Compounds for inhibiting conversion of alpha-KG into D-2-HG and preparation and application thereof

Abstract

The invention provides a compound for inhibiting conversion of alpha-KG into D-2-HG as well as preparation and application thereof, particularly, the compound has a structure as shown in a formula (I), and the compound can inhibit the activity of an IDH2 mutant and inhibit conversion of alpha-ketoglutaric acid (alpha-KG) into D-2-hydroxyglutaric acid (D-2-HG). The compounds are useful in the treatment of diseases associated with the conversion of alpha-KG to D-2-HG, preferably diseases associated with IDH2 mutants, including cancer. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medicine, and particularly to a class of IDH2 mutant inhibitors, their preparation methods and uses. The compounds can inhibit the activity of mutant IDH2 and inhibit the conversion of α-ketoglutaric acid (α-KG) to D-2-hydroxyglutaric acid (D-2-HG). The compounds can be used to prepare a pharmaceutical composition containing the compounds as active ingredients; the compounds and the pharmaceutical composition can be used to treat diseases related to the conversion of α-KG to D-2-HG. Background Art

[0002] Intracellular metabolic reprogramming plays a crucial role in the occurrence and development of tumors (Hanahan D et al., Cell, 2011, 144: 646-74; Pavlova NN et al., Cell Metab, 2016, 23: 27-47). As a key enzyme in the central carbon metabolic pathway, isocitrate dehydrogenase 2 (IDH2) catalyzes the conversion of isocitrate to α-ketoglutaric acid (α-KG), while converting nicotinamide adenine dinucleotide phosphate (NADP +)It is reduced to NADPH. However, once the arginine at the active center mutates (such as: R140Q or R172K), IDH2 will lose its original catalytic function and instead use NADPH to reduce α-KG to D-2-hydroxyglutaric acid (D-2-HG) (Gross S et al., J Exp Med, 2010, 207:339-44; Ward PS et al., Cancer Cell, 2010, 17:225-34). The accumulated D-2-HG in cells can competitively inhibit α-KG-dependent dioxygenases (including histone demethylases and methylcytosine dioxygenases that regulate the epigenetic state of cells), and the resulting epigenetic dysregulation often leads to cell differentiation disorders, thereby promoting the occurrence and development of various tumors (Figueroa ME et al., Cancer Cell, 2010, 18:553-67; Xu W et al., Cancer Cell, 2011, 19:17-30; Chowdhury R et al., EMBO Rep, 2011, 12:463-9; Koivunen P et al., Nature, 2012, 483:484-8; Turcan S et al., Nature, 2012, 483:479-83; Lu C et al., Nature, 2012, 483:474-8; Losman JA et al., Science, 2013, 339:1621-5; Wang F et al., Science, 2013, 340:622-6; Lu C et al., Genes Dev, 2013, 27:1986-98; Saha SK et al., Nature, 2014, 513:110-4). Therefore, in theory, one can inhibit the activity of IDH2 mutants, reduce the level of D-2-HG in tumor cells, thereby reversing cell differentiation disorders, and ultimately achieve the goal of treating tumors. It is worth mentioning that, as the first specific inhibitor of IDH2 mutants, AG-221 was approved by the US Food and Drug Administration (FDA) in 2017 for the treatment of relapsed or refractory acute myeloid leukemia (AML) with IDH2 mutations.

[0003] However, in addition to acute myeloid leukemia, IDH2 mutations have actually been found in a variety of other tumors, including gliomas, myelodysplastic syndromes (MDS), cholangiocarcinomas, and chondrosarcomas, etc. (Parsons DW et al., Science, 2008, 321:1807-12; Yan H et al., N Engl J Med, 2009, 360:765-73; Kosmider O et al., Leukemia, 2010, 24:1094-6; Amary MF et al., J Pathol, 2011, 224:334-43; Borger DR et al., Oncologist, 2012, 17:72-9). Therefore, there is still a large clinical demand for novel IDH2 mutant inhibitors that can be used for other tumors. SUMMARY OF THE INVENTION

[0004] The object of the present invention is to provide a class of compounds that inhibit the conversion of α-KG to 2-HG, and their preparation and uses.

[0005] In a first aspect of the present invention, there is provided a compound or a pharmaceutically acceptable salt thereof, which has a structure shown in the following formula (I)

[0006]

[0007] Wherein:

[0008] Ring A and ring B are each independently selected from a substituted or unsubstituted C6-C 10 aryl ring, a substituted or unsubstituted 5-12-membered heteroaryl ring;

[0009] R 1 and R 2 are independently selected from the group consisting of: hydrogen, a substituted or unsubstituted C1-C6 alkyl, a substituted or unsubstituted C3-C6 cycloalkyl, a substituted or unsubstituted 3-6-membered heterocycloalkyl, a halogen, a hydroxyl group, an amino group, a carboxyl group, a nitro group, a cyano group, a substituted or unsubstituted C2-C4 alkanoyl group, a substituted or unsubstituted C2-C4 amide group, a C2-C4 sulfonyl group, a C2-C4 sulfinyl group, a mercapto group;

[0010] m is selected from 1 or 2; is a single bond or a double bond, provided that is an aromatic ring;

[0011] n is 0, 1 or 2;

[0012] Unless otherwise specified, the substitution mentioned herein means that each of the said groups is independently substituted by 1 - 3 substituents selected from the group consisting of: halogen, hydroxyl, amino, cyano, -C1-C4 alkyl, -C1-C4 haloalkyl, -C1-C4 hydroxyalkyl, -C1-C4 alkoxy, -C1-C4 alkylamino, -C1-C4 cycloalkyl, -C1-C4 halocycloalkyl, -C1-C4 hydroxycycloalkyl, -C1-C4 cycloalkoxy, -C1-C4 cycloalkylamino, C2-C4 amido, C2-C4 sulfonyl, C2-C4 sulfinyl;

[0013] Unless otherwise specified, each of the said heterocycloalkyl or heteroaryl independently contains 1, 2, 3 or 4 heteroatoms selected from O, S and N.

[0014] In another preferred embodiment, the compound of formula (I) has the structure shown in formula IA or IB as follows:

[0015]

[0016] In another preferred embodiment, ring A is selected from the group consisting of: substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl, substituted or unsubstituted pyrimidinyl, substituted or unsubstituted pyrazinyl; preferably, the said ring A is 2-pyridyl substituted with trifluoromethyl.

[0017] In another preferred embodiment, ring B is selected from the group consisting of: substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl, substituted or unsubstituted pyrimidinyl, substituted or unsubstituted pyrazinyl; preferably, the said ring B is 2-pyridyl substituted with trifluoromethyl, or 4-pyridyl substituted with trifluoromethyl.

[0018] In another preferred embodiment, the compound is selected from the group consisting of

[0019]

[0020] In the second aspect of the present invention, there is provided the use of the compound as described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof, for preparing a pharmaceutical composition for treating diseases related to blocking the conversion of α-KG to D-2-HG, wherein the said diseases are preferably diseases related to IDH2 mutants, including cancer.

[0021] In the third aspect of the present invention, there is provided a pharmaceutical composition which comprises a compound of formula (I) as described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0022] In a fourth aspect of the present invention, there is provided a method for treating a disease with D-2-HG accumulation in a patient, comprising the step of administering to an individual a therapeutically effective amount of a compound of formula (I) as described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition as described in the third aspect of the present invention, wherein the disease is preferably cancer.

[0023] In a fifth aspect of the present invention, there is provided a method for inhibiting the conversion of α-KG to D-2-HG, comprising the step of administering to an individual a therapeutically effective amount of a compound of formula (I) as described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition as described in the third aspect of the present invention.

[0024] In a sixth aspect of the present invention, there is provided a method for inhibiting mutant IDH2, wild-type IDH2 or both of the above, comprising the step of administering to an individual a therapeutically effective amount of a compound of formula (I) as described in the first aspect of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition as described in the third aspect of the present invention.

[0025] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be repeated one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Representative reactions catalyzed by wild-type and mutant IDH1 / 2. DETAILED DESCRIPTION OF THE INVENTION

[0027] Through extensive and in-depth research, and through a large number of screenings and tests, the present inventors have provided a class of IDH2 mutant inhibitors, their preparation methods and uses. The compounds of the present invention can inhibit the conversion of α-ketoglutaric acid (α-KG) to D-2-hydroxyglutaric acid (D-2-HG), thereby reducing the level of D-2-HG in cells and further reversing the cell differentiation disorder. On this basis, the present invention has been completed.

[0028] TERMS

[0029] The compounds provided herein are described with reference to general chemical formulas and specific compounds. In addition, the compounds of the present disclosure can exist in many different forms or derivatives, all of which are within the scope of the present disclosure. These different forms or derivatives include, for example, tautomers, stereoisomers, racemic mixtures, salts, prodrugs, solvated forms, different crystal forms or polymorphs, and active metabolites.

[0030] Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention pertains.

[0031] As used herein, when referring to a specifically recited numerical value, the term "about" means that the value can vary by no more than 1% from the recited value. For example, as used herein, the expression "about 100" includes all values between 99 and 101 (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0032] As used herein, the term "comprising" or "including" can be open-ended, semi-closed, and closed. In other words, the term also includes "consisting essentially of" or "consisting of".

[0033] As used herein, the term "room temperature" or "ambient temperature" refers to a temperature of 4 - 40 °C, preferably 25 ± 5 °C.

[0034] As used herein, the term "isomer" includes any and all geometric isomers and stereoisomers. By way of example, within the scope of the present invention, "isomer" includes cis and trans isomers, E-isomers and Z-isomers, R-enantiomers and S-enantiomers, diastereomers, (D)-isomers, (L)-isomers, their racemic mixtures, and other mixtures thereof.

[0035] The compounds of the present disclosure also include prodrugs, active metabolite derivatives (active metabolites), active intermediates, and pharmaceutically acceptable salts thereof.

