Fused ring compounds as wee-1 inhibitors and methods of making and using the same

Abstract

Fused ring compounds as wee-1 inhibitors, and preparation method and use thereof. Specifically, a compound represented by general formula (1) and a preparation method thereof, and use of the compound represented by general formula (1) and each isomer, each crystal form, a pharmaceutically acceptable salt, a hydrate or a solvate thereof as a wee-1 inhibitor in preparation of an antitumor drug.

Description

[0001] This application claims priority to Chinese application CN202110485597.3, filed on April 30, 2021. The entire contents of the aforementioned Chinese application are incorporated herein by reference. TECHNICAL FIELD

[0002] The present application relates to the field of medicinal chemistry, more particularly, to compounds having Wee-1 kinase inhibitory effect, and the preparation method thereof and the use of the compounds in the preparation of anti-tumor drugs. BACKGROUND

[0003] Wee-1 protein kinase is an important negative regulatory protein in cell cycle checkpoint. Cell cycle checkpoint includes G1 checkpoint for the transition from G1 (cell resting phase) to S phase (DNA synthesis phase), G2 checkpoint for the transition from G2 (cell division preparation phase) to M (cell division phase), and spindle checkpoint for the transition from metaphase (cell division phase) to anaphase (cell division phase). Wee-1 protein kinase plays an important role in G2 checkpoint. The entry of cells into M phase depends on the activity of CDK1 kinase, and Wee-1 inhibits the activity of CDK1 by phosphorylating Tyr15 of CDK1 protein, thereby preventing the entry of cells into M phase (cell division phase). Polo kinase phosphorylates Wee-1, activates the degradation of Wee-1 protein, and promotes the entry of cells into M phase. Therefore, the activity of Wee-1 kinase determines the activity of G2 checkpoint, and further regulates the transition from G2 to M phase.

[0004] Cell cycle checkpoint is mainly activated after DNA damage, and plays an important role in the repair of DNA in cells. Normal activation of cell cycle checkpoint blocks cell cycle to promote DNA repair. Inhibition of checkpoint function, DNA damage cannot be repaired, and cells undergo apoptosis. Compared with normal cells, various tumor cells mainly rely on the activation of G2 checkpoint to repair DNA damage and evade apoptosis due to the impairment of the function of p53 protein, an important protein in G1 checkpoint. Therefore, inhibition of G2 checkpoint can selectively kill tumor cells. The important role of Wee-1 kinase activity in G2 checkpoint suggests that Wee-1 kinase determines the repair or death of tumor cells after DNA damage, and inhibition of Wee-1 activity can promote the entry of un-repaired tumor cells into M phase after DNA damage, thereby inducing apoptosis.

[0005] Studies have shown that in addition to the role in G2 checkpoint, Wee-1 is also involved in DNA synthesis, DNA homologous repair, chromatin histone post-translational modification and other functions closely related to tumor occurrence and development. In a large number of tumors including liver cancer, breast cancer, cervical cancer, melanoma and lung cancer, the expression of Wee-1 is greatly increased. High expression of Wee-1 is positively correlated with tumor development and poor prognosis, suggesting that Wee-1 kinase may be involved in tumor occurrence and development. Studies in vitro cell models and in vivo animal models have shown that inhibition of Wee-1 activity can significantly inhibit the growth of various tumors while inducing DNA damage.

[0006] Therefore, the development of specific small molecule inhibitors of high-activity Wee-1 kinase has important clinical value for tumor treatment, especially for targeting tumors with impaired G1 checkpoint such as P53 deletion.

[0007] Currently, AstraZeneca's Wee-1 inhibitor AZD1775 (MK-1775, Adavosertib) has entered the clinical phase 2 research stage, and more than 30 clinical trials are being developed. Patents related to AZD1775 include US20070254892, WO2007126122, EP2213673, WO2008133866, WO2011034743, etc. Abbott and Abbvie have also conducted research on Wee-1 inhibitors, and the related patents mainly include US2012220572, WO2013126656, WO2013012681, WO2013059485, WO2013013031, etc. Almac's patents on Wee-1 inhibitors include WO2014167347, WO2015019037, WO2015092431, WO2018011570, WO2018062932, WO2019138227, etc. Girafpharma's Wee-1 patents include WO2019074979 and WO2019074981. Zeno's patents on Wee-1 research include WO2018028008 and WO2019173082.

[0008] There are still some problems in the Wee-1 inhibitors under research. The therapeutic effect of single drug is poor, and the cell activity of combination with other chemotherapeutic drugs is not strong enough, so the clinical combination effect is not ideal. Targeted therapy usually produces drug resistance in the later stage, and chemotherapy becomes a common means of treatment for advanced tumors. The use of chemotherapeutic drugs alone often produces greater side effects, and patients have poor tolerance, so the Wee-1 inhibitor with good combination effect with chemotherapeutic drugs has very important significance. SUMMARY

[0009] The present application provides a compound represented by general formula (1) or each isomer, each crystal form, a pharmaceutically acceptable salt, a hydrate or a solvate thereof:

[0010]

[0011] In general formula (1):

[0012] m is 0 or 1;

[0013] R 1 is H or halogen;

[0014] R 2 is 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 membered) heterocycloalkyl;

[0015] R 3 is H or C1-C3 alkyl;

[0016] A is aryl or heteroaryl, which aryl and heteroaryl can be optionally substituted with 1-3 R 6 , each R 6 is independently H, halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, OH-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, halogen-substituted C3-C6 cycloalkyl, hydroxyl-substituted C3-C6 cycloalkyl, cyano-substituted C3-C6 cycloalkyl, CF3-substituted C3-C6 cycloalkyl, -NR 7a R 7b , -N=S(O)R 7a R 7b , -P(O)R 7a R 7b , -S(O)2R 7a , -S(O)2NR 7a R 7b , -NR 8 P(O)R 7a R 7b , -NR 8 S(O)2R 7a , -NR 8 C(O)R 7a , -N=S(=NR 8 )R 7a R 7b or pyridonyl, wherein R 7a and R 7bindependently C1-C3alkyl, deuterated C1-C3alkyl, C2-C6alkenyl, C2-C6alkynyl, or C3-C6cycloalkyl, or R 7a and R 7b together with the nitrogen, sulfur, or phosphorus atom to which they are attached form a (3-10 membered)heterocycloalkyl group, R 8 is H or C1-C3alkyl, or R 8 and R 7a together with the nitrogen and sulfur atom or the nitrogen and carbon atom to which they are attached form a (3-10 membered)heterocycloalkyl group;

[0017] B is partially unsaturated C5-C7cycloalkyl or partially unsaturated (5-7 membered)heterocycloalkyl.

