Diphenylaminopyrimidines for inhibiting kinase activity

Abstract

The present application provides a pharmaceutical composition of substituted diphenylamino pyrimidine compounds and uses thereof, said compounds are represented by formula (I), or pharmaceutically acceptable salts, prodrugs, hydrates or solvates, crystal forms, N-oxides and various diastereomers thereof. The compounds of the present application are useful in the treatment of diseases that can be treated with JAK2 kinase inhibitors.

Description

[0001] This application is a divisional application of the invention patent application filed on September 30, 2018, with application number 201811161081.8 and invention title "Diphenylaminopyrimidine Compounds for Inhibiting Kinase Activity". Technical Field

[0002] This invention belongs to the field of pharmaceutical technology, and particularly relates to a substituted diphenylaminopyrimidine compound, compositions comprising the compound, and their uses. More specifically, this invention relates to certain deuterium-substituted N-(tert-butyl)-3-((5-methyl-2-((4-(2-(pyrrolidone-1-yl)ethoxy)phenyl)amino)pyrimidin-4-yl)amino)benzenesulfonamides, which exhibit JAK2 protein tyrosine kinase inhibition and can be used to treat JAK2 kinase-mediated diseases, and these deuterium-substituted compounds have superior pharmacokinetic properties. Background Technology

[0003] The JAK family of cellular protein tyrosinases includes four kinase isoforms: JAK 1–JAK 3 and TYK 2. These four family members share high homology. They have seven homology domains (JAKhomology domain, JH) from the C-terminus to the N-terminus, which are usually further divided into four functional domains. JH1 is the active kinase catalytic domain, while the JH2 domain is unique to the JAK family and is called a pseudokinase domain. Although it has no catalytic function, it plays an essential regulatory role through cooperation. The SH2 domain, composed of JH3 and JH4, mainly plays a role in stabilizing the JAK kinase structure, while the JH5–JH7 regions (FERM domains) directly interact with the intracellular structures of cytokine receptors and also interact with the JH1 domain.

[0004] JAK2 possesses a unique cytokine spectrum within the JAK family. Besides being activated by GC family cytokines, it can also be activated by many other cytokines (such as BC family cytokines, erythropoietin, and thrombopoietin, among other blood-related cytokines). Its activity is essential for hematopoiesis. Therefore, JAK2 can influence some fundamental functions regulated by erythropoietin (EPO) and granulocyte-macrophage colony-stimulating factor (GM-CSF), and is closely related to blood disorders such as polycythemia vera (PV), anemia, and essential thrombocytosis (ET). Severe damage to JAK2 can pose a lethal threat to the organism.

[0005] The JAK pathway has been demonstrated to be complemented in cell survival and proliferation. For example, in the case of Philadelphia chromosome-positive cells that cause chronic myeloid leukemia (CML), there is evidence that the JAK2 pathway is complemented in constitutive activation. Therefore, JAK2 inhibitors can be used in CML, where the Philadelphia chromosome has been shown to produce hybrid Bcr-Abl, thereby maintaining constitutive cell activity.

[0006] More significantly, for resistance mutations based on BCR-ABL specific inhibitors, such as the T315I gatekeeper gene mutation or any other mutation, JAK2 inhibitors based on the JAK2 pathway (such as the BCR-ABL(T315I) mutation) may be used. Therefore, JAK2 inhibitors can be used to treat patients resistant to known therapies, those directly targeting BCR-ABL, and for whom resistance has now been confirmed as the dominant (50%-90%) of all resistances in patients who have failed existing therapies.

[0007] Therefore, due to the lack of therapies available to treat diseases that are directly or indirectly complemented by the above-mentioned JAK2 signaling pathway disorders, there is a need to develop compounds that act as inhibitors of kinases, particularly JAK2 kinases.

[0008] Fedratinib (SAR-302503, TG-101348), developed by TargeGene, was acquired by Sanofi in 2010, which advanced Fedratinib to late-stage clinical trials. Later, the drug was taken over by the startup Impact Biopharmaceuticals, which has since been acquired by Celgene. Fedratinib is a highly selective oral inhibitor of JAK2 protein kinase, chemically named (N-(tert-butyl)-3-((5-methyl-2-((4-(2-(pyrrolidone-1-yl)ethoxy)phenyl)amino)pyrimidin-4-yl)amino)benzenesulfonamide. Its clinical indications are myelofibrosis (MF) and polycythemia vera (PV). Early clinical trials showed excellent clinical benefits; however, in 2013, serious neurological side effects were discovered.

[0009] Therefore, there is still a need in the art to develop compounds with selective inhibitory activity or better pharmacodynamics / pharmacokinetics for JAK2 kinase-mediated diseases suitable as therapeutic agents. This invention provides a novel JAK2 inhibitor derived from Fedratinib through deuteration modification, which reduces or eliminates unwanted metabolites through a deuteration strategy; increases the half-life of the parent compound; reduces the number of doses required to achieve the desired effect; increases the formation of active metabolites (if any); reduces the production of harmful metabolites in specific tissues; and produces a more effective and / or safer drug for multidrug therapy (whether or not the multidrug therapy is intended). Invention Overview

[0010] To address the above technical problems, this invention discloses a novel deuterium-substituted diphenylpyrimidine compound, its composition, and its uses, which exhibits better JAK2 kinase and JAK2 / V617F kinase inhibitory activity, lower side effects, and better pharmacodynamic / pharmacokinetic performance, and can be used to treat JAK2 kinase-mediated diseases.