[0036] As used herein, the term "prodrug" refers to a compound or a pharmaceutically acceptable salt thereof that produces the desired active compound when metabolized under physiological conditions or when converted by solvolysis. Prodrugs include, but are not limited to, esters, amides, carbamates, carbonates, acylureas, solvates, or hydrates of the active compound. Typically, prodrugs are inactive or less active than the active compound, but may provide one or more of favorable disposition, use, and / or metabolic characteristics. For example, some prodrugs are esters of the active compound; during metabolism, the ester group is cleaved to produce the active drug. In addition, some prodrugs are activated by enzymes to produce the active compound, or are compounds that produce the active compound after further chemical reactions. Prodrugs can be developed from the prodrug form to the active form in a single step, or may have one or more intermediate forms that may themselves be active or inactive. The preparation and use of prodrugs are discussed in T. Higuchi and V. Stella, "Pro-drugs as Novel Delivery Systems", A.C.S. Symposium Series Vol. 14, and "Bioreversible Carriers in Drug Design", edited by Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987, both of which are hereby incorporated by reference in their entireties.

[0037] As used herein, the term "metabolite" (e.g., active metabolite) overlaps with the prodrug as described above. Thus, the metabolite is a pharmacologically active compound or a compound that is further metabolized to a pharmacologically active compound, which is a derivative produced by a metabolic process in an individual's body. For example, such metabolites can be produced by oxidizing, reducing, hydrolyzing, amidating, deamidating, esterifying, deesterifying, enzymatically cleaving, etc. of the administered compound or salt or prodrug. Among them, the active metabolite is the pharmacologically active derivative compound. For prodrugs, the prodrug compound is generally inactive or less active than the metabolite. For active metabolites, the parent compound can be an active compound or can be an inactive prodrug.

[0038] Prodrugs and active metabolites can be identified using conventional techniques known in the art. See, for example, Bertolini et al., 1997, Journal of Medicinal Chemistry 40:2011-2016; Shan et al., Journal of Pharmaceutical Sciences 86:756-757; Bagshawe, 1995, Drug Development Research 34:220-230; Wermuth, ibid.

[0039] As used herein, the term "active intermediate" refers to an intermediate compound in a synthesis process that exhibits the same or substantially the same biological activity as the ultimately synthesized compound.

[0040] The compounds of the present disclosure can be formulated into pharmaceutically acceptable salts or in the form of pharmaceutically acceptable salts. Unless otherwise indicated, the compounds provided herein include pharmaceutically acceptable salts of such compounds.

[0041] As used herein, the term "pharmaceutically acceptable" indicates that the substance or composition is chemically and / or toxicologically compatible with the other components of the formulation and / or the individual being treated therewith.

[0042] As used herein, unless otherwise indicated, the term "pharmaceutically acceptable salt" includes salts that retain the biological effectiveness of the free acids and bases of the designated compounds and are not otherwise undesirable biologically. Pharmaceutically acceptable salt forms contemplated include, but are not limited to, mono-, di-, tri-, tetra-salts, etc. Pharmaceutically acceptable salts are non-toxic at the amounts and concentrations at which they are administered. The preparation of such salts can facilitate pharmacological use by altering the physical characteristics of the compound without interfering with its ability to exert its physiological effects. Useful alterations in physical properties include lowering the melting point to facilitate transmucosal administration and increasing solubility to facilitate the use of higher drug concentrations.

[0043] Pharmaceutically acceptable salts include acid addition salts, such as acid addition salts containing: sulfate, chloride, hydrochloride, fumarate, maleate, phosphate, sulfamate, acetate, citrate, lactate, tartrate, mesylate, esylate, benzenesulfonate, p-toluenesulfonate, cyclohexylsulfamate, and quinate. Pharmaceutically acceptable salts can be obtained from acids such as: hydrochloric acid, maleic acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylsulfamic acid, fumaric acid, and quinic acid.

[0044] When acidic functional groups (such as carboxylic acid or phenol) are present, pharmaceutically acceptable salts also include base addition salts, such as base addition salts containing the following: benzathine, chloroprocaine, choline, diethanolamine, ethanolamine, tert-butylamine, ethylenediamine, meglumine, procaine, aluminum, calcium, lithium, magnesium, potassium, sodium, ammonium, alkylamine, and zinc. For example, see Remington's Pharmaceutical Sciences, 19th Edition, Mack Publishing Co., Easton, PA, Volume 2, page 1457, 1995; Stahl and Wermuth's "Handbook of Pharmaceutical Salts: Properties, Selection, and Use", Wiley-VCH, Weinheim, Germany, 2002. Such salts can be prepared using the appropriate corresponding base.

[0045] Pharmaceutically acceptable salts can be prepared by standard techniques. For example, the free base form of the compound can be dissolved in a suitable solvent (such as an aqueous solution or a water-alcohol solution containing the appropriate acid), and then separated by evaporating the solution. Thus, if the particular compound is a base, the desired pharmaceutically acceptable salt can be prepared by any suitable method available in the art, for example, treating the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., or with an organic acid such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosyl acids (such as glucuronic acid or galacturonic acid), α-hydroxy acids (such as citric acid or tartaric acid), amino acids (such as aspartic acid or glutamic acid), aromatic acids (such as benzoic acid or cinnamic acid), sulfonic acids (such as p-toluenesulfonic acid or ethanesulfonic acid), etc.

[0046] Similarly, if the particular compound is an acid, the desired pharmaceutically acceptable salt can be prepared by any suitable method, for example, treating the free acid with an inorganic or organic base such as an amine (primary, secondary, or tertiary), an alkali metal hydroxide, or an alkaline earth metal hydroxide, etc. Illustrative examples of suitable salts include: organic salts derived from amino acids (such as L-glycine, L-lysine, and L-arginine), ammonia, primary, secondary, and tertiary amines, and cyclic amines (such as hydroxyethylpyrrolidine, piperidine, morpholine, or piperazine); and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.

[0047] It should also be understood that the compounds of the present disclosure can exist in non-solvated form, solvated form (e.g., hydrated form), and solid form (e.g., crystalline or polymorphic form), and the present disclosure intends to cover all such forms.

[0048] As used herein, the terms "solvate" or "solvated form" refer to an addition form containing a stoichiometric or non-stoichiometric amount of a solvent. Some compounds tend to trap a fixed molar ratio of solvent molecules when in the crystalline solid state, thus forming a solvate. If the solvent is water, the resulting solvate is a hydrate; and if the solvent is an alcohol, the resulting solvate is an alcoholate. Hydrates are formed by combining one or more water molecules with a molecular substance, where the water retains its molecular state as H2O. Examples of solvents that form solvates include, but are not limited to, water, isopropyl alcohol, ethanol, methanol, DMSO, ethyl acetate, acetic acid, and ethanolamine.

[0049] As used herein, the terms "crystal form", "crystalline form", "polymorphic form", and "polymorph" are used interchangeably and refer to crystal structures in which a compound (or its salt or solvate) can crystallize in different crystal packing arrangements, all of which have the same elemental composition. Different crystal forms generally have different X-ray diffraction patterns, infrared spectra, melting points, densities, hardnesses, crystal shapes, optical and electrical properties, stability, and solubility. The recrystallization solvent, crystallization rate, storage temperature, and other factors can cause one crystal form to predominate. Polymorphs of a compound can be prepared by crystallization under different conditions.

[0050] The present disclosure also intends to include all isotopes of the atoms in the compounds. Isotopes of an atom include atoms having the same atomic number but different mass numbers. By way of example, unless otherwise stated, hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine, or iodine in the compounds of the present disclosure are meant to also include their isotopes, such as, but not limited to 1 H, 2 H, 3 H, 11 C, 12 C, 13 C, 14 C, 14 N, 15 N, 16 O, 17 O, 18 O, 31 P, 32 P, 32 S, 33 S, 34 S, 36 S, 17 F, 19 F, 35 Cl,37 Cl, 79 Br, 81 Br, 127 I, and 131 I. In some embodiments, hydrogen includes protium, deuterium, and tritium. In some embodiments, carbon includes 12 C, and 13 C.

[0051] Treatment method

[0052] The present disclosure provides a method for treating a disease associated with an IDH2 mutant, the method comprising administering to a subject an effective amount of one or more compounds, pharmaceutically acceptable salts, hydrates, solvates or stereoisomers thereof, or a pharmaceutical composition disclosed herein.

[0053] In some embodiments, the one or more compounds, pharmaceutically acceptable salts, hydrates, solvates or stereoisomers thereof, or the pharmaceutical composition disclosed herein, are administered via a parenteral or non-parenteral route. In some embodiments, the one or more compounds, pharmaceutically acceptable salts, hydrates, solvates or stereoisomers thereof, or the pharmaceutical composition, are administered by: oral administration, oral administration, buccal administration, nasal administration, intranasal administration, transmucosal administration, epidermal administration, transdermal administration, cutaneous administration, ocular administration, pulmonary administration, sublingual administration, rectal administration, vaginal administration, topical administration, subcutaneous administration, intravenous administration, intramuscular administration, intra-arterial administration, intrathecal administration, intracapsular administration, intraperitoneal administration, intracardiac administration, intradermal administration, intraperitoneal administration, intratracheal administration, subepidermal administration, intra-articular administration, subcapsular administration, subarachnoid administration, intraspinal or intrasternal administration.

[0054] The compounds provided herein can be administered in pure form, in combination with other active ingredients, or in the form of the pharmaceutical compositions of the present disclosure. In some embodiments, the compounds provided herein can be administered to a subject in need thereof simultaneously or sequentially in combination with one or more anti-cancer agents known in the art. In some embodiments, the administration is carried out once a day, twice a day, three times a day, or once every two days, once every three days, once every four days, once every five days, once every six days, once a week.

[0055] In certain embodiments, the present disclosure provides the use of the compounds, their pharmaceutically acceptable salts, hydrates, solvates or stereoisomers, or the pharmaceutical compositions of the present disclosure, in the preparation of a medicament for the treatment of a disease associated with the conversion of α-KG to D-2-HG. In certain embodiments, the present disclosure provides the use of the compounds, their pharmaceutically acceptable salts, hydrates, solvates or stereoisomers, or the pharmaceutical compositions of the present disclosure, in the preparation of a medicament for the treatment of a disease associated with an IDH2 mutant.

[0056] In certain embodiments, a disease associated with the conversion of α-KG to D-2-HG is a disease associated with an IDH2 mutant, including cancer.

