Pyrazolo[3,4-d]pyrimidin-3-one derivatives as Wee-1 inhibitors

Abstract

The present invention relates to novel compounds of general formula (1) and / or pharmaceutically acceptable salts thereof, compositions containing compounds of general formula (1) and / or pharmaceutically acceptable salts thereof, processes for their preparation, and their use as Wee-1 inhibitors in the preparation of antitumor drugs. [Formula 1] TIFF2023531431000120.tif44168

Description

[Technical Field]

[0001] This application claims priority to Chinese Patent Application No. 202010557580.X, filed on June 17, 2020, which is incorporated herein by reference in its entirety.

[0002] The present invention relates to the field of medicinal chemistry, and in particular to novel compounds having inhibitory activity against Wee-1 kinase, a process for their preparation, and the use of the compounds in the preparation of antitumor drugs. [Background technology]

[0003] Wee-1 protein kinase is an important negative regulatory protein in cell cycle checkpoints. These checkpoints include the G1 checkpoint, which marks the transition from G1 (resting phase) to S (DNA synthesis phase), the G2 checkpoint, which marks the transition from G2 (preparatory phase for cell division) to M (mitotic phase), and the spindle checkpoint, which marks the transition from metaphase (metaphase) to anaphase (anaphase). Wee-1 protein kinase plays an important role in the G2 checkpoint. Transition to M phase depends on CDK1 kinase activity, and Wee-1 phosphorylates Tyr15 of the CDK1 protein, inhibiting CDK1 activity and preventing cells from entering M phase (mitotic phase). Meanwhile, polo kinase phosphorylates Wee-1, activating its degradation and promoting cell transition to M phase. Thus, the kinase activity of Wee-1 determines the activity of the G2 checkpoint, which regulates the transition of cells from G2 to M phase.

[0004] Cell cycle checkpoints are activated primarily after DNA damage and play a critical role in intracellular DNA repair. When cell cycle checkpoints are activated normally, the cell cycle is inhibited and DNA repair is promoted. When checkpoint function is inhibited, DNA damage cannot be repaired and cells undergo apoptosis. Compared to normal cells, several tumor cells rely primarily on activation of the G2 checkpoint to repair DNA damage and avoid apoptosis due to impaired function of p53, a key protein in the G1 checkpoint. Therefore, inhibiting the G2 checkpoint can selectively kill tumor cells. The crucial role of Wee-1 kinase activity in the G2 checkpoint suggests that Wee-1 kinase determines whether tumor cells repair or die after DNA damage, and that inhibiting Wee-1 activity induces tumor cells that fail to repair DNA damage to transition to the M phase and undergo apoptosis.

[0005] In addition to its role in the G2 checkpoint, studies have shown that Wee-1 is involved in functions closely related to tumor initiation and progression, such as DNA synthesis, DNA homologous repair, and post-translational modification of chromosomal histones. Wee-1 expression is highly elevated in many tumors, including liver cancer, breast cancer, cervical cancer, melanoma, and lung cancer. High Wee-1 expression is positively correlated with tumor development or poor prognosis, suggesting that Wee-1 kinase may be involved in tumor initiation and progression. Studies using in vitro cell models and in vivo animal models have shown that inhibiting Wee-1 activity while inducing DNA damage significantly inhibits the growth of various tumors.

[0006] Therefore, the development of specific and highly active small molecule inhibitors of Wee-1 kinase would have significant clinical value for tumor therapy, particularly targeting tumors with compromised G1 checkpoints, such as those with p53 deletion.

[0007] AstraZeneca's Wee-1 inhibitor AZD-1775 has entered Phase II clinical trials, with over 30 clinical trials underway and demonstrating favorable therapeutic effects. Patents related to AZD-1775 include U.S. Patent Application Publication No. 20070254892, International Publication No. 2007126122, European Patent Application Publication No. 2213673, International Publication No. 2008133866, and International Publication No. 2011034743. Abbott and Abbvie are also conducting research on Wee-1 inhibitors. Related patents include U.S. Patent Application Publication No. 2012220572, International Publication No. 2013126656, International Publication No. 2013012681, International Publication No. 2013059485, and International Publication No. 2013013031. Almac's patents related to Wee-1 inhibitors include WO 2014167347, WO 2015019037, WO 2015092431, WO 2018011570, WO 2018062932, and WO 2019138227. Girafpharma's patents related to Wee-1 include WO 2019074979 and WO 2019074981. Zeno's patents related to Wee-1 research include WO 2018028008 and WO 2019173082. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] U.S. Patent Application Publication No. 20070254892 [Patent Document 2] International Publication No. 2007126122 [Patent Document 3] European Patent Application Publication No. 2213673, [Patent Document 4] International Publication No. 2008133866 [Patent Document 5] International Publication No. 2011034743 [Patent Document 6] U.S. Patent Application Publication No. 2012220572 [Patent Document 7] International Publication No. 2013126656 [Patent Document 8] International Publication No. 2013012681 [Patent Document 9] International Publication No. 2013059485 [Patent Document 10] International Publication No. 2013013031 [Patent Document 11] International Publication No. 2014167347 [Patent Document 12] International Publication No. 2015019037 [Patent Document 13] International Publication No. 2015092431 [Patent Document 14] International Publication No. 2018011570 [Patent Document 15] International Publication No. 2018062932 [Patent Document 16] International Publication No. 2019138227 [Patent Document 17] International Publication No. 2019074979 [Patent Document 18] International Publication No. 2019074981 [Patent Document 19] International Publication No. 2018028008 [Patent Document 20] International Publication No. 2019173082 Summary of the Invention [Problem to be solved by the invention]