[0011] To address this, the present invention adopts the following technical solution:

[0012] In a first aspect, the present invention provides a compound of formula (I):

[0013]

[0014] in,

[0015] R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 Y 1 Y 2 Y 3 Y 4 Y 5 Y 6 Y 7 Y 8 and Y 9 Each is independently selected from hydrogen or deuterium;

[0016] X 1 X 2 X 3 and X 4Each is independently selected from CH3, CD3, CHD2, and CH2D;

[0017] The condition is if X 1 X 2 X 3 and X 4 Each one is CH3, so R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 Y 1 Y 2 Y 3 Y 4 Y 5 Y 6 Y 7 Y 8 and Y 9 At least one of them is deuterium.

[0018] In another aspect, the present invention provides pharmaceutical compositions comprising the compounds of the present invention and pharmaceutically acceptable excipients. In a specific embodiment, the compounds of the present invention are provided in the pharmaceutical composition in an effective amount. In a specific embodiment, the compounds of the present invention are provided in a therapeutically effective amount. In a specific embodiment, the compounds of the present invention are provided in a preventatively effective amount.

[0019] In another aspect, the present invention provides a method for preparing a pharmaceutical composition as described above, comprising the steps of: mixing a pharmaceutically acceptable excipient with a compound of the present invention to form a pharmaceutical composition.

[0020] In another aspect, the present invention also relates to a method of treating at least a portion of a JAK2-mediated disease, comprising administering a therapeutically effective dose of the compound of the present invention to a subject in need. Specific embodiments include the use of the compound of the present invention in the preparation of a medicament for treating a JAK2-mediated disease in a subject in need. Specific embodiments include the use of the compound of the present invention in the preparation of a medicament for treating a JAK2 / V617F-mediated disease in a subject in need. Specific embodiments include oral, subcutaneous, intravenous, or intramuscular administration of the compound. Specific embodiments include prolonged administration of the compound. Specific embodiments include JAK2-mediated diseases selected from myeloproliferative disorders, polycythemia vera, idiopathic thrombocythemia, myelofibrosis, any other bone marrow-related disorders, proliferative diabetic retinopathy, cancer, eye diseases, inflammation, psoriasis, any disease related to angiogenesis, or viral infections.

[0021] Other objects and advantages of the invention will become apparent to those skilled in the art from the following detailed description, embodiments, and claims. Invention Details

[0022] Terms and Definitions

[0023] In this document, unless otherwise specified, "deuterated" means that one or more hydrogen atoms in a compound or group are replaced by deuterium; deuteration can be monosubstituted, disubstituted, polysubstituted, or total substituted. The terms "one or more deuterated" and "one or more deuterated" are used interchangeably.

[0024] Unless otherwise specified, "non-deuterated compounds" in this article refer to compounds containing a deuterium atom ratio not higher than the natural deuterium isotope content (0.015%).

[0025] This invention also includes isotopically labeled compounds, equivalent to the original compounds disclosed herein. Examples of isotopes that can be included as compounds of this invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine isotopes, respectively as follows: 2 H, 3 H, 13 C, 14 C, 15 N, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F and 36 Cl. The compounds of this invention, or enantiomers, diastereomers, isomers, or pharmaceutically acceptable salts or solvates, wherein the isotopes or other isotopic atoms of the aforementioned compounds are all within the scope of this invention. Certain isotopically labeled compounds of this invention, for example...3 H and 14 Radioactive isotopes of carbon are also included, and are useful in tissue distribution experiments of drugs and substrates. Tritium, i.e. 3 H and carbon-14, i.e. 14 C, these are relatively easy to prepare and detect, making them the preferred isotopes. Isotope-labeled compounds can be prepared using common methods by replacing non-isotopic reagents with readily available isotope-labeling reagents, as illustrated in the example.

[0026] The compounds of this invention may include one or more asymmetric centers and therefore may exist in a variety of "stereoisomer" forms, such as enantiomers and / or diastereomers. For example, the compounds of this invention may be individual enantiomers, diastereomers, or geometric isomers (e.g., cis and trans isomers), or may be in the form of mixtures of stereoisomers, including racemic mixtures and mixtures rich in one or more stereoisomers. The isomers may be separated from the mixture by methods known to those skilled in the art, including chiral high-performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or preferred isomers may be prepared by asymmetric synthesis.

[0027] As used herein, the term "compound of the invention" refers to a compound of formula (I). The term also includes pharmaceutically acceptable salts, prodrugs, hydrates or solvents, N-oxides, crystal forms, stereoisomers, isotopic variants or various diastereomers of compounds of formula (I).

[0028] As used herein, the term "pharmaceutically acceptable salt" refers to those salts that, to the extent of reliable medical judgment, are suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., and in proportion to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19.

