eq) was added to tetrahydrofuran (18.0 mL) at 0 °C, followed by the slow addition of NaH (1.35 g, 33.7 mmol, 60% purity, 4.0 eq). The reaction was carried out at 0 °C for 0.5 h. Then, compound 4-4 (3.00 g, 8.42 mmol, 1.00 eq) was dissolved in tetrahydrofuran (18.0 mL) and added dropwise to the reaction system. The reaction was carried out at 60 °C for 12 h. At room temperature, the reaction system was quenched with ammonium chloride solution (40.0 mL) and extracted three times with ethyl acetate (100 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (150 × 3 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 4-6. LCMS: m / z = 327.8 (M+H)+.

[0215] Step 5:

[0216] Compound 4-6 (3.56 g, 8.35 mmol, 1.00 eq) was dissolved in HCl / MeOH (30.0 mL) at room temperature and stirred at 20 °C for 4 hours. The pH was adjusted to 9 with sodium bicarbonate solution, and the solution was filtered to give compound 4-7. LCMS: m / z = 281.8(M+H)+ 1 HNMR: (400MHz, DMSO-d6)δ12.19-11.71(m,1H),8.46(br d,J=8.8Hz,1H),8.26(d,J=1.6Hz,1H),8.09(d,J=8.4Hz,1H),7.63(dd,J=1.6,8.4Hz,1H),6.68(br d,J=8.8Hz,1H).

[0217] Step 6:

[0218] Compound 4-7 (1.30 g, 4.64 mmol, 1.00 eq) was dissolved in DMF (15.0 mL) at 0 °C, and MeI (988 mg, 6.96 mmol, 433 μL, 1.50 eq) and K₂CO₃ (1.28 g, 9.28 mmol, 2.00 eq) were added. The mixture was stirred at 50 °C for 3 hours. The reaction mixture was diluted with water (20.0 mL) and extracted three times with ethyl acetate (40.0 × 3 mL). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (30.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by column chromatography (SiO₂, petroleum ether / ethyl acetate = 2 / 1 to 0 / 1) to obtain compound 4-8. LCMS: m / z = 293.8 (M+H)⁺. 1 HNMR: (400MHz, CDCl3) δ8.34(d,J=1.6Hz,1H),8.30(d,J=9.6Hz,1H),8.03(d, J=8.4Hz, 1H), 7.64 (dd, J=1.6, 8.4Hz, 1H), 6.53 (d, J=9.6Hz, 1H), 3.62 (s, 3H).

[0219] Step 7:

[0220] Compounds 4-8 (200 mg, 2.67 μmol, 1.00 eq), compounds 1-7 (218 mg, 1.02 mmol, 1.50 eq), XPhosPdG3 (57.5 mg, 67.9 μmol, 0.100 eq), and Cs2CO3 (664 mg, 2.04 mmol, 3.00 eq) were dissolved in DMSO (2.00 mL) at room temperature, purged three times with N2, and reacted at 100 °C under a nitrogen atmosphere for 12 hours. The reaction mixture was diluted with water (10.0 mL) and extracted three times with ethyl acetate (15.0 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (20.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by prep-HPLC (FA condition) to obtain compounds 4-9. LCMS: m / z = 428.0 (M+H)+.

[0221] Step 8:

[0222] Compound 4-9 (60.0 mg, 140 μmol, 1.00 eq), BnNCO (37.3 mg, 280 μmol, 34.2 μL, 2.00 eq), and DMAP (1.71 mg, 14.0 μmol, 0.100 eq) were dissolved in tetrahydrofuran (1.00 mL) at room temperature and reacted at 50 °C for 12 hours. The reaction mixture was concentrated to remove the solvent, yielding compound 4-10. LCMS: m / z = 561.0 (M+H)+.

[0223] Step 9:

[0224] Compound 4-10 (78.0 mg, 139 μmol, 1.00 eq) was dissolved in HCl / EtOAc (2.00 mL) at room temperature and reacted at 20 °C for 2 hours. The reaction mixture was concentrated to remove the solvent, yielding compound 4-11. LCMS: m / z = 460.9(M+H)+.

