Pyrrolopyrimidine derivatives containing a pyrazine structure

Abstract

Pyrrolopyrimidine derivatives containing a pyrazine structure. Specifically, a compound as shown in general formula (1) and / or a pharmaceutically acceptable salt thereof, a composition containing the compound as shown in general formula (1) and / or a pharmaceutically acceptable salt thereof, a preparation method and the use of the compound as a Wee-1 inhibitor in the preparation of an antitumor drug are disclosed.

Description

[0001] This application claims priority to Chinese application CN202110518869.5, filed on May 12, 2021. The full text of the aforementioned Chinese application is incorporated herein by reference. Technical Field

[0002] This invention relates to the field of medicinal chemistry, and more specifically, to a class of compounds with Wee-1 kinase inhibitory activity, their preparation methods, and the use of such compounds in the preparation of antitumor drugs. Background Technology

[0003] Wee-1 protein kinase is an important negative regulator of cell cycle checkpoints. Cell cycle checkpoints include the G1 checkpoint (transition from G1 (resting phase) to S (DNA synthesis phase), the G2 checkpoint (transition from G2 (preparation phase) to M (mitosis phase), and the spindle checkpoint (transition from M phase metaphase to anaphase). Wee-1 protein kinase plays a crucial role in the G2 checkpoint. Cell entry into M phase depends on CDK1 kinase activity. Wee-1 inhibits CDK1 activity by phosphorylating Tyr15 of the CDK1 protein, thus preventing cell entry into M phase. Conversely, Polo kinase phosphorylates Wee-1, activating its degradation and promoting M phase entry. Therefore, Wee-1 kinase activity determines the activity of the G2 checkpoint, thereby regulating the transition from G2 to M phase.

[0004] Cell cycle checkpoints are primarily activated after DNA damage, playing a crucial role in DNA repair within cells. Normal activation of cell cycle checkpoints arrests the cell cycle and promotes DNA repair. Inhibiting checkpoint function prevents DNA damage repair, leading to apoptosis. Compared to normal cells, many tumor cells, due to impaired function of the p53 protein (a key protein in the G1 phase checkpoint), primarily rely on G2 phase checkpoint activation to repair DNA damage and avoid apoptosis. Therefore, inhibiting the G2 phase checkpoint can selectively kill tumor cells. The important role of Wee-1 kinase activity in the G2 phase checkpoint suggests that Wee-1 kinase determines the repair or death of tumor cells after DNA damage. Inhibiting Wee-1 activity can promote the entry of unrepaired tumor cells into the M phase after DNA damage, inducing apoptosis.

[0005] Studies have shown that, in addition to its role at the G2 checkpoint, Wee-1 is also involved in functions closely related to tumorigenesis and development, such as DNA synthesis, DNA homology repair, and post-translational modifications of chromosomal histones. Wee-1 expression is significantly elevated in a large number of tumors, including liver cancer, breast cancer, cervical cancer, melanoma, and lung cancer. High Wee-1 expression is positively correlated with tumor development and poor prognosis, suggesting that Wee-1 kinase may be involved in tumorigenesis and development. In vitro cell models and in vivo animal models have demonstrated that inhibiting Wee-1 activity while inducing DNA damage can significantly inhibit the growth of various tumors.

[0006] Therefore, developing specific, highly active small molecule inhibitors of Wee-1 kinase is of significant clinical value for tumor therapy, especially for targeting tumors with impaired G1 checkpoints such as those lacking P53.

[0007] Currently, AstraZeneca's Wee-1 inhibitor AZD1775 (MK-1775, Adavosertib) has entered Phase II clinical trials, with over 30 clinical trials under development. Patents related to AZD1775 include US20070254892, WO2007126122, EP2213673, WO2008133866, and WO2011034743. Abbott and Abbvie have also conducted research on Wee-1 inhibitors, with related patents including US2012220572, WO2013126656, WO2013012681, WO2013059485, and WO2013013031. Almac's patents related to Wee-1 inhibitors include WO2014167347, WO2015019037, WO2015092431, WO2018011570, WO2018062932, and WO2019138227. Girafpharma's Wee-1 patents include WO2019074979 and WO2019074981. Zeno's patents related to Wee-1 research include WO2018028008 and WO2019173082.

[0008] The Wee-1 inhibitors in this study still have some limitations. Their monotherapy efficacy is poor, and their cellular activity is insufficient when combined with other chemotherapy drugs, resulting in less than ideal clinical outcomes. Targeted therapy often leads to drug resistance in later stages, making chemotherapy a common treatment for advanced cancers. However, chemotherapy alone often produces significant side effects and poor patient tolerance. Therefore, developing Wee-1 inhibitors that offer excellent combination therapy with chemotherapy drugs is of great importance. Summary of the Invention

[0009] This invention provides a compound of general formula (1) or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates:

[0010]

[0011] In general formula (1):

[0012] m is 0 or 1;

[0013] n is 0 or 1;

[0014] v is 1 or 2;

[0015] X is CR 8 or NR 9 R 10 ;

[0016] R 1 It is H or halogen;

[0017] R 2a and R 2b Independently H or C1-C3 alkyl, or R 2a and R 2b Together with the C atoms to which it is attached, it forms a C3-C6 cycloalkyl group;

[0018] R 3a and R 3b Independently H or C1-C3 alkyl, or R 3a and R 3b Together with the C atoms to which it is attached, it forms a C3-C6 cycloalkyl group;

[0019] R 4 The compounds are halogens, CN, C1-C6 alkyl, C1-C6 alkoxy, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C6 alkylthio, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, or (4-12-membered) heterocyclic alkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, C3-C6 cycloalkyl, and (4-12-membered) heterocyclic alkyl are optional. It is substituted by 1-3 of the following groups: H, halogen, OH, CN, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, CN-substituted C1-C3 alkyl, OH-substituted C1-C3 alkyl, C3-C6 cycloalkyl-substituted C1-C3 alkyl, C1-C3 alkoxy-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy, or (4-7) heterocyclic alkyl;

[0020] R 5 and R 6Independently classified as C1-C3 alkyl, deuterated C1-C3 alkyl, C2-C6 alkenyl, C2-C6 ynyl, and C3-C6 cycloalkyl, or R 5 and R 6 Together with the atoms to which it is attached, it forms (3-10) heterocyclic alkyl groups;

[0021] Each R 7 Independently, it can be H, halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, halogen-substituted C1-C6 alkyl, or halogen-substituted C1-C6 alkoxy;

[0022] R 8 It can be H, C1-C6 alkyl, C3-C6 cycloalkyl, deuterated C1-C6 alkyl, halogen-substituted C1-C6 alkyl, CN-substituted C1-C6 alkyl, OH-substituted C1-C6 alkyl, C1-C3 alkoxy-substituted C1-C6 alkyl, C3-C6 cycloalkyl-substituted C1-C6 alkyl, or (4-7) heterocyclic alkyl;

[0023] R 9 and R 10 Independently, it can be H, C1-C6 alkyl, or C3-C6 cycloalkyl, or R. 9 and R 10 Together with the N atom to which it is attached, it forms a (4-7) heterocyclic alkyl group, which may be substituted by 1-3 of the following groups: H, halogen, OH, CN or C1-C3 alkyl.

