IDH variant inhibitors and their use

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 China Application No. 202110661394.5, filed on 15 June 2021, which is incorporated herein by reference in its entirety.

[0002] This invention relates to the field of medicinal chemistry, and more particularly to a novel class of IDH variant inhibitors, methods for preparing the same, and the use of the same. [Background technology]

[0003] Isocitrate dehydrogenase (IDH) is an important enzyme involved in the tricarboxylic acid cycle. It catalyzes the rate-limiting step of the tricarboxylic acid cycle, the conversion of isocitrate to α-ketoglutarate (α-KG). Humans have three different isocitrate dehydrogenases: 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 oncogene mutations. IDH mutations are found in low-grade gliomas, secondary gliomas, melanoma, angioimmunoblastic T-cell lymphoma, 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 the loss of function of the wild-type IDH protein, and instead, the ability to convert α-KG to the oncogenic metabolite D-2-hydroxyglutarate (D-2HG) is gained. The oncogenic metabolite D-2HG inhibits DNA or histone demethylases, leading to hypermethylation of DNA and histones and promoting cancer development. IDH inhibitors reduce the oncogenic metabolite D-2HG in vivo by inhibiting the activity of proteins with IDH1 / R132, IDH2 / R172, or IDH2 / R140 mutations, inducing histone H3K9me3 demethylation and suppressing tumor development. Therefore, targeting mutant IDH1 and IDH2 (mIDH1 and mIDH2) could be a promising approach to cancer treatment.

[0005] To date, several IDH small molecule inhibitors have been commercially available. For example, Enasidenib and Ivosidenib developed by Agios Pharmaceuticals Inc. are both non-covalent inhibitors. Covalent inhibitors of IDH have been reported by Eli Lilly and Company in WO 2017 / 019429, WO 2017 / 213910 and WO 2018 / 111707, and have better selectivity for mutant IDH1 and IDH2 compared to wild-type IDH1 and IDH2. However, since there is no IDH covalent inhibitor in the clinical stage, it is necessary to research and explore IDH covalent inhibitors with better activity and drug discovery potential.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

[0007] The present invention provides a compound of general formula (1), or an isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate thereof.

Chemical Formula

Chemical Formula

Chem.

Chem.

Chem.

[0008] In another preferred embodiment, in general formula (1), R 2 and R 3 are independently H, or Me, or R 2 and R 3 together with the carbon atom to which they are attached,

Chem.

[0009] In another preferred embodiment, in general formula (1), R 4 and R 5 are independently H, Me, Et, -CH2CH2CH3, -CH(CH3)2, -CH2CH(CH3)2,

Chem.

[0010] In another specific embodiment of the present invention, the compound of general formula (1) has the following structure: [ka] It has one of the following: TIFF0007884019000010.tif220168.

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

[0012] A further object of the present invention is intended to provide the use of a compound of general formula (1) of the present invention, or its isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate, or the pharmaceutical composition thereof, in the preparation of a pharmacopoeia for treating, modulating or preventing diseases associated with IDH mutant proteins.

[0013] Both the above general description and the following detailed description of the present invention are illustrative and descriptive, and should be understood as being intended to provide a further description of the claimed invention.

[0014] Compound synthesis

[0015] The following describes in detail the preparation methods for the compound of general formula (1) of the present invention, but these specific methods do not limit the present invention in any way.

[0016] The compound of formula (1) above can be synthesized using standard synthetic techniques combined with the methods described herein, or using well-known techniques. Furthermore, the solvent, temperature, and other reaction conditions described herein may vary. Starting materials for the synthesis of the compound can be obtained synthetically or commercially. The compound described herein and other related compounds with different substituents are referenced 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 ORGANICSYNTHESIS, 3 rd The compounds can be synthesized using well-known techniques and starting materials, including the method found in Ed., (Wiley 1999). General methods for preparing the compounds can be modified by using appropriate reagents and conditions for introducing 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 method, such as reactants, solvents, bases, amounts of compounds used, reaction temperature, and reaction time, are not limited to those described below. Furthermore, the compounds of the present invention can be easily prepared by any combination of various synthesis methods described herein or known in the art, and such combinations can be readily determined by those skilled in the art to whom the present invention relates. In one embodiment, the present invention further provides a method for preparing the compound of general formula (1) prepared by the following general reaction scheme 1.

