IDH mutant inhibitors and uses thereof

Abstract

The present invention discloses a class of IDH mutant inhibitors and their uses. In particular, the present invention relates to a class of compounds of general formula (1), their preparation methods, and the use of the compounds of general formula (1) or their optical isomers, crystalline forms, or pharma- ceutically acceptable salts as irreversible inhibitors of IDH mutants in the preparation of antitumor drugs. [Formula 1] TIFF2024524933000043.tif42168

Description

[Technical field]

[0001] This application claims priority to Chinese Application No. 202110661394.5, filed on June 15, 2021, which is incorporated by reference in its entirety herein.

[0002] The present invention relates to the field of medicinal chemistry, and in particular to a novel class of IDH mutant inhibitors, methods for their preparation, and uses thereof. [Background technology]

[0003] Isocitrate dehydrogenase (IDH) is a key enzyme involved in the tricarboxylic acid cycle. It catalyzes the conversion of isocitrate to α-ketoglutarate (α-KG), which is the rate-limiting step of the tricarboxylic acid cycle. There are three different isocitrate dehydrogenases in humans: IDH1, IDH2, and IDH3. IDH1 is mainly localized in the cytoplasm and peroxisomes, while IDH2 and IDH3 are mainly distributed in mitochondria.

[0004] IDH1 and IDH2 are the most common metabolic genes in the identification of human cancer genetic mutations, and IDH mutations are found in low-grade gliomas, secondary malignant gliomas, melanomas, angioimmunoblastic T-cell lymphomas, myeloproliferative neoplasms, myelodysplastic syndromes (MDS), and acute myelocytic leukemia (AML). The IDH mutation sites in tumor cells are IDH1 Arg132 (R132), IDH2 Arg172 (R172), or IDH2 Arg140 (R140). These mutations result in loss of function of the wild-type IDH protein and instead confer the ability to convert α-KG to the tumorigenic metabolite D-2-hydroxyglutarate (D-2HG). The oncogenic metabolite 2-HG inhibits DNA or histone demethylases, leading to DNA and histone hypermethylation and promoting cancer development. IDH inhibitors can reduce oncogenic metabolite D-2HG in vivo by inhibiting the activity of proteins with IDH1 / R132, IDH2 / R172 or IDH2 / R140 mutations, and induce histone H3K9me3 demethylation, thus suppressing tumor development. Therefore, targeting mutant IDH1 and IDH2 (mIDH1 and mIDH2) could be a promising approach for cancer therapy.

[0005] So far, several IDH small molecule inhibitors are commercially available, such as Enasidenib and Ivosidenib developed by Agios Pharmaceuticals Inc., both of which are non-covalent inhibitors. Covalent inhibitors of IDH have been reported by Eli Lilly and Company in WO 2017019429, WO 2017213910 and WO 2018111707, and have better selectivity for mutant IDH1 and IDH2 compared with wild-type IDH1 and IDH2. However, no IDH covalent inhibitors have entered the clinical stage, so there is a need to study and explore IDH covalent inhibitors with better activity and druggability. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] International Publication No. 2017019429 [Patent Document 2] International Publication No. 2017213910 [Patent Document 3] International Publication No. 2018111707 Summary of the Invention

[0007] The present invention provides a compound of general formula (1), or an isomer, crystalline form, pharma- ceutically acceptable salt, hydrate or solvate thereof: [ka] (In general formula (1), L is [ka] where " * " indicates the site of attachment to the carbonyl group, X is NH or NMe; R 1is Me, Et, -CH2CH2CH3, -CH(CH3)2, [ka] and R 2 and R 3 are independently H, Me or Et, or R 2 and R 3 along with the carbon atoms attached to them, [ka] Forming R 4 and R 5 are independently H, Me, Et, -CH2CH2CH3, -CH(CH3)2, -CH2CH(CH3)2, [ka] or R 4 and R 5 together with the carbon atom to which they are bonded, form a C3-C7 cycloalkyl, wherein the C3-C7 cycloalkyl is optionally substituted with halogen or C1-C3 alkyl.

[0008] In another preferred embodiment, in the general formula (1), R 2 and R 3 are independently H or Me, or R 2 and R 3 along with the carbon atoms attached to them, [ka] Form.

[0009] In another preferred embodiment, in the general formula (1), R 4 and R 5 are independently H, Me, Et, -CH2CH2CH3, -CH(CH3)2, -CH2CH(CH3)2, [ka] or R 4 and R 5 along with the carbon atoms attached to them, [ka] Form.