[0009] Currently, there are several issues with the Wee-1 inhibitors under investigation, such as the metabolic properties of AZD-1775, which leaves room for further optimization. [Means for solving the problem]

[0010] (overview) The present invention provides a compound having a structure represented by general formula (1), or an isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate thereof. [ka] During the ceremony, m is an integer of 1, 2, or 3; X is N or CH; A is a divalent or higher aryl, a divalent or higher heteroaryl, a divalent or higher cycloalkyl-aryl, a divalent or higher heterocycloalkyl-aryl, or a divalent or higher heterocycloalkyl-heteroaryl; R 1 is C1-C6 alkyl, halogen-substituted C1-C3 alkyl, C3-C6 cycloalkyl, —CH2(C3-C6)cycloalkyl, or C3-C5 alkenyl, R 2 is a C1-C6 alkyl, a C3-C6 cycloalkyl, or a (4- to 6-membered) heterocycloalkyl, wherein said alkyl, said cycloalkyl, and said heterocycloalkyl are optionally substituted with 1 to 3 of the following groups: H, halogen, OH, Me, or OMe; R 3 is H, halogen, CN, C1-C3 alkyl, halogen-substituted C1-C3 alkyl or C1-C3 alkoxy, Each R 4 are independently H, halogen, C1-C3 alkyl, halogen-substituted C1-C3 alkyl, hydroxyl-substituted C1-C3 alkyl, C1-C3 alkoxy, NMe2-substituted C1-C3 alkyl, NMe2-substituted C1-C3 alkoxy, [ka] NMe2, C3-C6 cycloalkyl, (4-12 membered)heterocycloalkyl or —CH2(4-12 membered)heterocycloalkyl, wherein the (4-12 membered)heterocycloalkyl is selected from the group consisting of 1 to 3 R 5 and each R 5are independently H, halogen, CN, OH, C1-C3 alkyl, halogen-substituted C1-C3 alkyl, hydroxyl-substituted C1-C3 alkyl, cyano-substituted C1-C3 alkyl, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, [ka] NR 6 R 7 or -(C1-C3 alkyl)-NR 6 R 7 where R 6 and R 7 are independently H or C1-C3 alkyl, or R 6 and R 7 together with the N atom to which they are both attached form a 4- to 7-membered heterocycloalkyl, where two R 4 can form a C2-C3 alkylene together with the C atom, where R 4 or R 5 When attached to a heteroatom, cannot be a halogen.

[0011] In some embodiments of the present invention, R 1 Me, Et, [ka] is.

[0012] In some embodiments of the present invention, R 2 Me, Et, [ka] is.

[0013] In some embodiments of the present invention, R 3 H, F, Me, Et, [ka] CF3, OMe or OEt.

[0014] In some embodiments of the present invention, [ka] is the following group: [ka] wherein m is an integer of 1, 2, or 3, and each R 4 are independently H, halogen, C1-C3 alkyl, halogen-substituted C1-C3 alkyl, hydroxyl-substituted C1-C3 alkyl, C1-C3 alkoxy, NMe2-substituted C1-C3 alkyl, NMe2-substituted C1-C3 alkoxy, [ka] NMe2, C3-C6 cycloalkyl, (4-12 membered)heterocycloalkyl or —CH2(4-12 membered)heterocycloalkyl, wherein the (4-12 membered)heterocycloalkyl is selected from the group consisting of 1 to 3 R 5 and R 5 are independently H, halogen, CN, OH, C1-C3 alkyl, halogen-substituted C1-C3 alkyl, hydroxyl-substituted C1-C3 alkyl, cyano-substituted C1-C3 alkyl, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, [ka] NR 6 R 7 or -(C1-C3 alkyl)-NR 6 R 7 where R 6 and R 7 are independently H or C1-C3 alkyl, or R 6 and R 7 together with the N atom to which they are both attached form a 4- to 7-membered heterocycloalkyl, where two R 4 can form a C2-C3 alkylene together with the C atom, where R 4 or R 5When connected to a heteroatom, it cannot be a halogen.