[0057] In particular, the cancer includes but is not limited to leukemia, glioblastoma, melanoma, chondrosarcoma, cholangiocarcinoma, osteosarcoma, lymphoma, lung cancer, adenoma, myeloma, hepatocellular carcinoma, adrenocortical carcinoma, pancreatic cancer, breast cancer, prostate cancer, liver cancer, gastric cancer, colon cancer, colorectal cancer, ovarian cancer, cervical cancer, brain cancer, esophageal cancer, bone cancer, testicular cancer, skin cancer, kidney cancer, mesothelioma, neuroblastoma, thyroid cancer, head and neck cancer, esophageal cancer, eye cancer, prostate cancer, nasopharyngeal cancer or oral cancer. In some embodiments, the cancer is leukemia, glioblastoma or cholangiocarcinoma.

[0058] The compounds of the present disclosure and their pharmaceutical compositions can be used to prevent or treat the onset or development of any disease or disorder associated with the conversion of α-KG to D-2-HG in a mammal (especially a human). In some embodiments, the compounds of the present disclosure and their pharmaceutical compositions can be used to prevent or treat the onset or development of any disease or disorder associated with a mutant IDH in a mammal (especially a human).

[0059] In such a case, the present invention also provides a method for screening a patient suitable for treatment with the compounds or pharmaceutical compositions of the present disclosure alone or in combination with other ingredients (such as a second active ingredient, such as an anti-cancer agent). The method includes sequencing a tumor sample from the patient and detecting the accumulation of D-2-HG in the patient or detecting the mutation status of IDH in the patient.

[0060] Use of the compound

[0061] In one aspect, the compound of formula (I) or its pharmaceutically acceptable salt can inhibit the conversion of α-KG to D-2-HG.

[0062] In another aspect, the compound of formula (I) or a pharmaceutically acceptable salt thereof can inhibit mutant IDH2. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof can inhibit wild-type IDH2. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof can inhibit both mutant IDH and wild-type IDH2. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof can selectively inhibit mutant IDH2 but not wild-type IDH2.

[0063] As used herein, the term "selectively inhibits mutant IDH2" means that the provided compound inhibits mutant IDH more than wild-type IDH in at least one assay described herein.

[0064] Pharmaceutical composition

[0065] The present disclosure provides pharmaceutical compositions comprising at least one compound disclosed herein. In some embodiments, the pharmaceutical composition comprises more than one compound disclosed herein. In some embodiments, the pharmaceutical composition comprises one or more compounds disclosed herein and a pharmaceutically acceptable carrier.

[0066] A pharmaceutically acceptable carrier is a conventional pharmaceutical carrier in the art and can be prepared in a manner well known in the pharmaceutical art. In some embodiments, the compounds disclosed herein can be mixed with a pharmaceutically acceptable carrier to prepare a pharmaceutical composition.

[0067] As used herein, the phrase "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that are suitable for use in contact with the tissues of humans and animals within the scope of sound medical judgment, without excessive toxicity, irritation, allergic response, or other problems or complications, and commensurate with a reasonable benefit / risk ratio. In some embodiments, pharmaceutically acceptable compounds, materials, compositions, and / or dosage forms refer to those compounds, materials, compositions, and / or dosage forms that are approved by a regulatory agency (such as the U.S. Food and Drug Administration, the China Food and Drug Administration, or the European Medicines Agency) or listed in a recognized pharmacopoeia (such as the U.S. Pharmacopoeia, the China Pharmacopoeia, or the European Pharmacopoeia) for use in animals and particularly humans.

[0068] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or vehicle involved in carrying or transporting the compounds provided herein from one location, body fluid, tissue, organ (internal or external), or part of the body to another location, body fluid, tissue, organ, or part of the body, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. A pharmaceutically acceptable carrier can be a vehicle, diluent, excipient, or other material that can be used in contact with animal tissue without excessive toxicity or adverse effects. Exemplary pharmaceutically acceptable carriers include sugars, starches, celluloses, malt, tragacanth, gelatin, Ringer's solution, alginic acid, isotonic saline, buffers, and the like. Pharmaceutically acceptable carriers useful in the present disclosure include those generally known in the art, such as those disclosed in Remington Pharmaceutical Sciences, Mack Pub. Co., New Jersey (1991), which is incorporated herein by reference.

[0069] Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) alcohols, such as ethanol and propanol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances used in pharmaceutical formulations, such as acetone.

[0070] The pharmaceutical composition may optionally contain pharmaceutically acceptable auxiliary substances to approximate physiological conditions, such as pH regulators and buffers, toxicity regulators, etc., such as sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, and the like.

[0071] The form of the pharmaceutical composition depends on a variety of criteria, including but not limited to the route of administration, the degree of the disease, or the dose to be administered.

[0072] The pharmaceutical composition can be formulated for oral, nasal, rectal, transdermal, intravenous or intramuscular administration. Depending on the desired route of administration, the pharmaceutical composition can be formulated in the form of tablets, capsules, pills, dragees, powders, granules, sachets, cachets, lozenges, suspensions, emulsions, solutions, syrups, aerosols (in solid form or in a liquid medium), sprays, ointments, pastes, creams, lotions, gels, patches, inhalants or suppositories.

[0073] In some embodiments, the pharmaceutical composition comprises one or more of the compounds disclosed herein as a first active ingredient and further comprises a second active ingredient. The second active ingredient can be any anti-cancer agent known in the art. Representative examples of anti-cancer agents for treating cancer or tumors can include, but are not limited to, cell signal transduction inhibitors (e.g., imatinib, gefitinib, bortezomib, erlotinib, sorafenib, sunitinib, dasatinib, vorinostat, lapatinib, temsirolimus, nilotinib, everolimus, pazopanib, trastuzumab, bevacizumab, cetuximab, ranibizumab, pegaptanib, panitumumab, etc.), mitotic inhibitors (e.g., paclitaxel, vincristine, vinblastine, etc.), alkylating agents (e.g., cisplatin, cyclophosphamide, chlorambucil, carmustine, etc.), antimetabolites (e.g., methotrexate, 5-FU, etc.), intercalating anti-cancer agents (e.g., actinomycin, anthracycline, bleomycin, mitomycin-C, etc.), topoisomerase inhibitors (e.g., irinotecan, topotecan, teniposide, etc.), immunotherapeutic agents (e.g., interleukin, interferon, etc.) and anti-hormonal agents (e.g., tamoxifen, raloxifene, etc.).

[0074] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions noted in the following examples are generally carried out under conventional conditions or according to the conditions recommended by the manufacturers. Unless otherwise specified, percentages and parts are calculated by weight.

[0075] The structures of the compounds shown in the following examples were characterized by nuclear magnetic resonance (NMR) or / and mass spectrometry (ESI). The chemical shifts (δ) of NMR were given in units of 10 -6 (ppm). 1H-NMR spectra were recorded on a Varian Mercury VX 400 spectrometer using tetramethylsilane (TMS) as an internal standard in dimethyl sulfoxide-d6 (DMSO-d6) or CDCl3. 1 H-NMR spectra.

[0076] ESI-MS measurements were carried out using an Agilent 1260-6230 TOF LC-MS mass spectrometer.

[0077] High performance liquid chromatography (HPLC) measurements were carried out on an Agilent 1200 LC using a Phenomen C18 column (4.6 mm * 150 mm, 0.4 μm).

[0078] Thin layer chromatography analysis was carried out using HSGF254 silica gel plates from Yantai Huanghai. The silica gel plates used for thin layer chromatography (TLC) were 0.15 mm to 0.2 mm. The silica gel plates used for separating and purifying the product by TLC were 0.4 mm to 0.5 mm.

[0079] The purified chromatographic column used the silica gel as the carrier (200 - 300 mesh, produced by Yantai Huanghai Company).

[0080] The known starting materials of the present disclosure can be synthesized by using or according to methods known in the art, or can be purchased from Alfa Aesar, Langcaster, TCI, Aldrich, Bepharm, and Scochem.

[0081] Unless otherwise specified, the reactions in the examples were carried out under an argon or nitrogen atmosphere. The argon or nitrogen atmosphere means that the reaction flask was connected to an argon or nitrogen balloon with a volume of about 1 L. Hydrogenation was usually carried out under vacuum, filled with hydrogen, and repeated three times. Unless otherwise specified, the reaction temperature in the examples was ambient temperature, i.e., 20 °C to 30 °C.

[0082] The progress of the reactions shown in the examples was monitored by TLC. The eluent systems used for the reactions included dichloromethane - methanol system and petroleum ether - ethyl acetate system. The volume ratio of the solvents was adjusted according to the different polarities of the compounds.

[0083] The column chromatography elution system for purifying compounds and the elution system for TLC included dichloromethane - methanol system and petroleum ether - ethyl acetate system. The volume ratio of the solvents was adjusted according to the different polarities of the compounds. A small amount of basic or acidic reagents such as triethylamine and acetic acid could be added for adjustment.