[0024] In some embodiments of the present invention, compounds of general formula (1) have a structure as shown in general formula (1A):

[0025]

[0026] In general formula (1A):

[0027] v is 1 or 2;

[0028] R 1 It is H or halogen;

[0029] R 2a and R 2b Independently H or C1-C3 alkyl, or R 2a and R 2b The C atoms connected together form C3-C6 cycloalkyl groups;

[0030] R 4The compounds are halogens, CN, C1-C6 alkyl, C1-C6 alkoxy, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C6 alkylthio, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, or (4-12-membered) heterocyclic alkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, C3-C6 cycloalkyl, and (4-12-membered) heterocyclic alkyl are optional. It is substituted by 1-3 of the following groups: H, halogen, OH, CN, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, CN-substituted C1-C3 alkyl, OH-substituted C1-C3 alkyl, C3-C6 cycloalkyl-substituted C1-C3 alkyl, C1-C3 alkoxy-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy, or (4-7) heterocyclic alkyl;

[0031] R 5 and R 6 Independently classified as C1-C3 alkyl, deuterated C1-C3 alkyl, C2-C6 alkenyl, C2-C6 ynyl, and C3-C6 cycloalkyl, or R 5 and R 6 Together with the atoms to which it is attached, it forms (3-10) heterocyclic alkyl groups;

[0032] Each R 7 Independently, it can be H, halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, halogen-substituted C1-C6 alkyl, or halogen-substituted C1-C6 alkoxy;

[0033] R 8 It can be H, C1-C6 alkyl, C3-C6 cycloalkyl, deuterated C1-C6 alkyl, halogen-substituted C1-C6 alkyl, CN-substituted C1-C6 alkyl, OH-substituted C1-C6 alkyl, C1-C3 alkoxy-substituted C1-C6 alkyl, C3-C6 cycloalkyl-substituted C1-C6 alkyl, or (4-7) heterocyclic alkyl.

[0034] In some embodiments of the present invention, compounds of general formula (1) have structures as shown in general formula (1B):

[0035]

[0036] In general formula (1B):

[0037] m is 0 or 1;

[0038] n is 0 or 1;

[0039] v is 1 or 2;

[0040] R 1 It is H or halogen;

[0041] R 4 The compounds are halogens, CN, C1-C6 alkyl, C1-C6 alkoxy, N(C1-C6 alkyl)2, NH(C1-C6 alkyl), C1-C6 alkylthio, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, or (4-12-membered) heterocyclic alkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylthio, C3-C6 cycloalkyl, and (4-12-membered) heterocyclic alkyl are optional. It is substituted by 1-3 of the following groups: H, halogen, OH, CN, C1-C3 alkyl, C3-C6 cycloalkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, CN-substituted C1-C3 alkyl, OH-substituted C1-C3 alkyl, C3-C6 cycloalkyl-substituted C1-C3 alkyl, C1-C3 alkoxy-substituted C1-C3 alkyl, halogen-substituted C1-C3 alkoxy, or (4-7) heterocyclic alkyl;

[0042] R 5 and R 6 Independently classified as C1-C3 alkyl, deuterated C1-C3 alkyl, C2-C6 alkenyl, C2-C6 ynyl, and C3-C6 cycloalkyl, or R 5 and R 6 Together with the atoms to which it is attached, it forms (3-10) heterocyclic alkyl groups;

[0043] Each R 7 Independently, it can be H, halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, halogen-substituted C1-C6 alkyl, or halogen-substituted C1-C6 alkoxy;

[0044] R 9 and R 10 Independently, it can be H, C1-C6 alkyl, or C3-C6 cycloalkyl, or R. 9 and R 10 Together with the N atom to which it is attached, it forms a (4-7) heterocyclic alkyl group, which may be substituted by 1-3 of the following groups: H, halogen, OH, CN or C1-C3 alkyl.

[0045] In some embodiments of the present invention, wherein the general formula (1), general formula (1A) or general formula (1B), for Preferred More preferably

[0046] In some embodiments of the present invention, in the general formula (1), general formula (1A) or general formula (1B), R 4For Me, Et, CF3, CHF2, F, Cl, Br, I, OMe, OEt, CN, SMe, SEt, OCF3, NMe2, NHMe, R 4 Preferred ions are Me, Et, F, Cl, OMe, OEt, CN, and NMe2.

[0047] In some embodiments of the present invention, in the general formula (1) or general formula (1A), R 8 For H, Me, Et, CD3, R 8 Preferably H, Me, Et,

[0048] In some embodiments of the present invention, in said general formula (1) or general formula (1B), NR 9 R 10 for NR 9 R 10 Preferred

[0049] In some embodiments of the present invention, the compound of general formula (1) has one of the following structures:

[0050]

[0051]

[0052]

[0053]

[0054] Another object of the present invention is to provide a pharmaceutical composition comprising a pharmaceutically acceptable carrier, a diluent and / or an excipient, and a compound of general formula (1) of the present invention, or its isomers, crystal forms, pharmaceutically acceptable salts, hydrates or solvates as active ingredients.

[0055] Another object of the present invention is to provide the use of the compound of general formula (1) of the present invention, or any isomer thereof, crystal form thereof, pharmaceutically acceptable salt, hydrate or solvate thereof, or the pharmaceutical composition thereof, in the preparation of a medicament for the treatment, regulation or prevention of diseases mediated by Wee-1.

[0056] Another object of the present invention is to provide a method for treating, modulating or preventing diseases mediated by Wee-1, comprising administering to a subject a therapeutically effective amount of a compound of general formula (1) of the present invention, or any isomer thereof, crystal form thereof, pharmaceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition thereof.

[0057] Through thorough research, the inventors have discovered that compounds with structures as shown in general formula (1) have strong Wee-1 inhibitory activity and co-administration activity with the chemotherapy drug gemcitabine (GMC). The above results indicate that the compounds of the present invention may have better clinical effects when used in combination with chemotherapy drugs.