[0225] Step 10:

[0226] Compound 4-11 (64.0 mg, 138 μmol, 1.00 eq) and compound 1-11 (35.3 mg, 138 μmol, 1.00 eq) were dissolved in DMF (1.00 mL) at room temperature, followed by the addition of DIEA (53.8 mg, 416 μmol, 72.6 μL, 3.00 eq). The reaction mixture was then reacted at 20 °C for 12 hours. The reaction mixture was diluted with water (3.00 mL) and extracted three times with ethyl acetate (5.00 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (8.00 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by prep-HPLC (FA condition) to obtain the formate of compound 4. LCMS: m / z = 678.9 [M+H]+. 1HNMR: (400MHz, DMSO) δ8.35 (dd, J=2.4, 9.6Hz, 1H), 8.31-8.26 (m, 1H), 8.16 (dd, J=8.4,10.8Hz,1H),8.00-7.68(m,2H),7.33-7.25(m,3H),7.20-7.14(m,3H),6. 54(d,J=9.4Hz,1H),5.95(td,J=6.0,16.4Hz,1H),5.66-5.45(m,1H),4.85(t,J= 6.8Hz,1H),4.69(t,J=7.2Hz,1H),4.62-4.47(m,2H),4.37-4.24(m,1H),4.14(br d,J=6.0Hz,2H),3.64(s,3H),3.48(br s,1H),1.93-1.79(m,4H),1.49-1.32(m,2H),1.24-1.08(m,2H).

[0227] Example 5

[0228] This embodiment provides compound 5, whose structural formula is as follows:

[0229]

[0230] The reaction route is shown below:

[0231]

[0232] The preparation process is as follows:

[0233] Step 1:

[0234] Compound 5-1 (1.00 g, 4.61 mmol, 1.00 eq), DIEA (1.19 g, 9.22 mmol, 1.61 mL, 2.00 eq), and compound 5-2 (404 mg, 5.53 mmol, 1.20 eq) were added to 10.0 mL of THF at room temperature, and the mixture was stirred at 0 °C for 7 hours. The reaction mixture was diluted with 50.0 mL of H₂O, extracted three times with EtOAc (15.0 mL × 3), washed three times with saturated sodium chloride solution (15.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. Compound 5-3 was purified by preparative-high performance liquid chromatography (FA conditions). LCMS:MS(ESI) m / z = 253.9 [M+H]⁺. 1H NMR: CDCl3, 400MHz δ = 8.40-8.31 (m, 1H), 5.95-5.79 (m, 1H), 5.32-5.21 (m, 1H), 5.04 (t, J = 7.2Hz, 2H), 4.59 (t, J = 6.4Hz, 2H).

[0235] Step 2:

[0236] Compounds 1-10 (100 mg, 224 μmol, 1.00 eq), 5-3 (68.4 mg, 269 μmol, 1.20 eq), and DIEA (87.2 mg, 674 μmol, 117 μL, 3.00 eq) were added to 1.00 mL of DMF at room temperature and stirred at 25 °C for 10 hours. The reaction solution was filtered to remove insoluble matter, and the mixture was purified by preparative high performance liquid chromatography (FA conditions: column: Phenomenexluna C18 150 × 25 mm × 10 μm; mobile phase: [water (FA)-ACN]; B%: 15%-45%, 9 min). The obtained 5-10 compound was LCMS (ESI) m / z = 622.3 [M+H]+. 1 H NMR: DMSO-d6, 400MHz δ = 8.93-8.86 (m, 1H), 8.50-8.21 (m, 1H), 8.01 (q, J = 8.0Hz, 2H), 7.48-7.41 (m, 1H), 7.29-7.0 7(m,9H),6.31-6.25(m,1H),5.77-5.62(m,1H),4.78-4.64(m,1H),4.59-4.47(m,2H),4.39-4.26(m,1H),4.14(br d,J=6.3Hz,2H),3.57(s,3H),1.91-1.82(m,4H),1.46-1.32(m,2H),1.15-1.06(m,2H).

[0237] Example 6

[0238] This embodiment provides compound 6, whose structural formula is as follows:

[0239]

[0240] The reaction route is shown below:

[0241]

[0242] The preparation process is as follows:

[0243] Step 1:

[0244] Compound 6-1 (1.00 g, 5.39 mmol, 1.00 eq), compound 6-2 (3.00 g, 10.7 mmol, 2.00 eq), Pd(Amphos)₂Cl₂ (114 mg, 161 μmol, 114 μL, 0.03 eq), and K₂CO₃ (1.49 g, 10.7 mmol, 2.00 eq) were dissolved in dioxane (10.0 mL) and water (2.00 mL) at room temperature. The mixture was substituted with N₂ three times and reacted at 90.0 °C under a nitrogen atmosphere for 12 hours. The reaction mixture was then diluted with water (10.0 mL) and extracted three times with ethyl acetate (50.0 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (80.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 6-3. LCMS: m / z = 218.0 (M + H)⁺. 1 H NMR: (400MHz, CDCl3) δ 8.80 (s, 1H), 7.70 (d, J = 2.0Hz, 1H), 7.22 (d, J = 2.0Hz, 1H), 6.22 (dd, J = 2.4, 9.2Hz, 1H), 2.66 (s, 3H).