[0010] In another specific embodiment of the present invention, the compound of general formula (1) has the following structure: [ka] I have one of the following: TIFF2024524933000011.tif220168.

[0011] Another object of the present invention is to provide a pharmaceutical composition comprising a pharma- ceutically acceptable carrier, diluent and / or excipient and, as an active ingredient, a compound of general formula (1) of the present invention or an isomer, crystalline form, pharma- ceutically acceptable salt, hydrate or solvate thereof.

[0012] Yet another object of the present invention is to provide the use of a compound of general formula (1) of the present invention, or an isomer, crystalline form, pharma- ceutically acceptable salt, hydrate or solvate thereof, or said pharmaceutical composition, in the preparation of a medicament for treating, regulating or preventing a disease associated with an IDH mutant protein.

[0013] 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.

[0014] Synthesis of compounds

[0015] Methods for preparing the compound of the general formula (1) of the present invention are specifically described below, but these specific methods do not limit the present invention in any way.

[0016] The compounds of formula (1) above can be synthesized using standard synthetic techniques in combination with the methods described herein, or well-known techniques. Additionally, 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 different 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 may be modified by using appropriate reagents and conditions to introduce various groups into the molecular formulas provided herein.

[0017] 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, the amount of compounds used, reaction temperature, and reaction time, are not limited to the following description. 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 further provides a method for preparing a compound of general formula (1), which is prepared according to the following general reaction scheme 1.

[0018] General reaction scheme 1 [ka]

[0019] PG represents a protecting group for an amine group, and R1 , R 2 , R 3 , R 4 , R 5 and L is as defined above. As shown in General Reaction Scheme 1, starting material A1 is subjected to substitution reaction to obtain compound A2; compound A2 and compound A3 are reacted under basic conditions to obtain compound A4; compound A4 is reduced at the nitro group to obtain compound A5; compound A5 is cyclized to obtain compound A6; compound A6 is removed with the protecting group PG (e.g., Boc) to obtain compound A7; compound A7 is reacted with acryloyl chloride to obtain target compound A8.

[0020] Further forms of the compound

[0021] As used herein, "pharmaceutical acceptable" refers to a relatively non-toxic substance, such as a carrier or diluent, that does not cause the loss of biological activity or properties of a compound. For example, when a substance is administered to an individual, the substance does not cause undesired biological effects or adverse interactions with any of its components.

[0022] The term "pharmaceutical acceptable salt" refers to a form of a compound that does not cause significant irritation to the organism receiving the compound and does not eliminate the biological activity and properties of the compound.In certain embodiments, the pharmaceutical acceptable salt is 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, carbonic acid, 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, and acidic amino acids such as aspartic acid and glutamic acid.

[0023] It is to be understood that pharma- ceutically acceptable salts include solvent addition forms or crystal forms, particularly solvates or polymorphs. Solvates contain stoichiometric or non-stoichiometric amounts of solvent and are selectively formed during crystallization in pharma- ceutical 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 in a water / organic solvent mixture, and the organic solvents used include, but are not limited to, tetrahydrofuran, acetone, ethanol, or methanol. Furthermore, compounds described herein may exist in either unsolvated or solvated forms. In general, solvated forms are considered equivalent to unsolvated forms for the purposes of the compounds and methods provided herein.

[0024] In other specific examples, the compound of general formula (1) is prepared in different forms, including but not limited to amorphous, crushed, and nanoparticle forms. Furthermore, the compound of formula (1) may be a polymorph, including crystalline forms. A polymorph includes different lattice arrangements of the same elements of a compound. The polymorphs generally have different X-ray diffraction spectra, infrared spectra, melting points, density, hardness, crystal morphology, optical properties, electrical properties, stability, and solubility. Various factors, such as recrystallization solvent, crystallization rate, and storage temperature, may result in a single predominant crystal system.

[0025] In another embodiment, the compounds of general formula (1) may have chiral centers and / or axial asymmetry and therefore may exist in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds, single diastereomers and cis-trans isomers. Each chiral center or axial asymmetry independently produces two optical isomers, and all possible optical isomers, diastereomeric mixtures and pure or partially pure compounds are included within the scope of the present invention. The present invention is meant to include all such isomers of these compounds.

[0026] The compounds of the present invention may also contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute such compounds. For example, the compounds may contain tritium ( 3 H), iodine-125( 125 I) and C-14( 14 The deuterated pharmaceuticals may be labeled with radioisotopes such as 1,2-dihydro-1,2-triazolidinediamine (DHA) and 1,2-diazolidinediamine (C). As another example, deuterium may be used to replace hydrogen atoms to form deuterated compounds. The bond formed by deuterium and carbon is stronger than the bond formed by general hydrogen and carbon. Compared to non-deuterated pharmaceuticals, deuterated pharmaceuticals generally have the advantages of reduced toxic effects and side effects, improved pharmaceutical stability, enhanced efficacy, and extended pharmaceutical in vivo half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are within the scope of the present invention.