[0029] Pharmaceutically acceptable salts of the compounds of this invention include salts derived from suitable inorganic and organic acids and inorganic and organic bases. Examples of pharmaceutically acceptable, non-toxic acid addition salts are salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or salts formed with organic acids, such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid. Salts formed using methods conventional in the art are also included, such as ion exchange methods. Other pharmaceutically acceptable salts include: adipic acid salts, alginate salts, ascorbate salts, aspartate salts, benzenesulfonate salts, benzoate salts, bisulfate salts, borate salts, butyrate salts, camphorate salts, camphor sulfonate salts, citrate salts, cyclopentylpropionate salts, diglucuronate salts, dodecyl sulfate salts, ethanesulfonate salts, formate salts, fumarate salts, gluconate salts, glyceryl phosphate salts, glucuronate salts, hemisulfate salts, heptarate salts, hexanoate salts, hydroiodate salts, 2-hydroxy-ethanesulfonate salts, lactobionate salts, lactate salts, laurate salts, lauryl sulfate salts, malate salts, maleate salts, malonate salts, methanesulfonate salts, 2-naphthalenesulfonate salts, nicotinate salts, nitrate salts, oleate salts, oxalate salts, palmitate salts, dihydroxynaphthalate salts, pectin ester salts, persulfate salts, 3-phenylpropionate salts, phosphate salts, picrate salts, p-pentanoate salts, propionate salts, stearate salts, succinate salts, sulfate salts, tartrate salts, thiocyanate salts, p-toluenesulfonate salts, undecanoate salts, valerate salts, etc. Pharmaceutically acceptable salts derived from suitable bases include alkali metals, alkaline earth metals, ammonium, and nitrogen. + (C 1-4 Alkyl)4 salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Other pharmaceutically acceptable salts, if appropriate, include non-toxic ammonium salts, quaternary ammonium salts, and amine cations that form with counterions such as halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, lower alkyl sulfonates, and aryl sulfonates.

[0030] The term "solvate" refers to a complex formed by the coordination of the compound of the present invention with solvent molecules in a specific ratio. "Hydrate" refers to a complex formed by the coordination of the compound of the present invention with water.

[0031] The term "prodrug" includes compounds that are themselves biologically active or inactive, and which, when taken in a suitable manner, are metabolized or chemically reacted in the human body to form compounds of formula (I), or salts or solutions of compounds of formula (I). Prodrugs include (but are not limited to) compounds consisting of amino acid residues or polypeptide chains consisting of one or more (e.g., 2, 3, or 4) amino acid residues covalently linked by amide or ester bonds to the free amino, hydroxyl, or carboxyl group of the compound of the present invention. Amino acid residues include, but are not limited to, 20 naturally occurring amino acids typically represented by three-letter symbols, and also include 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, valine, ornithine, and sulfones. Other types of prodrugs are also included. For example, the free carboxyl group may be derived into an amide or alkyl ester. As described in Advanced Drug Delivery Reviews 1996, 19, 115, free hydroxyl groups are derivatized using groups including, but not limited to, hemisuccinates, phosphates, dimethylaminoacetate, and phosphoryloxymethoxycarbamates. Carbamate prodrugs of hydroxyl and amino groups, as well as carbonate prodrugs of hydroxyl groups, sulfonates, and sulfates, are also included. Derivatizations of hydroxyl groups such as (acyloxy)methyl and (acyloxy)ethyl ethers are also included, wherein the acyl group may be an alkyl ester, optionally substituted with groups including, but not limited to, ether, amine, and carboxylic acid functional groups, or wherein the acyl group is an amino acid ester as described above. This type of prodrug is described in J. Med. Chem. 1996, 39, 10. Free hydroxyl groups can also be derivatized as amides, sulfonamides, or phosphoramides. All these other portions may be incorporated with groups including, but not limited to, ether, amine, and carboxylic acid functional groups.

[0032] The term "crystal form" refers to the different arrangements of chemical drug molecules, generally manifested as the form in which the drug raw material exists in a solid state. A drug can exist in multiple crystal forms, and different crystal forms of the same drug may have different solubility and absorption in the body, thus affecting the dissolution and release of the formulation.

[0033] As used herein, the term "subject" includes, but is not limited to: humans (i.e., men or women of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or older adults)) and / or non-human animals, such as mammals, e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In some embodiments, the subject is a human. In other embodiments, the subject is a non-human animal.

[0034] The terms “disease,” “disorder,” and “symptom” are used interchangeably in this article.

[0035] Unless otherwise stated, the term “treatment” as used herein includes effects that occur when a subject has a specific disease, disorder, or condition, which reduce the severity of the disease, disorder, or condition, or delay or slow the development of the disease, disorder, or condition (“therapeutic treatment”), and also includes effects that occur before a subject begins to have a specific disease, disorder, or condition (“preventive treatment”).

[0036] Generally, the "effective amount" of a compound refers to the amount sufficient to elicit a response in the target organism. As will be understood by those skilled in the art, the effective amount of the compounds of the present invention can vary depending on factors such as the biological target, the pharmacokinetics of the compound, the disease being treated, the administration method, and the age, health status, and symptoms of the subject. Effective amounts include both therapeutic and prophylactic effective amounts.