[0058] It should be understood that the foregoing general description of the invention and the following detailed description are exemplary and illustrative, and are intended to provide further explanation of the claimed invention.

[0059] Compound Synthesis

[0060] The preparation method of the compound of general formula (1) of the present invention is described in detail below, but these specific methods do not constitute any limitation on the present invention.

[0061] The compounds of general formula (1) described above can be synthesized using standard synthetic techniques or known techniques combined with the methods described herein. Furthermore, the solvents, temperatures, and other reaction conditions mentioned herein can be varied. Starting materials used for the synthesis of the compounds can be obtained synthetically or from commercial sources. The compounds described herein and other related compounds with different substituents can be synthesized using known techniques and starting materials, including those discovered in March, ADVANCED ORGANIC CHEMISTRY 4. th Ed., (Wiley 1992); Carey and Sundberg, ADVANCED ORGANIC CHEMISTRY 4 th Ed., Vols.A and B (Plenum 2000, 2001), Green and Wuts, PROTECTIVE GROUPS IN ORGANIC SYNTHESIS 3 rd The method described in Ed. (Wiley 1999) can be used to prepare compounds by employing appropriate reagents and by introducing different groups into the molecular formulas provided herein.

[0062] On the one hand, the compounds described herein are prepared according to methods known in the art. However, the conditions of the method, such as reactants, solvents, bases, amounts of compounds used, reaction temperatures, and reaction times, are not limited to the explanations below. The compounds of the present invention can also be conveniently prepared by combining various synthetic methods described in this specification or known in the art, such combinations being readily performed by those skilled in the art. On the other hand, the present invention also provides a method for preparing the compound of general formula (1), wherein the compound of general formula (1) can be prepared by the following method A:

[0063] Method A includes the following steps: First, compounds A1 and A2 undergo a coupling reaction to generate compound A3. Compound A3 and compound A4 react to further generate target compound (1). When compound (1) contains an amino or hydroxyl protecting group, the protecting group needs to be further removed.

[0064]

[0065] In the above reaction equation, X, Y, and R 1 R 2a R 2b R 3a R 3b R 4 R 5 R 6 R 7 R 8 R 9 The definitions of m, n, and v are as described above, and Z is Br, I, or -B(OH)2. Compound A4 includes compounds B3, C6, and other commercially available amines described below.

[0066] Further forms of the compound

[0067] "Pharmaceutical acceptable" here means that a substance, such as a carrier or diluent, will not destroy the biological activity or properties of a compound and is relatively non-toxic. For example, when given to an individual, a substance will not cause unwanted biological effects or interact with any of its components in a harmful manner.

[0068] The term "pharmaceutically acceptable salt" refers to a form of a compound that does not cause significant irritation to the administered organism and does not diminish the compound's biological activity and properties. In some specific respects, pharmaceutically acceptable salts are obtained by reacting compounds of general formula (1) with acids, such as inorganic acids like hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, and carbonic acid; organic acids like formic acid, acetic acid, propionic acid, oxalic acid, trifluoroacetic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, and p-toluenesulfonic acid; and acidic amino acids like aspartic acid and glutamic acid.

[0069] It should be understood that references to pharmaceutically acceptable salts include solvent-added forms or crystalline forms, especially solvates or polymorphs. Solvates contain stoichiometric or non-stoichiometric solvents and are selectively formed during crystallization with pharmaceutically acceptable solvents such as water, ethanol, etc. A hydrate is formed when the solvent is water, or an alcohol is formed when the solvent is ethanol. Solvates of compounds of general formula (1) are readily prepared or formed according to the methods described herein. For example, hydrates of compounds of general formula (1) are readily prepared by recrystallization from a mixture of water and organic solvents, including but not limited to tetrahydrofuran, acetone, ethanol, or methanol. Furthermore, the compounds mentioned herein can exist in both solvated and non-solvated forms. In summary, for the purposes of the compounds and methods provided herein, the solvated form is considered equivalent to the non-solvated form.

[0070] In other specific embodiments, compounds of general formula (1) are prepared in various forms, including but not limited to amorphous, pulverized, and nano-particle forms. Furthermore, compounds of general formula (1) include crystalline forms and can also be polymorphic. Polymorphs comprise different lattice arrangements of the same elemental composition of the compound. Polymorphs typically have different X-ray diffraction spectra, infrared spectra, melting points, densities, hardness, crystal forms, optical and electrical properties, stability, and solubility. Different factors such as recrystallization solvents, crystallization rates, and storage temperatures may cause a single crystal form to dominate.

[0071] In another aspect, compounds of general formula (1) may possess a chiral center and / or axial chirality, and thus appear as racemates, racemic mixtures, single enantiomers, diastereomers, and single diastereomers, and cis-trans isomers. Each chiral center or axial chirality will independently produce two optical isomers, and all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds, are included within the scope of this invention. This invention means including all such isomeric forms of these compounds.

[0072] 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) and C-14 14 C). For example, deuterium can be used to replace hydrogen atoms to form deuterated compounds. The bond between deuterium and carbon is stronger than that between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs generally have advantages such as reduced toxicity, increased drug stability, enhanced efficacy, and prolonged drug half-life in vivo. All isotopic variations of the compounds of this invention, regardless of radioactivity, are included within the scope of this invention.

[0073] the term

[0074] Unless otherwise specified, the terms used in this application, including the specification and claims, are defined as follows. It must be noted that in the specification and appended claims, unless otherwise clearly indicated, the singular form "a" includes the plural meaning. Unless otherwise specified, conventional methods such as mass spectrometry, nuclear magnetic resonance, HPLC, protein chemistry, biochemistry, recombinant DNA techniques, and pharmacology are used. In this application, unless otherwise specified, "or" or "and" refers to "and / or".

[0075] Unless otherwise specified, "alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched groups with 1 to 6 carbon atoms. Lower alkyl groups containing 1 to 4 carbon atoms are preferred, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, and tert-butyl. As used herein, "alkyl" includes unsubstituted and substituted alkyl groups, especially alkyl groups substituted with one or more halogens. Preferred alkyl groups are selected from CH3, CH3CH2, CF3, CHF2, CF3CH2, CF3(CH3)CH, etc. i Pr、 n Pr、 i Bu、 n Bu or t Bu.

[0076] Unless otherwise specified, "alkenyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon double bond, including straight-chain or branched groups with 1 to 14 carbon atoms. Lower alkenyl groups containing 1 to 4 carbon atoms are preferred, such as vinyl, 1-propenyl, 1-butenyl, or 2-methylpropenyl.

[0077] Unless otherwise specified, "alkynyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon triple bond, including straight-chain and branched groups with 1 to 14 carbon atoms. Lower alkynyl groups containing 1 to 4 carbon atoms are preferred, such as ethynyl, 1-propynyl or 1-butynyl.