[0245] Step 2:

[0246] Compound 6-3 (300 mg, 1.38 mmol, 1.00 eq) was dissolved in acetonitrile (5.00 mL) at 0 °C. SO₂Cl₂ (559 mg, 4.14 mmol, 414 μL, 3.00 eq) was added dropwise to the system, and the reaction was carried out at 20 °C for 12 hours. The reaction mixture was diluted with water (5.00 mL) and extracted three times with ethyl acetate (10.0 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (15.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 6-4. LCMS: m / z = 240.0 (M + H) + 1 HNMR: (400MHz, CDCl3) δ8.97 (s, 1H), 7.80 (s, 1H).

[0247] Step 3:

[0248] Compounds 1-10 (10.0 mg, 22.5 μmol, 1.00 eq) and 6-4 (5.40 mg, 22.5 μmol, 1.00 eq) were dissolved in DMF (1.00 mL) at room temperature, followed by the addition of K2CO3 (9.33 mg, 67.5 μmol, 3.00 eq). The reaction mixture was then reacted at 60 °C for 12 hours. The reaction mixture was diluted with water (3.00 mL) and extracted three times with ethyl acetate (5.00 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (8.00 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by prep-HPLC (FA condition) to obtain compound 6. LCMS: m / z = 648.3 [M+H]+. 1 H NMR: (400MHz, DMSO) δ13.84-13.62(m,1H),8.96-8.84(m,1H),8.68-8.60(m,1H),8.32-8.19(m,1H),8.12-8.06(m,1H),8.01(d,J=8.4Hz,1H), 7.48-7.41(m,1H),7.33-7.22(m,2H),7.20-7.14(m,3H),7.12-7.07(m ,1H),6.40-6.21(m,1H),5.79-5.63(m,1H),4.47-4.24(m,1H),4.14(br d,J=5.6Hz,2H),3.56(s,3H),2.02-1.95(m,2H),1.89(br d,J=9.2Hz,2H),1.51-1.40(m,2H),1.23(br s,1H),1.16-1.04(m,2H).

[0249] Example 7

[0250] This embodiment provides compound 7, whose structural formula is as follows:

[0251]

[0252] The reaction route is shown below:

[0253]

[0254] The preparation process is as follows:

[0255] Step 1:

[0256] At room temperature, compounds 7-1 (500 mg, 2.69 mmol, 1.00 eq), 7-2 (1.24 g, 4.04 mmol, 1.50 eq), and sodium carbonate (2.0 M, 2.69 mL, 2.00 eq) were dissolved in dimethyl ether (10.0 mL). Under N2 protection, dichlorobis[di-tert-butyl-(4-dimethylaminophenyl)phosphine]palladium(II) (190 mg, 269 μmol, 190 μL, 0.10 eq) was added, and the mixture was stirred at 80 °C for 10 hours. The mixture was poured into water (10 mL) and extracted three times with ethyl acetate (10.0 mL x 3). The organic phases were combined, washed twice with saturated brine (10.0 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by reversed-phase high-performance liquid chromatography (0.1%, formic acid conditions) to obtain compound 7-3. LCMS: MS (ESI) m / z=231.7[M+H]+.

[0257] Step 2:

[0258] Compound 7-3 (20.0 mg, 86.5 μmol, 1.00 eq) was dissolved in acetonitrile (2.00 mL), and the air was purged three times with nitrogen. Sulfonyl chloride (46.7 mg, 346 μmol, 34.6 μL, 4.00 eq) was slowly added under nitrogen. The reaction was carried out at 25 °C for 10 hours. The mixture was concentrated to obtain the crude product. The crude product was purified by reversed-phase high-performance liquid chromatography (0.1%, formic acid conditions) to obtain compound 7-4. LCMS:MS(ESI) m / z = 219.7 [M+H]+. 1 HNMR: CDCl3, 400MHz δ9.15(s,1H),8.41(s,1H),2.55(s,3H).