[0027] Explanation of terms

[0028] Unless otherwise specified, the terms used in the present specification and claims are defined as follows. Please note that in the present specification and the appended claims, the singular forms "a" and "an" include the plural meaning unless otherwise specified in the context. Conventional methods such as mass spectrometry, nuclear magnetic resonance spectroscopy, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacology are used unless otherwise specified. In the present specification, "or" or "and" means "and / or" unless otherwise specified.

[0029] Unless otherwise specified, "alkyl" refers to saturated aliphatic hydrocarbon groups, including straight-chain and branched groups, having 1 to 6 carbon atoms. Lower alkyl groups having 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 CH3, CH3CH2, CF3, CHF2, CF3CH2, CF3(CH3)CH, i Pr, n Pr, i Bu, n Bu or t is selected from Bu.

[0030] Unless otherwise specified, "cycloalkyl" refers to a non-aromatic hydrocarbon ring system (monocyclic, bicyclic, or polycyclic); partially unsaturated cycloalkyls may be referred to as "cycloalkenyls" if the carbocyclic ring contains at least one double bond, or "cycloalkynyls" if the carbocyclic ring contains at least one triple bond. Cycloalkyls may include monocyclic or polycyclic groups (e.g., having 2, 3, or 4 fused rings) and spirocycles. In some embodiments, cycloalkyls are monocyclic. In some embodiments, cycloalkyls are monocyclic or bicyclic. The ring carbon atoms of a cycloalkyl may be optionally oxidized to form an oxo or sulfide group. Cycloalkyls further include cycloalkylene. In some embodiments, cycloalkyls include 0, 1, or 2 double bonds. In some embodiments, cycloalkyls include 1 or 2 double bonds (partially unsaturated cycloalkyls). In some embodiments, cycloalkyls may be fused with aryls, heteroaryls, cycloalkyls, and heterocycloalkyls. In some embodiments, cycloalkyl may be fused with aryl, cycloalkyl, and heterocycloalkyl. In some embodiments, cycloalkyl may be fused with aryl and heterocycloalkyl. In some embodiments, cycloalkyl may be fused with aryl and cycloalkyl. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norcamphanyl, norpinanyl, norcaranyl, bicyclo[1.1.1]pentyl, bicyclo[2.1.1]hexyl, and the like.

[0031] Unless otherwise specified, "halogen" (or halo) refers to fluorine, chlorine, bromine, or iodine. The term "halo" (or "halogenated") before a radical name indicates that the radical is partially or fully halogenated, i.e., substituted by F, Cl, Br or I, preferably F or Cl, in any combination.

[0032] "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 the event or circumstance does not occur.

[0033] The substituent "-O-CH2-O-" means that the two oxygen atoms in the substituent are attached to two adjacent carbon atoms in a heterocycloalkyl, aryl, or heteroaryl, for example: [ka]

[0034] When the number of a linker group is 0, such as -(CH2)0-, it means that the linker group is a single bond.

[0035] When one of the variables is selected from a chemical bond, it means that the two groups linked by this variable are directly linked, for example, when L in XLY represents a chemical bond, it means that the structure is actually XY.

[0036] 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, pyridinyl, pyranyl, and thiopyranyl are 6-membered rings, and cyclopentyl, pyrrolyl, furanyl, and thienyl are 5-membered rings.

[0037] 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.

[0038] TIFF2024524933000014.tif30168

[0039] Specific pharmaceutical and medical terms

[0040] The term "acceptable" as used herein means that the formulation or active ingredient does not have an excessive and deleterious effect on the general health of the treated subject.

[0041] The terms "treatment", "course of treatment" and "treatment" as used herein include alleviating, inhibiting or ameliorating a disease symptom or condition, inhibiting the occurrence of a complication, improving or preventing the underlying metabolic syndrome, inhibiting the occurrence of a disease or condition (e.g., controlling the progression of a disease or condition), relieving a disease or condition, regressing a disease or condition, and alleviating complications caused by a disease or condition, or preventing or treating symptoms caused by a disease or condition. As used herein, a compound or pharmaceutical composition, when administered, can improve a disease, symptom or condition, and in particular can improve the severity, delay the onset, delay the progression, or shorten the duration of a disease. Fixed or episodic administration, or continuous or intermittent administration, can result from or relate to administration.