[0078] Unless otherwise specified, "cycloalkyl" refers to a non-aromatic hydrocarbon ring system (monocyclic, bicyclic, or polycyclic). A partially unsaturated cycloalkyl group may be referred to as "cycloalkenyl" if the carbon ring contains at least one double bond, or as "cycloynyl" if the carbon ring contains at least one triple bond. Cycloalkyl groups may include monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) groups and spirocyclic groups. In some embodiments, the cycloalkyl group is monocyclic. In some embodiments, the cycloalkyl group is monocyclic or bicyclic. The cycloforming carbon atom of the cycloalkyl group may optionally be oxidized to form an oxo or thio group. Cycloalkyl groups also include cycloalkylene groups. In some embodiments, the cycloalkyl group contains 0, 1, or 2 double bonds. In some embodiments, the cycloalkyl group contains 1 or 2 double bonds (partially unsaturated cycloalkyl). In some embodiments, the cycloalkyl group may be fused with aryl, heteroaryl, cycloalkyl, and heterocyclic alkyl groups. In some embodiments, the cycloalkyl group may be fused with aryl, cycloalkyl, and heterocyclic alkyl groups. In some embodiments, the cycloalkyl group may be fused with aryl and heterocyclic alkyl groups. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cyclohepttrienyl, norcamphenyl, norpinel, norcarel, bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, and so on.

[0079] Unless otherwise specified, "alkoxy" refers to an alkyl group bonded to the remainder of the molecule via an ether oxygen atom. Representative alkoxy groups are those having 1-6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, and tert-butoxy. As used herein, "alkoxy" includes unsubstituted and substituted alkoxy groups, particularly those substituted with one or more halogens. Preferred alkoxy groups are selected from OCH3, OCF3, CHF2O, CF3CH2O, etc. i- PrO, n- PrO, i- BuO、 n- BuO or t- BuO.

[0080] Unless otherwise specified, "alkithioyl" refers to an alkyl group bonded to the remainder of the molecule via an ether sulfide atom. Representative alkithioyl groups are those having 1-6 carbon atoms, such as methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, sec-butylthio, and tert-butylthio. As used herein, "alkithioyl" includes both unsubstituted and substituted alkithioyl groups, especially those substituted by one or more halogens.

[0081] Unless otherwise specified, "heterocyclic alkyl" refers to a non-aromatic ring or ring system that may optionally contain one or more alkenyl groups as part of a ring structure, having at least one heteroatom ring member independently selected from boron, phosphorus, nitrogen, sulfur, oxygen, and phosphorus. If a heterocyclic alkyl contains at least one double bond, then a partially unsaturated heterocyclic alkyl may be referred to as a "heterocyclic alkenyl," or if a heterocyclic alkyl contains at least one triple bond, then a partially unsaturated heterocyclic alkyl may be referred to as a "heterocyclic ynyl." Heterocyclic alkyl can include monocyclic, bicyclic, spirocyclic, or polycyclic (e.g., having two fused or bridging rings) ring systems. In some embodiments, a heterocyclic alkyl is a monocyclic group having one, two, or three heteroatoms independently selected from nitrogen, sulfur, and oxygen. The cyclic carbon atom and heteroatom of a heterocyclic alkyl may optionally be oxidized to form an oxo or thio ion group or other oxidized bond (e.g., C(O), S(O), C(S), or S(O)₂, N-oxide, etc.), or the nitrogen atom may be quaternized. Heterocyclic alkyl may be linked via cyclic carbon atoms or cyclic heteroatoms. In some embodiments, the heterocyclic alkyl group contains 0 to 3 double bonds. In some embodiments, the heterocyclic alkyl group contains 0 to 2 double bonds. The definition of heterocyclic alkyl group also includes a portion having one or more aromatic rings fused with (i.e., sharing bonds with) the heterocyclic alkyl ring, such as benzo[a] derivatives of piperidine, morpholine, aziridine-heptadiene, or thiophene. Heterocyclic alkyl groups containing fused aromatic rings can be linked via any cyclizing atom, including the cyclizing atom of the fused aromatic ring. Examples of heterocyclic alkyl groups include, but are not limited to, azirrobutyl, azirroheptyl, dihydrobenzofuranyl, dihydrofuranyl, dihydropyranyl, N-morpholinyl, 3-oxa-9-azaspiro[5.5]undecyl, 1-oxa-8-azaspiro[4.5]decyl, piperidinyl, piperazinyl, oxoperazinyl, pyranyl, pyrrolidinyl, quininyl, tetrahydrofuranyl, tetrahydropyranyl, 1,2,3,4-tetrahydroquinolinyl, scopolamine, 4,5,6,7-tetrahydrothiazo[5,4-c]pyridinyl, and 4,5,6,7-tetrahydro-1H-imidazolium. Azo[4,5-c]pyridine, N-methylpiperidinyl, tetrahydroimidazolyl, pyrazolyl, butyrolactam, valproic acid, imidazolinone, hydantoin, dioxolane, phthalimide, pyrimidin-2,4(1H,3H)-diketoyl, 1,4-dioxane, morpholinyl, thiomorpholinyl, thiomorpholin-S-oxide, thiomorpholin-S,S-oxide, piperazine, pyranyl, pyridinone, 3-pyrrololinyl, thiaranyl, pyranone, tetrahydrothiophene, 2-azaspiro[3,3]heptyl, indololinyl,

[0082] Unless otherwise specified, "halogen" (or halogenated group) means fluorine, chlorine, bromine or iodine. The term "halogenated" (or "halogen substituted") appearing before the group name indicates that the group is partially or completely halogenated, that is, substituted by F, Cl, Br or I in any combination, preferably substituted by F or Cl.

[0083] "Optional" or "optionally" means that the event or condition described below may, but is not required, occur, and the description includes both the scenario in which the event or condition occurs and the scenario in which the event or condition does not occur.

[0084] When one of the variables is selected as a chemical bond, it means that the two groups connected are directly linked. For example, when L in XLY represents a chemical bond, it means that the structure is actually XY.

[0085] The term "membered ring" includes any ring structure. The term "membered" refers to the number of skeleton atoms that make up the ring. For example, cyclohexyl, pyridyl, pyranyl, and thioranyl are six-membered rings, while cyclopentyl, pyrroleyl, furanyl, and thiophenyl are five-membered rings.

[0086] The term "fragment" refers to a specific part or functional group of a molecule. Chemical fragments are generally considered to be chemical entities contained in or attached to a molecule.