[0259] Step 3:

[0260] Compounds 1-10 (30.0 mg, 67.5 μmol, 1.00 eq) and 7-4 (14.8 mg, 67.5 μmol, 1.00 eq) were dissolved in N,N-dimethylformamide (0.50 mL). Under nitrogen atmosphere, N,N-diisopropylethylamine (26.2 mg, 202 μmol, 35.3 μL, 3.00 eq) was added to the reaction solution, and the reaction was carried out at 20 °C for 12 hours. The reaction solution was slowly poured into water (5.00 mL), and extracted three times with ethyl acetate (3.00 mL x 3). The organic phases were combined, washed once with saturated brine (2.00 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the product. The crude product was purified by reversed-phase high-performance liquid chromatography (0.1%, formic acid conditions) to obtain compound 7. LCMS:MS(ESI) m / z = 628.3 [M+H]+. 1H NMR: DMSO-d6, 400MHz δ12.91-13.26(m,1H),8.84-8.89(m,1H),8.52-8.55(m,1H),7.95-8.08(m,2H),7.38(s,1H),7. 23-7.30(m,2H),7.15–7.17(m,3H),7.07-7.10(m,1H),6.26(d,J=8.0Hz,1H),5.70-5.83(m,1H),4.33(s,1H),4.16(br d,J=5.2Hz,2H),3.54(s,3H),2.54(s,3H),2.33(s,2H),1.88(s,4H),1.44(d,J=12.0Hz,2H),1.23(s,1H),1.00-1.18(m,2H).

[0261] Example 8

[0262] This embodiment provides compound 8, whose structural formula is as follows:

[0263]

[0264] The reaction route is shown below:

[0265]

[0266] The preparation process is as follows:

[0267] Step 1:

[0268] Compound 8-1 (500 mg, 2.30 mmol, 1.00 eq) and compound 8-2 (242 mg, 2.30 mmol, 1.00 eq) were dissolved in tetrahydrofuran (10.0 mL) and acetonitrile (10.0 mL) at room temperature and reacted at 25 °C for 10 hours. The reaction was concentrated under reduced pressure to obtain the crude product, which was purified by prep-HPLC (FA condition) to obtain compound 8-3. LCMS:MS (ESI) m / z = 285.9 [M+H]+.

[0269] Step 2:

[0270] Compound 8-3 (12.9 mg, 45.2 μmol, 1.00 eq) and compound 1-10 (20.0 mg, 45.1 μmol, 1.00 eq) were dissolved in DMF (1.00 mL) at room temperature, followed by the addition of DIEA (17.5 mg, 135 μmol, 23.6 μL, 3.00 eq). The reaction mixture was then reacted at 25 °C for 12 hours. The reaction mixture was diluted with water (3.00 mL) and extracted three times with ethyl acetate (5.00 mL × 3). The organic phases were combined, washed three times with saturated sodium chloride aqueous solution (8.00 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by prep-HPLC (FA condition) to obtain compound 8. LCMS: m / z = 694.3 [M+H]+. 1 HNMR: (400MHz, DMSO-d6) δ9.91(s,1H),7.95(s,1H),7.01(s,1H),6.02(s,1H),4.99(br s,1H),4.37(s,2H),3.62-3.53(m,1H),2.89(s,3H),2.73(s,3H),2.43-2.32(m,1H),1.81-1.61(m,6H),1.51(s,9H),1.03(br d,J=6.4Hz,6H).

[0271] In vitro activity test

[0272] Experimental Example 1: In vitro CDK2 / Cyclin E1 enzyme activity assay

[0273] Experimental materials:

[0274] CDK2 / Cyclin E1 was purchased from Syngenes. MLight-4E-BP1 peptide, Eu-anti-phospho-tyrosine antibody, and 1X assay buffer were purchased from PerkinElmer. High-purity ATP was purchased from Promega. EDTA was purchased from Sigma. A Nivo multilabel analyzer (PerkinElmer) was used.

[0275] Experimental methods:

[0276] Kinase buffer preparation: The kinase buffer contains 50mM HEPES, 1mM EDTA, 10mM MgCl2, 0.01% Brij-35, pH 7.4. Add 2.38g HEPES, 58mg EDTA, 406mg MgCl2, and 20mg Brij-35 to 200ml of buffer and adjust the pH to 7.4.