[0042] "Active ingredient" refers to compounds of general formula (1) and pharma- ceutically acceptable inorganic or organic salts of compounds of general formula (1). The compounds of the present invention may contain one or more asymmetric centers (chiral centers or axial asymmetry) and therefore may exist in the form of racemates, racemic mixtures, single enantiomers, diastereomeric compounds, and single diastereomers. The asymmetric centers that may exist depend on the properties of the various substituents on the molecule. Each such asymmetric center independently produces two optical isomers, and all possible optical isomers, diastereomeric mixtures, and pure or partially pure compounds are included within the scope of the present invention. The present invention is meant to include all such isomeric forms of these compounds.

[0043] As used herein, terms such as "compound," "composition," "agent," or "medicine or pharmaceutical agent" are used interchangeably and all refer to a compound or composition that, when administered to an individual (human or animal), is capable of eliciting a desired pharmacological and / or physiological response through local and / or systemic action.

[0044] The terms "administered, administering, or administration" as used herein refer to direct administration of a compound or composition, or to administration of a prodrug, derivative, analog, etc. of an active compound.

[0045] Although the numerical ranges and parameters defining the broad scope of the present invention are approximations, the relevant values ​​set forth in certain embodiments are set forth herein as precisely as possible. However, any numerical value inherently contains a standard deviation that necessarily results from certain testing methods. Here, "about" generally means that the actual value is within a particular value or range ±10%, 5%, 1%, or 0.5%. Alternatively, the term "about" indicates that the actual numerical value is within an acceptable standard error of the mean value, as would be understood by one of ordinary skill in the art. Except in experimental examples or unless otherwise indicated, all ranges, amounts, values, and percentages used herein (e.g., to describe amounts of materials, lengths of time, temperatures, operating conditions, proportions of amounts, etc.) are understood to be modified by the term "about". Thus, unless otherwise indicated, all numerical parameters set forth in this specification and the appended claims are approximations that may be varied as desired. At the very least, these numerical parameters should be interpreted as numerical values ​​obtained using the significant digits given or conventional rounding rules.

[0046] Scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art, unless otherwise defined herein. Furthermore, singular nouns used herein include their plurals, unless otherwise contradicted by context, and plural nouns used herein also include their singulars.

[0047] therapeutic use

[0048] The present invention provides methods of treating diseases, including but not limited to conditions associated with IDH mutant proteins (e.g., cancer), using a compound of general structural formula (1) or a pharmaceutical composition of the present invention.

[0049] In some embodiments, a method of treating cancer is provided, comprising administering to an individual in need thereof an effective amount of any of the aforementioned pharmaceutical compositions comprising a compound of general structural formula (1). In some embodiments, the cancer is mediated by an IDH mutant protein. In other embodiments, the cancer is a hematological cancer and solid tumor, including but not limited to leukemia, breast cancer, lung cancer, pancreatic cancer, colon cancer, bladder cancer, brain cancer, urothelial cancer, prostate cancer, liver cancer, ovarian cancer, head and neck cancer, gastric cancer, mesothelioma, or all cancer metastases.

[0050] Route of administration

[0051] The compound of the present invention and its pharmaceutically acceptable salts can be prepared into various formulations containing a safe and effective amount of the compound of the present invention or its pharmaceutically acceptable salts and a pharmaceutically acceptable excipient or carrier, where "safe and effective amount" means an amount of the compound sufficient to significantly improve the condition without causing serious adverse 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.

[0052] "Pharmaceutically acceptable excipient or carrier" refers to one or more compatible solid or liquid fillers or gel substances that must be suitable for human use and have sufficient purity and low toxicity. As used herein, "compatible" means that the components of the composition are capable of being mixed with the compounds of the present invention without significantly reducing the pharmaceutical efficacy of the compounds. 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, flavors, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0053] The compounds of the present invention may be administered orally, rectally, parenterally (intravenous, intramuscular, or subcutaneous) or topically.

[0054] Solid dosage forms for oral administration include capsules, tablets, pills, pulvises, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier) such as sodium citrate or dicalcium phosphate, or the following ingredients: (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; (c) humectants 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 retarding agents such as paraffin; (f) absorption accelerators such as quaternary ammonium compounds; (g) wetting agents such as cetyl alcohol and glycerol monostearate; (h) adsorbents such as kaolin; and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, and sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, said dosage forms may further comprise buffering agents.