[0018] In some embodiments of the application, wherein the general formula (1) is is wherein R 2 is H, Me, Et, CD3,

[0019] In some embodiments of the application, wherein the general formula (1) is is is preferably

[0020] In some embodiments of the application, wherein the general formula (1) has the structure of general formula (1A):

[0021]

[0022] In general formula (1A):

[0023] n is 1, 2, or 3;

[0024] X is CH2, O, or S;

[0025] m, A, R 1 and R 2 are as previously described.

[0026] In some embodiments of the application, wherein the general formula (1) or general formula (1A), A is phenyl, pyridyl, pyrimidinyl, or pyrazinyl, which phenyl, pyridyl, pyrimidinyl, and pyrazinyl can be optionally substituted with 1-3 R 6 each R 6independently H, halogen, CN, C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, halogen- substituted C1-C6alkyl, halogen-substituted C1-C6alkoxy, hydroxy-substituted C1-C6alkyl, cyano-substituted C1-C6alkyl, halogen-substituted C3-C6cycloalkyl, OH- substituted C3-C6cycloalkyl, cyano-substituted C3-C6cycloalkyl, CF3-substituted C3-C6cycloalkyl, -NR 7a R 7b , -N=S(O)R 7a R 7b , -P(O)R 7a R 7b , -S(O)2R 7a , -S(O)2NR 7a R 7b , -NR 8 P(O)R 7a R 7b , -NR 8 S(O)2R 7a , -NR 8 C(O)R 7a , -N=S(=NR 8 )R 7a R 7b or pyridonyl, wherein R 7a and R 7b are independently C1-C3alkyl, deuterated C1-C3alkyl, C2-C6alkenyl, C2-C6alkynyl or C3-C6cycloalkyl, or R 7a and R 7b together with the nitrogen, sulfur or phosphorus atom to which they are attached form a (3-10 membered)heterocycloalkyl group, R 8 is H or C1-C3alkyl, or R 8 and R 7a together with the nitrogen and sulfur atom or the nitrogen and carbon atom to which they are attached form a (3-10 membered)heterocycloalkyl group.

[0027] In some embodiments of the application, wherein said general formula (1) or general formula (1A), A is wherein v is 1, 2 or 3, and each R 6 is independently H, F, Cl, Br, I, Me, Et,

[0028] In some embodiments of the application, wherein said general formula (1) or general formula (1A), A is

[0029]

[0030] A preferably is A more preferably is

[0031] In another embodiment of the present application, the compound of the present application has one of the following structures:

[0032]

[0033]

[0034]

[0035]

[0036] Another object of the present application is to provide a pharmaceutical composition containing a pharmaceutically acceptable carrier, diluent and / or excipient, and a compound of the present application of general formula (1) or general formula (1A), or each isomer, each crystalline form, a pharmaceutically acceptable salt, hydrate or solvate thereof as an active ingredient.

[0037] Still another object of the present application is to provide use of a compound of the present application of general formula (1), or each isomer, each crystalline form, a pharmaceutically acceptable salt, hydrate or solvate thereof or the above-mentioned pharmaceutical composition for the manufacture of a medicament for treating, regulating or preventing a disease mediated by Wee-1.

[0038] Still another object of the present application is to provide a method for treating, regulating or preventing a disease mediated by Wee-1, comprising administering to a subject a therapeutically effective amount of a compound of the present application of general formula (1), or each isomer, each crystalline form, a pharmaceutically acceptable salt, hydrate or solvate thereof or the above-mentioned pharmaceutical composition.

[0039] The present inventors have found, through intensive research, that a fused ring compound having a structure as shown in formula (1) has strong Wee-1 inhibitory activity, and combined administration activity with a chemotherapeutic drug gemcitabine (GMC), and the above results show that the compound of the present application can have a better effect in combination with a chemotherapeutic drug in a clinical setting.

[0040] It is to be understood that both the foregoing general description and the following detailed description of the present application are exemplary and explanatory, and are intended to provide further explanation of the application as claimed.

[0041] Synthesis of compounds

[0042] The preparation method of the compound of general formula (1) of the present application is specifically described below, but these specific methods do not constitute any limitation on the present application.

[0043] The compounds of general formula (1) described above can be synthesized using standard synthetic techniques or known techniques in combination with the methods described herein. In addition, the solvents, temperatures, and other reaction conditions mentioned herein can be varied. The starting materials used in the synthesis of the compounds can be obtained by synthesis or from commercial sources. The compounds described herein and other related compounds having different substituents can be synthesized using known techniques and starting materials, including those found 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 Ed., (Wiley 1999). The general methods of compound preparation can be varied by using appropriate reagents and conditions to introduce the different groups in the formula provided herein.

[0044] In one aspect, the compounds described herein are prepared according to methods known in the art. However, the conditions of the methods, such as the reactants, solvents, bases, amounts of compounds used, reaction temperatures, times required for the reactions, etc., are not limited to the explanations below. The compounds of the present application can also be prepared conveniently by optionally combining various synthetic methods described in this specification or known in the art, such combinations being readily made by those skilled in the art. In one aspect, the present application also provides a method for preparing a compound represented by general formula (1) described herein, wherein the compound of general formula (1) is prepared by the following method A:

[0045] Method A comprises the following steps: first, compounds A1 and A2 are coupled to form compound A3, and compound A3 and compound B7 are further reacted to form the target compound A5.

[0046]

[0047] In the above reaction equation, A, B, R 1 , R 2 , R 3 and m are as defined above, and Y is Br, I or -B(OH)2.

[0048] Further forms of compounds

[0049] "Pharmaceutically acceptable" means a substance, such as a carrier or diluent, which does not itself induce the production of antibodies harmful to the individual who receives the substance, and which is relatively non-toxic, i.e., the substance can be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the mixture in which it is contained.

[0050] The term "pharmaceutically acceptable salt" means a salt of a compound that does not cause significant irritation to an organism to which it is administered and does not abrogate the biological activity and properties of the compound. In certain embodiments, the pharmaceutically acceptable salts are obtained by reaction of a compound of Formula (1) with an acid such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, and the like, and organic acids such as 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, p-toluenesulfonic acid, and the like, and acidic amino acids such as aspartic acid and glutamic acid.