[0087] 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 Or straight dashed key

[0088] Unless otherwise stated, use Indicates a single bond or a double bond.

[0089] Specific pharmaceutical and medical terms

[0090] The term “acceptable,” as used herein, means that a prescription component or active ingredient does not have an excessively harmful effect on health for general therapeutic purposes.

[0091] The terms “treatment,” “treatment process,” or “therapy” as used herein include alleviating, suppressing, or improving symptoms or conditions of a disease; suppressing the development of complications; improving or preventing underlying metabolic syndromes; suppressing the development of a disease or symptom, such as controlling the progression of a disease or condition; reducing a disease or symptom; alleviating a disease or symptom; reducing complications arising from a disease or symptom; or preventing or treating signs arising from a disease or symptom. As used herein, a compound or pharmaceutical composition, when administered, may improve a disease, symptom, or condition, particularly by improving its severity, delaying its onset, slowing its progression, or reducing its duration. Whether administered regularly or intermittently, continuously or intermittently, it may be attributable to or related to the administration.

[0092] "Active ingredient" refers to the compound represented by general formula (1), and pharmaceutically acceptable inorganic or organic salts of compounds of general formula (1). The compounds of the present invention may contain one or more asymmetric centers (chiral centers or axial chirality), and thus appear as racemates, racemic mixtures, single enantiomers, diastereomers, and single diastereomers. The asymmetric centers that may exist depend on the nature of the various substituents on the molecule. Each such asymmetric center will independently produce two optical isomers, and all possible optical isomers and diastereomer mixtures, as well as pure or partially pure compounds, are included within the scope of the present invention. The present invention means including all such isomeric forms of these compounds.

[0093] The terms “compound,” “composition,” “agent,” or “medicine or medicament” may be used interchangeably here, and all refer to a compound or composition that, when applied to an individual (human or animal), can induce a desired pharmaceutical and / or physiological response through local and / or systemic action.

[0094] The term “administered, administering, or administration” here refers to the direct application of the compound or composition described herein, or the application of a prodrug, derivative, or analog of the active compound.

[0095] While the numerical ranges and parameters used to define the broader scope of this invention are approximate values, the relevant values ​​in the specific embodiments have been presented as precisely as possible. However, any value inevitably contains standard deviations due to individual test methods. Here, "approximately" generally means that the actual value is within plus or minus 10%, 5%, 1%, or 0.5% of a particular value or range. Alternatively, the term "approximately" may mean that the actual value falls within the acceptable standard error of the mean, as determined by those skilled in the art. Except for experimental examples, or unless explicitly stated otherwise, it is understood that all ranges, quantities, values, and percentages used herein (e.g., to describe material usage, duration, temperature, operating conditions, quantity ratios, and others similar) are modified with "approximately". Therefore, unless otherwise stated, the numerical parameters disclosed in this specification and the accompanying claims are approximate values ​​and are subject to change as needed. At a minimum, these numerical parameters should be understood as the indicated significant digits and values ​​obtained using general rounding.

[0096] Unless otherwise defined in this specification, scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art. Furthermore, unless conflicting with the context, singular nouns used herein include their plural forms, and vice versa.

[0097] Therapeutic uses

[0098] The present invention provides that compounds or pharmaceutical compositions of general formula (1) of the present invention are generally used to inhibit Wee-1 kinase and are therefore used to treat one or more conditions associated with Wee-1 kinase activity. Therefore, in some embodiments, the present invention provides a method for treating Wee-1 kinase-mediated conditions, the method comprising the step of administering a compound of the present invention, or a pharmaceutically acceptable composition thereof, to a patient in need.

[0099] Cancers that can be treated with the compounds of this invention include, but are not limited to, hematologic malignancies (leukemia, lymphoma, myeloma including multiple myeloma, myelodysplastic syndrome and myeloproliferative syndrome) and solid tumors (cancers such as prostate, breast, lung, colon, pancreas, kidney, ovary and soft tissue cancers and osteosarcoma, as well as stromal tumors).

[0100] route of administration

[0101] The compounds of this invention and their pharmaceutically acceptable salts can be formulated into various preparations, comprising, within a safe and effective range, the compounds of this invention or their pharmaceutically acceptable salts and pharmacologically acceptable excipients or carriers. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. The safe and effective range of the compound is determined based on the age, condition, and duration of treatment of the patient.

[0102] "Pharmaceutically acceptable excipients or carriers" refers to one or more compatible solid or liquid fillers or gelling substances that are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with and with the compounds of the present invention without significantly reducing the efficacy of the compounds. Examples of pharmacologically acceptable excipients or carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), and emulsifiers (such as Tween). Wetting agents (such as sodium dodecyl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0103] When applying the compounds of this invention, they can be administered orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously), or topically.

[0104] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following components: (a) fillers or compatibilizers, such as starch, lactose, sucrose, glucose, mannitol, and silica; (b) binders, such as hydroxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants, such as glycerin; (d) disintegrants, such as agar, calcium carbonate, potato starch or cassava starch, alginate, certain complex silicates, and sodium carbonate; (e) slowing agents, such as paraffin; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glyceryl monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. Buffers may also be included in capsules, tablets, and pills.

[0105] Solid dosage forms such as tablets, sugar pills, capsules, pellets, and granules can be prepared using coatings and shells, such as casings and other materials known in the art. They may contain opacifying agents, and the release of the active compound or compound from such compositions can be delayed in a portion of the digestive tract. Examples of encapsulating components that can be used are polymeric substances and waxes. If necessary, the active compound may also be formed into microcapsules with one or more of the excipients described above.

[0106] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0107] In addition to these inert diluents, the composition may also contain auxiliaries such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and fragrances.

[0108] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances.

[0109] Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0110] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays, and inhalers. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary.

[0111] The compounds of this invention can be administered alone or in combination with other pharmaceutically acceptable compounds. When using the pharmaceutical composition, a safe and effective amount of the compound of this invention is applied to the mammal (such as a human) requiring treatment, wherein the dose administered is a pharmaceutically considered effective dose. For a person weighing 60 kg, the daily dose is typically 1–2000 mg, preferably 50–1000 mg. Of course, the specific dosage should also consider factors such as the route of administration and the patient's health condition, which are all within the scope of a skilled physician's expertise.

[0112] The features mentioned above in this invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this specification can be used in any compositional form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features. Detailed Implementation

[0113] The following description will elaborate on the specific aspects, characteristics, and advantages of the aforementioned compounds, methods, and pharmaceutical compositions, making the content of this invention readily apparent. It should be understood that the detailed descriptions and examples described below are specific embodiments and are for reference only. After reading this description, those skilled in the art can make various alterations or modifications to this invention, and these equivalent forms also fall within the scope defined by this application.