[0037] Unless otherwise stated, the term "therapeuticly effective amount" of a compound as used herein is a quantity sufficient to provide therapeutic benefit in the treatment of a disease, disorder, or condition, or to delay or minimize one or more symptoms associated with the disease, disorder, or condition. A therapeutically effective amount of a compound refers to the quantity of a therapeutic agent, used alone or in combination with other therapies, that provides therapeutic benefit in the treatment of a disease, disorder, or condition. The term "therapeuticly effective amount" may include quantities that improve overall treatment, reduce or prevent symptoms or causes of a disease or condition, or enhance the therapeutic efficacy of other therapeutic agents.

[0038] Unless otherwise stated, the term "preventively effective amount" of a compound as used herein is a quantity sufficient to prevent a disease, disorder, or condition, or a quantity sufficient to prevent one or more symptoms associated with a disease, disorder, or condition, or a quantity sufficient to prevent recurrence of a disease, disorder, or condition. The preventively effective amount of a compound refers to the quantity of a therapeutic agent, used alone or in combination with other agents, that provides preventive benefit in the prevention of a disease, disorder, or condition. The term "preventively effective amount" may include quantities that improve overall prevention or enhance the preventive efficacy of other preventive agents.

[0039] The term "combination" and related terms refer to the simultaneous or sequential administration of the therapeutic agents of the present invention. For example, the compounds of the present invention may be administered simultaneously or sequentially with another therapeutic agent in separate unit dosage forms, or simultaneously with another therapeutic agent in a single unit dosage form.

[0040] Specific embodiments of the present invention

[0041] compound

[0042] This invention provides compounds of formula (I), or pharmaceutically acceptable salts, prodrugs, hydrates or solvents thereof, crystal forms, N-oxides and various diastereomers:

[0043]

[0044] in,

[0045] R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 Y 1 Y 2 Y 3 Y 4 Y 5 Y 6 Y 7 Y 8 and Y 9 Each is independently selected from hydrogen or deuterium;

[0046] X 1 X 2 X 3 and X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D;

[0047] The condition is if X 1 X 2 X 3 and X 4 Each one is CH3, so R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 Y 1 Y 2 Y 3 Y 4 Y 5 Y 6 Y 7 Y 8 and Y 9 At least one of them is deuterium;

[0048] As a preferred embodiment of the present invention, the compound in formula (I) contains at least one deuterium atom, more preferably one deuterium atom, more preferably two deuterium atoms, more preferably three deuterium atoms, more preferably four deuterium atoms, more preferably five deuterium atoms, more preferably six deuterium atoms, more preferably seven deuterium atoms, more preferably eight deuterium atoms, more preferably nine deuterium atoms, more preferably ten deuterium atoms, more preferably eleven deuterium atoms, more preferably twelve deuterium atoms, more preferably thirteen deuterium atoms, and more preferably fourteen deuterium atoms.

[0049] As a preferred embodiment of the present invention, the deuterium isotope content at the deuterated position is at least greater than 0.015% of the natural deuterium isotope content, preferably greater than 30%, more preferably greater than 50%, more preferably greater than 75%, more preferably greater than 95%, and more preferably greater than 99%.

[0050] Specifically, in this invention R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 Y 1 Y 2 Y 3 Y 4 Y 5 Y 6 Y 7 Y 8 Y 9 X 1 X 2 X 3 and X 4 The deuterium isotope content at each deuteration position is at least 5%, preferably greater than 10%, more preferably greater than 15%, more preferably greater than 20%, more preferably greater than 25%, more preferably greater than 30%, more preferably greater than 35%, more preferably greater than 40%, more preferably greater than 45%, more preferably greater than 50%, more preferably greater than 55%, more preferably greater than 60%, more preferably greater than 65%, more preferably greater than 70%, more preferably greater than 75%, more preferably greater than 80%, more preferably greater than 85%, more preferably greater than 90%, more preferably greater than 95%, and more preferably greater than 99%.

[0051] In another specific embodiment, the R of the compound in formula (I) 1 R 2 R 3 R 4 R 5 R6 R 7 R 8 R 9 R 10 R 11 R 12 Y 1 Y 2 Y 3 Y 4 Y 5 Y 6 Y 7 Y 8 Y 9 X 1 X 2 X 3 and X 4 At least one contains deuterium, preferably two, preferably three, preferably four, preferably five, preferably six, preferably seven, preferably eight, preferably nine, preferably ten, preferably eleven, preferably twelve, preferably thirteen, preferably fourteen, preferably fifteen, preferably sixteen, preferably seventeen, preferably eighteen. Containing deuterium, preferably nineteen, preferably twenty, preferably twenty-one, preferably twenty-two, preferably twenty-three, preferably twenty-four, preferably twenty-five, preferably twenty-six, preferably twenty-seven, preferably twenty-eight, preferably twenty-nine, preferably thirty, preferably thirty-one, preferably thirty-two, preferably thirty-three. Specifically, the compound in formula (I) contains at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, or thirty-three deuterium atoms.

[0052] In one implementation of general formula (I), R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Each is independently selected from hydrogen or deuterium.

[0053] In another preferred embodiment, R 1 and R 2 It is deuterium.