[0051] It is understood that references to a pharmaceutically acceptable salt includes solvent addition form or crystal form, especially solvate or polymorph, of such salts. Solvates contain either stoichiometric or non-stoichiometric amounts of the solvent, and are formed either by contact with such solvent during crystallization, either in the solvent of the reaction medium, or by contact with such solvent in the crystallization process. When the solvent is water, the solvates are hydrates, or when the solvent is ethanol, the solvates are alcoholates. Solvates of the compounds of Formula (1) are readily prepared as described herein, and are conveniently prepared by methods known to those skilled in the art. For example, the hydrates of the compounds of Formula (1) are conveniently prepared by recrystallization from a mixture of water and an organic solvent, including but not limited to, tetrahydrofuran, acetone, ethanol or methanol. In addition, the compounds referred to herein can exist in unsolvated and solvated forms. In general, the solvated forms are considered equivalent to the unsolvated forms for the purposes of the compounds and methods provided herein.

[0052] In other embodiments, the compounds of Formula (1) are prepared in different forms, including but not limited to, amorphous, pulverized and nano-particle size forms. In addition, the compounds of Formula (1) include crystalline forms, which can also exist as polymorphs. Polymorphs include different crystal lattice arrangements of the same elemental composition of the compound. Polymorphs often have different X-ray diffraction patterns, infrared spectra, melting points, density, hardness, crystal shape, optical and electrical properties, stability, and solubility. Different factors such as recrystallization solvent, rate of crystallization, and storage temperature can cause a single crystal form to dominate.

[0053] In another aspect, the compounds of general formula (1) can exist as chiral centers and / or axial chirality, and thus occur as racemates, racemic mixtures, single enantiomer, diastereomeric compounds and single diastereomers, and as cis and trans isomers. Each chiral center or axial chirality will independently give rise to two optical isomers, and all possible optical isomers and diastereomeric mixtures as well as pure or partially purified compounds are included within the scope of the present application. The present application is meant to include all such isomeric forms of these compounds.

[0054] The compounds of the present application can contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds can be labeled with radioactive isotopes, such as for example tritium ( 3 H), iodine-125 ( 125 I) and carbon-14 ( 14 C). For example, deuterium can be substituted for hydrogen, i.e., deuterium-

[0055] The term

[0056] If not otherwise specified, the terms as used in the present application, including the specification and claims, are defined as follows. It must be noted that, as used in the specification and the appended claims, the singular forms "a," "an" and "the" include plural referents unless the context clearly dictates otherwise. If not otherwise specified, conventional methods of mass spectroscopy, nuclear magnetic resonance, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology are used. In the present application, "or" or "and" as used herein means "and / or" unless otherwise stated.

[0057] Unless otherwise specified, "alkyl" means a saturated aliphatic hydrocarbon group, including straight-chain and branched-chain groups having from one to six carbon atoms. Preferred are lower alkyl groups containing from one to four carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, t-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, i Pr, n Pr, i Bu, n Bu or t Bu.

[0058] Unless otherwise specified, "alkylene" refers to a divalent alkyl group as defined above. Examples of alkylene groups include, but are not limited to, methylene and ethylene.

[0059] Unless otherwise specified, "alkenyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon double bond, including straight-chain and branched-chain groups. Preferred are lower alkenyl groups containing from 1 to 4 carbon atoms, such as ethenyl, 1 -propenyl, 1 -butenyl, or 2-methylpropenyl.

[0060] Unless otherwise specified, "alkynyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon triple bond, including straight-chain and branched-chain groups. Preferred are lower alkynyl groups containing from 1 to 4 carbon atoms, such as ethynyl, 1 -propynyl, or 1 -butynyl.

[0061] Unless otherwise specified, "cycloalkyl" refers to a non-aromatic hydrocarbon ring system (monocyclic, bicyclic, or polycyclic), and partially unsaturated cycloalkyl groups can be referred to as "cycloalkenyl" if the carbon ring contains at least one double bond, or "cycloalkynyl" if the carbon ring contains at least one triple bond. Cycloalkyl groups can include single ring or multiple ring (e.g., with 2, 3, or 4 fused rings) groups and spiro rings. In some embodiments, the cycloalkyl group is monocyclic. In some embodiments, the cycloalkyl group is monocyclic or bicyclic. The ring-forming carbon atoms of the cycloalkyl group can optionally be oxidized to form oxo or thiono groups. 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 groups). In some embodiments, the cycloalkyl group can be fused to aryl, heteroaryl, cycloalkyl, and heterocycloalkyl groups. In some embodiments, the cycloalkyl group can be fused to aryl, cycloalkyl, and heterocycloalkyl groups. In some embodiments, the cycloalkyl group can be fused to aryl and heterocycloalkyl groups. In some embodiments, the cycloalkyl group can be fused to aryl and cycloalkyl groups. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norbornyl, norpinyl, norcaryl, bicyclo[1.1.1]pentan-yl, bicyclo[2.1.1]hexan-yl, and the like.

[0062] Unless otherwise specified, "alkoxy" refers to an alkyl group bonded to the rest of the molecule through an ether oxygen atom. Representative alkoxys are alkoxys having from 1 to 6 carbon atoms, such as methoxy, ethoxy, propyloxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, and t-butoxy. As used herein, "alkoxy" includes unsubstituted and substituted alkoxys, especially alkoxys substituted with one or more halogens. Preferred alkoxys are selected from the group consisting of OCH3, OCF3, CHF2O, CF3CH2O, i- PrO, n- PrO, i-BuO、 n- BuO or t- BuO.

[0063] 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,

[0064] Unless otherwise specified, "aryl" means a carbocyclic aromatic group, aryl groups being monocyclic or polycyclic, e.g., a monocyclic aryl ring is fused to one or more carbocyclic aromatic groups. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, and phenanthryl.

[0065] Unless otherwise specified, "aryloxy" means an aryl group bonded to the rest of the molecule through an ether oxygen atom. Examples of aryloxy groups include, but are not limited to, phenoxy and naphthoxy.

[0066] Unless otherwise specified, "arylene" means a divalent aryl group as defined above. Examples of arylene groups include, but are not limited to, phenylene, naphthylene, and phenanthrylene.