[0277] Preparation of the stop solution:

[0278] The stop solution was prepared by mixing 100 μL of 1M EDTA stock solution with 0.625 μL of 1X detection buffer and 1725 μL of distilled water. The enzyme, MLight-4E-BP1 peptide, ATP, and inhibitor were diluted using kinase buffer. The Eu-anti-phospho-tyrosine antibody was diluted to 8 nM / L using detection buffer. The test compound was diluted 5-fold to the 8th concentration using a multi-channel pipette, i.e., from 40 μM to 0.512 nM, with a final DMSO concentration of 4%, and a double-dose assay was performed. 2.5 μL of each inhibitor concentration gradient, 5 μL of 10 ng of DK2 / Cyclin E1 enzyme, and 2.5 μL of a mixture of substrate and ATP (4 mM ATP, 100 nM MLight-4E-BP1 peptide) were added to the microplate. At this point, the final compound concentration gradient was 10 μM diluted to 0.128 nM, and the final concentrations of ATP and substrate were 1 mM and 25 nM, respectively. The reaction system was incubated at 25°C for 120 minutes. After the reaction, 5 μL of stop solution was added to each well, and the reaction was continued at 25°C for 5 minutes. After the reaction was completed, 5 μL of Eu-anti-phospho-tyrosine antibody dilution buffer was added to each well, and the reaction was continued at 25°C for 60 minutes. Data acquisition was performed using a PerkinElmerNivo multilabel analyzer in TR-FRET mode (excitation wavelength 320 nm, emission wavelengths 615 nm and 665 nm).

[0279] Data Analysis:

[0280] The original data were converted into inhibition rate using the equation (Sample-Min) / (Max-Min)×100%. The IC50 value can then be obtained by curve fitting using four parameters (obtained in GraphPad Prism using log(inhibitor) vs. response--Variable slope mode). Table 1 provides the inhibitory activity of the compounds of this invention against CDK2 / CyclinE1.

[0281] Experimental Example 2: In vitro CDK12 / CyclinK enzyme activity assay

[0282] Experimental materials:

[0283] CDK12 / CyclinK was purchased from Biotus. MLight-4E-BP1 peptide, Eu-anti-phospho-tyrosine antibody, and 1X assay buffer were purchased from PerkinElmer. High-purity ATP was purchased from Promega. EDTA was purchased from Sigma. A Nivo multilabel analyzer (PerkinElmer) was used.

[0284] Experimental methods:

[0285] Kinase buffer preparation: The kinase buffer contains 50mM HEPES, 1mM EDTA, 10mM MgCl2, 0.01% Brij-35, pH 7.4. Add 2.38g HEPES, 58mg EDTA, 406mg MgCl2, and 20mg Brij-35 to 200ml of buffer and adjust the pH to 7.4.

[0286] Preparation of the stop solution:

[0287] The stop solution was prepared by mixing 100 μL of 1M EDTA stock solution with 0.625 μL of 1X detection buffer and 1725 μL of distilled water. The enzyme, M Light-4E-BP1 peptide, ATP, and inhibitor were diluted using kinase buffer. The Eu-anti-phospho-tyrosine antibody was diluted to 8 nM / L using detection buffer. The test compound was diluted 5-fold to the 8th concentration using a multi-channel pipette, i.e., from 40 μM to 0.512 nM, with a final DMSO concentration of 4%, and a double-dose assay was performed. 2.5 μL of each inhibitor concentration gradient, 5 μL of M Light-4E-BP1 peptide (50 ng), and 2.5 μL of a mixture of substrate and ATP (800 μM ATP, 100 nM M Light-4E-BP1 peptide) were added to the microplate. At this point, the final compound concentration gradient was 10 μM diluted to 0.128 nM, and the final ATP and substrate concentrations were 200 μM and 25 nM, respectively. The reaction system was incubated at 25°C for 180 minutes. After the reaction, 5 μL of stop solution was added to each well, and the reaction was continued at 25°C for 5 minutes. After the reaction was completed, 5 μL of Eu-anti-phospho-tyrosine antibody dilution buffer was added to each well, and the reaction was continued at 25°C for 60 minutes. Data acquisition was performed using a PerkinElmerNivo multilabel analyzer in TR-FRET mode (excitation wavelength 320 nm, emission wavelength 665 nm).

[0288] Data Analysis:

[0289] The original data were converted into inhibition rate using the equation (Sample-Min) / (Max-Min)×100%, IC 50 The values ​​can be obtained by curve fitting using four parameters (obtained in GraphPad Prism using log(inhibitor) vs. response -- Variable slope mode). Table 1 provides the inhibitory activity of the compounds of this invention against CDK12 / CyclinK enzymatic activity.