[0084] The abbreviations used in the following examples and elsewhere in this article are:

[0085]

[0086]

[0087] Preparation of Compound 1 in Example 1

[0088] Step 1, Compound 1 - 2

[0089] 2 - Chloro - N-(2-(trifluoromethyl)pyridin - 4 - yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H - pyrrolo[2,3 - d]pyrimidin - 4 - amine

[0090] 2-chloro-N-(2-(trifluoromethyl)pyridin-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0091]

[0092] At 0 °C, sodium hydride (148.0 mg, 3.70 mmol, 60% purity, 1.2 eq) was added to a solution of 2-(trifluoromethyl)pyridin-4-amine (500.0 mg, 3.09 mmol, 1.0 eq) in tetrahydrofuran (20 mL), and the mixture was stirred for 15 minutes. Then, 1-1 (983.4 mg, 3.09 mmol, 1.0 eq) was added to the mixed solution, and the reaction was stirred at room temperature for 2 hours. LCMS showed the appearance of the molecular ion peak of the target compound. The reaction solution was diluted with water (50 mL), and the mixture was extracted with EtOAc (30 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by flash acidic chromatography column (PE / EtOAc = 10:1) to obtain 1-2 as a colorless oil (841.7 mg, 1.90 mmol, 61.4% yield). LCMS: (ESI) m / z: 444.2 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ: 8.37 (d, J = 5.2 Hz, 1H), 7.27 (brs, 1H), 6.70 - 6.40 (m, 2H), 6.12 (brs, 1H), 5.80 (s, 2H), 3.51 (t, J = 7.7 Hz, 2H), -0.21 (t, J = 7.7 Hz, 2H), -0.95 (s, 9H). Step 2, Compound 1-3

[0093] 2-(6-(trifluoromethyl)pyridin-2-yl)-N-(2-(trifluoromethyl)pyridin-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0094] 2-(6-(trifluoromethyl)pyridin-2-yl)-N-(2-(trifluoromethyl)pyridin-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0095]

[0096] At room temperature, s-Phos (27.9 mg, 0.068 mmol, 0.1 eq), sodium carbonate (216.2 mg, 2.04 mmol, 3.0 eq) and palladium acetate (15.4 mg, 0.068 mmol, 0.1 eq) were added to a mixed solution of 1-2 (300.0 mg, 0.68 mmol, 1.0 eq), (6-(trifluoromethyl)pyridin-2-yl)boronic acid (390.0 mg, 2.04 mmol, 3.0 eq) in dioxane (20 mL) and water (2 mL). The reaction solution was purged with nitrogen three times and reacted at 100 °C for 12 hours. LCMS showed that the main peak was the target compound. The reaction solution was filtered, poured into ice water (50 mL) for dilution, and extracted with EtOAc (30 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by flash acidic chromatography column (PE / EtOAc = 10:1) to obtain 1-3 as a white solid (171.8 mg, 0.31 mmol, 45.5% yield). LC-MS: (ESI) m / z: 555.2 [M+H] + . 1 H NMR (400 MHz, CDCl3) δ: 8.47 - 8.21 (m, 2H), 7.85 (m, 1H), 7.40 - 7.15 (m, 2H), 6.69 (brs, 1H), 6.70 (brs, 1H), 6.2 (d, J = 3.5 Hz, 1H), 5.68 (s, 2H), 3.51 (d, J = 9.3 Hz, 2H), -0.19 (d, J = 9.3 Hz, 2H), -0.89 (s, 9H).

[0097] Step 3, Compound 1

[0098] 2-(6-(trifluoromethyl)pyridin-2-yl)-N-(2-(trifluoromethyl)pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0099] 2-(6-(trifluoromethyl)pyridin-2-yl)-N-(2-(trifluoromethyl)pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0100]

[0101] At room temperature, a solution of compound 1-3 (100.0 mg, 0.18 mmol, 1.0 eq) in trifluoroacetic acid (5 mL) was stirred for 1 hour. LCMS showed that 1-3 was consumed completely and the intermediate was formed. The trifluoroacetic acid was removed by rotary evaporation under reduced pressure, and the mixture was dissolved in a mixed solution of THF (4 mL) and aqueous sodium hydroxide solution (86.4 mg, 2.16 mmol, 12 eq, 1 mL). The mixture was stirred at room temperature for 3 hours. LCMS showed the appearance of the molecular ion peak of the target compound. The reaction solution was poured into saturated ammonium chloride (30 mL) for dilution and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v) - ACN]; B%: 40% - 100%, 11 min), and compound 1 as a yellow solid was obtained (20.1 mg, 0.047 mmol, 26.1%).

[0102] LC-MS: (ESI) m / z: 425.2 [M+H] + .

[0103] 1 1H NMR (400 MHz, CDCl3) δ: 8.47 - 8.21 (m, 2H), 7.90 (m, 1H), 7.42 - 7.18 (m, 2H), 6.69 (brs, 1H), 6.85 (brs, 1H), 6.21 (d, J = 3.6 Hz, 1H).

[0104] Example 2 Preparation of Compound 2

[0105] Step 1, Compound 2

[0106] 7-isopropyl-2-(6-(trifluoromethyl)pyridin-2-yl)-N-(2-(trifluoromethyl)pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0107] 7-isopropyl-2-(6-(trifluoromethyl)pyridin-2-yl)-N-(2-(trifluoromethyl)pyridin-4-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0108]

[0109] At 0 °C, isopropyl iodide (33.0 mg, 0.20 mmol, 1.0 eq.) and Cs2CO3 (130.3 mg, 0.40 mmol, 2.0 eq.) were added to a DMF solution (5 mL) of compound 1 (84.8 mg, 0.20 mmol, 1.0 eq.). Under nitrogen protection at 0 °C, the mixture was stirred and reacted for 2 hours. LCMS showed the appearance of the molecular ion peak of the target compound. The reaction solution was poured into water (30 mL) for dilution and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v) - ACN]; B%: 40% - 100%, 11 min) to obtain compound 2 as a yellow solid (16.1 mg, 0.034 mmol, 17.0% yield).

[0110] LC-MS: (ESI) m / z: 467.2 [M+H] + .

[0111] 1 1H NMR (400 MHz, CDCl3) δ: 8.45 - 8.17 (m, 2H), 7.92 (m, 1H), 7.42 - 7.22 (m, 2H), 7.03 (brs, 1H), 6.85 (brs, 1H), 6.11 (d, J = 3.9 Hz, 1H), 5.30 (d, J = 7.2 Hz, 1H), 1.50 (d, J = 7.2 Hz, 6H).

[0112] Example 3 Preparation of Compound 3

[0113] Step 1, Compound 3

[0114] 2-methyl-1-(2-(6-(trifluoromethyl)pyridin-2-yl)-4-((2-(trifluoromethyl)pyridin-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propan-2-ol

[0115] 2-methyl-1-(2-(6-(trifluoromethyl)pyridin-2-yl)-4-((2-(trifluoromethyl)pyridin-4-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propan-2-ol

[0116]

[0117] Under nitrogen protection, sodium hydride (18.4 mg, 0.46 mmol, 60% purity, 2.0 eq.) was added to a dry DMF solution (5 mL) of compound 1 (100.0 mg, 0.23 mmol, 1.0 eq.). After stirring at room temperature for 15 minutes, ((1-bromo-2-methylpropyl)oxy)tert-butyldimethylsilane (93.1 mg, 0.35 mmol, 1.5 eq.) was added to the reaction system, and the reaction was stirred at 50 °C for 20 hours under nitrogen protection to form a brown reaction solution. LCMS showed that the main peak was the target compound. The mixture was poured into water (30 mL) for dilution, and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to obtain compound 3 as a pale yellow solid (13.5 mg, 0.027 mmol, 11.7% yield).

[0118] LC-MS: (ESI) m / z: 497.1 [M+H] + .

[0119] 1 H NMR (400 MHz, CDCl3) δ: 8.47 - 8.20 (m, 2H), 7.90 (m, 1H), 7.37 - 7.16 (m, 2H), 6.95 (brs, 1H), 6.70 (brs, 1H), 6.06 (d, J = 3.8 Hz, 1H), 4.30 (s, 2H), 1.44 (s, 6H).

[0120] Example 4, Compound 4

[0121] Step 1, Compound 4-2

[0122] 6-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0123] 6-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0124]

[0125] At 0 °C, sodium hydride (222.0 mg, 5.55 mmol, 60% purity, 1.5 eq) was added to a solution of 4-(trifluoromethyl)pyridin-2-amine (600.0 mg, 3.70 mmol, 1.0 eq) in tetrahydrofuran (30 mL), and the mixture was stirred for 15 minutes. Then, 4-1 (1.22 g, 3.70 mmol, 1.0 eq) was added to the mixed solution, and the reaction was stirred at room temperature for 2 hours. The LCMS of the reaction solution showed the appearance of the molecular ion peak of the target compound. The reaction solution was diluted with water (80 mL), and the mixture was extracted with EtOAc (30 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by flash acidic chromatography column (PE / EtOAc = 10:1) to obtain 4-2 as a colorless oil (800.1 mg, 1.74 mmol, 47.0% yield).

[0126] LCMS: (ESI) m / z: 458.2 [M+H] + .

[0127] 1 H NMR (400 MHz, CDCl3) δ: 7.84 (d, J = 5.5 Hz, 1H), 6.80 - 6.62 (m, 2H), 6.12 (brs, 1H), 5.90 (s, 2H), 3.42 (t, J = 7.5 Hz, 2H), 2.30 (s, 3H), -0.18 (t, J = 7.5 Hz, 2H), -0.92 (s, 9H).

[0128] Step 2, Compound 4-3

[0129] 2-(6-(trifluoromethyl)pyridin-2-yl)-N-(2-(trifluoromethyl)pyridin-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0130] 2-(6-(trifluoromethyl)pyridin-2-yl)-N-(2-(trifluoromethyl)pyridin-4-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0131]

[0132] At room temperature, s-Phos (27.0 mg, 0.066 mmol, 0.1 eq.), sodium carbonate (209.6 mg, 1.98 mmol, 3.0 eq.), and palladium(II) acetate (14.9 mg, 0.066 mmol, 0.1 eq.) were added to a mixed solution of 4-2 (300.0 mg, 0.66 mmol, 1.0 eq.) and (6-(trifluoromethyl)pyridin-2-yl)boronic acid (378.2 mg, 1.98 mmol, 3.0 eq.) in dioxane (20 mL) and water (2 mL). The reaction mixture was purged with nitrogen three times and then reacted at 100 °C for 12 h. LCMS showed that the main peak was the target compound. The reaction mixture was filtered, poured into ice water (50 mL) for dilution, and extracted with EtOAc (30 mL × 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by flash acidic chromatography column (PE / EtOAc = 10:1) to give 4-3 as a white solid (171.8 mg, 0.30 mmol, 45.4% yield).

[0133] LC-MS: (ESI) m / z: 569.2 [M+H] + .

[0134] 1 H NMR (400 MHz, CDCl3) δ: 8.40 - 8.20 (m, 2H), 7.80 (m, 1H), 7.40 (m, 1H), 6.78 - 6.66 (m, 2H), 6.55 (brs, 1H), 5.80 (s, 2H), 3.62 (d, J = 9.0 Hz, 2H), 2.21 (s, 3H), -0.21 (d, J = 9.0 Hz, 2H), -0.88 (s, 9H).