[0067] Unless otherwise specified, "heteroaryl" means an aromatic group containing one or more heteroatoms (O, S, or N), heteroaryl groups being monocyclic or polycyclic, e.g., a monocyclic heteroaryl ring is fused to one or more carbocyclic aromatic groups or other monocyclic heterocycloalkyl groups. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyridazinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, quinolinyl, isoquinolinyl, furanyl, thiophenyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, benzothiophenyl, benzoxazolyl, benzopyridinyl, pyrrolopyrimidinyl, 1H-pyrrolo[3,2-b]pyridinyl, 1H-pyrrolo[2,3-c]pyridinyl, 1H-pyrrolo[3,2-c]pyridinyl, 1H-pyrrolo[2,3-b]pyridinyl,

[0068] Unless otherwise specified, "halogen" (or halo) means fluorine, chlorine, bromine, or iodine. The term "halo" (or "halogen substituted") appearing before a group denotes that the group is partially or fully halogenated, that is, substituted with F, Cl, Br, or I, in any combination, preferably F or Cl.

[0069] "Optional" or "optionally" means that the subsequently described event or circumstance can or can not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0070] The substituent "-O-CH2-O-" means that the two oxygen atoms and the two adjacent carbon atoms of the heterocycloalkyl, aryl, or heteroaryl group are connected in this substituent, such as:

[0071] When the number of a linking group is 0, such as -(CH2)0-, it means that the linking group is a single bond.

[0072] When one of the variables is selected from a chemical bond, it means that the two groups to which it is attached are directly connected, such as L represents a chemical bond in X-L-Y means that the structure is actually X-Y.

[0073] The term "membered ring" includes any ring structure. The term "member" means to indicate the number of skeletal atoms that make up the ring. For example, cyclohexyl, pyridyl, pyranyl, thiopyranyl are six-membered rings, and cyclopentyl, pyrrolyl, furanyl, and thienyl are five-membered rings.

[0074] The term "moiety" refers to a specific portion or functional group of a molecule. A chemical moiety is generally considered to be a chemical entity that is included in or attached to a molecule.

[0075] Unless otherwise indicated, the use of a wavy line and a dashed wavy line indicates the absolute configuration of a stereogenic center, the use of a straight line and a dashed straight line indicates the relative configuration of a stereogenic center, and the use of a wavy line indicates a wavy line or a dashed wavy line or a straight line or a dashed straight line or a dashed straight line

[0076] Unless otherwise indicated, the use of indicates a single or double bond.

[0077] Specific Pharmaceutical and Medical Terms

[0078] The term "acceptable," as used herein, means that a prescription component or active ingredient has no excessive deleterious effect on the health of the general treatment goal.

[0079] The terms "treatment," "treatment regime," or "therapy," as used herein, include alleviating, inhibiting or ameliorating a disease symptom or condition; inhibiting the development of complications; ameliorating or preventing a latent metabolic syndrome; inhibiting the development of a disease or symptom, such as controlling the progression of a disease or condition; reducing a disease or symptom; causing regression of a disease or symptom; reducing complications resulting from a disease or symptom, or preventing or treating an indication resulting from a disease or symptom. As used herein, a compound or pharmaceutical composition, upon administration to a subject, can result in the improvement of a disease, symptom, or condition, particularly the severity thereof, delay in onset, slowing of disease progression, or reduction in the duration of the disease. The condition can be attributable to or associated with the administration, whether fixed or contingent, continuous or intermittent.

[0080] "Active ingredient" means a compound of Formula (1), as well as pharmaceutically acceptable inorganic or organic salts of the compounds of Formula (1). The compounds of the present application can contain one or more asymmetric centers (chiral centers or axes of chirality) and therefore occur as racemates, racemic mixtures, single enantiomers, diastereomeric mixtures, and as individual diastereomers. Asymmetric centers can exist, depending 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 stereoisomeric mixtures are intended, while all geometric isomeric forms of the compounds are also intended, for example, (E) and (Z) isomers. The present application is meant to include all such isomeric forms of these compounds.

[0081] "Compound," "composition," "agent," or "medicine" and the like are used interchangeably herein and refer to a compound or composition that, when administered to a subject (human or animal), is capable of eliciting the desired pharmacological and / or physiologic effect, either locally or systemically.

[0082] The term "administered," "administering" or "administration" refers to the direct administration of the compound or composition, or the administration of a prodrug, derivative, or analog of the active compound.

[0083] Notwithstanding that the numerical ranges and parameters setting forth the broadest scope of the application are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Herein, "about" or "approximately" means within 10%, 5%, 1%, or 0.5% of a given value or range. Alternatively, "about" or "approximately" means within an acceptable standard error of the mean when considered by one of ordinary skill in the art. Other than in the operating examples, or unless otherwise expressly specified, all of the numerical ranges, amounts, values and percentages such as amounts of materials, durations, temperatures, concentrations of components, amounts of uses and other recited numerical values, are intended to be read as "about" or "approximately." It is also understood that all ranges, amounts, values and percentages stated in this specification are before conversion to the desired unit, and that special unit conversions, if any, are made to provide the desired unit. At the very least, therefore, each numerical parameter it is understood as being preceded by the word "about" or "approximately." It is further understood that the endpoints of all ranges, amounts, values and percentages are closer to the value of the range, amount, value or percentage than is indicated when the term "about" or "approximately" is used.

[0084] As used herein, the terms "about" and "comprising" are used in their broadest context. The terms "about" and "comprising" are used in their broadest context. Unless otherwise defined, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the application pertains. Furthermore, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. The use herein of "about" to modify a recited numeric value, parameter, or other numerical property, means that the indicated value, parameter, or other numerical property can vary from a stated (or recited) value by as much as 1%, 2%, 5%, 10%, 15%, 20%, 25%, or 50%, or any range or value therein. The use herein of "comprising" means "including, but not limited to."

[0085] Therapeutic uses

[0086] The compounds or compositions provided herein are generally useful for inhibiting Wee-1 kinase, and thus are useful for treating one or more conditions associated with Wee-1 kinase activity. Accordingly, in certain embodiments, the present application provides methods for treating a Wee-1 kinase-mediated condition, comprising the step of administering to a patient in need thereof a compound of the present application, or a pharmaceutically acceptable composition thereof.

[0087] Cancers that can be treated with the compounds of the present application include, but are not limited to, hematological malignancies (leukemias, lymphomas, myelomas including multiple myeloma, myelodysplastic syndromes, and myeloproliferative syndromes) and solid tumors (carcinomas such as prostate, breast, lung, colon, pancreatic, renal, ovarian, and soft tissue carcinomas and osteosarcomas, as well as stromal tumors), among others.