[0055] Solid dosage forms such as tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other materials known in the art.These may contain opacifying agents, and the active compound or compounds in such compositions can be released in a certain part of the digestive tract in a delayed manner.The examples of embedding components that can be used include polymeric materials and wax-based materials.If necessary, the active compound can be formed into microcapsules with one or more of the above-mentioned excipients.

[0056] Liquid dosage forms for oral administration include pharma- ceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs, etc. In addition to the active compound, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing and emulsifying agents, e.g., ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0057] Besides such inert diluents, the composition can also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0058] In addition to the active compound, suspensions may contain suspending agents such as ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methylate, and agar-agar, or mixtures of these substances.

[0059] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0060] Dosage forms for topical administration of the compounds of the present invention 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 that may be required.

[0061] The compound of the present invention may be administered alone or in combination with other pharma- ceutical acceptable compounds. When the pharmaceutical composition of the present invention is used, a safe and effective amount of the compound of the present invention is administered to the mammal (such as a human) to be treated, and the dosage is a pharma- ceutical effective dose. For a 60 kg human, the daily dosage is usually 1 to 2000 mg, preferably 50 to 1000 mg. In addition, when determining the specific dosage, the administration route, the health condition of the patient, etc. are also taken into consideration, but these are well known to those skilled in the art.

[0062] The above features described in the present invention or the features described in the embodiments above can be combined in any combination.All features disclosed herein can be used in any composition, and various features disclosed herein can be replaced with any alternative features that provide the same, equivalent or similar purpose.Thus, unless otherwise specified, the features disclosed herein are merely generic examples of equivalent or similar features. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0063] Various specific aspects, features and advantages of the above compounds, methods and pharmaceutical compositions are described in detail as follows, which will make the contents of the present invention very clear. It should be understood that the following detailed description and examples are for reference purposes only. 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 application as defined herein.

[0064] In all examples, 1 H-NMR spectra were recorded on a Varian 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.

[0065] In the present invention, the following abbreviations are used: CDCl3 represents deuterated chloroform, DCM represents dichloromethane, dioxane represents 1,4-dioxane, DIPEA represents diisopropylethylamine, DMSO represents dimethylsulfoxide, EA represents ethyl acetate, EtOH represents ethanol, h represents hour, H2 represents hydrogen, KOH represents potassium hydroxide, LC-MS represents liquid chromatography mass spectrometry, min represents minute, mL represents milliliter, MS represents mass spectrometry, n-BuLi represents n-butylaluminum, NaBH(OAc)3 represents sodium triacetoxyborohydride, NHCl represents ammonium chloride, NMR represents nuclear magnetic resonance, Pd / C represents palladium on carbon, PE represents petroleum ether, THF represents tetrahydrofuran, Ti(O i -Pr)4 represents titanium tetraisopropoxide.

[0066] Preparation 1: 2-Chloro-N-ethyl-5-nitropyrimidin-4-amine [ka]

[0067] 2,4-Dichloro-5-nitropyrimidine (2 g, 10.31 mmol) was dissolved in THF (30 mL). Under an argon atmosphere, an aqueous solution of ethylamine (1.33 g, 70%) was added dropwise at -70°C. After the addition, the mixture was warmed to room temperature and reacted for about 0.5 hours. After confirming the completion of the reaction by LC-MS, water (50 mL) was added, and the mixture was extracted twice with EA (30 mL x 2). The organic phases were combined and concentrated. The residue was purified by column chromatography to obtain a yellow solid product A2-1 (1.763 g, 84% yield). ESI-MS m / z: 203 [M+H] + .

[0068] Following the synthesis method of intermediate A2-1, the target intermediates A2-2 to A2-6 were obtained using different starting materials. [Table 1]

[0069] Preparation Example 2: Synthesis of tert-butyl 4-(1-(4-((S)-1-aminoethyl)phenyl)-2-cyclopropylethyl)piperazine-1-carboxylate (A3-1) [ka]

[0070] Synthesis of (S)-N-(1-(4-(2-cyclopropylacetyl)phenyl)ethyl)-2,2,2-trifluoroacetamide

[0071] (S)-N-(1-(4-bromophenyl)ethyl)-2,2,2-trifluoroacetamide (10.7 g, 36.2 mmol) was dissolved in anhydrous THF (100 mL), and n-BuLi (2.5 M, 30 mL, 72.3 mmol) was added dropwise under argon atmosphere at -78 °C. After the addition was completed, the mixture was incubated at -78 °C to -60 °C for about 1 h to react, and then a solution of 2-cyclopropyl-N-methoxy-N-methylacetamide (5.7 g, 39.8 mmol) in anhydrous THF (50 mL) was slowly added dropwise. After the addition, the mixture was incubated at a constant temperature for another 0.5 h to react. After the completion of the reaction was detected by LC-MS, the mixture was quenched with saturated aqueous NH4Cl (100 mL) and extracted twice with EA (50 mL × 2). The organic phase was concentrated. The residue was purified by column chromatography to give a white solid product (6.06 g, 56% yield). ESI-MS m / z: 300 [M+H] + .