[0015] In some embodiments of the present invention, R 4 are independently H, F, Cl, Me, Et, [ka] CF3, CH2CF3, CH2OH, CH2CH2OH, OMe, OEt, [ka] NMe2, [ka] where two R 4 together with the C atom, spirocyclopropyl [ka] or spirocyclobutyl [ka] where R 5 is H, halogen, CN, OH, C1-C3 alkyl, halogen-substituted C1-C3 alkyl, hydroxyl-substituted C1-C3 alkyl, cyano-substituted C1-C3 alkyl, C3-C6 cycloalkyl, halogen-substituted C3-C6 cycloalkyl, NH2, NHMe, NMe2, [ka] is.

[0016] In some embodiments of the present invention, [ka] is the following group: [ka] TIFF0007796674000019.tif248168TIFF0007796674000020.tif248168TIFF0007796674000021.tif248168TIFF0007796674000022.tif91168.

[0017] In some embodiments of the invention, the compound, isomer or pharmaceutically acceptable salt is [ka] TIFF0007796674000024.tif248168TIFF0007796674000025.tif248168TIFF0007796674000026.tif248168TI FF0007796674000027.tif248168TIFF0007796674000028.tif248168TIFF0007796674000029.tif248168TIFF Selected from 0007796674000030.tif248168TIFF0007796674000031.tif248168TIFF0007796674000032.tif248168TIFF0007796674000033.tif248168TIFF0007796674000034.tif248168TIFF0007796674000035.tif248169.

[0018] The present invention further contemplates providing a pharmaceutical composition comprising, as an active ingredient, a pharmaceutically acceptable excipient or carrier and a compound of general formula (1) of the present invention or an isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate thereof.

[0019] The present invention further contemplates the use of a compound of the present invention, or an isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate thereof, in preparing a medicament for treating a related disease mediated by Wee-1.

[0020] It is to be understood that both the foregoing general description and the following detailed description of the invention are exemplary and explanatory and are intended to provide further explanation of the invention as claimed. DETAILED DESCRIPTION OF THE INVENTION

[0021] Definitions and Explanations As used herein, the following terms and phrases are intended to have the following meanings unless otherwise specified: A particular term or phrase should not be considered uncertain or unclear and should be construed according to its common definition unless specifically defined otherwise.

[0022] Reference herein to a trade name is intended to refer to the corresponding product name or its active ingredient. As used herein, the term "pharmaceutically acceptable" refers to those compounds, compositions and / or formulations that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0023] The term " pharmaceutically acceptable salt " refers to the form of a compound that does not cause significant stimulation to the living body due to drug administration or does not eliminate the biological activity and properties of the compound.In certain embodiments, pharmaceutically acceptable salts can be obtained by reacting the compound of general formula (1) with an acid, for example, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, phosphoric acid, etc., 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 and p-toluenesulfonic acid, acidic amino acids such as aspartic acid and glutamic acid.

[0024] It should be understood that pharmaceutically acceptable salts include solvent addition forms or crystalline forms, particularly solvates or polymorphs. Solvates contain stoichiometric or non-stoichiometric amounts of solvent and are selectively formed upon crystallization with pharmaceutically acceptable solvents such as water and ethanol. Hydrates are formed when the solvent is water, and alcoholates are formed when the solvent is ethanol. Solvates of compounds of general formula (1) are conveniently prepared or formed according to the methods described herein. For example, hydrates of compounds of general formula (1) are conveniently prepared by recrystallization from a water / organic solvent mixture, where the organic solvent used includes, but is not limited to, acetonitrile, tetrahydrofuran, ethanol, or methanol. Furthermore, the compounds referred to herein can exist in both unsolvated and solvated forms. In general, solvated forms are considered equivalent to unsolvated forms for the purposes of the compounds and methods provided herein.

[0025] In other specific examples, compounds of general formula (1) are prepared in various forms, including, but not limited to, amorphous, crushed, and nanoparticle forms. Furthermore, compounds of general formula (1) may include crystalline forms and may also be polymorphs. Polymorphs contain different lattice arrangements of the same elements of a compound. Polymorphs typically have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardnesses, crystalline forms, optical properties, electrical properties, stability, and solubility. Various factors, such as recrystallization solvents, crystallization rates, and storage temperatures, may result in the predominance of single crystals.

[0026] In another embodiment, the compounds of general formula (1) have one or more stereocenters and therefore occur in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds, and single diastereomers. The possible asymmetric centers are determined by the nature of the various substituents on the molecule. These asymmetric centers independently give rise to two optical isomers, and all possible optical isomers, diastereomeric mixtures, and pure or partially pure compounds are within the scope of the present invention. The present invention is meant to include all such isomers of these compounds.

[0027] Unless otherwise indicated, the absolute configuration of the stereocenter is determined by the wedge bond. [ka] and broken linear combinations [ka] and is expressed as a wedge or broken linear combination. [ka] is a wavy line [ka] It is expressed as:

[0028] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute said compounds. For example, the compounds may contain tritium ( 3 H), iodine-125( 125 I) and C-14( 14 C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.