[0018] General reaction scheme 1 [ka]

[0019] PG represents a protecting group for amine groups, R1 , R 2 , R 3 , R 4 , R 5 And L are as defined above. As shown in general reaction scheme 1, starting material A1 is subjected to a 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; the protecting group PG (e.g., Boc) of compound A6 is removed to obtain compound A7; and compound A7 is reacted with acryloyl chloride to obtain the target compound A8.

[0020] Further forms of compounds

[0021] In this specification, "pharmaceutically acceptable" means a relatively non-toxic substance, such as a carrier or diluent, that does not cause the loss of the biological activity or properties of a compound. For example, when a substance is administered to an individual, the substance does not cause undesirable biological effects or harmful interactions with any of its components.

[0022] The term "pharmaceutically acceptable salt" refers to a form of a compound that does not cause significant irritation to the organism to which it is administered and does not eliminate the biological activity and properties of the compound. In certain embodiments, the pharmaceutically acceptable salt is obtained by reacting the compound of general formula (1) with an acid, such as an inorganic acid like hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, phosphoric acid, nitric acid, or carbonic acid; an organic acid like formic acid, acetic acid, propionic acid, oxalic acid, trifluoroacetic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzenesulfonic acid, or p-toluenesulfonic acid; or an acidic amino acid like aspartic acid or glutamic acid.

[0023] It should be understood that pharmaceutically acceptable salts include solvated or crystalline forms, particularly solvates or polymorphs. Solvates are formed selectively upon crystallization in pharmaceutically acceptable solvents such as water and ethanol, containing stoichiometric or nonstoichiometric amounts of solvent. Hydrates are formed when the solvent is water, and alkoxides 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, the organic solvents used including, but not limited to, tetrahydrofuran, acetone, ethanol, or methanol. Furthermore, the compounds described herein may exist in either a non-solvated or solvated form. Generally, the solvated form is considered equivalent to the non-solvated form for the purposes of the compounds and methods provided herein.

[0024] In other specific examples, compounds of general formula (1) may be prepared in different forms, including but not limited to amorphous, pulverized, and nanoparticle forms. Furthermore, compounds of formula (1) may also be in crystalline form and may be polymorphs. Polymorphs contain different lattice arrangements of the same elements of the compound. The polymorphs generally have different X-ray diffraction spectra, infrared spectra, melting points, densities, hardness, crystalline forms, optical properties, electrical properties, stability, and solubility. A single dominant crystalline system may be formed depending on various factors such as the recrystallization solvent, crystallization rate, and storage temperature.

[0025] In another embodiment, compounds of general formula (1) may have chiral centers and / or axial chirality, and thus may exist in the form of racemates, racemic mixtures, single enantiomers, diastereomer compounds, single diastereomers, and cis-trans isomers. Each chiral center or axial chirality independently produces two optical isomers, and all possible optical isomers, diastereomer mixtures, and pure or partially pure compounds are included in the scope of the present invention. The present invention means that it includes all such isomers of these compounds.

[0026] The compounds of the present invention may contain unnatural proportions of atomic isotopes in one or more of the atoms constituting such compounds. For example, the compound may contain tritium ( 3 H), Iodine-125( 125 I) and C-14( 14 They can be labeled with radioactive isotopes such as C). As another example, deuterated compounds can be formed by substituting hydrogen atoms with deuterium. The bond formed by deuterium and carbon is stronger than the bond formed by ordinary hydrogen and carbon. Compared with non-deuterated pharmaceuticals, deuterated pharmaceuticals generally have advantages such as reduced toxicity and side effects, improved pharmaceutically stable, enhanced potency, and extended pharmaceutically in vivo half-life. All isotopic variations of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.

[0027] Explanation of terms

[0028] Unless otherwise specified, terms used herein and in the claims are defined as follows: Note that in this specification and in the appended claims, the singular forms “a” and “an” have plural meanings unless otherwise specified in the context. Unless otherwise specified, conventional methods such as mass spectrometry, nuclear magnetic resonance spectroscopy, HPLC, protein chemistry, biochemistry, recombinant DNA technology and pharmacology are used. In this specification, “or” or “and” means “and / or” unless otherwise specified.