[0072] Synthesis of tert-butyl 4-(2-cyclopropyl-1-(4-((S)-1-(2,2,2-trifluoroacetylamino)ethyl)phenyl)ethyl)piperazine-1-carboxylate

[0073] (S)-N-(1-(4-(2-cyclopropylacetyl)phenyl)ethyl)-2,2,2-trifluoroacetamide (3.068 g, 10.25 mmol) and tert-butyl piperazine-1-carboxylate (3.82 g, 20.5 mmol) were dissolved in anhydrous THF (50 mL), and Ti(i-PrO)4 (15 mL, 51.25 mmol) was added dropwise under argon atmosphere. The mixture was reacted at 60 °C overnight and then cooled to room temperature. MeOH (20 mL) and NaBH(OAc)3 (1.288 g, 20.5 mmol) were added. The mixture was reacted at room temperature for 10 h. A small amount of starting material remained was detected by LC-MS. The mixture was quenched with water (100 mL) and extracted with EA (50 mL × 2). The organic phase was concentrated and the residue was purified by column chromatography to recover the unreacted starting material (2.2 g) while giving a white solid product (957 mg, 20% yield). ESI-MS m / z: 470 [M+H] + .

[0074] Synthesis of tert-butyl 4-(1-(4-((S)-1-aminoethyl)phenyl)-2-cyclopropylethyl)piperazine-1-carboxylate (A3-1)

[0075] tert-Butyl 4-(2-cyclopropyl-1-(4-((S)-1-(2,2,2-trifluoroacetylamino)ethyl)phenyl)ethyl)piperazine-1-carboxylate (3.74 g, 7.97 mmol) was dissolved in EtOH / HO (100 mL / 20 mL), KOH (2.24 g, 39.87 mmol) was added in portions in an ice bath, and the mixture was heated to 50° C. under an argon atmosphere and reacted for 3 h. After confirming the completion of the reaction by LC-MS, the mixture was concentrated under reduced pressure to leave about 30 mL. The remaining mixture was diluted with water (50 mL) and extracted with DCM (50 mL×2). The organic phases were combined, washed with saturated aqueous sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the desired compound A3-1 (3.854 g, 100% yield). ESI-MS m / z: 374 [M+H] + .

[0076] Following the synthesis method of intermediate A3-1, the target intermediates A3-2 to A3-20 were obtained using different starting materials.

[0077] [Table 2] TIFF2024524933000019.tif46168

[0078] Example 1: Synthesis of 2-(((1S)-1-(4-(1-(4-acryloylpiperazin-1-yl)-2-cyclopropylethyl)phenyl)ethyl)amino)-9-ethyl-7,9-dihydro-8H-purin-8-one (Compound 1) [ka]

[0079] Step 1: Synthesis of compound A4-1 Compound A3-1 (200 mg, 0.535 mmol) and compound A2-1 (130 mg, 0.642 mmol) were dissolved in DMSO (5 mL). DIPEA (207 mg, 1.605 mmol) was added. The reaction was carried out at 80° C. for about 3 hours. After the completion of the reaction was confirmed by LC-MS, water (30 mL) was added and the mixture was extracted with EA (20 mL×2). The organic phases were combined and concentrated. The residue was purified by column chromatography to obtain a yellow solid product A4-1 (290 mg, 91% yield). ESI-MS m / z: 540 [M+H] + .

[0080] Step 2: Synthesis of compound A5-1 Compound A4-1 (290 mg, 0.537 mmol) was dissolved in MeOH (10 mL) and Pd / C (40 mg, 10%) was added. The mixture was purged with hydrogen and then reacted at room temperature overnight. After confirming the completion of the reaction by LC-MS, the mixture was filtered and concentrated to give purple solid A5-1 (236 mg, 86% yield). ESI-MS m / z: 510 [M+H] + .

[0081] Step 3: Synthesis of compound A6-1 Compound A5-1 (236 mg, 0.463 mmol) was dissolved in DCM (10 mL) and CDI (150 mg, 0.926 mmol) was added. The mixture was reacted at room temperature overnight. After the completion of the reaction was confirmed by LC-MS, the mixture was concentrated under reduced pressure. The residue was purified by Flash to give a dark purple solid A6-1 (151 mg, 61% yield). ESI-MS m / z: 536 [M+H] + .