[0088] Routes of administration

[0089] The compounds of the present application and their pharmaceutically acceptable salts can be formulated into various pharmaceutical forms for administration purposes. These include solid, semisolid, and liquid forms such as tablets, capsules, powders, granules, creams, lotions, solutions, suspensions, and the like. The term "pharmaceutically acceptable" means a substance that is not biologically or otherwise undesirable, i.e., the substance can be administered to a subject without causing any undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of

[0090] The term "pharmaceutically acceptable" means a substance that is not biologically or otherwise undesirable, i.e., the substance can be administered to a subject without causing any undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained. The term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of arachis oil, olive oil, sesame oil, etc.), polyols (such as propylene glycol, glycerine, mannitol, sorbitol, etc.), emulsifying agents (such as Tween®, lecithin, etc.), wetting agents, coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0091] When the compounds of the present application are administered, they can be administered orally, rectally, parenterally (intravenous, intramuscular, or subcutaneous), topically.

[0092] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert excipient (or carrier) such as sodium citrate or dicalcium phosphate, or with such other ingredients as are known in the art of compounding, including binders, (a) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) humectants, such as hydroxymethylcellulose, alginic acid, gelatin, polyvinylpyrrolidone, sucrose, and acacia, (c) moisturizing agents, such as glycerol, (d) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate, (e) solution retarders, such as paraffin, (f) absoφtion accelerators, such as quaternary ammonium compounds, (g) wetting agents, such as cetyl alcohol and glycerol monostearate, (h) adsorbents, such as kaolin and bentonite, and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage forms also can comprise buffering agents.

[0093] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings and shells known in the art. They can optionally contain opacifying agents, and can also be of a composition that they release the active compound or compounds in a certain part of the digestive tract. Examples of embedding compositions that can be used are polymeric substances and waxes. The active compounds can also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0094] Liquid dosage forms for oral administration include pharmaceutically-acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage forms can contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, and the like, either with or without the addition of such

[0095] Besides such inert diluents, the composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0096] Suspensions, in addition to the active compounds, can contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, and sodium carbomate, among others.

[0097] Compositions for parenteral injection can include physiologically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0098] Dosage forms for topical administration of a compound of this application include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants as can be required.

[0099] The compounds of the present application can be administered alone or in combination with other pharmaceutically acceptable compounds. In using pharmaceutical compositions, a safe and effective amount of a compound of the present application is administered to a mammal (e.g., human) in need of treatment, wherein the dosage is that amount which is pharmaceutically considered an effective amount for administration, and for a 60 kg body weight human, the daily amount is usually 1-2000 mg, preferably 50-1000 mg. Of course, the specific dose will also take into account a route of administration, the patient's health status, and the like, which are within the skill of the skilled practitioner.

[0100] The above features mentioned in the summary or the features mentioned in the embodiments can be combined in any combination. All features disclosed in the specification can be used in any combination, and each individual feature disclosed in the specification can be replaced by any alternative feature which provides the same, equivalent or similar functionality. Thus, unless specifically stated otherwise, the features disclosed in the specification are merely exemplary of the generic type of feature they represent. DETAILED DESCRIPTION

[0101] The foregoing compounds, methods, and pharmaceutical compositions will be described in greater detail in the following specification with specific reference being made to the various figures in detail. It is to be understood that the description and examples described herein are illustrative of specific embodiments of the application and are not intended to be limiting. Changes can be made to the application in light of the teachings provided herein, which changes are intended to be encompassed by the scope of the application.

[0102] In all examples, melting points were determined using a X-4 melting point apparatus, and the thermometers were not calibrated; 1 H-NMR was recorded on a Varian Mercury 400 NMR spectrometer, and chemical shifts are expressed in δ (ppm); silica gel used for separation was 200-300 mesh unless otherwise stated, and the ratio of eluent was volume ratio.

[0103] The following abbreviations are used in the present application: CDCl3 represents deuterated chloroform; Cul represents copper iodide; DCM represents dichloromethane; dioxane represents 1,4-dioxane; DMF represents N,N-dimethylformamide, EA represents ethyl acetate; EtOH represents ethanol; h represents hour(s); H2 represents hydrogen; H2SO4 represents sulfuric acid; K2CO3 represents potassium carbonate; KNO3 represents potassium nitrate; LC-MS represents liquid chromatography-mass spectrometry; LiAlH4 represents lithium aluminum hydride; mL represents milliliter(s); MeOH represents methanol; min represents minute(s); MS represents mass spectrometry; n BuLi represents n-butyllithium; NMR represents nuclear magnetic resonance; °C represents degrees Celsius; Pd2(dba)3 represents tris(dibenzylideneacetone)dipalladium; PE represents petroleum ether; r.t. represents room temperature; Xantphos represents 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene; TEA represents triethylamine; TFA represents trifluoroacetic acid; THF represents tetrahydrofuran, T3P represents propylphosphonic anhydride.

[0104] Preparation of N-(6-(2-chloro-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyridin-2- yl)methanesulfonamide

[0105]

[0106] Into a 100 mL round bottom flask, 2-chloro-5-fluoro-7H-pyrrolo[2,3-d]pyrimidine (200 mg, 1.17 mmol), N-(6-bromopyridin-2-yl)methanesulfonamide (310 mg, 1.17 mmol), Cul (230 mg, 1.17 mmol), K2CO3 (240 mg, 1.75 mmol), 1,4-dioxane (12 mL), N 1 , N 2 -dimethylcyclohexane-1,2-diamine (190 mg, 1.29 mmol), Ar protection, 100 °C for 2 h, LC-MS monitoring, reaction was completed. The solid product was obtained by reversed column (308 mg, yield 77%), LC-MS: 342.0 [M+H] + .

[0107] Using different starting materials, the synthesis of intermediate A3-1 can be used to obtain intermediates A3-2 to A3-27.