[0029] Unless otherwise specified, “alkyl” means saturated aliphatic hydrocarbon groups including linear and branched groups having 1 to 6 carbon atoms. Lower alkyl groups having 1 to 4 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, or tert-butyl, are preferred. As used herein, “alkyl” includes unsubstituted and substituted alkyl groups, in particular alkyl groups substituted with one or more halogens. Preferred alkyl groups include CH3, CH3CH2, CF3, CHF2, CF3CH2, CF3(CH3)CH, i Pr, n Pr, i Bu, n Bu or t 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 called “cycloalkenyl” if the carbocyclic ring contains at least one double bond, or “cycloalkynyl” 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 spiro rings. In some embodiments, cycloalkyls are monocyclic. In some embodiments, cycloalkyls are monocyclic or bicyclic. The ring carbon atoms of cycloalkyls may optionally be oxidized to form oxo or sulfide groups. Cycloalkyls further include cycloalkylenes. In some embodiments, cycloalkyls contain 0, 1, or 2 double bonds. In some embodiments, cycloalkyls contain 1 or 2 double bonds (partially unsaturated cycloalkyls). In some embodiments, cycloalkyls may be condensed with aryl, heteroaryl, cycloalkyl, and heterocycloalkyl groups. In some embodiments, the cycloalkyl may be condensed with aryl, cycloalkyl, and heterocycloalkyl. In some embodiments, the cycloalkyl may be condensed with aryl and heterocycloalkyl. In some embodiments, the cycloalkyl may be condensed with aryl and cycloalkyl. Examples of cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopentenyl, cyclohexenyl, cyclohexadienyl, cycloheptatrienyl, norcamphanyl, norpinanil, norcalanil, bicyclo[1.1.1]pentyl, and bicyclo[2.1.1]hexyl.

[0031] Unless otherwise specified, “halogen” refers to fluorine, chlorine, bromine, or iodine. The term “halogen” (or “halogenated”) preceding a group name indicates that the group is partially or completely halogenated, i.e., substituted with any combination of F, Cl, Br, or I, preferably F or Cl.

[0032] "Optional" or "optional" means that the event or situation described thereafter may occur, but does not necessarily occur, and that description includes examples in which the event or situation may occur and examples in which it does not occur.

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

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

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

[0036] The term "membered ring" includes any cyclic structure. The term "member" 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, while cyclopentyl, pyrrolyl, furanyl, and thienyl are 5-membered rings.

[0037] The term "part" refers to a specific part or functional group of a molecule. A chemical part is generally considered to refer to a chemical substance that is contained within or bonded to a molecule.

[0038] TIFF0007884019000013.tif30168

[0039] Specific pharmaceutical and medical terms

[0040] As used herein, the term “acceptable” means that the formulation ingredient or active ingredient does not have an excessive or harmful effect on the health of the general subject being treated.

[0041] As used herein, the terms “treatment,” “treatment course,” and “therapy” include alleviating, inhibiting, or improving the symptoms or condition of a disease; inhibiting the development of complications; improving or preventing an underlying metabolic syndrome; inhibiting the development of a disease or condition (e.g., controlling the progression of a disease or condition); alleviating a disease or symptoms; regressing a disease or symptoms; alleviating complications caused by a disease or symptoms; or preventing or treating signs caused by a disease or symptoms. As used herein, a compound or pharmaceutical composition, when administered, may improve a disease, symptoms, or condition, particularly by improving the severity of a disease, delaying its onset, slowing its progression, or shortening its duration. A fixed or single dose, or a continuous or intermittent dose, may be caused by or related to the administration.

[0042] "Active ingredient" refers to the compound of general formula (1) and a pharmaceutically acceptable inorganic or organic salt of the compound of general formula (1). The compounds of the present invention may contain one or more chiral centers (chiral centers or axial chiral centers) and therefore may exist in the form of racemates, racemic mixtures, single enantiomers, diastereomer compounds, and single diastereomers. The possible chiral centers depend on the properties of various substituents on the molecule. Each such chiral center independently produces two optical isomers, and all possible optical isomers, diastereomer mixtures, and pure or partially pure compounds are included in the scope of the present invention. The present invention means that it includes all such isomeric forms of these compounds.

[0043] In this specification, terms such as “compound,” “composition,” “drug,” or “pharmaceutical or medicinal product” are used interchangeably and all refer to compounds or compositions that, when administered to an organism (human or animal), can induce a desired pharmacological and / or physiological response through local and / or systemic effects.

[0044] The terms “administered, given, or administered” refer, in this specification, to the direct administration of a compound or composition, or to the administration of a prodrug, derivative, analog, etc., of an active compound.