[0054] In another preferred embodiment, R 3 and R 4 It is deuterium.

[0055] In another preferred embodiment, R 1 R 2 R 3 and R 4 It is deuterium.

[0056] In another preferred embodiment, R 5 R 6 R 7 and R 8 It is deuterium.

[0057] In another preferred embodiment, R 9 R 10 R 11 and R 12 It is deuterium.

[0058] In another preferred embodiment, R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 It is deuterium.

[0059] In one implementation of general formula (I), X 1 X 2 X 3 and X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0060] In another preferred embodiment, X 1 It's CD3.

[0061] In another preferred embodiment, X 2 It's CD3.

[0062] In another preferred embodiment, X 3 It's CD3.

[0063] In another preferred embodiment, X 4 It's CD3.

[0064] In another preferred embodiment, X 1 and X 2 Both are CD3.

[0065] In another preferred embodiment, X 1 X 2 and X 3 Both are CD3

[0066] In one implementation of general formula (I), Y 1 Y 2 Y 3 Y 4 Y 5 Y 6 Y 7 Y 8 and Y 9 Each is independently selected from hydrogen or deuterium.

[0067] In another preferred embodiment, Y 5 It is deuterium.

[0068] One embodiment of the present invention provides a compound represented by general formula (II):

[0069]

[0070] in,

[0071] R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 Y 5 X 1 X 2 X 3 and X 4 The definition is as described above;

[0072] Or its pharmaceutically acceptable salts, prodrugs, hydrates or solvent compounds, crystal forms, N-oxides and various diastereomers.

[0073] In one implementation of general formula (II), R 1 R 2 Selected from deuterium, R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R11 R 12 Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0074] In another preferred embodiment, R 1 R 2 Selected from deuterium, R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X 4 Each is independently selected from CH3 or CD3.

[0075] In another preferred embodiment, R 1 R 2 R 3 R 4 Selected from deuterium, R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X 4 Each is independently selected from CH3 or CD3.

[0076] In one implementation of general formula (II), Y 5 Selected from deuterium, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R12 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0077] In another preferred embodiment, Y 5 Selected from deuterium, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X 4 Each is independently selected from CH3 or CD3.

[0078] In one implementation of general formula (II), Y 5 Selected from deuterium, X 4 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Each is independently selected from hydrogen or deuterium, X 1 X 2 and X 3 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0079] In another preferred embodiment, Y 5 Selected from deuterium, X 4 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11and R 12 Each is independently selected from hydrogen or deuterium, X 1 X 2 and X 3 Each is independently selected from CH3 and CD3.

[0080] In one embodiment of general formula (II), X 4 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 and X 3 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0081] In another preferred embodiment, X 4 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 and X 3 Each is independently selected from CH3 and CD3.

[0082] In one embodiment of general formula (II), X 1 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5Each is independently selected from hydrogen or deuterium, X 2 X 3 and X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0083] In another preferred embodiment, X 1 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5 Each is independently selected from hydrogen or deuterium, X 2 X 3 and X 4 Each is independently selected from CH3 and CD3.

[0084] In one embodiment of general formula (II), X 1 and X 3 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5 Each is independently selected from hydrogen or deuterium, X 2 and X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0085] In another preferred embodiment, X 1 and X 3 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5Each is independently selected from hydrogen or deuterium, X 3 and X 4 Each is independently selected from CH3 and CD3.

[0086] In one embodiment of general formula (II), X 1 X 2 and X 3 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5 Each is independently selected from hydrogen or deuterium, X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0087] In another preferred embodiment, X 1 X 2 and X 3 Selected from CD3, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 and Y 5 Each is independently selected from hydrogen or deuterium, X 4 Each is independently selected from CH3 and CD3.

[0088] In one implementation of general formula (II), R 5 R 6 R 7 Or R 8 Selected from deuterium, R 1 R 2 R 3 R 4 R 9 R 10 R 11 R 12 Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 X3 and X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0089] In another preferred embodiment, R 5 R 6 R 7 Or R 8 Selected from deuterium, R 1 R 2 R 3 R 4 R 9 R 10 R 11 R 12 Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X 4 Each is independently selected from CH3 and CD3.

[0090] In one implementation of general formula (II), R 9 R 10 R 11 and R 12 Selected from deuterium, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 Or Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0091] In another preferred embodiment, R 9 R 10 R 11 and R 12 Selected from deuterium, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 Or Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X4 Each is independently selected from CH3 and CD3.

[0092] In one implementation of general formula (II), R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Selected from deuterium, R 1 R 2 R 3 R 4 , or Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X 4 Each is independently selected from CH3, CD3, CHD2, and CH2D.

[0093] In another preferred embodiment, R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Selected from deuterium, R 1 R 2 R 3 R 4 and Y 5 Each is independently selected from hydrogen or deuterium, X 1 X 2 X 3 and X 4 Each is independently selected from CH3 and CD3.

[0094] In a preferred embodiment of the present invention, the compound is selected from the group consisting of compounds or pharmaceutically acceptable salts thereof:

[0095]

[0096]

[0097]

[0098]

[0099]

[0100] In another preferred embodiment, the compound does not include non-deuterated compounds.