[0029] The compounds of the present invention and their pharmaceutically acceptable salts can be prepared into various formulations containing the compounds disclosed herein or their pharmaceutically acceptable salts and pharmaceutically acceptable excipients or carriers in a safe and effective amount, where "safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious side effects. The safe and effective amount of the compound is determined according to the age, condition, course of treatment, and other specific conditions of the subject to be treated.

[0030] "Pharmaceutically acceptable excipient or carrier" refers to one or more compatible solid or liquid fillers or gel substances that are suitable for human use and must have sufficient purity and low toxicity. Examples of pharmaceutically acceptable excipients or carriers include cellulose and its derivatives (e.g., sodium carboxymethylcellulose, sodium ethylcellulose, or cellulose acetate), gelatin, talc, solid lubricants (e.g., stearic acid or magnesium stearate), calcium sulfate, vegetable oils (e.g., soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (e.g., Tween®), wetting agents (e.g., sodium lauryl sulfate), colorants, flavorings, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0031] When administered, the compounds of the present invention may be administered orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously) or topically.

[0032] Unless otherwise specified, "alkyl" refers to saturated aliphatic hydrocarbon groups, including straight-chain and branched groups, containing 1 to 6 carbon atoms. Lower alkyls containing 1 to 4 carbon atoms are preferred, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, or tert-butyl. As used herein, "alkyl" includes unsubstituted and substituted alkyls, particularly alkyls substituted with one or more halogens. Preferred alkyls are selected from CH3, CH3CH2, CF3, CHF2, CF3CH2, i-Pr, n-Pr, i-Bu, c-Pr, n-Bu, and t-Bu.

[0033] Unless otherwise specified, "alkylene" refers to a divalent alkyl as defined above. Alkylene also includes spirocycloalkyl. Examples of alkylene include methylene, ethylene, and the like. [ka] Spirocyclopropyl [ka] Spirocyclobutyl [ka] These include, but are not limited to:

[0034] Unless otherwise specified, "cycloalkyl" refers to a 3- to 14-membered all-carbon monocyclic aliphatic hydrocarbon group in which one or more rings may contain one or more double bonds, but none of them has a completely conjugated pi-electron system; examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexane, and cyclohexadiene.

[0035] Unless otherwise specified, the term "heterocycloalkyl" refers to a saturated or partially saturated non-aromatic cyclic group consisting of carbon atoms and heteroatoms selected from nitrogen, oxygen, and sulfur. The cyclic group may be monocyclic or polycyclic. In the present invention, the number of heteroatoms in the heterocycloalkyl is preferably 1, 2, 3, or 4, and the nitrogen atom, carbon atom, or sulfur atom in the heterocycloalkyl may optionally be oxidized. In addition, the nitrogen atom may optionally be further substituted with other groups to form a tertiary amine or a quaternary ammonium salt. Examples of heterocycloalkyl include, but are not limited to, aziridinyl, azetidin-1-yl, N-alkylazetidin-3-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, morpholin-4-yl, thiomorpholin-4-yl, thiomorpholin-S-oxide-4-yl, piperidin-1-yl, N-alkylpiperidin-4-yl, pyrrolidin-1-yl, N-alkylpyrrolidin-2-yl, piperazin-1-yl, 4-alkylpiperazin-1-yl, and the like.

[0036] Unless otherwise specified, "alkoxy" refers to an alkyl group attached to the remainder of the molecule via an ether oxygen atom. Representative alkoxy groups are those containing 1 to 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, particularly alkoxy substituted with one or more halogens. Preferred alkoxy is selected from OCH3, OCF3, CHF2O, CF3CHO, i-PrO, n-PrO, i-BuO, n-BuO, and t-BuO.

[0037] Unless otherwise specified, "aryl" refers to a monocyclic or polycyclic aromatic hydrocarbon group. For example, a monocyclic aryl ring may be fused with one or more carbocyclic aromatic groups. Examples of aryl include, but are not limited to, phenyl, naphthyl, and phenanthryl.

[0038] Unless otherwise specified, "heteroaryl" refers to an aromatic group containing one or more heteroatoms (O, S, or N), which may be monocyclic or polycyclic; for example, a monocyclic heteroaryl ring may be fused to one or more carbocyclic aromatic groups or other monocyclic heterocyclyl groups. Examples of heteroaryl include, but are not limited to, pyridyl, pyridazinyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, quinolinyl, isoquinolinyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, indolyl, benzimidazolyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, benzopyridyl, and pyrrolopyrimidinyl.

[0039] Unless otherwise specified, "alkenyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon double bond, including straight-chain or branched groups containing 1 to 14 carbon atoms. Lower alkenyl containing 1 to 4 carbon atoms, such as vinyl, 1-propenyl, 1-butenyl, or 2-methylpropenyl, is preferred.