[0135] Step 3, Compound 4

[0136] 6-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0137] 6-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0138]

[0139] At room temperature, a solution of compound 4-3 (102.3 mg, 0.18 mmol, 1.0 eq.) in trifluoroacetic acid (5 mL) was stirred for 1 hour. LCMS showed that 4-3 was consumed completely and the intermediate was formed. The trifluoroacetic acid was removed by rotary evaporation under reduced pressure. The mixture was dissolved in a mixed solution of THF (4 mL) and aqueous sodium hydroxide solution (86.4 mg, 2.16 mmol, 1 mL, 12.0 eq.). The mixture was stirred at room temperature for 3 hours. LCMS showed the appearance of the molecular ion peak of the target compound. The reaction solution was poured into saturated ammonium chloride (30 mL) for dilution and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to obtain compound 4 as a pale yellow solid (23.0 mg, 0.052 mmol, 28.8%).

[0140] LC-MS: (ESI) m / z: 439.1 [M+H] + .

[0141] 1 H NMR (400 MHz, CDCl3) δ: 8.25 - 7.80 (m, 3H), 7.90 (m, 1H), 6.72 - 6.58 (m, 2H), 5.87 (s, 1H), 2.20 (s, 3H).

[0142] Example 5, Compound 5

[0143] Step 1, Compound 5-2

[0144] 2-chloro-N-(4-(trifluoromethyl)pyridin-2-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0145] 2-chloro-N-(4-(trifluoromethyl)pyridin-2-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0146]

[0147] At 0 °C, sodium hydride (222.0 mg, 5.55 mmol, 60% purity, 1.5 eq) was added to a solution of 4-(trifluoromethyl)pyridin-2-amine (600.0 mg, 3.70 mmol, 1.0 eq) in tetrahydrofuran (30 mL), and the mixture was stirred for 15 minutes. Then, 1-1 (1.17 g, 3.70 mmol, 1.0 eq) was added to the mixed solution, and the reaction was stirred at room temperature for 2 hours. The LCMS of the reaction solution showed the appearance of the molecular ion peak of the target compound. The reaction solution was diluted with water (80 mL), and the mixture was extracted with EtOAc (30 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by flash acidic chromatography column (PE / EtOAc = 10:1) to obtain 5-2 as a colorless oil (802.2 mg, 1.81 mmol, 48.9% yield).

[0148] LCMS: (ESI) m / z: 444.1 [M+H] + .

[0149] 1 H NMR (400 MHz, CDCl3) δ: 7.90 (m, 1H), 7.27 (brs, 1H), 6.71 - 6.58 (m, 2H), 6.01 (brs, 1H), 5.58 (s, 2H), 3.40 (t, J = 9.6 Hz, 2H), -0.19 (t, J = 9.6 Hz, 2H), -0.93 (s, 9H).

[0150] Step 2, Compound 5-3

[0151] N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0152] N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7-((2-(trimethylsilyl)ethoxy)methyl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0153]

[0154] At room temperature, to a mixed solution of 5-2 (300.0 mg, 0.68 mmol, 1.0 eq), (6-(trifluoromethyl)pyridin-2-yl)boronic acid (389.6 mg, 2.04 mmol, 3.0 eq) in dioxane (20 mL) and water (2 mL) were added s-phos (27.9 mg, 0.068 mmol, 0.1 eq), sodium carbonate (216 mg, 2.04 mmol, 3.0 eq) and palladium acetate (15.4 mg, 0.068 mmol, 0.1 eq). The reaction solution was purged with nitrogen three times and reacted at 100 °C for 12 hours. LCMS showed that the main peak was the target compound. The reaction solution was filtered, poured into ice water (50 mL) for dilution, and extracted with EtOAc (30 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by flash acidic chromatography column (PE / EtOAc = 10:1) to obtain 5-3 as a white solid (171.8 mg, 0.31 mmol, 45.5% yield).

[0155] LC-MS: (ESI) m / z: 555.2 [M+H] + .

[0156] 1 H NMR (400 MHz, CDCl3) δ: 8.16 - 7.93 (m, 2H), 7.40 - 7.25 (m, 2H), 7.40 - 7.15 (m, 2H), 6.70 (brs, 1H), 6.60 (s, 1H), 5.92 (s, 2H), 3.40 (d, J = 9.3 Hz, 2H), -0.23 (d, J = 9.3 Hz, 2H), -0.92 (s, 9H).

[0157] Step 3, Compound 5

[0158] N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0159] N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0160]

[0161] At room temperature, a solution of compound 5-3 (100.0 mg, 0.18 mmol, 1.0 eq.) in trifluoroacetic acid (5 mL) was stirred for 1 hour. LCMS showed that 5-3 was consumed completely and the intermediate was formed. The trifluoroacetic acid was removed by rotary evaporation under reduced pressure, and the mixture was dissolved in a mixed solution of THF (4 mL) and aqueous sodium hydroxide solution (86.4 mg, 2.16 mmol, 1 mL, 12.0 eq.). The mixture was stirred at room temperature for 3 hours. LCMS showed the appearance of the molecular ion peak of the target compound. The reaction solution was poured into saturated ammonium chloride (30 mL) for dilution and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v) - ACN]; B%: 40% - 100%, 11 min) to obtain compound 5 as a white solid (15.8 mg, 0.037 mmol, 20.5%).

[0162] LC-MS: (ESI) m / z: 425.2 [M+H] + .

[0163] 1 H NMR (400 MHz, CDCl3) δ: 8.05 - 7.77 (m, 4H), 7.40 (m, 1H), 6.80 (brs, 1H), 6.71 (m, 1H), 6.60 (s, 1H).

[0164] Example 6, Compound 6

[0165] Step 1, Compound 6

[0166] 7-isopropyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0167] 7-isopropyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0168]

[0169] At 0 °C, isopropyl iodide (32.3 mg, 0.19 mmol, 1.0 eq.) and Cs2CO3 (123.9 mg, 0.38 mmol, 2.0 eq.) were added to a DMF solution (5 mL) of compound 5 (80.0 mg, 0.19 mmol, 1.0 eq.). Under nitrogen protection, the mixture was stirred at 0 °C for 2 h. The molecular ion peak of the target compound appeared as shown by LCMS. The reaction solution was poured into water (30 mL) for dilution and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to give compound 6 as a pale yellow solid (16.1 mg, 0.035 mmol, 18.4% yield).

[0170] LC-MS: (ESI) m / z: 467.2 [M+H] + .

[0171] 1 1H NMR (400 MHz, CDCl3) δ: 8.15 - 7.80 (m, 3H), 7.45 - 7.25 (m, 2H), 6.77 - 6.65 (m, 2H), 6.06 (d, J = 3.9 Hz, 1H), 5.36 (m, 1H), 1.52 (d, J = 7.5 Hz, 6H).

[0172] Example 7, Compound 7

[0173] Step 1, Compound 7

[0174] 2-methyl-1-(2-(trifluoromethyl)pyridin-2-yl)-4-((4-(trifluoromethyl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propan-2-ol

[0175] 2-methyl-1-(2-(trifluoromethyl)pyridin-2-yl)-4-((4-(trifluoromethyl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)propan-2-ol

[0176]

[0177] Under nitrogen protection, sodium hydride (24 mg, 0.6 mmol, 60% purity, 2.5 eq.) was added to a dry DMF solution (4 mL) of compound 5 (100.0 mg, 0.24 mmol, 1.0 eq.). After stirring at room temperature for 15 minutes, ((1-bromo-2-methyl-propan-2-yl)oxy)tert-butyldimethylsilane (127.6 mg, 0.48 mmol, 2.0 eq.) was added to the reaction system. The reaction was stirred at 50 °C for 20 hours under nitrogen protection to form a brown reaction solution. LCMS showed that the main peak was the target compound. The mixture was poured into saturated NH4Cl solution (30 mL), and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*255u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to obtain compound 7 as a pale yellow solid (13.4 mg, 0.027 mmol, 11.2% yield).

[0178] LC-MS: (ESI) m / z: 497.2 [M+H] + .

[0179] 1 H NMR (400 MHz, CDCl3) δ: 8.12 - 7.83 (m, 3H), 7.40 - 7.21 (m, 2H), 6.77 - 6.65 (m, 2H), 6.10 (d, J = 3.9 Hz, 1H), 4.10 (s, 2H), 1.32 (s, 6H).

[0180] Example 8, compound 8

[0181] Step 1, compound 8

[0182] 6,7-dimethyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0183] 6,7-dimethyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0184]

[0185] At 0 °C, isopropyl iodide (30.6 mg, 0.18 mmol, 1.0 eq.) and Cs2CO3 (117.3 mg, 0.36 mmol, 2.0 eq.) were added to a DMF solution (5 mL) of compound 4 (80.0 mg, 0.18 mmol, 1.0 eq.). Under nitrogen protection, the mixture was stirred at 0 °C for 2 hours. LCMS showed the appearance of the molecular ion peak of the target compound. The reaction solution was poured into water (30 mL) for dilution and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v) - ACN]; B%: 40% - 100%, 11 min) to obtain compound 8 as a pale yellow solid (13.6 mg, 0.030 mmol, 16.6% yield).

[0186] LC-MS: (ESI) m / z: 453.2 [M+H] + .

[0187] 1 1H NMR (400 MHz, CDCl3) δ: 8.12 - 7.83 (m, 3H), 7.36 (m, 1H), 6.75 - 6.60 (m, 2H), 5.76 (s, 1H), 3.65 (s, 3H), 2.25 (s, 3H).

[0188] Example 9, Compound 9

[0189] Step 1, Compound 9

[0190] 2-(6-Methyl-2-(6-(trifluoromethyl)pyridin-2-yl)-4-((4-(trifluoromethyl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethanol

[0191] 2-(6-methyl-2-(6-(trifluoromethyl)pyridin-2-yl)-4-((4-(trifluoromethyl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-ol

[0192]

[0193] Under nitrogen protection, sodium hydride (23.0 mg, 0.58 mmol, 60% purity, 2.5 eq.) was added to a dry DMF solution (4 mL) of compound 4 (100.0 mg, 0.23 mmol, 1.0 eq.). After stirring at room temperature for 15 minutes, (2-bromoethoxy)tert-butyldimethylsilane (83.3 mg, 0.35 mmol, 1.5 eq.) was added to the reaction system, and the reaction was stirred at 50 °C for 20 hours under nitrogen protection to form a brown reaction solution. LCMS showed that the main peak was the target compound. The mixture was poured into saturated NH4Cl solution (30 mL), and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to obtain compound 9 as a white solid (18.5 mg, 0.038 mmol, 16.5% yield).