[0045] While the numerical ranges and parameters defining the broad scope of this invention are approximations, the relevant values ​​shown in specific embodiments are presented herein as accurately as possible. However, any numerical value inherently includes a standard deviation that inevitably arises from certain test methods. Here, “approximately” generally means that the actual value is within a specific value or range of ±10%, 5%, 1%, or 0.5%. Alternatively, the term “approximately” indicates that the actual numerical value is within the acceptable standard error of the mean, as those skilled in the art would understand. Except for experimental examples or unless otherwise specifically stated, all ranges, quantities, values, and percentages used herein (e.g., to describe quantities of material, lengths of time, temperatures, operating conditions, proportions of quantities, etc.) are understood to be modified by the term “approximately.” Thus, unless otherwise specifically stated, all numerical parameters described herein and in the appended claims are approximations that may vary as desired. At a minimum, these numerical parameters should be interpreted as numerical values ​​obtained using the indicated significant figures or conventional rounding rules.

[0046] The scientific and technical terms used herein have the same meanings as those generally understood by those skilled in the art, unless otherwise defined herein. Furthermore, singular nouns used herein include their plural forms unless inconsistent with the context, and plural nouns used include their singular forms.

[0047] therapeutic use

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

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

[0050] Route of administration

[0051] The compounds of the present invention and their pharmaceutically acceptable salts can be prepared into various formulations comprising a safe and effective amount of the compound or its pharmaceutically acceptable salt, 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 being treated.

[0052] "Pharmacopoeia-acceptable excipients or carriers" means one or more compatible solid or liquid fillers or gels that are suitable for human use and must be of sufficient purity and low toxicity. In this specification, "compatible" means that the components of the composition can be mixed with the compounds of the present invention without significantly reducing the pharmaceutically active properties 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, fragrances, stabilizers, antioxidants, preservatives, and water free of pyrogens.

[0053] The compounds of the present invention can be administered orally, rectally, parenterally (intravenously, intramuscularly, or subcutaneously), 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 any of the following components: (a) fillers or bulking agents such as starch, 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) disintegrants such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solution retarders such as paraffin; (f) absorption enhancers 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 glycol, and sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may further contain a buffering agent.

[0055] Solid dosage forms such as tablets, sugar-coated tablets, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. These may contain opacifying agents, and the active compound or compound in such compositions may be released with delay in specific parts of the gastrointestinal tract. Examples of embedding components that can be used include polymers and waxes. If necessary, the active compound may form microcapsules with one or more of the above excipients.

[0056] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, the liquid dosage form may contain water or other solvents, solubilizers and emulsifiers, such as 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 thereof, which are commonly used in the art.

[0057] In addition to such inert diluents, the composition may further contain adjuvants such as wetting agents, emulsifiers, suspending agents, sweeteners, flavoring agents, and fragrances.

[0058] In addition to the active compound, the suspension may contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methylate, and agar, or mixtures thereof.

[0059] Parenteral injection compositions may include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, and sterile powders for reconstitution into sterile injection 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 sprays as required.

[0061] The compounds of the present invention can be administered alone or in combination with other pharmaceutically 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 target mammal (such as a human), where the dose is a pharmaceutically effective dose. For a 60 kg human, the daily dose is usually 1 to 2000 mg, preferably 50 to 1000 mg. When determining the specific dose, the route of administration, the patient's health condition, etc., should also be considered, but these are well known to those skilled in the art.

[0062] The features described above in this invention, or the features described above in the embodiments, can be combined in any way. All features disclosed herein can be used in the form of 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 general examples of equivalent or similar features. [Modes for carrying out the invention]

[0063] Various specific embodiments, features, and advantages of the above-mentioned compounds, methods, and pharmaceutical compositions are described in detail below, which will make the scope of the present invention very clear. Please understand that the following detailed description and examples describe specific examples for reference 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 this application as defined herein.

[0064] In all embodiments, 1 ¹H-NMR spectra were recorded using a Varian Mercury 400 nuclear magnetic resonance spectrometer, and chemical shifts were expressed as δ (ppm). Unless otherwise specified, 200-300 mesh silica gel was used for separation, and the eluent ratio was expressed as a volume ratio.

[0065] In this invention, the following abbreviations are used: CDCl3 represents deuterated chloroform, DCM represents dichloromethane, dioxane represents 1,4-dioxane, DIPEA represents diisopropylethylamine, DMSO represents dimethyl sulfoxide, EA represents ethyl acetate, EtOH represents ethanol, h represents time, 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, NH4Cl represents ammonium chloride, NMR represents nuclear magnetic resonance, Pd / C represents palladium-carbon, PE represents petroleum ether, THF represents tetrahydrofuran, Ti(O i -Pr)4 represents titanium tetraisopropoxide.