[0290] Experiment Example 3: In vitro A2780 cell viability test

[0291] Experimental materials:

[0292] 1640 culture medium was purchased from Vivacell, fetal bovine serum from Biosera, and penicillin / streptomycin antibiotics from Progen. CellTiter-Glo (a chemiluminescent cell viability assay) reagent was purchased from Promega. A2780 cell line was purchased from Kebai. Envision multilabel analyzer (PerkinElmer).

[0293] Experimental methods:

[0294] A2780 cells were seeded in white 96-well plates, with 80 μL of cell suspension per well, containing 3000 A2780 cells. The plates were incubated overnight in a CO2 incubator. The test compound was diluted 5-fold to the 8th concentration using a multi-channel pipette, i.e., from 2000 μM to 0.0256 nM, in duplicate. 78 μL of culture medium was added to the intermediate plate, and then 2 μL of serially diluted compound was transferred to each well of the intermediate plate according to the corresponding position. After mixing, 20 μL of the compound was transferred to each well of the cell plate. The concentration range of the compound transferred to the cell plate was 10 μM to 0.128 nM. The cell plates were incubated in a CO2 incubator for 3 days. A separate cell plate was prepared, and the signal value was read on the day of drug addition as the maximum value (Max value in the equation below) for data analysis. 25 μL of chemiluminescent cell viability assay reagent was added to each well of this cell plate, and the plate was incubated at room temperature for 10 minutes to stabilize the luminescence signal. The readings were taken using a multi-label analyzer. Add 25 μL of chemiluminescent cell viability assay reagent to each well of the cell plate and incubate at room temperature for 10 minutes to stabilize the luminescence signal. Read the values ​​using a multi-label analyzer.

[0295] Data Analysis:

[0296] The original data were converted into inhibition rate using the equation (Sample-Min) / (Max-Min)×100%, IC 50 The values ​​can be obtained by curve fitting using four parameters (obtained in the "log(inhibitor) vs. response--Variable slope" mode in GraphPad Prism). Table 1 provides the inhibitory activity of the compounds of this invention on the proliferation of A2780 cells.

[0297] Table 1

[0298]

[0299] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0300] The embodiments described above are merely illustrative of several implementations of the present invention, facilitating a detailed understanding of the technical solutions of the present invention, but should not be construed as limiting the scope of protection of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the scope of protection of the present invention. It should be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided by the present invention through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this invention patent should be determined by the content of the appended claims, and the specification can be used to interpret the content of the claims.

Claims

1. A compound of formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof: in: R1 is selected from any one of cyano, -CF3, -OCF3, -Cl, -Br, -F, hydroxy, and nitro groups; R2 is selected from any of the following structures: ； R3 and R4 are each independently selected from -H, -D, or unsubstituted aromatic groups of 6 to 15 ring atoms; Ar1 is selected from any of the following structures: ； R5 and R6 are each independently selected from -H, -D, straight-chain alkyl with 1 to 10 carbon atoms, alkoxy with 1 to 10 carbon atoms, branched alkyl with 3 to 10 carbon atoms, cyclic alkyl with 3 to 10 carbon atoms, branched alkoxy with 3 to 10 carbon atoms, and cyclic alkoxy with 3 to 10 carbon atoms.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, Compound (I) has the structure shown in formula (II):

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R3 and R4 are independently selected from -H, -D, or unsubstituted phenyl groups, respectively.

4. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, Compound (I) has the structure shown in formula (III):

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R5 and R6 are each independently selected from -H, -D, straight-chain alkyl with 1 to 5 carbon atoms, alkoxy with 1 to 5 carbon atoms, branched alkyl with 3 to 5 carbon atoms, cyclic alkyl with 3 to 5 carbon atoms, branched alkoxy with 3 to 5 carbon atoms, and cyclic alkoxy with 3 to 5 carbon atoms.

6. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, Ar1 is selected from any of the following structures:

7. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, R1 is selected from any one of cyano, -CF3, -Cl, -Br, and -F.

8. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, characterized in that, The compound of formula (I) is any one of the following compounds:

9. The use of the compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, in the preparation of a medicament for treating and / or preventing diseases associated with or mediated by CDK12 activity.

10. The application according to claim 9, characterized in that, The diseases associated with or mediated by CDK12 activity are cancers.

11. A pharmaceutical composition, characterized in that, It includes the compound as described in any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier.

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