[0194] LC-MS: (ESI) m / z: 483.1 [M+H] + .

[0195] 1 H NMR (400 MHz, MeOD-d4) δ: 8.24 - 7.91 (m, 3H), 7.36 (m, 1H), 6.81 - 6.70 (m, 2H), 5.83 (s, 1H), 4.52 (t, J = 6.5 Hz, 2H), 3.30 (t, J = 6.5 Hz, 2H), 2.49 (s, 3H).

[0196] Example 10, Compound 10

[0197] Step 1, Compound 10

[0198] 6-methyl-7-(oxetan-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0199] 6-methyl-7-(oxetan-3-yl)-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0200]

[0201] At 90 °C, a DMF solution (10 mL) of compound 4 (200.0 mg, 0.46 mmol, 1 eq.), 3-iodooxetane (101.2 mg, 0.55 mmol, 1.2 eq.) and Cs2CO3 (300.0 mg, 0.92 mmol, 2 eq.) was stirred for 3 h. LCMS showed the appearance of the molecular ion peak of the target compound. The mixture was diluted with water (50 mL), extracted with EtOAc (30 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to give compound 10 as a pale yellow solid (24.5 mg, 0.050 mmol, 10.8% yield).

[0202] LC-MS: (ESI) m / z: 495.0 [M+H] + .

[0203] 1 1H NMR (400 MHz, MeOD-d4) δ: 9.43 (s, 1H), 8.80 (d, J = 8.0 Hz, 1H), 8.52 (d, J = 5.2 Hz, 1H), 8.16 (t, J = 7.6 Hz, 1H), 7.87 (d, J = 6.8 Hz, 1H), 7.29 (d, J = 4.0 Hz, 1H), 6.67 (d, J = 0.8 Hz, 1H), 5.95 - 5.87 (m, 1H), 5.68 (t, J = 6.8 Hz, 2H), 5.17 - 5.14 (dd, J = 8.0, 6.8 Hz, 2H), 2.55 (d, J = 0.8 Hz, 3H).

[0204] Example 11, Compound 11

[0205] Step 1, Compound 11

[0206] 6-Methyl-7-(2,2,2-trifluoroethyl)-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0207] 6-methyl-7-(2,2,2-trifluoroethyl)-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0208]

[0209] At 80 °C, a DMF solution (5 mL) of compound 4 (100 mg, 0.23 mmol, 1.0 eq.), 2,2,2-trifluoroethyl trifluoromethanesulfonate (65.0 mg, 0.28 mmol, 1.2 eq.) and Cs2CO3 (150.0 mg, 0.46 mmol, 2.0 eq.) was stirred and reacted for 0.5 h. LCMS showed the molecular ion peak of the target compound. The mixture was diluted with water (30 mL), extracted with EtOAc (20 mL x 3), washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to obtain compound 11 as a pale yellow solid (10.8 mg, 0.020 mmol, 8.6% yield).

[0210] LC-MS: (ESI) m / z: 521.1 [M+H] + .

[0211] 1 1H NMR (400 MHz, MeOD-d4) δ: 9.34 (s, 1H), 8.77 (d, J = 8.0 Hz, 1H), 8.54 (d, J = 5.2 Hz, 1H), 8.18 (t, J = 8.0 Hz, 1H), 7.90 (d, J = 8.0 Hz, 1H), 7.31 (d, J = 4.4 Hz, 1H), 6.70 (d, J = 1.2 Hz, 1H), 5.25 - 5.18 (q, J = 8.8 Hz, 2H), 2.57 (s, 3H).

[0212] Example 12, Compound 12

[0213] Step 1, Compound 12

[0214] 7-Ethyl-N-(4-(trifluoroethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0215] 7-ethyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0216]

[0217] At 0°C, ethyl iodide (35.9 mg, 0.23 mmol, 1.0 eq.) and Cs2CO3 (150.0 mg, 0.46 mmol, 2.0 eq.) were added to a DMF solution (5 mL) of compound 5 (100.0 mg, 0.23 mmol, 1.0 eq.). The mixture was stirred and reacted for 2 hours under nitrogen protection at 0°C. LCMS showed that the molecular ion peak of the target compound appeared. The reaction solution was poured into saturated ammonium chloride (30 mL) and diluted, and extracted with EtOAc (20 mL x 2). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to give compound 12 (22.5 mg, 0.049 mmol, 21.3% yield) as a pale yellow solid.

[0218] LC-MS: (ESI) m / z: 453.2 [M+H] + .

[0219] 1 H NMR (400MHz, CDCl3) δ: 8.15-7.85 (m, 3H), 7.40-7.29 (m, 2H), 6.80-6.62 (m, 2H), 6.02 (d, J = 3.8Hz, 1H), 4.33 (t, J = 7.3Hz, 2H), 1.22 (t, J = 7.3Hz, 3H).

[0220] Example 13, Compound 13

[0221] Step 1, Compound 13

[0222] 7-propyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0223] 7-propyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0224]

[0225] At 0 °C, iodopropane (33.0 mg, 0.23 mmol, 1.0 eq.) and Cs2CO3 (150.0 mg, 0.46 mmol, 2.0 eq.) were added to a DMF solution (5 mL) of compound 5 (100.0 mg, 0.23 mmol, 1.0 eq.). Under nitrogen protection at 0 °C, the mixture was stirred for 2 hours. LCMS showed the appearance of the molecular ion peak of the target compound. The reaction solution was poured into saturated NH4Cl (30 mL) for dilution and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v) - ACN]; B%: 40% - 100%, 11 min) to obtain compound 13 as a pale yellow solid (8.90 mg, 0.012 mmol, 5.2% yield).

[0226] LC-MS: (ESI) m / z: 467.0 [M+H] + .

[0227] 1 1H NMR (400 MHz, CDCl3) δ: 8.18 - 7.80 (m, 3H), 7.42 - 7.25 (m, 2H), 6.75 - 6.62 (m, 2H), 6.05 (d, J = 3.6 Hz, 1H), 4.22 (t, J = 7.5 Hz, 2H), 1.52 (m, 2H), 1.01 (t, J = 8.0 Hz, 3H).

[0228] Example 14, Compound 14

[0229] Step 1, Compound 14-2

[0230] 1-(2-chloro-4-((4-(trifluoromethyl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-one

[0231] 1-(2-chloro-4-((4-(trifluoromethyl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyri midin-7-yl)ethan-1-one

[0232]

[0233] At 0 °C, sodium hydride (74.0 mg, 1.85 mmol, 60% purity, 1.5 eq) was added to a solution of 4-(trifluoromethyl)pyridin-2-amine (200.0 mg, 1.23 mmol, 1.0 eq) in tetrahydrofuran (20 mL), and the mixture was stirred for 15 minutes. Then, 14-1 (281.7 mg, 1.23 mmol, 1.0 eq) was added to the mixed solution, and the reaction was stirred at room temperature for 2 hours. The LCMS of the reaction solution showed the appearance of the molecular ion peak of the target compound. The reaction solution was diluted with saturated ammonium chloride solution (50 mL), and the mixture was extracted with EtOAc (30 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by flash acidic chromatography column (PE / EtOAc = 10:1) to obtain the compound 14-2 as a white solid (128.2 mg, 0.56 mmol, 45.5% yield).

[0234] LCMS: (ESI) m / z: 356.0 [M+H] + .

[0235] 1 H NMR (400 MHz, CDCl3) δ: 7.80 (d, J = 8.0 Hz, 1H), 7.35 (d, J = 4.0 Hz, 1H), 6.82 - 6.61 (m, 2H), 6.40 (brs, 1H), 2.75 (s, 3H).

[0236] Step 2, Compound 14

[0237] 1-(2-(6-(trifluoromethyl)pyridin-2-yl)-4-((4-(trifluoromethyl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-one

[0238] 1-(2-(6-(Trifluoromethyl)pyridin-2-yl)-4-((4-(trifluoromethyl)pyridin-2-yl)amino)-7H-pyrrolo[2,3-d]pyrimidin-7-yl)ethan-1-one

[0239]

[0240] At room temperature, to a mixed solution of 14-2 (80.0 mg, 0.23 mmol, 1.0 eq), (6-(trifluoromethyl)pyridin-2-yl)boronic acid (131.8 mg, 0.69 mmol, 3.0 eq) in dioxane (10 mL) and water (1 mL) were added s-phos (9.4 mg, 0.023 mmol, 0.1 eq), sodium carbonate (65.8 mg, 0.69 mmol, 3.0 eq) and palladium acetate (5.2 mg, 0.023 mmol, 0.1 eq). The reaction solution was purged with nitrogen three times and reacted at 100 °C for 12 hours. LCMS showed that the main peak was the target compound. The reaction solution was filtered, poured into ice water (30 mL) for dilution, and extracted with EtOAc (20 mL x 2). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to obtain compound 14 as a pale yellow solid (9.2 mg, 0.019 mmol, 8.2% yield).

[0241] LC-MS: (ESI) m / z: 467.1 [M+H] + .

[0242] 1 1H NMR (400 MHz, CDCl3) δ: 8.31 - 7.90 (m, 3H), 7.40 - 7.25 (m, 2H), 6.78 - 6.67 (m, 2H), 6.34 (d, J = 3.8 Hz, 1H), 2.75 (s, 3H).

[0243] Example 15, Compound 15

[0244] Step 1, Compound 15

[0245] 7-cyclopropyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0246] 7-cyclopropyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0247]

[0248] Under nitrogen protection at 0 °C, DEAD (48.7 mg, 0.28 mmol, 1.2 eq.) was added dropwise to a solution of compound 5 (100 mg, 0.23 mmol, 1.0 eq.) and triphenylphosphine in anhydrous THF (5 mL), and the mixture was stirred for 10 minutes. Under nitrogen protection, a solution of cyclopropanol (26.7 mg, 0.46 mmol, 2.0 eq.) in tetrahydrofuran (5 mL) was added dropwise to the mixture, and the reaction was carried out at room temperature for 3 hours. LCMS showed the appearance of the molecular ion peak of the target compound. The reaction solution was poured into saturated ammonium chloride (30 mL) for dilution, and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*255u; mobile phase: [water (0.05% ammonia hydroxide v / v) - ACN]; B%: 40% - 100%, 11 min) to obtain compound 15 as a white solid (11.2 mg, 0.024 mmol, 10.4% yield).