[0114] In all embodiments, the melting point was determined using an X-4 melting point apparatus, and the thermometer was not calibrated. 1 H-NMR was recorded using a Varian Mercury 400 NMR spectrometer, and chemical shifts are expressed as δ (ppm). Unless otherwise specified, the silica gel used for separation was 200-300 mesh, and all eluent ratios were by volume.

[0115] The following abbreviations are used in this invention: Ar represents argon; ClCH2CH2Cl represents 1,2-dichloroethane; Cs2CO3 represents cesium carbonate; CuI represents cuprous iodide; DCM represents dichloromethane; dioxane represents 1,4-dioxane; EA represents ethyl acetate; h represents hours; K2CO3 represents potassium carbonate; LC-MS represents liquid chromatography-mass spectrometry; mL represents milliliters; MeOH represents methanol; min represents minutes; MS represents mass spectrometry; NaBH3CN represents boron cyanocyanate. Sodium hydride; NaBH(OAc)3 represents sodium triacetoxyborohydride; NIS represents N-chlorosuccinimide; NMR represents nuclear magnetic resonance; ℃ represents degrees Celsius; Pd2(dba)3 represents tris(dibenzylideneacetone)palladium; Pd(dppf)Cl2 represents [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride; PE represents petroleum ether; rt represents room temperature; Xantphos represents 4,5-bisdiphenylphosphine-9,9-dimethyloxanthracene; TEA represents triethylamine.

[0116] Preparation Example 1 (((6-(2-chloro-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl]pyrazin-2-yl)imino)dimethyl-λ 6 Preparation of thioketones (intermediate A3-1)

[0117]

[0118] 2-chloro-5-fluoro-7H-pyrrolo[2,3-d]pyrimidine (688 mg, 4 mmol), ((6-bromopyrazin-2-yl)imino)dimethyl-λ 6 β-sulfone (1.0 g, 4 mmol) was added to a 100 mL single-necked flask, dissolved in 40 mL of 1,4-dioxane, and then K₂CO₃ (1.1 mg, 8 mmol), CuI (764 mg, 4 mmol), and N₂ were added. 1 N 2 -Dimethylcyclohexane-1,2-diamine (568 mg, 4 mmol), Ar gas was replaced 5 times, and the mixture was stirred at 100 °C for 5 h. The reaction was monitored by LC-MS until complete. The mixture was filtered, concentrated, and the residue was subjected to silica gel column chromatography (DCM / MEOH = 100 / 1 to 80 / 1) to give a pale yellow solid (820 mg, yield 51.2%). LC-MS: 343.1 [M+H] + .

[0119] By using different raw materials, similar to the synthesis of intermediate A3-1, intermediates A3-2 to A3-9 can be obtained.

[0120] Table 1. Structural formulas of intermediates A3-2 to A3-9

[0121]

[0122]

[0123] Preparation Example 2: Preparation of 6-(pyrrolidone-1-yl)-5,6,7,8-tetrahydronaphthyl-2-amine (intermediate B3-1)

[0124]

[0125] Step 1: Synthesis of compound B2-1

[0126] 6-Nitro-3,4-dihydronaphthyl-2(1H)-one (1.7 g, 8.9 mmol) was dissolved in ClCH2CH2Cl (40 mL), and pyrrolidine (1.264 g, 17.8 mmol) and HOAc (1.6 g, 26.7 mmol) were added. Under argon protection and in an ice-water bath, NaBH3CN (1.675 g, 26.7 mmol) was added. The reaction was allowed to proceed overnight at room temperature, and the reaction was monitored by LC-MS until completion. The solution was passed through a reverse-phase column to give a pink solid product B2-1 (1.1 g, 50% yield), LC-MS: 247.3 [M+H]. + .

[0127] Step 2: Synthesis of compound B3-1

[0128] B2-1 (492 mg, 2.0 mmol) was dissolved in methanol (50 mL), Pd / C was added, hydrogen gas was bubbled through, and the reaction was carried out overnight at room temperature. The reaction was monitored by LC-MS until completion. The solution was filtered and concentrated to give a brown solid B3-1 (372 mg, yield 86%), ESI-MS m / z: 217.2 [M+H]. + .

[0129] By using different raw materials, similar to the synthesis of intermediate B3-1, intermediates B3-2 to B3-18 can be obtained.

[0130] Table 2. Structural formulas of intermediates B3-2 to B3-18

[0131]

[0132]

[0133] Preparation Example 3: Preparation of 2,5-dimethyl-1,2,3,4-tetrahydroisoquinoline-7-amine (intermediate C6-1)

[0134]

[0135] Step 1: Synthesis of compound C1-1

[0136] 7-Nitro-1,2,3,4-Tetrahydroisoquinoline (10.0 g, 46.10 mmol, HCl) was dissolved in TFA (50.0 mL) under argon protection and cooled to 0°C in an ice bath. NIS (15.7 g, 69.88 mmol) was added, and the reaction was allowed to proceed overnight at room temperature until the solution turned black. LC-MS monitoring showed that the reaction was complete. The solution was then poured into ice water, and the pH was adjusted to 8-9 with NaOH, causing a solid to precipitate. The solid was filtered, and dried to obtain a black solid Cl-1 (14 g, used directly in the next reaction). ESI-MS m / z: 305.0 [M+H] + .

[0137] Step 2: Synthesis of compound C2-1

[0138] C1-1 (14.0 g, 46.0 mmol) and (Boc)2O (25.1 g, 115 mmol, 26.4 mL) were dissolved in DCM (200 mL), and TEA (14.0 g, 138 mmol, 19.2 mL) was added. The mixture was reacted overnight at room temperature. LC-MS showed the reaction was complete. Water (100 mL) was added to the reaction mixture, and the mixture was shaken well. The aqueous phase was extracted with DCM (150 mL x 3). The organic phases were combined, dried, filtered, and concentrated. The residue was subjected to silica gel column chromatography (PE / EA = 100 / 1 to 10 / 1) to give a white solid C2-1 (10.0 g, yield 53.7%), ESI-MS m / z: 405.0 [M+H]. + .

[0139] Step 3: Synthesis of compound C3-1

[0140] C2-1 (6.00 g, 14.8 mmol), methylboric acid (8.90 g, 148.4 mmol), Pd(dppf)Cl2.CH2Cl2 (1.2 g, 1.5 mmol), and Cs2CO3 (2 M aqueous solution, 14.8 mL) were dissolved in dioxane (100 mL). The mixture was heated to 100 °C for 5 h under argon protection. The reaction was monitored by LC-MS until completion. Water (100 mL) was added to the reaction solution, and the mixture was extracted with EA (100 mL x 3). The organic phases were combined, dried, filtered, and concentrated. The residue was subjected to silica gel column chromatography (PE / EA = 100 / 1 to 10 / 1) to give a white solid C3-1 (2.5 g, yield 57.6%), ESI-MS m / z: 293.1 [M+H]. + .