[0108] Table 1. Structural formula of intermediates A3-2 to A3-27

[0109]

[0110]

[0111]

[0112] Preparation Example 2: Preparation of 2-methyl-2,3,7,8,9,9a-hexahydro-1H-phenylpropane[de]isoquinoline-5-amine (intermediate B7-1)

[0113]

[0114] Step 1: Synthesis of compound B2-1:

[0115] B1-1 (50 g, 284 mmol), methylamine hydrochloride (57.5 g, 851 mmol), and TEA (144 g, 1.42 mol, 197 mL) were dissolved in acetonitrile (600 mL). T3P (217 g, 341 mmol, 203 mL, 50% purity) was added dropwise at room temperature. After the addition was complete, the mixture was heated to 50 °C and reacted for 16 hours. The reaction solution was diluted with 1500 mL of ethyl acetate and washed with three 400 mL solutions of NaHCO3. The organic phase was dried over anhydrous sodium sulfate. The organic phase was filtered and distilled under reduced pressure to give a crude product as a white solid (50 g, 264 mmol, yield: 93.1%). The crude product could be used directly in the next reaction step.

[0116] 1 H NMR: (400MHz, CDCl3) δ: 7.12-7.01 (m, 3H), 6.10-5.71 (m, 1H), 2.93 (d, J = 4.9Hz, 3H), 2.83 (br s,2H),2.79-2.71(m,2H),1.84-1.65(m,4H),MS(ESI):190.1[M+H] + .

[0117] Step 2: Synthesis of compound B3-1:

[0118] B2-1 (50g, 264mmol) was dissolved in THF (500mL) and slowly added dropwise at -23°C under nitrogen protection. n BuLi (2.5M, 275mL). DMF (48.3g, 660mmol, 50.8mL) was then slowly added dropwise at -23℃. HCl solution (6M, 300mL) was then slowly added dropwise at 20℃. The reaction mixture was diluted with water (100mL), extracted with ethyl acetate (500mL x 3), and the organic phase was dried over anhydrous sodium sulfate. The organic phase was filtered and concentrated under reduced pressure to give a yellow solid (55g, crude product). The crude product can be used directly in the next reaction step.

[0119] 1H NMR: (400 MHz, CDC13) δ: 8.36-8.20 (m, 1H), 7.46-7.32 (m, 2H), 6.84 (s, 1H), 3.63-3.52 (m, 3H), 2.99-2.92 (m, 3H), 2.75-2.69 (m, 2H), 2.01-1.90 (m, 2H), MS (ESI): 200.1 [M+H] + .

[0120] Step 3: Synthesis of compound B4-1:

[0121] B3-1 (55 g, 276 mmol) and 20 g of palladium on carbon were suspended in methanol (800 mL) and stirred at 30 °C under hydrogen pressure (50 psi) overnight. The palladium on carbon was removed by filtration and the filtrate was concentrated under reduced pressure. Column chromatography (Si02, PE / EtOAc = 1 / 0 to 3 / 1) gave a yellow solid (38.5 g, yield: 69.3%).

[0122] 1 H NMR: (400 MHz, DMSO-d6) δ: 7.71-7.62 (m, 1H), 7.28-7.20 (m, 2H), 3.42 (dd, J = 5.6, 11.9 Hz, 1H), 3.25 (t, J = 12.5 Hz, 1H), 3.13-2.99 (m, 4H), 2.87-2.69 (m, 2H), 2.06-1.90 (m, 2H), 1.75-1.61 (m, 1H), 1.41-1.22 (m, 1H), MS (ESI): 202.1 [M+H] + .

[0123] Step 4: Synthesis of compound B5-1:

[0124] B4-1 (3.1 g, 19.2 mmol) was dissolved in H2SO4(300 mL) and KNO3(17.9 g, 177 mmol) was added slowly at 0 °C over 3 hours. After the addition was completed, the reaction was stirred at room temperature for 2 hours. TLC detection showed that the reaction was complete. The reaction solution was diluted with 500 mL of water, and a large amount of solid precipitated. The precipitate was filtered to obtain a yellow solid (79 g, crude product). The crude product can be directly used in the next step reaction, MS (ESI): 247.1 [M+H] + .

[0125] Step 5: Synthesis of compound B6-1:

[0126] B5-1 (4.9, 19.9 mmol) and palladium on carbon (2 g, 19.9 mmol, 10% purity) were suspended in methanol (100 mL) and reacted under hydrogen pressure (50 psi) at 25 °C for 16 h. The palladium on carbon was removed by filtration and the filtrate was concentrated under reduced pressure and column chromatography (SiO2, PE / EA = 1 / 0 to 1 / 2) to give yellow solid B6-1 (1.44 g, yield: 33.5%).

[0127] 1 H NMR: (400 MHz, CDC13) δ: 7.24 (d, J = 2.3 Hz, 1H), 6.56 (d, J = 2.0 Hz, 1H), 3.67 (br s, 2H), 3.32-3.25 (m, 2H), 3.19-3.13 (m, 3H), 3.09-2.97 (m, 1H), 2.81-2.65 (m, 2H), 2.07-1.90 (m, 2H), 1.78-1.62 (m, 1H), 1.37-1.23 (m, 1H), MS (ESI): 217.2 [M+H] + .

[0128] Step 6: Synthesis of compound B7-1:

[0129] B6-1 (7 g, 32.4 mmol) was dissolved in anhydrous tetrahydrofuran (300 ml) and LiAlH4(6.14 g, 162 mmol) was added at 0 °C. The mixture was warmed to 25 °C for 2 h under nitrogen protection. The reaction was quenched by slowly adding water to the reaction mixture, keeping the temperature of the reaction mixture at 0-10 °C during the process. The reaction mixture was diluted with 800 mL of ethyl acetate and washed with water (100 mL*3). The organic phase was dried over anhydrous sodium sulfate. The organic phase was filtered and distilled under reduced pressure to obtain the crude product. The crude product was column chromatographed (SiO2, DCM / (MeOH+1% NH4OH) = 10 / 0 to 10 / 1) to obtain yellow oil (6.25 g, yield: 95.5%).

[0130] 1 H NMR: (400 MHz, CDC13) δ: 6.31 (s, 1H), 6.21 (s, 1H), 3.88 (d, J = 15.1 Hz, 1H), 3.62-3.34 (br s, 2H), 3.26 (d, J = 15.1 Hz, 1H), 2.98-2.69 (m, 4H), 2.42 (s, 3H), 2.03 (t, J = 10.7 Hz, 1H), 1.92 (tdd, J = 3.4, 6.5, 13.1 Hz, 1H), 1.88-1.75 (m, 2H), 1.34-1.17 (m, 1H), MS (ESI): 203.2 [M+H] + .