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

[0067] 2,4-Dichloro-5-nitropyrimidine (2 g, 10.31 mmol) was dissolved in THF (30 mL). Under an argon atmosphere, aqueous ethylamine solution (1.33 g, 70%) was added dropwise at -70°C. After addition, the mixed solution was warmed to room temperature and reacted for approximately 0.5 hours. After confirming the completion of the reaction by LC-MS, water (50 mL) was added, and the mixed solution 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, yield 84%). ESI-MS m / z: 203 [M+H] + .

[0068] Following the synthesis method for intermediate A2-1, we obtained the desired intermediates A2-2 to A2-6 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 an argon atmosphere at -78°C. After the addition was complete, the mixed solution was incubated at -78°C to -60°C for approximately 1 hour to allow the reaction to proceed, 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 mixed solution was incubated at a constant temperature for a further 0.5 hours to allow the reaction to proceed. After the completion of the reaction was detected by LC-MS, the mixed solution was quenched with saturated NH4Cl aqueous solution (100 mL) and extracted twice with EA (50 mL x 2). The organic phase was concentrated. The residue was purified by column chromatography to obtain 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-butylpiperazine-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 an argon atmosphere. The mixture was reacted overnight at 60 °C 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 hours. A small amount of residual starting material was detected by LC-MS. The mixture was quenched with water (100 mL) and extracted with EA (50 mL x 2). The organic phase was concentrated, and the residue was purified by column chromatography to recover the unreacted starting material (2.2 g) while obtaining a white solid product (957 mg, yield 20%). 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 / H2O (100 mL / 20 mL), and KOH (2.24 g, 39.87 mmol) was added in several portions in an ice bath. The mixed solution was heated to 50°C under an argon atmosphere and reacted for 3 hours. After confirming the completion of the reaction by LC-MS, the mixed solution was concentrated under reduced pressure, leaving approximately 30 mL. The remaining mixture was diluted with water (50 mL) and extracted with DCM (50 mL x 2). The organic phases were combined, washed with saturated sodium chloride aqueous solution (50 mL), dried on anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the target compound A3-1 (3.854 g, yield 100%). ESI-MS m / z: 374 [M+H] + .

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

[0077] [Table 2] TIFF0007884019000018.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-purine-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 approximately 3 hours. After confirming completion of the reaction by LC-MS, water (30 mL) was added, and the mixed solution was extracted with EA (20 mL x 2). The organic phases were combined and concentrated. The residue was purified by column chromatography to obtain the yellow solid product A4-1 (290 mg, yield 91%). 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. After purging the mixed solution with hydrogen, the reaction was allowed to proceed overnight at room temperature. After confirming the completion of the reaction by LC-MS, the mixed solution was filtered and concentrated to obtain the purple solid A5-1 (236 mg, yield 86%). 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 mixed solution was reacted overnight at room temperature. After confirming the completion of the reaction by LC-MS, the mixed solution was concentrated under reduced pressure. The residue was purified by flash to obtain 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), and 4 M HCl / Diox (1 mL, 4 mmol) was added. The mixed solution was stirred at room temperature for approximately 3 hours. After confirming the completion of the reaction by LC-MS, the mixed solution was concentrated under reduced pressure to obtain the 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 mixed solution 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 mixed solution was reacted in an ice bath for approximately 10 minutes. After confirming the completion of the reaction 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, yield 72%).

[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 2-40: Synthesis of Compounds 2-40

[0086] Using a procedure similar to that for the synthesis of compound 1, and employing different intermediates as starting materials, the target compounds 2-40 in Table 3 can be obtained. [Table 3] TIFF0007884019000021.tif226168TIFF0007884019000022.tif226168

[0087] Example 41: Preparation of chiral isomers of the compound 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, 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 method described above: [ka]

[0090] Compounds 3, 4, 5, 12, 29, 30, 33, and 34 were chiral-cleared using the same synthetic or preparation method to obtain chiral isomer 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] TIFF0007884019000025.tif68168