[0141] Step 4: Synthesis of compound C4-1

[0142] C3-1 (485 mg, 2 mmol) was dissolved in EA (50 mL), and HCl / dioxane (4 M, 10 mL) was added. The mixture was reacted at room temperature for 1.5 h. The reaction was monitored by LC-MS until completion. The solution was directly concentrated for the next reaction. ESI-MS m / z: 193.1 [M+H] + .

[0143] Step 5: Synthesis of compound C5-1

[0144] The C4-1 obtained in the previous step (theoretical amount, 2 mmol) was dissolved in a mixture of DCM and MeOH (DCM / MeOH = 10 / 1, 50 mL), formaldehyde aqueous solution (0.5 mL), and TEA (607 mg, 6 mmol). Under argon protection, the mixture was stirred in an ice bath for 30 minutes. Sodium triacetoxyborohydride (848 mg, 4 mmol) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by LC-MS until complete. The reaction solution was washed with sodium bicarbonate aqueous solution, dried, and concentrated. The residue was subjected to silica gel column chromatography (DCM / MeOH = 100 / 1 to 20 / 1) to give a pale yellow solid C5-1 (300 mg, yield 72.7%), ESI-MS m / z: 207.1 [M+H]. + .

[0145] Step 6: Synthesis of compound C6-1

[0146] C5-1 (300 mg, 1.45 mmol) was dissolved in methanol (50 mL), Pd / C was added, hydrogen gas was bubbled through, and the reaction was carried out overnight at room temperature. The reaction was monitored by LC-MS until completion. The solution was filtered and concentrated to give a brown solid C6-1 (230 mg, 90% yield), ESI-MS m / z: 177.2 [M+H]. + .

[0147] By using different raw materials, similar to the synthesis of intermediate C6-1, intermediates C6-2 to C6-32 can be obtained.

[0148] Table 3. Structural formulas of intermediates C6-2 to C6-32

[0149]

[0150]

[0151] Example 1 ((6-(5-fluoro-2-((2-methyl-1,2,3,4-tetrahydroisoquinoline-7-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyrazin-2-yl)imino)dimethyl-λ 6 Synthesis of thioketones (compound 1)

[0152]

[0153] 2-Methyl-1,2,3,4-tetrahydroisoquinoline-7-amine (228 mg, 1.2 mmol), A3-1 (204 mg, 0.6 mmol), and Cs2CO3 (288 mg, 0.66 mmol) were added to a 100 mL single-necked flask. 1,4-dioxane (10 mL) was added, and the mixture was substituted three times with Ar. Pd2(dba)3 (27 mg, 0.03 mmol) and Xantphos (41 mg, 0.07 mmol) were then added. The mixture was heated to 100 °C and stirred for 5 hours under argon protection. After the reaction was complete as monitored by LC-MS, the reaction solution was evaporated to dryness and purified by reverse-phase column chromatography to give a pale yellow solid, compound 1 (72 mg, 25.7%).

[0154] 1 H NMR (400MHz, DMSO-d6) δ9.77(s,1H),9.34(s,1H),8.91(d,J=0.8Hz,1H),7.96(s,1H),7.71(d,J=1.9Hz,1H),7.62(d,J=2.2Hz,1H),7.43-7.38( m,1H),7.05(d,J=8.4Hz,1H),3.53(d,J=2.4Hz,2H),3.50(s,6H),2.79(t,J=6.0Hz,2H),2.63(d,J=6.0Hz,2H),2.36(s,3H),LC-MS:467.2[M+H] + .

[0155] Example 2-66 Synthesis of Compound 2-66

[0156] Similar to the synthesis of compound 1, intermediates A3-1 to A3-9 react with B3-1 to B3-18, C6-1 to C6-32 or other commercially available amines to obtain target compounds 2-66 in Table 4. When the target compounds contain amino or hydroxyl protecting groups, further removal of protecting groups is required.

[0157] Table 4 Structure of Compound 2-66

[0158]

[0159]

[0160]

[0161]

[0162]

[0163] Example 67 Assay of the inhibitory activity of the compound of the present invention on Wee-1 enzyme

[0164] After serially diluted compounds and enzymes were mixed, incubated at room temperature (25°C) for 15 minutes, centrifuged at 1000 rpm for 1 minute to mix, and then 5 μL of substrate was added to initiate the reaction. After reacting at room temperature for 60 minutes, 5 μL of ADP-GLO reagent was added, centrifuged at 1000 rpm for 1 minute to mix, and then incubated at room temperature for another 60 minutes. Then, 10 μL of kinase detection reagent was added and incubated for another 60 minutes, followed by chemiluminescence detection. The percentage of enzyme activity inhibited by the compounds was calculated compared to the DMSO group, and the IC50 was then calculated. 50 .

[0165] Table 5. IC50 of the compounds of this invention inhibiting Wee-1 kinase activity 50 (nM)

[0166] compound <![CDATA[IC 50 (nM)]]> compound <![CDATA[IC 50 (nM)]]> compound <![CDATA[IC 50 (nM)]]> 1 2.03 2 2.78 3 2.38 4 2.41 5 2.59 6 2.75 7 2.12 11 3.12 17 2.26 18 2.35 19 2.67 20 2.32 21 2.02 22 2.14 24 2.57 34 1.99 36 2.17 37 2.11 49 2.05 59 2.33 60 1.84 61 3.19 MK-1775 4.32

[0167] As can be seen from the data in Table 5, the compounds of the present invention have a strong inhibitory effect on Wee-1 kinase. All compounds of formula (1) have a strong inhibitory activity on Wee-1 kinase, such as compound 1 and compound 34, whose activity is about twice that of the control drug MK-1775.

[0168] Example 68: In vitro antiproliferative activity of the compounds of the present invention against MIA PaCa-2 cells

[0169] 3000 MIA PaCa-2 cells / well were seeded into 384-well plates and allowed to adhere overnight. Then, DMSO or a compound at a maximum concentration of 5 μM, serially diluted 1:5, was added. Cell viability was evaluated by measuring intracellular ATP levels 72 hours after drug addition. The percentage of cell viability inhibition by the compound was calculated compared to the DMSO group, and the IC50 was calculated. 50 The values ​​are shown in Table 6 below.