[0040] Unless otherwise specified, "alkynyl" refers to an unsaturated aliphatic hydrocarbon group containing a carbon-carbon triple bond, and includes straight-chain and branched groups containing 1 to 14 carbon atoms. Lower alkynyl groups containing 1 to 4 carbon atoms, such as ethynyl, 1-propynyl, or 1-butynyl, are preferred.

[0041] Unless otherwise stated, the terms "halogen-substituted" or "halogen," by themselves or as part of another substituent, refer to a fluorine, chlorine, bromine, or iodine atom. Additionally, "haloalkyl" is intended to include monohaloalkyl or polyhaloalkyl. For example, "halogenated C1-C3 alkyl" is intended to include, but is not limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 2-chloropropyl, 3-bromopropyl, and the like.

[0042] The term "membered ring" includes any cyclic structure. The term "membered" is intended to refer to the number of main chain atoms that form the ring. For example, cyclohexyl, pyridyl, pyranyl, and thiopyranyl are six-membered rings, and cyclopentyl, pyrrolyl, furanyl, and thienyl are five-membered rings.

[0043] The term "moiety" refers to a specific portion or functional group of a molecule. A chemical moiety is generally considered to refer to a chemical substance contained in or attached to a molecule.

[0044] "Optional" or "optionally" means that the subsequently described event or circumstance may occur, but does not necessarily occur, and the description includes instances in which the event or circumstance occurs and instances in which it does not occur.

[0045] Compound synthesis Methods for preparing the compounds of the general formula (1) of the present invention are specifically described below, but these specific methods do not limit the present invention.

[0046] The compounds of formula (1) above can be synthesized using standard synthetic techniques, well-known techniques, in combination with the methods described herein. Furthermore, the solvents, temperatures, and other reaction conditions described herein may vary. Starting materials for the synthesis of the compounds can be obtained synthetically or commercially. The compounds described herein and other related compounds with various substituents are described 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), and Green and Wuts, Protective Groups in Organic Synthesis, 3 rd Ed., (Wiley 1999). General methods for preparing compounds can be modified by using appropriate reagents and conditions to introduce various groups into the formulae described herein.

[0047] In one embodiment, the compounds described herein are prepared according to methods well known in the art. However, the conditions of the methods, such as reactants, solvents, bases, amounts of compounds used, reaction temperatures, and reaction times, are not limited to those described below. In addition, the compounds of the present invention can be easily prepared by any combination of various synthetic methods described herein or known in the art, and such combinations can be easily determined by those skilled in the art to which the present invention pertains. In one embodiment, the present invention also provides a method for preparing a compound of general formula (1) prepared using the following method A.

[0048] Method A comprises the following steps: first, reacting compound A1 with R 2 -Y to produce compound A2; coupling compound A2 with compound A3 to produce compound A4; and further reacting compound A4 with compound A5 to produce target compound A6; and [ka] However, if the compound contains protecting groups for the primary and secondary amines, it is necessary to further remove the protecting groups to obtain the target compound.

[0049] [ka]

[0050] In the above reaction formula, A and R 1 , R 2 , R 3 , R 4 and m is as defined above, Y is OH, Br or I, and Q is CH3S, CH3SO, CH3SO2, Br, Cl, I, etc.

[0051] therapeutic use The compounds or compositions described herein are generally useful for inhibiting Wee-1 kinase and may therefore be useful for treating one or more disorders associated with Wee-1 kinase activity. Accordingly, in certain embodiments, the present invention provides a method for treating a Wee-1 kinase-mediated disorder, comprising administering to a patient in need thereof a compound of the present invention or a pharmaceutically acceptable composition thereof.

[0052] Cancers that can be treated with the compounds of the invention include, but are not limited to, hematological malignancies (leukemia, lymphoma, myeloma including multiple myeloma, myelodysplastic syndromes, and myeloproliferative disorders), solid tumors (carcinomas such as prostate, breast, lung, colon, pancreatic, renal, ovarian, and soft tissue cancers, osteosarcoma, and stromal tumors).

[0053] (Detailed explanation) Various specific aspects, features and advantages of the above compounds, methods and pharmaceutical compositions are described in detail below, thereby clarifying the present invention. It should be understood that the following detailed description and examples describe specific embodiments for reference. After reading the description of the present invention, those skilled in the art can make various changes or modifications to the present invention, and such equivalents also fall within the scope of the present invention as defined herein.

[0054] In all examples, 1 H-NMR spectra were recorded on a Vian Mercury 400 nuclear magnetic resonance spectrometer, and chemical shifts were expressed in δ (ppm). Unless otherwise specified, 200–300 mesh silica gel was used for separation, and the ratio of eluents was by volume.