[0091] Other compounds of the present invention can also be prepared by similar synthesis or preparation methods to obtain their 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 the mIDH1 R132H mutation, or HT1080 cells possessing IDH1 R132C, were seeded at 50,000 cells / well and 10,000 cells / well in 48-well and 96-well plates, respectively. After incubation overnight to allow adhesion, the supernatant was removed. Cell culture medium containing serially diluted compounds was added, and the cells were incubated for 72 hours. After 72 hours, the medium was collected and diluted 10-fold and 20-fold with water, respectively. Metabolites were extracted by adding acetonitrile. The 2-HG content in the culture medium was analyzed by LC-MS-MS. The inhibition rate and IC50 of the compounds against 2-HG in the supernatant were analyzed. 50 This was calculated in comparison to the control group. [Table 5]

[0095] A indicates an inhibition rate exceeding 90%. B indicates an inhibition rate of over 60% and under 90%. C indicates that the inhibition rate is above 30% and below 60%. D indicates an inhibition rate of 30% or less.

[0096] [Table 6]

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

[0098] After incubating a 1 μM compound with 500 μg / ml human or mouse liver microsomes and an NADPH Regeneration System for different durations at 37°C, the remaining amount of the compound was analyzed using LC-MS-MS. 1 / 2 The result was calculated.

[0099] [Table 7]

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

[0101] 1 x 10 nude mice 6 Numerous HT1080 cells were subcutaneously inoculated. Tumor volume was 100-150 mm². 3 At the point of administration, mice were randomly divided into a solvent control group and a group administered 20 mg / kg of compound 1, 3, 29, 33, or LY-3410738. Tumors were collected 3 and 7 days after continuous administration, weighed, digested with digestive fluid, and then homogenized. 2-HG concentrations in tumor tissue were measured by LC-MS-MS. The inhibition rate of each compound against 2-HG in tumor tissue was calculated compared to the control group. [Table 8]

[0102] The activity data in Tables 5-8 demonstrates that the compound of general formula (1) of the present invention exhibits stronger inhibitory activity against 2-HG levels in the supernatant of U87-IDH R132H and HT1080 cells, as well as in tumor tissue, compared to the commercially available IDH inhibitor ivosidenib. This indicates that these compounds have a stronger ability to inhibit IDH1 R132H and IDH1 R132C mutant proteins. On the other hand, these compounds exhibit equivalent or stronger activity compared to LY-3410738 (compound 2 in International Publication No. 2018111707).

[0103] Although specific embodiments of the present invention have been described above, these embodiments are merely illustrative, and those skilled in the art will understand that many changes or modifications can be made to these embodiments without departing from the principles and spirit of the present invention. Accordingly, the scope of protection of the present invention is defined by the appended claims.

Claims

1. Compounds of general formula (1), or their optical isomers, crystalline forms, pharmaceutically acceptable salts, hydrates, or solvates. 【Chemistry 1】 (In general formula (1), L is 【Chemistry 2】 And here, ＊ " indicates the site that binds to the carbonyl group. X is either NH or NMe, R １ は、Me、Et、-CH ２ CH ２ CH ３ ,-EH(EH ３ ) ２ 、 【Transformation 3】 And, R ２ and R ３ are each independently H, Me or Et, or R ２ and R ３ together with the carbon atoms to which they are attached 【Chemistry 4】 Forming, R ４ H is, R 5 は、Me、Et、-CH ２ CH ２ CH ３ ,-EH(EH ３ ) ２ 、-CH ２ HH(H) ３ ) ２ 、 【Transformation 5】 (That is the case.)

2. In general formula (1), R ２ and R ３ However, independently, it is either H or Me, or R ２ and R ３ However, along with the carbon atoms bonded to them, 【Transformation 6】 The compound according to claim 1, or its optical isomer, crystalline form, pharmaceutically acceptable salt, hydrate, or solvate, which forms [the specified compound].

3. The compound has the following structure: 【Chemistry 7-1】 【Chemistry 7-2】 【Transformation 7-3】 A compound according to claim 1 or 2, having one of the above, or an optical isomer, crystalline form, pharmaceutically acceptable salt, hydrate or solvate thereof.

4. A pharmaceutical composition comprising a pharmaceutically acceptable excipient or carrier, and as an active ingredient, the compound described in claim 1, or its optical isomer, crystalline form, pharmaceutically acceptable salt, hydrate, or solvate.

5. Use of the compound according to claim 1, or its optical isomer, crystalline form, pharmaceutically acceptable salt, hydrate, or solvate, in the preparation of a pharmaceutical for treating related diseases mediated by IDH mutant proteins.

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