[0082] Step 4: Synthesis of compound A7-1 Compound A6-1 (151 mg, 0.282 mmol) was dissolved in DCM (5 mL), 4M HCl / Diox (1 mL, 4 mmol) was added, and the mixture was stirred at room temperature for about 3 hours. After confirming the completion of the reaction by LC-MS, the mixture was concentrated under reduced pressure to obtain yellow solid product A7-1 (170 mg, 100% yield). ESI-MS m / z: 436 [M+H] + .

[0083] Step 5: Synthesis of Compound 1 Compound A7-1 (170 mg, 0.282 mmol) and DIPEA (183 mg, 1.41 mmol) were dissolved in DCM (5 mL). The mixture was cooled to 0° C. in an ice bath under an argon atmosphere. A solution of acryloyl chloride (23 mg, 0.254 mmol) in DCM (3 mL) was added dropwise. The mixture was reacted in an ice bath for about 10 minutes. After the completion of the reaction was confirmed by LC-MS, water (10 mL) was added, followed by stirring and liquid separation, and the aqueous phase was extracted with DCM (10 mL). The organic phases were combined. The concentrated residue was purified by pre-TLC to obtain a pale yellow solid product (100 mg, 72% yield).

[0084] 1H NMR (400 MHz, CDCl3) δ: 7.81 (s, 1H), 7.32 (d, J = 7.9 Hz, 2H), 7.18 (d, J = 8.0 Hz, 2H), 6.49 (dd, J = 16.9, 10.5 Hz, 1H), 6.23 (dd, J = 16.8, 1.9 Hz, 1H), 5.64 (dd, J = 10.5, 1.9 Hz, 1H), 5.26 (s, 1H), 5.10 (p, J = 6.9 Hz, 1H), 3.86 (q, J = 7.2 Hz, 2H), 3.63 (m, 2H), 3.49 (m, 2H), 3.44-3.36 (m, 1H), 2.38 (m, 4H), 1.89-1.79 (m, 1H), 1.65 (m, 2H), 1.55 (d, J = 6.8 Hz, 3H), 1.29-1.25 (m, 3H), 0.40 (q, J = 6.4, 4.8 Hz, 1H), 0.36-0.24 (m, 2H), -0.01-0.09 (m, 2H); ESI-MS m / z: 490 [M+H] + .

[0085] Examples 2 to 40: Synthesis of Compounds 2 to 40

[0086] Following the same procedure as in the synthesis of compound 1, target compounds 2 to 40 in Table 3 can be obtained using different intermediates as starting materials. [Table 3] TIFF2024524933000022.tif226168TIFF2024524933000023.tif226168

[0087] Example 41: Preparation of chiral isomers of compounds of the present invention

[0088] The compounds of the present invention contain one or more chiral centers. Various optically pure isomers of the compounds of the present invention can be prepared by using optically pure intermediates as starting materials. Alternatively, the optically pure isomers of the compounds of the present invention can be prepared using chiral HPLC or achiral HPLC.

[0089] Compound 1 of the present invention can be obtained as two optically pure isomers 1-1 and 1-2 of compound 1 by using the above method: [ka]

[0090] Compounds 3, 4, 5, 12, 29, 30, 33 and 34 were chirally resolved by the same synthetic or preparative methods to give the chiral isomeric pairs 3-1 / 3-2, 4-1 / 4-2, 5-1 / 5-2, 12-1 / 12-2, 29-1 / 29-2, 30-1 / 30-2, 33-1 / 33-2 and 34-1 / 34-2, respectively: [ka] TIFF2024524933000026.tif68168

[0091] Other compounds of the present invention can be prepared by similar synthetic or preparative methods to give the corresponding chiral isomers.

[0092] [Table 4] [Table 4-2]

[0093] Example 42: Detection of 2-HG in the supernatant of U87-IDH1 R132H cells

[0094] U87MG cells overexpressing mIDH1 R132H mutation or HT1080 cells harboring IDH1 R132C were seeded in 48-well and 96-well plates at 50,000 and 10,000 cells / well, respectively. The cells were incubated overnight to allow attachment, and the supernatant was removed. Cell culture medium containing serially diluted compounds was added, and the cells were incubated for 72 h. After 72 h, the medium was collected and diluted 10-fold and 20-fold with water, respectively. Acetonitrile was added to extract metabolites. The 2-HG content in the culture medium was analyzed by LC-MS-MS. The inhibition rate and IC of compounds against 2-HG in the supernatant were 50 was calculated in comparison with the control group. [Table 5]

[0095] A indicates that the inhibition rate is more than 90%. B indicates that the inhibition rate is more than 60% and less than 90%. C indicates that the inhibition rate is more than 30% and less than 60%. D indicates that the inhibition rate is 30% or less.