[0055] In the present invention, the following abbreviations are used: CDCl3 = deuterated chloroform; CuI = cuprous iodide; DCM = dichloromethane; DIPEA = diisopropylethylamine; DMF = dimethylformamide; EA = ethyl acetate; h = hour; K2CO3 = potassium carbonate; LC-MS = liquid chromatography mass spectrometry; m-CPBA = m-chloroperoxybenzoic acid; MeI (CHI) = methyl iodide; mL = milliliter; MeOH = methanol; min = minute; MS = mass spectrum; NaHCO3 = sodium bicarbonate; Na2SO4 = sodium sulfate; NMR = nuclear magnetic resonance; °C = degrees Celsius; PE = petroleum ether; rt = room temperature; TFA = trifluoroacetic acid; and toluene = methylbenzene. [Example]

[0056] Preparation Example 1: Preparation of 2-allyl-1-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Intermediate B1)

[0057] [ka]

[0058] Step 1: Synthesis of Compound A-1 In a 50 mL reaction flask, 6-bromo-3-pyridazinol (826 mg, 4.72 mmol) and K2CO3 (1.3 g, 9.44 mmol) were added to DMF (10 mL), followed by the addition of MeI (0.6 mL, 9.44 mmol). The mixture was stirred at room temperature, and the reaction was monitored by TLC (PE / EA = 1 / 1). After completion of the reaction, water (50 mL) was added to quench the reaction. The mixture was extracted with EA (50 mL × 2) to obtain an organic phase. The organic phase was washed with saturated brine (30 mL), dried over anhydrous Na2SO4, and distilled under reduced pressure to remove EA. Cold hydrazine was added to remove DMF. The residue was purified by column chromatography (PE / EA = 3 / 1) to obtain compound A-1 (705 mg, 79% yield). ESI-MS m / z: 189 [M+H] + .

[0059] Step 2: Synthesis of compound B-1 In a 50 mL reaction flask, compound A-1 (621 mg, 3.55 mmol) and 2-propenyl-6-(methylthio)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (607 mg, 2.73 mmol, see U.S. Patent No. 2019106427 for synthesis) were dissolved in dioxane (20 mL), followed by the addition of CuI (520 mg, 2.73 mmol) and KCO (528 mg, 2.73 mmol). The mixture was heated to 80 °C under a nitrogen atmosphere. N,N'-Dimethylethylenediamine (0.59 mL, 5.46 mmol) was added. The reaction was heated to 95 °C and stirred. The reaction was monitored by TLC (PE / EA = 1 / 1). After completion of the reaction, the reaction flask was cooled to room temperature. The mixture was distilled under reduced pressure and extracted with EA (50 mL x 2) to obtain an organic phase. The organic phase was washed with saturated brine (50 mL), dried over anhydrous Na2SO4, and distilled under reduced pressure. The residue was purified by column chromatography (PE / EA = 2 / 1) to obtain compound B-1 (309 mg, yield 34%). ESI-MS m / z: 333 [M+H]+.

[0060] The following intermediates B2 to B45 were obtained by the same procedure as in the synthesis of compound B-1.

[0061] [Table 1] TIFF0007796674000047.tif248168TIFF0007796674000048.tif248168TIFF0007796674000049.tif81168

[0062] Example 1: Synthesis of 2-allyl-1-(1-methyl-6-oxo-1,6-dihydropyridazin-3-yl)-6-((4-(4-methylpiperazin-1-yl)phenyl)amino)-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one (Compound 1)

[0063] [ka]

[0064] In a 50 mL reaction flask, compound B-1 (100 mg, 0.3 mmol) was dissolved in toluene (10 mL), and m-CPBA (76 mg, 0.33 mmol) was added. The mixture was stirred at room temperature for 1 hour, and DIPEA (0.2 mL, 1.58 mmol) and 4-(4-methylpiperazine)aniline (74.6 mg, 0.39 mmol) were added. The mixture was stirred at room temperature for 3 hours, and the reaction was monitored by TLC (DCM / MeOH = 10 / 1). After completion of the reaction, the mixture was extracted with EA (30 mL × 2) to obtain an organic phase. The organic phase was washed with saturated brine (30 mL), dried over anhydrous Na2SO4, and distilled under reduced pressure. The residue was purified by column chromatography (DCM / MeOH = 100 / 1) to obtain compound 1 (75 mg, 54% yield).

[0065] 1H NMR(400 MHz, CDCl3) δ 8.80 (s, 1H), 7.99 (s, 1H), 7.87 (d, J = 9.8 Hz, 1H), 7.43-7.31 (m, 2H), 7.03 (d, J = 9.9 Hz, 1H), 6.96-6.84 (m, 2H), 5.68 (ddt, J = 16.6, 10.1, 6.3 Hz, 1H), 5.15-4.95 (m, 2H), 4.60 (d, J = 6.3 Hz, 2H), 3.79 (s, 3H), 3.25-3.13 (m, 4H), 2.64-2.56 (m, 4H), 2.36 (s, 3H); ESI-MS m / z: 474 [M+H] + .

[0066] Examples 2 to 30: Synthesis of Compounds 2 to 30 Following the same procedure as in the synthesis of compound 1, target compounds 2-30 in Table 2 can be obtained using B2 to B30 as starting materials.