[0131] Step 7: Preparation of compound B7-2 and compound B7-3:

[0132]

[0133] B7-1 (1.5 g, 7.41 mmol) was subjected to chiral resolution with prep supercritical fluid chromatography (prep SFC) (SFC chiral resolution conditions: instrument: Waters SFC350; column: DAICEL CHIRALPAK AD (250 mm*50 mm, 10 um); mobile phase: A: CO2, B: IPA (0.1% NH3H2O); gradient: B%: 50%-50%; flow rate: 200 ml / min; column temperature: 40 °C), and the fractionated liquids were concentrated under reduced pressure, lyophilized to obtain yellow oil B7-2 (peak 1, 438 mg, yield: 29.20%) and yellow oil B7-3 (peak 2, 450 mg, yield: 30.00%).

[0134] B7-2: 1 H NMR: (400 MHz, CDCl3) δ: 6.32 (s, 1H), 6.22 (s, 1H), 3.88 (d, J = 15.1 Hz, 1H), 3.48 (br s, 2H), 3.26 (br d, J = 15.1 Hz, 1H), 2.93 (dd, J = 4.6, 10.5 Hz, 1H), 2.90-2.80 (m, 1H), 2.79-2.64 (m, 2H), 2.42 (s, 3H), 2.02 (t, J = 10.7 Hz, 1H), 1.92 (dtd, J = 3.6, 6.5, 9.8 Hz, 1H), 1.87-1.79 (m, 2H), 1.36-1.15 (m, 1H)

[0135] MS (ESI): 203.2 [M+H] + .

[0136] B7-3: 1H NMR: (400 MHz, CDC13) δ: 6.32 (s, 1H), 6.22 (s, 1H), 3.87 (d, J = 15.3 Hz, 1H), 3.47 (br s, 2H), 3.26 (d, J = 15.1 Hz, 1H), 2.93 (dd, J = 4.8, 10.6 Hz, 1H), 2.89-2.80 (m, 1H), 2.80-2.65 (m, 2H), 2.42 (s, 3H), 2.02 (t, J = 10.7 Hz, 1H), 1.97-1.88 (m, 1H), 1.87-1.75 (m, 2H), 1.35-1.17 (m, 1H), MS (ESI): 203.2 [M+H] + .

[0137] Using different starting materials, the synthesis of similar intermediate B7-1 can give intermediates B7-4 to B7-39.

[0138] Table 2. Structural formulas of intermediates B7-4 to B7-39

[0139]

[0140]

[0141] Example 1 (6-(5-fluoro-2-((2-methyl-2,3,7,8,9,9a-hexahydro-1H- benzo[de]isoquinolin-5-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyridin-2- yl)iminio)dimethyl-λ 6 Synthesis of (6-(5-fluoro-2-((2-methyl-2,3,7,8,9,9a-hexahydro-1H- benzo[de]isoquinolin-5-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyridin-2- yl)iminio)dimethyl-λ

[0142]

[0143] ((6-(2-chloro-5-fluoro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyridin-2- yl)iminio)dimethyl-λ 6- sulfonamide ketone (200 mg, 0.59 mmol), cesium carbonate (289 mg, 0.89 mmol) were weighed into a 100 mL single neck flask, 1,4-dioxane (15 mL) was added, argon was purged for three times, Pd2(dba)3(27 mg, 0.03 mmol) and Xantphos (41 mg, 0.07 mmol) were added, the flask was heated to 100 °C under argon protection for 5 hours. LC-MS monitoring, the reaction was completed. The reaction solution was rotary dried, and purified by reverse phase column to obtain a light yellow solid ((6-(5-fluoro-2-((2-methyl-2,3,7,8,9,9a-hexahydro-1H-benzo[de] isoquinolin-5-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)pyridin-2-yl)imino)dimethyl-λ 6 - sulfonamide ketone (50 mg, 17%).

[0144] 1 H NMR (400 MHz, CDCl3) δ: 8.63 (s, 1H), 8.17 (s, 1H), 7.63 (d, J = 27.8 Hz, 2H), 7.13 (s, 2H), 6.60 (s, 1H), 3.96-3.92 (m, 1H), 3.34 (s, 7H), 2.88-2.84 (m, 4H), 2.41 (s, 3H), 2.03-1.82 (m, 3H), 1.23-1.21 (m, 2H), LC-MS: 506.3 [M+H] + .

[0145] Example 2 - Synthesis of compound 2-70

[0146] Similar to the synthesis of compound 1, using intermediates A3-1 to A3-27 and B7-1 to B7-39 as reaction materials, the target compounds 2-70 in Table 3 can be obtained.

[0147] Table 3 Structure of compound 2-70

[0148]

[0149]

[0150]

[0151]

[0152]

[0153] Example 71 Determination of the inhibition of Wee-1 enzyme activity by compounds of the present application

[0154] After the gradient dilution of the compound and enzyme mixture, incubate at room temperature (25°C) for 15 minutes, mix by centrifugation at 1000 rpm for 1 minute, add 5 μL of substrate to start the reaction. After 60 minutes of reaction at room temperature, add 5 μL of ADP-GLO reagent, mix by centrifugation at 1000 rpm for 1 minute, continue to incubate at room temperature for 60 minutes, then add 10 μL of kinase detection reagent and incubate for 60 minutes, and detect chemiluminescence. Compared with the DMSO group, calculate the percentage of inhibition of enzyme activity of the compound, and then calculate the IC 50 .

[0155] Table 4. IC of the compound of the present application for inhibiting Wee-1 kinase activity 50 (nM)

[0156] Compound IC 50 (nM) Compound IC 50 (nM) Compound IC 50 (nM) 1 1.52 2 1.96 3 2.48 4 2.47 5 2.79 14 2.71 19 2.08 21 1.67 23 2.05 24 2.49 28 2.33 63 3.47 64 3.36 65 1.56 66 1.19 67 1.86 68 1.97 69 1.54 70 1.84 MK-1775 4.38

[0157] As can be seen from the data in Table 4, the compounds of the present application have strong inhibitory effect on Wee-1 kinase, for example, the IC 50 values of compounds 1, 2, 21, 65, 66, 67, 68, 69 and 70, etc. on Wee-1 kinase are all less than 2.0 nM, which is about 2 times higher than the control drug MK-1775.