[0101] Pharmaceutical Compositions and Administration

[0102] In another aspect, the present invention provides pharmaceutical compositions comprising the compound of the present invention (also referred to as the "active ingredient") and pharmaceutically acceptable excipients. In some embodiments, the pharmaceutical composition comprises an effective amount of the active ingredient. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the active ingredient. In some embodiments, the pharmaceutical composition comprises a preventatively effective amount of the active ingredient.

[0103] The pharmaceutical compositions of the present invention comprise, within a safe and effective range, the compound of the present invention or a pharmacologically acceptable salt thereof, and a pharmacologically acceptable excipient or carrier. "Safe and effective range" refers to an amount of the compound sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 0.5-2000 mg of the compound of the present invention per dose, more preferably, 1-500 mg of the compound of the present invention per dose. Preferably, "one dose" is one capsule or tablet.

[0104] "Pharmaceutically acceptable excipients" refer to non-toxic carriers, adjuvants, or mediators that do not impair the pharmacological activity of the compounds formulated together. Pharmaceutically acceptable carriers, adjuvants, or mediators that can be used in the compositions of this invention include (but are not limited to) ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (such as human serum albumin), buffering substances (such as phosphates), glycine, sorbic acid, potassium sorbate, mixtures of saturated vegetable fatty acid metaglycerides, water, salts or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and lanolin.

[0105] The pharmaceutical compositions of the present invention can be prepared by combining the compounds of the present invention with suitable pharmaceutically acceptable excipients, for example, in solid, semi-solid, liquid or gaseous formulations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, solutions, suppositories, injections, inhalers, gels, microspheres and aerosols.

[0106] Typical routes of administration of the compounds or pharmaceutical compositions thereof of the present invention include, but are not limited to, oral, rectal, transmucosal, enteral, or topical, transdermal, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intradermal, subcutaneous, and intravenous administration.

[0107] The pharmaceutical compositions of the present invention can be manufactured using methods well known in the art, such as conventional mixing, dissolving, granulation, sugar-coated pill making, grinding, emulsification, freeze drying, etc.

[0108] For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present invention to be formulated into tablets, pills, lozenges, sugar-coated tablets, capsules, liquids, gels, pastes, suspensions, etc., for oral administration to patients.

[0109] Solid oral compositions can be prepared using conventional mixing, filling, or tableting methods. For example, they can be obtained by mixing the active compound with a solid excipient, optionally milling the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain a tablet or sugar-coated core. Suitable excipients include, but are not limited to: binders, diluents, disintegrants, lubricants, glidants, sweeteners, or flavoring agents. Examples include microcrystalline cellulose, glucose solution, gum arabic, gelatin solution, sucrose, and starch paste; talc, starch, calcium stearate, or stearic acid; lactose, sucrose, starch, mannitol, sorbitol, or dicalcium phosphate; silica; croscarmellose sodium, precrossed starch, starch-hydroxyacetic acid sodium, alginate, corn starch, potato starch, methylcellulose, agar, croscarmellose, croscarmellose, etc. The sugar-coated core can be optionally coated using methods known in general pharmaceutical practice, particularly enteric coating.

[0110] The pharmaceutical composition may also be suitable for parenteral administration, such as in suitable unit dosage forms of sterile solutions, suspensions, or lyophilized products. Appropriate excipients, such as fillers, buffers, or surfactants, can be used.

[0111] The compounds of the present invention can be administered by any route and method of use, such as oral or parenteral (e.g., intravenous) administration. The therapeutically effective amount of the compounds of the present invention is from about 0.0001 to 20 mg / kg body weight / day, for example from 0.001 to 10 mg / kg body weight / day.

[0112] The dosage frequency of the compounds of the present invention is determined by the individual patient's needs, for example, once or twice daily, or more times daily. Dosing may be intermittent, for example, wherein the patient receives a daily dose of the compounds of the present invention for a period of several days, followed by a period of several or more days during which the patient does not receive a daily dose of the compounds of the present invention.

[0113] Therapeutic indications of the compounds of this invention

[0114] The compounds of this invention exhibit JAK2 protein tyrosine kinase inhibition and are used, alone or in combination with other active agents (e.g., chemotherapeutic agents or protein therapeutic agents described below), to treat a variety of diseases, including, but not limited to: myeloproliferative disorders, proliferative diabetic retinopathy and other angiogenesis-related conditions, including solid tumors and other types of cancer, eye diseases, inflammation, psoriasis, and viral infections. Types of cancer that can be treated include, but are not limited to, digestive / gastrointestinal cancers, colon cancer, liver cancer, skin cancer, breast cancer, ovarian cancer, prostate cancer, lymphoma, leukemia (including acute myeloid leukemia and chronic myeloid leukemia), kidney cancer, lung cancer, muscle cancer, bone cancer, bladder cancer, or brain cancer.