[0067] [Table 2] TIFF0007796674000052.tif248168

[0068] Example 31

[0069] [ka]

[0070] Step 1: Synthesis of compound C-1 In a 50 mL reaction flask, compound B-3 (107 mg, 0.3 mmol) was dissolved in toluene (10 mL), and M-CPBA (76 mg, 0.33 mmol) was added. The mixture was stirred at room temperature for 1 hour, and DIPEA (0.2 mL, 1.58 mmol) and tert-butyl 4-(4-aminobenzene)piperazine-1-carboxylate (100 mg, 0.36 mmol) were added. The mixture was stirred at room temperature for 3 hours, and the reaction was monitored by TLC (DCM / MeOH = 20 / 1). After completion of the reaction, the mixture was extracted with EA (30 mL × 2) to obtain an organic phase. The organic phase was washed with saturated brine (30 mL), dried over anhydrous Na2SO4, and distilled under reduced pressure. The residue was purified by column chromatography (DCM / MeOH = 100 / 1) to obtain compound C1 (135 mg, 77% yield). ESI-MS m / z: 588 [M+H] + .

[0071] Step 1: Synthesis of compound 31 In a 20 mL reaction flask, compound C-1 (117 mg, 0.2 mmol) was dissolved in DCM (5 mL), and TFA (1 mL) was added while cooling in an ice-salt bath. After the addition, the mixture was stirred at room temperature for 3 hours, and the reaction was monitored by TLC (DCM / MeOH = 20 / 1). After the reaction was completed, the mixture was diluted with DCM (50 mL), adjusted to alkaline with saturated NaHCO3 solution, and then the liquid was separated. The organic phase was washed with saturated brine (30 mL), dried over anhydrous Na2SO4, and distilled under reduced pressure. The residue was purified by column chromatography (DCM / MeOH = 50 / 1) to give compound 31 (58 mg, 59% yield).

[0072] 1H NMR (400 MHz, CDCl3) δ: 8.80 (s, 1H), 7.99 (s, 1H), 7.84 (d, J = 9.8 Hz, 1H), 7.43-7.34 (m, 2H), 7.00 (d, J = 9.8 Hz, 1H), 6.94-6.87 (m, 2H), 5.66 (ddt, J = 16.7, 10.1, 6.4 Hz, 1H), 5.40-5.28 (m, 1H), 5.08 (dd, J = 10.1, 1.3 Hz, 1H), 5.00 (dd, J = 17.1, 1.4 Hz, 1H), 4.66 (d, J = 6.4 Hz, 2H), 3.25-3.14 (m, 4H), 2.60 (t, J = 5.0 Hz, 4H), 1.38 (d, J = 6.7 Hz, 6H); ESI-MS m / z: 488 [M+H] + .

[0073] Examples 32 to 341: Synthesis of Compounds 32 to 341 By the same procedures as in the synthesis of Compound 1 and Compound 31, target Compounds 32 to 341 in Table 3 can be obtained using various intermediates in Table 1 as starting materials.

[0074] [Table 3] TIFF0007796674000055.tif248168TIFF0007796674000056.tif248168TIFF0007796 674000057.tif248168TIFF0007796674000058.tif248168TIFF0007796674000059.t if248168TIFF0007796674000060.tif248168TIFF0007796674000061.tif248168TIF F0007796674000062.tif248168TIFF0007796674000063.tif248168TIFF00077966740 00064.tif228168TIFF0007796674000065.tif248168TIFF0007796674000066.tif24 8168TIFF0007796674000067.tif248168TIFF0007796674000068.tif248168TIFF000 7796674000069.tif248168TIFF0007796674000070.tif248168TIFF00077966740000 71.tif248168TIFF0007796674000072.tif248168TIFF0007796674000073.tif248168

[0075] Example 342: Assay of inhibitory activity of compounds against Wee-1 kinase The inhibitory activity of compounds against Wee-1 kinase was measured using the Lanthra ScreenWee-1 kinase kit (Invitrogen). Five μL of compound diluted in a gradient with DMSO, 5 μL of Wee-1 kinase (final concentration 5 nM), 5 μL of a mixture of Eu-Anti-GST antibody (final concentration 2 nM), and 5 μL of kinase tracer 178 (final concentration 50 nM) were mixed thoroughly. The plate was incubated at room temperature for 1 hour and then read. The IC of the inhibitory activity of compounds against Wee-1 kinase was calculated in comparison with the DMSO solvent control group. 50 was calculated.

[0076] [Table 4] TIFF0007796674000075.tif248168TIFF0007796674000076.tif248168TIFF0007796674000077.tif248168

[0077] A is IC 50 indicates that the concentration is 30 nM or less. B is IC 50 indicates that the concentration is greater than 30 nM and less than 100 nM. C is IC 50 indicates that the concentration is greater than 100 nM.

[0078] As can be seen from the data in Table 4, the compounds of the present invention have strong inhibitory effects on Wee-1 kinase.