[0249] LC-MS: (ESI) m / z: 465.0 [M+H] + .

[0250] 1 1H NMR (400 MHz, CDCl3) δ: 8.25 - 7.83 (m, 3H), 7.37 - 7.25 (m, 2H), 6.70 - 6.53 (m, 2H), 6.22 (d, J = 3.9 Hz, 1H), 2.52 (m, 1H), 1.07 - 0.82 (m, 4H).

[0251] Example 16, Compound 16

[0252] Step 1, Compound 16

[0253] 7-(2-(dimethylamino)ethyl)-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0254] 7-(2-(dimethylamino)ethyl)-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0255]

[0256] At room temperature, 2-(dimethylamino)ethyl bromide hydrobromide (53.6 mg, 0.23 mmol, 1.0 eq.) and Cs2CO3 (150.0 mg, 0.46 mmol, 2.0 eq.) were added to a DMF solution (5 mL) of compound 5 (100.0 mg, 0.23 mmol, 1.0 eq.). Under nitrogen protection at 50 °C, the mixture was stirred and reacted for 1 hour. LCMS showed the appearance of the molecular ion peak of the target compound. The reaction solution was poured into saturated NH4Cl (30 mL) for dilution and extracted with EtOAc (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product. The crude product was purified by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v)-ACN]; B%: 40%-100%, 11 min) to obtain compound 16 as a white solid (22.0 mg, 0.044 mmol, 19.1% yield).

[0257] LC-MS: (ESI) m / z: 496.2 [M+H] + .

[0258] 1 1H NMR (400 MHz, CDCl3) δ: 8.16 - 7.88 (m, 3H), 7.31 - 7.20 (m, 2H), 6.68 - 6.53 (m, 2H), 6.15 (d, J = 4.0 Hz, 1H), 4.52 (t, J = 5.2 Hz, 2H), 2.92 (s, 6H), 2.67 (t, J = 5.2 Hz, 2H).

[0259] Example 17, Compound 17

[0260] Step 1, Compound 17-2

[0261] 2,4-dichloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine

[0262] 2,4-dichloro-7-isopropyl-6-methyl-7H-pyrrolo[2,3-d]pyrimidine

[0263]

[0264] Under nitrogen protection, isopropyl iodide (673.2 mg, 3.96 mmol, 2.0 eq.) and Cs2CO3 (1.94 g, 5.94 mmol, 3.0 eq.) were added to a solution of Compound 17-1 (400.0 mg, 1.98 mmol, 1.0 eq.) in ACN (40 mL). The reaction was stirred at 50 °C for 16 h. LCMS showed that the main peak was the target compound. The reaction solution was concentrated to obtain a crude product, which was purified by flash acidic chromatography column (PE / EtOAc = 8:1) to obtain Compound 17-2 as a yellow solid (417.1 mg, 1.71 mmol, 77.63% yield).

[0265] ESI-MS m / z = 244.1 [M+H] + .

[0266] 1 1H NMR (400 MHz, MeOD) δ 6.37 (d, J = 1.0 Hz, 1H), 4.80 (dd, J = 13.8, 6.9 Hz, 1H), 2.51 (d, J = 0.9 Hz, 3H), 1.66 (d, J = 6.9 Hz, 6H).

[0267] Step 2, Compound 17-3

[0268] 2-chloro-7-isopropyl-6-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0269] 2-chloro-7-isopropyl-6-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0270]

[0271] To a toluene solution (15 mL) of Compound 17-2 (376.0 mg, 1.54 mmol, 1.0 eq.) and 4-(trifluoromethyl)pyridin-2-amine (300.0 mg, 1.85 mmol, 1.2 eq.) was added Cs2CO3 (1.01 g, 3.09 mmol, 2.0 eq.) and palladium(II) acetate (69.6 mg, 0.31 mmol, 0.2 eq.). The reaction mixture was stirred at 50 °C for 2 h under a nitrogen atmosphere. The reaction solution was concentrated to obtain a crude product, which was purified by flash acidic chromatography column (DCM / MeOH = 15:1) to give Compound 17-3 as a white solid (297.2 mg, 0.80 mmol, 51.9% yield).

[0272] ESI-MS m / z = 370.0 [M+H] + .

[0273] 1 H NMR (400 MHz, DMSO-d6) δ 10.79 (s, 1H), 8.76 (d, J = 0.6 Hz, 1H), 8.61 (d, J = 5.1 Hz, 1H), 7.41 (dd, J = 5.1, 0.9 Hz, 1H), 6.81 (d, J = 0.9 Hz, 1H), 4.82 - 4.68 (m, 1H), 2.44 (s, 3H), 1.57 (d, J = 6.9 Hz, 6H).

[0274] Step 3, Compound 17

[0275] 7-Isopropyl-6-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0276] 7-isopropyl-6-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

[0277]

[0278] To a mixed solution of compound 17-3 (150.0 mg, 0.41 mmol, 1.0 eq.) and 2-(trifluoromethyl)pyridine-6-boronic acid ester (224.0 mg, 0.82 mmol, 2.0 eq.) in dioxane (5 mL) and water (1 mL), add K2CO3 (170.0 mg, 1.23 mmol, 3.0 eq.) and Pd(dppf)Cl2 (60.0 mg, 0.082 mmol, 0.2 eq.). Stir the reaction mixture at 90 °C for 2 hours under nitrogen protection. LCMS shows that the main peak is the target compound. Concentrate the reaction solution to obtain the crude product, and purify the crude product by preparative HPLC (column: Agela Durashell C18 150*25 5u; mobile phase: [water (0.05% ammonia hydroxide v / v) - ACN]; B%: 40% - 100%, 11 min) to obtain compound 17 as a white solid (27.9 mg, 0.058 mmol, 14.1% yield).

[0279] ESI-MS m / z = 481.1 [M+H] + .

[0280] 1 H NMR (400 MHz, MeOD) δ 8.73 (d, J = 8.0 Hz, 1H), 8.54 (s, 1H), 8.41 (s, 1H), 8.19 (s, 1H), 7.90 (s, 1H), 7.31 (s, 1H), 6.64 (s, 1H), 5.02 (dt, J = 13.7, 6.9 Hz, 1H), 2.55 (s, 3H), 1.77 (d, J = 6.9 Hz, 6H).

[0281] Example 18, Compound 18

[0282] Step 1, Compound 18-2

[0283] 7-Methylpyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione

[0284] 7-methylpyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione

[0285]

[0286] Under nitrogen protection at 110 °C, a DMSO solution (25 mL) of 4-trimethylsilyl-3-butyn-2-one (5 g, 35 mmol, 1 eq.), 18-1 (4.52 g, 35 mmol, 1 eq.) and zinc bromide (0.8 g, 3.5 mmol, 0.1 eq.) was stirred for reaction for 20 hours. LCMS showed that the main peak was the target compound. The reaction solution was diluted with water (80 mL), the mixture was filtered, and the filter cake was freeze-dried to obtain the compound 18-2 as a red solid (4.8 g, 27.09 mmol, 77.1% yield).

[0287] ESI-MS m / z = 178.2 [M+H] + .

[0288] Step 2, Compound 18-3

[0289] 2,4-dichloro-7-methylpyrido[2,3-d]pyrimidine

[0290] 2,4-dichloro-7-methylpyrido[2,3-d]pyrimidine

[0291]

[0292] At 80 °C, DIEA (1.094 g, 8.47 mmol, 3 eq.) was added to a phosphorus oxychloride solution (10 mL) of compound 18-2 (500 mg, 2.82 mmol, 1 eq.). The reaction was stirred at 80 °C for 3 hours. The reaction solution was poured into ice water (100 mL), and then saturated sodium bicarbonate (30 mL) was added to the ice water. The mixture was extracted with ethyl acetate (50 mL * 3). The combined organic phases were concentrated to obtain a crude product, and the crude product was purified by flash acidic chromatography column (EtOAc / PE = 1 / 1) to obtain the compound 18-3 as a yellow solid (297.2 mg, 1.38 mmol, 48.9% yield).

[0293] ESI-MS m / z = 214.1 [M+H] + .

[0294] Step 3, Compound 18-4

[0295] 2-chloro-7-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)pyrido[2,3-d]pyrimidin-4-amine

[0296] 2-chloro-7-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)pyrido[2,3-d]pyrimidin-4-amine

[0297]

[0298] Under nitrogen protection at 90 °C, a toluene solution (2 mL) of compound 18-3 (50 mg, 0.23 mmol, 1 eq.), 4-(trifluoromethyl)pyridin-2-amine (38 mg, 0.23 mmol, 1 eq.), RuPhos (33 mg, 0.07 mmol, 0.3 eq.), Pd2(dba)3 (43 mg, 0.0467 mmol, 0.2 eq.) and cesium carbonate (228 mg, 0.7008 mmol, 3 eq.) was stirred for 4 hours. The reaction mixture was diluted with water (40 mL) and extracted with ethyl acetate (40 mL × 3). The combined organic phases were concentrated to obtain a crude product, which was purified by flash acidic chromatography column (EtOAc / PE = 1:1) to give compound 18-4 as a yellow solid (60 mg, 0.17 mmol, 73.9% yield).

[0299] ESI-MS m / z = 340.1 [M+1] + .

[0300] 1 H NMR (400 MHz, CDCl3) δ 9.00 (s, 1H), 8.55 (s, 1H), 8.51 (d, J = 4.7 Hz, 1H), 8.27 (d, J = 8.3 Hz, 1H), 7.45 (d, J = 8.4 Hz, 1H), 7.34 (d, J = 4.4 Hz, 1H), 2.80 (s, 3H).