[0115] Some examples of treatable diseases and conditions include ocular neovascularization, infantile hemangioma; organ hypoxia, angiogenesis, organ graft rejection, lupus, multiple sclerosis, rheumatoid arthritis, psoriasis, type 1 diabetes and its complications, inflammatory diseases, acute pancreatitis, chronic pancreatitis, asthma, rhinitis, atopic dermatitis, autoimmune thyroid disease, ulcerative colitis, Crohn's disease, metastatic melanoma, Kaposi's sarcoma, multiple myeloma, cytokine-related disorders and other autoimmune gastritis, autoimmune hemolytic diseases, and autoimmune neutrophilia. Less severe conditions, thrombocytopenia, atopic diseases (e.g., allergic asthma, atopic dermatitis, or allergic rhinitis), chronic active hepatitis, myasthenia gravis, multiple sclerosis, inflammatory bowel disease, graft-versus-host disease, neurodegenerative diseases including motor neuron disease, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's disease, cerebral ischemia or neurodegenerative diseases caused by traumatic injury, stroke, glutamate neurotoxicity or hypoxia; ischemic / reperfusion injury in stroke, myocardial ischemia, renal ischemia, heart attack, cardiac hypertrophy, atherosclerosis and arteriosclerosis, organ hypoxia and platelet aggregation.

[0116] Examples of additional diseases and conditions that can be treated include cell-mediated hypersensitivity reactions (allergic contact dermatitis, allergic pneumonia), rheumatic diseases (e.g., systemic lupus erythematosus, rheumatoid arthritis, juvenile arthritis, Sjögren's syndrome, scleroderma, polymyositis, ankylosing spondylitis, psoriatic arthritis), viral diseases (EBV, hepatitis B, hepatitis C, HIV, HTLV1, varicella-zoster virus, human papillomavirus), food allergies, skin inflammation, and immunosuppression induced by solid tumors.

[0117] In some embodiments, the compounds or compositions of the present invention can be used to treat primary myelofibrosis. In some embodiments, the compounds or compositions of the present invention can be used to treat myelofibrosis following polycythemia vera. In some embodiments, the compounds or compositions of the present invention can be used to treat myelofibrosis following idiopathic thrombocythemia. In some embodiments, the compounds or compositions of the present invention can be used to treat high-risk myelofibrosis. In some embodiments, the compounds or compositions of the present invention can be used to treat intermediate-risk myelofibrosis (e.g., intermediate risk level 2). In some embodiments, the compounds of the present invention can be used to treat myelofibrosis with a positive mutation in JAK 2 valine 617 to phenylalanine (i.e., V617F). In some embodiments, the compounds of the present invention can be used to treat myelofibrosis with a negative mutation in JAK 2 valine 617 to phenylalanine (i.e., V617F).

[0118] combination therapy

[0119] The compounds of this invention are administered to subjects requiring such treatment in combination with anti-inflammatory drugs, antihistamines, chemotherapeutic agents, immunomodulators, therapeutic antibodies, or protein kinase inhibitors, such as tyrosine kinase inhibitors. While not limited, chemotherapeutic agents include antimetabolites such as methotrexate; DNA cross-linking agents such as cisplatin / carboplatin; alkylating agents such as canbusil; topoisomerase I inhibitors such as dextrin; and microtubule inhibitors such as paclitaxel. Other chemotherapeutic agents include, for example, vinca alkaloids, mitomycin-like antibiotics, bleomycin-like antibiotics, antifolate agents, colchicine, demecoline, etoposide, taxanes, anthracycline antibiotics, doxorubicin, daunorubicin, erythromycin, epirubicin, idarubicin, mithoxanthrone, 4-dimethoxy-daunorubicin, 11-deoxydaunorubicin, 13-deoxydaunorubicin, and doxorubicin. -14-benzoate, doxorubicin-14-octanoate, doxorubicin-14-naphthaleneacetate, acridine, carmustine, cyclophosphamide, cytarabine, etoposide, lovastatin, melphalan, topotecan, oxalaplatin, chlorambucil, methotrexate, lomustine, thioguanine, asparaginase, vinblastine, vindesin, tamoxifen, or nitrogen mustard. Although not restricted, therapeutic antibodies include antibodies targeting the HER2 protein, such as trastuzumab; antibodies targeting growth factors or growth factor receptors, such as bevacizumab targeting vascular endothelial growth factor and OSI-774 targeting epidermal growth factor; and antibodies targeting integrin receptors, such as Vitaxin (also known as MEDI-522).The types of anticancer drugs suitable for the compositions and methods of the present invention include, but are not limited to: 1) alkaloids, including microtubule inhibitors (e.g., vincristine, vinblastine, and vindesine, etc.), microtubule stabilizers (e.g., paclitaxel [Taxol] and docetaxel, Taxotere, etc.), and chromatin function inhibitors, including topoisomerase inhibitors, such as epipodophyllotoxin (e.g., etoposide [VP-16] and teniposide [VM-26], etc.) and activators targeting topoisomerase I (e.g., camptothecin and isirinotecan [CPT-11], etc.); 2) covalent DNA-binding agents [alkylating agents], including nitrogen mustard (e.g., nitrogen mustard, chlorambucil, cyclophosphamide, ifosfamide, and busulfan […]). 3) Non-covalent DNA-binding agents [antitumor antibiotics], including nucleic acid inhibitors (e.g., actinomycin D), anthracyclines (e.g., daunorubicin [daunorubicin and daunorubicin], doxorubicin [doxacin] and idarubicin [demethoxydaunorubicin], anthraquinones (e.g., anthracycline antibiotic analogs, such as [mitoxantrone]), bleomycin [bleomycin] and purcamycin [bleomycin], etc.); 4) Antimetabolites, including antifolates (e.g., methotrexate, iron phosphate, etc.). - Folic acid and methotrexate sodium, etc.), purine antimetabolites (e.g., 6-mercaptopurine [6-MP, mercaptopurine], 6-thioguanine [6-TG], azathioprine, acyclovir, ganciclovir, chlorodeoxyadenosine, 2-chlorodeoxyadenosine [CdA] and 2'-deoxyinterstitial mycotoxin [pentostatin], etc.), pyrimidine antagonists (e.g., fluoropyrimidines [e.g., 5-fluorouracil (Adrucil), 5-fluorodeoxyuridine (FdUrd) (fluorouridine)], etc.) and cytosine arabinoside (e.g., sedosa[ara-C] and fludarabine, etc.); 5) enzymes, including L-asparaginase; 6) hormones, including glucocorticoids, such as anti-estrogens (e.g., tamoxifen, etc.), nonsteroidal anti-inflammatory drugs (NSAIDs), etc. Androgen drugs (e.g., flutamide) and aromatase inhibitors (e.g., anastrozole [Renind]); 7) Platinum compounds (e.g., cisplatin and carboplatin); 8) Monoclonal antibodies conjugated to anticancer drugs, toxins and / or radionuclides; 9) Bioreactivity modifiers (e.g., interferons [e.g., IFN-α] and interleukins [e.g., IL-2]); 10) Adoptive immunotherapy; 11) Hematopoietic growth factors; 12) Active agents that induce tumor cell differentiation (e.g., all-trans retinoic acid); 13) Gene therapy technology; 14) Antisense therapy technology; 15) Tumor vaccines; 16) Therapies targeted at metastatic tumors (e.g., Batimistat); and 17) Angiogenesis inhibitors.