[0079] Example 343: Assay of antiproliferative activity against HT29 cells 3000 HT29 cells were seeded in a 384-well plate (Fisher 142762). After allowing the cells to adhere to the wall overnight, gradient diluted compounds were added. After 72 hours, CellTiter-Lumi (Beyotime C0068XL) was added to measure intracellular ATP content. Cell proliferation was assessed, and the IC of compound inhibition of cell proliferation was calculated. 50 was calculated.

[0080] [Table 5] TIFF0007796674000079.tif248168TIFF0007796674000080.tif248168TIFF0007796674000081.tif248168

[0081] A is IC 50 indicates that the concentration is 1 μM or less. B is IC 50 indicates that the concentration is greater than 1 μM and less than 3 μM. C is IC 50 indicates that the concentration is greater than 3 μM.

[0082] As can be seen from the data in Table 5, the compounds of the present invention have potent antiproliferative activity against HT-29 cells.

[0083] Example 344. Pharmacokinetic evaluation in mice The compound was administered by intravenous injection at a dose of 2 mg / kg and by oral gavage at a dose of 10 mg / kg (0.5% CMC-Na suspension). Fifteen male ICR mice were selected for each group, and blood samples were collected from three mice at three discrete time points: pre-dose, 5 min, 15 min, 30 min, 1 h, 3 h, 5 h, 8 h, 12 h, and 24 h post-dose. At each post-dose time point, 80 μL of blood was collected from the mouse orbit or heart. Whole blood samples were collected into tubes containing EDTA K2 and centrifuged (1500-1600 rpm) at 4°C for 10 min to separate plasma, which was then stored in a refrigerator at -90°C to -60°C for sample analysis. Plasma compound concentrations were measured by liquid chromatography-tandem mass spectrometry, and the corresponding pharmacokinetic parameters were obtained according to the plasma concentration-time curve.

[0084] [Table 6]

[0085] NA indicates no data available.

[0086] As can be seen from the table above, compound 3 has good oral absorption properties and its half-life (t 1 / 2 ), maximum plasma concentration (C max ), area under the drug-time curve (AUC 0-t ), oral bioavailability, metabolic parameters, etc. were all confirmed to be superior to those of the control drug AZD-1775. Good oral absorption is of great significance in terms of improving drug efficacy, reducing dosage, and reducing costs.

[0087] Further experiments have shown that other compounds of the present invention also have good oral absorption properties and their half-lives (t 1 / 2), maximum plasma concentration (C max ), area under the drug-time curve (AUC 0-t ), oral bioavailability and metabolic parameters were all proven to be superior to those of the control drug, AZD-1775.

Claims

1. A compound having a structure represented by general formula (1), or an optical isomer thereof, Acceptable salts, hydrates or solvates. 【Chemistry 1】 (In the formula, m is an integer of 1, 2, or 3; X is N or CH; A partial structure of the formula (1) shown below: 【Chemistry 2】 teeth, 【Transformation 3】 【change】 【change】 【change】 【change】 and R １ represents C1-C6 alkyl, halogen-substituted C1-C3 alkyl, C3-C6 cycloalkyl, —CH ２ (C3-C6)cycloalkyl or C3-C5 alkenyl; R ２ is a C1-C6 alkyl, a C3-C6 cycloalkyl, or a (4-6 membered) heterocycloalkyl, wherein said alkyl, said cycloalkyl, and said heterocycloalkyl are optionally substituted with 1 to 3 of the following groups: H, halogen, OH, Me, or OMe; and R ３ is H, halogen, CN, C1-C3 alkyl, halogen-substituted C1-C3 alkyl, or C1-C3 alkoxy.

2. R １ が、Me、Et、 【Chemistry 4】 or an optical isomer or a pharmaceutically acceptable salt thereof. Hydrates or solvates.

3. R ２ が、Me、Et、 【Transformation 5】 3. The compound of claim 1 or claim 2, wherein:

4. R ３ が、H、F、Me、Et、 【Transformation 6】 CF ３ 4. The compound according to any one of claims 1 to 3, wherein R is O, OMe, or OEt, or an optical isomer, pharmaceutically acceptable salt, hydrate, or solvate thereof.

5. The compound has the following structure: 【Transformation 7】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof, of the compound according to any one of claims 1 to 4, having one of the following formulas:

6. A pharmaceutical composition comprising a therapeutically effective amount of an active ingredient and a pharmaceutically acceptable adjuvant, wherein the active ingredient comprises the compound according to any one of claims 1 to 5, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof, and the pharmaceutically acceptable adjuvant is a pharmaceutically acceptable carrier, diluent and / or excipient.

7. Use of a compound according to any one of claims 1 to 5, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof, or use of a composition according to claim 6 in the preparation of a Wee-1 inhibitor.

8. Use of a compound according to any one of claims 1 to 5, or an optical isomer, pharmaceutically acceptable salt, hydrate or solvate thereof, or use of a composition according to claim 6, in preparing a medicament for treating a related disease mediated by Wee-1.

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