[0301] Step 4, compound 18

[0302] 7-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)pyrido[2,3-d]pyrimidin-4-amine

[0303] 7-methyl-N-(4-(trifluoromethyl)pyridin-2-yl)-2-(6-(trifluoromethyl)pyridin-2-yl)pyrido[2,3-d]pyrimidin-4-amine

[0304]

[0305] Under nitrogen protection at 90 °C, 18-4 (70 mg, 0.21 mmol, 1 eq.), 2-pinacolborate-6-trifluoromethylpyridine (68 mg, 0.25 mmol, 1.2 eq.), Pd(dppf)Cl2 (30 mg, 0.04 mmol, 0.2 eq.) and potassium carbonate (85 mg, 0.62 mmol, 3 eq.) were added to 1,4-dioxane (3 mL) and water (1.5 mL), and the mixture was stirred and reacted for 4 hours. The reaction solution was diluted with water (40 mL), and extracted with ethyl acetate (40 mL × 3). The combined organic phases were concentrated to obtain a large crude product, and the crude product was purified by preparative TLC (DCM / MeOH = 20 / 1) to obtain the yellow solid compound 18 (26 mg, 0.057 mmol, 27% yield).

[0306] ESI-MS m / z = 451.1 [M+H] + .

[0307] 1 1H NMR (400 MHz, DMSO) δ 11.12 (s, 1H), 9.42 (s, 1H), 9.19 (s, 1H), 8.81 (d, J = 7.9 Hz, 1H), 8.71 (s, 1H), 8.33 (s, 1H), 8.10 (s, 1H), 7.62 (d, J = 7.8 Hz, 1H), 7.54 (d, J = 4.9 Hz, 1H), 2.73 (s, 3H).

[0308] Biological assay

[0309] Determination of the biochemical activity and selectivity of the compound

[0310] The present disclosure provides a biochemical assay method for detecting the inhibitory activity and selectivity of a compound by detecting the enzyme activities of wild-type and mutant IDH2. Wild-type IDH2 can convert NADP + into NADPH, while its mutant IDH2-R140Q / R172K can convert NADPH into NADP + . Since NADPH itself is fluorescent (excitation 340 nm, emission 460 nm), the enzyme activities of wild-type IDH2 and mutant IDH2-R140Q / R172K can both be determined by detecting the change in the level of NADPH in the reaction system. Finally, the inhibitory activity IC 50 of the compound can be calculated from the change in enzyme activity. The specific operation steps are as follows:

[0311] The test compound was dissolved in DMSO to prepare a 50 mM compound stock solution. Then, the stock solution was further serially diluted with DMSO to obtain 100× compound working solutions with concentrations of 200 μM, 100 μM, 50 μM, 25 μM, 12.5 μM, 6.25 μM, and 3.125 μM, respectively.

[0312] Inhibitory effect on wild-type IDH2:

[0313] Five components, 1 M Tris-HCl pH 7.5 (200 μL), 5 M NaCl (300 μL), 1 M MgCl2 (100 μL), 1 M DTT (10 μL), and 20 mg / mL BSA (25 μL), were mixed and deionized water was added to a final volume of 10 mL to prepare the reaction buffer. Then, the reaction buffer (15 mL) was mixed with 107 μM wild-type IDH2 (0.37 μL) to prepare Solution A; the reaction buffer (5 mL) was mixed with 50 mM NADP+ (20 μL) and 80 mM isocitrate (20 μL) to prepare Solution B.

[0314] Solution A (148 μL / well) was added to a 96-well plate, followed by the addition of 2 μL of each concentration of the 100× compound working solution and 2 μL of DMSO (as a negative control). After incubation at room temperature for 1 h, 50 μL of Solution B was added to each well. Then, the 96-well plate was placed in a BioTek Synergy H4 microplate reader (BioTek Instruments Inc., USA) with the following program settings: 37 °C, shake the plate for 5 s, run in kinetic mode, total detection time of 40 minutes, detection interval of 60 s, and detection mode of fluorescence (excitation at 340 nm and emission at 460 nm). Finally, the data was exported, processed, and analyzed: the enzyme activity was represented by the absolute value of the slope of the fluorescence reading of each well relative to time, a dose-response curve was plotted, and the biochemical IC of the compound was calculated by fitting. 50 .

[0315] Inhibitory effect on mutant IDH2-R140Q / R172K:

[0316] Mix five components: 1M Tris-HCl pH7.5 (200 μL), 5M NaCl (300 μL), 1M MgCl2 (100 μL), 1M DTT (10 μL), and 20 mg / mL BSA (25 μL), and add deionized water to a final volume of 10 mL to prepare the reaction buffer. Then, mix the reaction buffer (15 mL) with mutant IDH2 at 107 μM (0.37 μL) to prepare Solution A; mix the reaction buffer (5 mL) with 50 mM NADPH (20 μL) and 1M α-KG (20 μL) to prepare Solution B. The remaining operations are the same as above.

[0317] Determination of the Cell Activity of Compounds

[0318] The present disclosure also provides a cell assay method for detecting the inhibitory activity of a compound by detecting the intracellular activity of mutant IDH2 enzyme. A large amount of D-2-HG can be produced and accumulated in 293T cells overexpressing IDH2-R140Q / R172K constructed by genetic engineering techniques. When the activity of IDH2-R140Q / R172K is inhibited, the level of D-2-HG in the cells will decrease significantly. Therefore, we can measure the inhibitory activity of the compound by detecting the decrease in the level of D-2-HG in IDH2-R140Q / R172K-293T cells. The specific operation steps are as follows:

[0319] Culture IDH2-R140Q / R172K-293T cells in DMEM containing 10% FBS and 1X penicillin & streptomycin. When the cell confluence reaches 80%, digest the cells with trypsin, perform cell counting, and adjust the cell density to 10,000 cells / ml. Then, inoculate the cells into a 96-well plate at a volume of 200 μL / well and culture them under the conditions of 37°C and 5% CO2.

[0320] The next day, gradient-dilute the compound stock solution with DMSO to obtain a 100× compound working solution. Take out 2 μL of each concentration of the 100× compound working solution and 2 μL of DMSO (as a negative control) and add them to the cell culture medium. After shaking well, continue to culture under the conditions of 37°C and 5% CO2.

[0321] Two days later, remove the cell culture medium, wash the cells once with PBS, then extract intracellular metabolites with 80% methanol, and detect the level of D-2-HG in each sample by LC-MS. Finally, represent the enzyme activity by the level of D-2-HG in the cells at different compound concentrations, draw a dose-response curve, and calculate the IC of the compound for the cells by fitting. 50 。

[0322] Activity of the Target Compound:

[0323]

[0324] It can be seen that the compounds of the present invention can selectively inhibit the activity of mutant IDH2 without affecting the activity of wild-type IDH2. The compounds of the present invention can exhibit good activity both at the enzyme level and at the cell level, especially compounds 3, 9, and 17.

[0325] All documents mentioned in the present invention are incorporated herein by reference as if each individual document was specifically and individually incorporated by reference. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, characterized in that, Has the structure shown in the following formula (I) Wherein: Ring A and Ring B are each independently selected from a substituted or unsubstituted C6-C 10 aryl ring having 6 members, or a substituted or unsubstituted heteroaryl ring having 5 to 12 members; R 1 and R 2 are independently selected from the group consisting of: hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, halogen, hydroxy, amino, carboxy, nitro, cyano, substituted or unsubstituted C2-C4 alkanoyl, substituted or unsubstituted C2-C4 acylamino, C2-C4 sulfonyl, C2-C4 sulfinyl, mercapto; m is selected from 1 or 2; is a single bond or a double bond, provided that is an aromatic ring; n is 0, 1 or 2; Unless otherwise specified, the substitution mentioned herein means that each of the said groups is independently substituted by 1-3 substituents selected from the following group: halogen, hydroxy, amino, cyano, -C1-C4 alkyl, -C1-C4 haloalkyl, -C1-C4 hydroxyalkyl, -C1-C4 alkoxy, -C1-C4 alkylamino, -C1-C4 cycloalkyl, -C1-C4 halocycloalkyl, -C1-C4 hydroxycycloalkyl, -C1-C4 cycloalkoxy, -C1-C4 cycloalkylamino, C2-C4 acylamino, C2-C4 sulfonyl, C2-C4 sulfinyl; Unless otherwise specified, each of the said heterocycloalkyl or heteroaromatic independently contains 1, 2, 3 or 4 heteroatoms selected from O, S and N.

2. The compound or a pharmaceutically acceptable salt thereof as described in claim 1, wherein The compound of formula (I) has the structure shown in the following formula IA or IB:

3. The compound or a pharmaceutically acceptable salt thereof as described in claim 1, wherein, In the said compound, ring A is selected from the following group: substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl, substituted or unsubstituted pyrimidinyl, substituted or unsubstituted pyrazinyl; preferably, the said ring A is 2-pyridyl substituted with trifluoromethyl.

4. The compound or a pharmaceutically acceptable salt thereof as described in claim 1, characterized in that, In the said compound, ring B is selected from the following group: substituted or unsubstituted phenyl, substituted or unsubstituted pyridyl, substituted or unsubstituted pyrimidinyl, substituted or unsubstituted pyrazinyl; preferably, the said ring B is 2-pyridyl substituted with trifluoromethyl, or 4-pyridyl substituted with trifluoromethyl.

5. The compound or a pharmaceutically acceptable salt thereof as described in claim 1, characterized in that, The said compound is selected from the following group:

6. Use of the compound according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof, characterized in that, Preparing a pharmaceutical composition for treating diseases related to the blockade of the conversion of α-KG to D-2-HG, wherein the said diseases, preferably diseases related to IDH2 mutants, include cancer.

7. A pharmaceutical composition, which comprises a compound of formula (I) according to any one of the preceding claims 1-6 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

8. A method for treating a disease characterized by D-2-HG accumulation in a patient, characterized in that, Comprising the step: administering to an individual a therapeutically effective amount of a compound of formula (I) according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 7, wherein the said disease is preferably cancer.

9. A method for inhibiting the conversion of α-KG to D-2-HG, characterized in that, Comprising the step: administering to an individual a therapeutically effective amount of a compound of formula (I) according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 7.

10. A method for inhibiting mutant IDH2, wild-type IDH2, or both of the above, characterized in that, Comprising the step: administering to an individual a therapeutically effective amount of a compound of formula (I) according to any one of claims 1-5 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 7.

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