[0158] Example 72 In vitro anti-proliferative activity of the compound of the present application on MIA PaCa-2 cells

[0159] 3000 cells / well of MIA PaCa-2 cells were plated in 384-well plates, after overnight attachment, DMSO or the compound diluted at a gradient of 1:5 with the highest concentration of 5 μM was added. 72 hours after drug addition, cell survival was evaluated by measuring intracellular ATP content. Compared with the DMSO group, the percentage of inhibition of cell survival of the compound was calculated, and the IC 50 value was calculated, and the results are shown in Table 5 below.

[0160] Example 73 In vitro anti-proliferative activity of the compound of the present application combined with Gemcitabine (GMC) on MIA PaCa-2 cells

[0161] 3000 cells / well of MIA PaCa-2 cells were plated in 384-well plates and 20 nM of Gemcitabine was added, after overnight attachment, DMSO or the compound diluted at a gradient of 1:5 with the highest concentration of 100 nM was added. 72 hours after drug addition, cell survival was evaluated by measuring intracellular ATP content. Compared with the DMSO group, the percentage of inhibition of cell survival of the compound was calculated, and the IC 50 value was calculated, and the results are shown in Table 5 below.

[0162] Table 5 Anti-proliferative activity of the compound of the present application alone or in combination with GMC on MIA PaCa-2 cells

[0163]

[0164]

[0165] As shown in Table 5, compared with the control drug MK-1775, the compounds of this invention exhibit stronger anti-proliferative activity against MIA PaCa-2 cells. Furthermore, the compounds of this invention demonstrate stronger activity when used in combination with GMC, such as the IC50 values ​​of compounds 66, 67, and 68. 50 Less than 1 nM. The compounds of this invention exhibit stronger combination activity with GMC, suggesting that they may have better efficacy when used in combination with chemotherapy drugs in clinical practice.

[0166] 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) or a pharmaceutically acceptable salt or optical isomer thereof: In general formula (1): m is 0 or 1; R 1 It is H or halogen; R 2 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; R 3 It is H or C1-C3 alkyl; A is phenyl, pyridyl, pyrimidinyl, pyridazinyl, or pyrazinyl, wherein the phenyl, pyridyl, pyrimidinyl, pyridazinyl, or pyrazinyl group is optionally surrounded by 1-3 R groups. 6 Replace, each R 6 Independently, H, halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, OH-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, halogen-substituted C3-C6 cycloalkyl, hydroxyl-substituted C3-C6 cycloalkyl, cyano-substituted C3-C6 cycloalkyl, CF3-substituted C3-C6 cycloalkyl, -NR 7a R 7b -N=S(O)R 7a R 7b -P(O)R 7a R 7b -S(O)2R 7a -S(O)2NR 7a R 7b -NR 8 P(O)R 7a R 7b -NR 8 S(O)2R 7a -NR 8 C(O)R 7a -N = S( = NR) 8 )R 7a R 7b Or pyridone group; R 7a and R 7b Independently, it is a C1-C3 alkyl, deuterated C1-C3 alkyl, C2-C6 alkenyl, C2-C6 ynyl or C3-C6 cycloalkyl, or R 7a and R 7b Together with the nitrogen, sulfur, or phosphorus atoms to which they are attached, they form (3-10) heterocyclic alkyl groups; R 8 It is H or C1-C3 alkyl, or R 8 and R 7a Together with the nitrogen and sulfur atoms or nitrogen and carbon atoms to which they are attached, they form (3-10) heterocyclic alkyl groups; B is a partially unsaturated C5-C7 cycloalkyl or a partially unsaturated (5-7 member) heterocycloalkyl.

2. The compound of claim 1 or a pharmaceutically acceptable salt or optical isomer thereof, wherein in the general formula (1), Where R 2 For H, Me, Et, CD3, 3. The compound of claim 2 or a pharmaceutically acceptable salt or optical isomer thereof, wherein in general formula (1), 4. The compound of claim 1 or a pharmaceutically acceptable salt or optical isomer thereof, wherein the compound has the structure of general formula (1A): In general formula (1A): n is 1, 2, or 3; X is CH2, O, or S; m, A, R 1 and R 2 The definition is as defined in claim 1.

5. The compound of claim 1 or 4, or a pharmaceutically acceptable salt or optical isomer thereof, wherein in general formula (1) or general formula (1A), A is phenyl, pyridyl, pyrimidinyl, or pyrazinyl, and the phenyl, pyridyl, pyrimidinyl, or pyrazinyl group is optionally surrounded by 1-3 R groups. 6 Replace, each R 6 Independently, H, halogen, CN, C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, halogen-substituted C1-C6 alkyl, halogen-substituted C1-C6 alkoxy, hydroxyl-substituted C1-C6 alkyl, cyano-substituted C1-C6 alkyl, halogen-substituted C3-C6 cycloalkyl, OH-substituted C3-C6 cycloalkyl, cyano-substituted C3-C6 cycloalkyl, CF3-substituted C3-C6 cycloalkyl, -NR 7a R 7b -N=S(O)R 7a R 7b -P(O)R 7a R 7b -S(O)2R 7a -S(O)2NR 7a R 7b -NR 8 P(O)R 7a R 7b -NR 8 S(O)2R 7a -NR 8 C(O)R 7a -N = S( = NR) 8 )R 7a R 7b Or pyridone group, where R 7a and R 7b Independently, it is a C1-C3 alkyl, deuterated C1-C3 alkyl, C2-C6 alkenyl, C2-C6 ynyl or C3-C6 cycloalkyl, or R 7a and R 7b Together with the nitrogen, sulfur, or phosphorus atoms to which it is attached, it forms a (3-10) heterocyclic alkyl group; R 8 It is H or C1-C3 alkyl, or R 8 and R 7a Together with the nitrogen and sulfur atoms or nitrogen and carbon atoms to which they are attached, they form (3-10) heterocyclic alkyl groups.

6. The compound of any one of claims 1-4, or a pharmaceutically acceptable salt or optical isomer thereof, wherein in general formula (1) or general formula (1A), A is... Where v is 1, 2, or 3, and each R 6 Independent of H, halogen, Me, Et, 7. The compound of claim 6 or a pharmaceutically acceptable salt or optical isomer thereof, wherein in general formula (1) or general formula (1A), A is...

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

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

10. The use of a compound as described in any one of claims 1-8, or a pharmaceutically acceptable salt or optical isomer thereof, or a pharmaceutical composition as described in claim 9, in the preparation of a medicament for treating Wee-1-mediated related diseases.

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