[0170] Example 69: In vitro antiproliferative activity of the compound of the present invention in combination with gemcitabine (GMC) against MIA PaCa-2 cells.

[0171] 3000 MIA PaCa-2 cells / well were seeded in 384-well plates and 20 nM Gemcitabine was added. After overnight adhesion, DMSO or a compound serially diluted 1:5 at a maximum concentration of 100 nM was added. Cell viability was evaluated by measuring intracellular ATP levels 72 hours after drug addition. The percentage of cell survival inhibition by the compound was calculated compared to the DMSO group, and the IC50 was then calculated. 50 The values ​​are shown in Table 6 below.

[0172] Table 6. Antiproliferative activity of the compounds of the present invention, alone or in combination with GCM, against MIA PaCa-2 cells.

[0173]

[0174] As can be seen from the data in Table 6, the compounds of the present invention all exhibit strong anti-proliferative activity against MIA PaCa-2 cells. For example, compounds 19 and 31 show an IC50 of anti-proliferative activity against MIA PaCa-2 cells. 50 The concentration is less than 100 nM, which is more than 10 times higher than that of the control drug MK-1775. In particular, the compounds of this invention exhibit stronger activity when used in combination with GMC, such as compounds 1, 2, 7, and 61, with IC50 values ​​of less than 100 nM. 50 Less than 2 nM. The compounds of this invention exhibit strong activity in combination with GMC, suggesting that they may have better efficacy when used in combination with chemotherapy drugs in clinical practice.

[0175] While specific embodiments of the present invention have been described above, those skilled in the art should understand that these are merely illustrative examples, and various changes or modifications can be made to these embodiments without departing from the principles and essence of the present invention. Therefore, the scope of protection of the present invention is defined by the appended claims.

Claims

1. A compound of general formula (1), its optical isomer, or a pharmaceutically acceptable salt thereof: ； In general formula (1): m is 0 or 1; n is 0 or 1; v is 1 or 2; X is NR 8 or CH-NR 9 R 10 ; R 1 It is H or halogen; R 2a and R 2b Independently defined as H or C1-C3 alkyl; R 3a and R 3b Independently defined as H or C1-C3 alkyl; R 4 The group is halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, or C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl are optionally substituted with 1 to 3 of the following groups: halogen, OH, CN, C1-C3 alkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, CN-substituted C1-C3 alkyl, OH-substituted C1-C3 alkyl, or halogen-substituted C1-C3 alkoxy; R 5 and R 6 Independently C1-C3 alkyl or deuterated C1-C3 alkyl, or R 5 and R 6 Together with the atoms to which it is attached, it forms 3-10 membered heterocyclic alkyl groups; Each R 7 Independently, it can be H, halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, or halogen-substituted C1-C6 alkoxy; R 8 It can be H, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen-substituted C1-C6 alkyl, CN-substituted C1-C6 alkyl, or OH-substituted C1-C6 alkyl; R 9 and R 10 It can be H or C1-C6 alkyl.

2. The compound of claim 1, its optical isomer, or a pharmaceutically acceptable salt thereof, wherein the general formula (1) has a structure as shown in general formula (1A): ； In general formula (1A): v is 1 or 2; R 1 It is H or halogen; R 2a and R 2b Independently defined as H or C1-C3 alkyl; R 4 The group is halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, or C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl are optionally substituted with 1 to 3 of the following groups: halogen, OH, CN, C1-C3 alkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, CN-substituted C1-C3 alkyl, OH-substituted C1-C3 alkyl, or halogen-substituted C1-C3 alkoxy; R 5 and R 6 Independently C1-C3 alkyl or deuterated C1-C3 alkyl, or R 5 and R 6 Together with the atoms to which it is attached, it forms 3-10 membered heterocyclic alkyl groups; Each R 7 Independently, it can be H, halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, or halogen-substituted C1-C6 alkoxy; R 8 It can be H, C1-C6 alkyl, deuterated C1-C6 alkyl, halogen-substituted C1-C6 alkyl, CN-substituted C1-C6 alkyl, or OH-substituted C1-C6 alkyl.

3. The compound of claim 1 or 2, its optical isomer, or a pharmaceutically acceptable salt, wherein in the general formula (1) or general formula (1A), R 8 For H, Me, Et, CD3, , , , , , , or .

4. The compound of claim 1, its optical isomer, or a pharmaceutically acceptable salt thereof, wherein the general formula (1) has a structure as shown in general formula (1B): ； In general formula (1B): m is 0 or 1; n is 0 or 1; v is 1 or 2; R 1 It is H or halogen; R 4 The group is halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, or C3-C6 cycloalkyl, wherein the C1-C6 alkyl, C1-C6 alkoxy, and C3-C6 cycloalkyl are optionally substituted with 1 to 3 of the following groups: halogen, OH, CN, C1-C3 alkyl, C1-C3 alkoxy, halogen-substituted C1-C3 alkyl, CN-substituted C1-C3 alkyl, OH-substituted C1-C3 alkyl, or halogen-substituted C1-C3 alkoxy; R 5 and R 6 Independently C1-C3 alkyl or deuterated C1-C3 alkyl, or R 5 and R 6 Together with the atoms to which it is attached, it forms 3-10 membered heterocyclic alkyl groups; Each R 7 Independently, it can be H, halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, halogen-substituted C1-C6 alkyl, or halogen-substituted C1-C6 alkoxy; R 9 and R 10 It can be H or C1-C6 alkyl.

5. The compound, its optical isomer, or pharmaceutically acceptable salt as claimed in any one of claims 1 or 4, wherein in general formula (1) or general formula (1B), -NR 9 R 10 for , or .

6. The compound of claim 1, 2, or 4, its optical isomer, or a pharmaceutically acceptable salt, wherein in general formula (1), general formula (1A), and general formula (1B), for , , , , , , , , or .

7. The compound of claim 1, 2 or 4, its optical isomer or pharmaceutically acceptable salt, wherein in the general formula (1), general formula (1A) and general formula (1B), R 4 For Me, Et, CF3, CHF2, F, Cl, Br, I, OMe, OEt, CN, OCF3, , , , , , , , , , , , , , , , , , , , , , , , or .

8. A compound, its optical isomer, or a pharmaceutically acceptable salt, wherein said compound has one of the following structures: or .

9. A pharmaceutical composition, characterized in that, It contains a pharmaceutically acceptable excipient or carrier, and a compound, its optical isomer or a pharmaceutically acceptable salt as any one of claims 1-8 as the active ingredient.

10. The use of a compound, an optical isomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in any one of claims 1-8, in the preparation of a medicament for treating Wee-1-mediated related diseases.

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