[0096] [Table 6]

[0097] Example 43: Stability studies of human and mouse liver microsomes

[0098] 1μM of the compound was incubated with 500μg / ml of human or mouse liver microsomes and NADPH Regeneration System at 37℃ for different times, and the remaining amount of the compound was analyzed by LC-MS-MS. 1 / 2 was calculated.

[0099] [Table 7]

[0100] Example 44: Detection of 2-HG in tumor tissue

[0101] 1 × 10 in nude mice 6 HT1080 cells were inoculated subcutaneously. 3 At the time of reaching 100 mg / kg, the mice were randomly divided into a vehicle control group and a group treated with 20 mg / kg of compounds 1, 3, 29, 33 or LY-3410738. After 3 and 7 days of continuous administration, the tumors were harvested, weighed, digested with digestive juice, and then homogenized. The concentration of 2-HG in the tumor tissue was measured by LC-MS-MS. The inhibition rate of the compound against 2-HG in the tumor tissue was calculated compared with that of the control group. [Table 8]

[0102] The activity data in Tables 5-8 demonstrate that the compounds of the general formula (1) of the present invention have stronger inhibitory activity against the levels of 2-HG in the supernatants of U87-IDH R132H and HT1080 cells and in tumor tissues compared to the commercially available IDH inhibitor Ivosidenib, indicating that these compounds have a stronger ability to inhibit IDH1 R132H and IDH1 R132C mutant proteins. Meanwhile, these compounds have equal or stronger activity compared to LY-3410738 (compound 2 in WO2018111707).

[0103] Although specific embodiments of the present invention have been described above, those skilled in the art will understand that these embodiments are merely illustrative and that many changes or modifications can be made to these embodiments without departing from the principles and spirit 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 an isomer, crystal form, pharma- ceutically acceptable salt, hydrate or solvate thereof. 【Chemistry 1】 (In general formula (1), L is, 【Chemistry 2】 where " ＊ " indicates the site of attachment to the carbonyl group, X is NH or NMe; R １ は、Me、Et、-CH ２ CH ２ CH ３ ,-EH(EH ３ ) ２ 、 【Chemistry 3】 and R ２ and R ３ are each independently H, Me or Et; or R ２ and R ３ along with the carbon atoms attached to them, 【Chemistry 4】 Forming R ４ and R ５ are each independently H, Me, Et, or -CH ２ CH ２ CH ３ , -CH(CH ３ ) ２ , -CH ２ CH (CH ３ ) ２ , 【Chemistry 5】 or R ４ and R ５ together with the carbon atom to which they are attached form a C3-C7 cycloalkyl, wherein said C3-C7 cycloalkyl is optionally substituted with halogen or C1-C3 alkyl.

2. In general formula (1), R ２ and R ３ is independently H or Me, or R ２ and R ３ together with the carbon atoms attached to them, 【Chemistry 6】 2. The compound of claim 1, or an isomer, crystalline form, pharma- ceutically acceptable salt, hydrate or solvate thereof, which forms:

3. In general formula (1), R ４ and R ５ are independently H, Me, Et, -CH ２ CH ２ CH ３ , -CH(CH ３ ) ２ , -CH ２ CH (CH ３ ) ２ , 【Chemistry 7】 or R ４ and R ５ together with the carbon atoms attached to them, 【Chemistry 8】 2. The compound of claim 1, or an isomer, crystalline form, pharma- ceutically acceptable salt, hydrate or solvate thereof, which forms:

4. The compound has the following structure: 【Chemistry 9】 【change】 4. The compound of claim 1, or an isomer, crystalline form, pharma- ceutically acceptable salt, hydrate or solvate thereof, having one of the following formula:

5. 13. A pharmaceutical composition comprising a pharma- ceutically acceptable excipient or carrier and, as an active ingredient, a compound according to any one of claims 1 to 4, or an isomer, crystalline form, pharma- ceutically acceptable salt, hydrate or solvate thereof.

6. Use of a compound according to any one of claims 1 to 4, or an isomer, crystalline form, pharma- ceutically acceptable salt, hydrate or solvate thereof, or a pharmaceutical composition according to claim 5, in the preparation of a medicament for treating a related disease mediated by an IDH mutant protein.

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