[0120] The pharmaceutical compositions and methods of the present invention may further include other therapeutically active compounds commonly used herein for treating the aforementioned pathological conditions. Examples of other therapeutic agents include: cyclosporins (e.g., cyclosporine A); CTLA4-Ig; antibodies, such as ICAM-3, anti-IL-2 receptor (anti-Tac), anti-CD45RB, anti-CD2, anti-CD3 (OKT-3), anti-CD4, anti-CD80, anti-CD86; active agents that block the interaction between CD40 and gp39, such as antibodies specific to CD40 and / or gp39 (i.e., CD154), fusion proteins constructed from CD40 and gp39 (CD40Ig and CD8gp39), NF-κB function inhibitors, such as nuclear translocation inhibitors, such as deoxyguanidin (DSG); cholesterol biosynthesis inhibitors, such as HMG. CoA reductase inhibitors (lovastatin and simvastatin); nonsteroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen and cyclooxygenase inhibitors, such as rofecoxib; steroids, such as prednisone or dexamethasone; gold compounds; antiproliferative drugs, such as methotrexate, FK506 (tacrolimus, prazolam), mycophenolate mofetil; cytotoxic drugs, such as azathioprine and cyclophosphamide; TNF-α inhibitors, such as tenidap; anti-TNF antibodies or soluble TNF receptors and rapamycin (sirolimus or rapamycin) or their derivatives.

[0121] Other active agents that can be co-administered with the compounds of the present invention include protein therapeutic agents, such as cytokines, immunomodulators, and antibodies. As used herein, the term "cytokine" includes chemokines, interleukins, lymphokines, monokines, colony-stimulating factors, and receptor-associated proteins and their functional fragments. As used herein, the term "functional fragment" refers to a polypeptide or peptide having a biological function or activity identified by a defined functional assay.

[0122] Cytokines include endothelial monocyte activation peptide II (EMAP-II), granulocyte-macrophage-CSF (GM-CSF), granulocyte-CSF (G-CSF), macrophage-CSF (M-CSF), IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-12 and IL-13, interferons, etc., and are associated with specific biological, morphological or phenotypic changes in cells or cellular mechanisms.

[0123] When other therapeutic agents are used in combination with the compounds of the present invention, they may be used, for example, in the amounts described in the Physician Desk Reference (PDR), or as determined by those skilled in the art.

[0124] In the treatment or prevention of diseases involving cell proliferation, appropriate dose levels are generally from about 0.01 to about 1000 mg / kg patient body weight / day, and can be administered in single or multiple doses. For example, this dose level can be from about 0.01 to about 250 mg / kg / day; more narrowly, it can be from about 0.5 to about 100 mg / kg / day. Appropriate dose levels can be from about 0.01 to about 250 mg / kg / day, from about 0.05 to about 100 mg / kg / day, or from about 0.1 to about 50 mg / kg / day, or from about 1.0 mg / kg / day. For example, within this range, the dose can be from about 0.05 to about 0.5 mg / kg / day, or from about 0.5 to about 5 mg / kg / day, or from about 5 to about 50 mg / kg / day. For oral administration, the composition is formulated as tablets containing about 1.0 to about 1,000 mg of the active ingredient, for example, about 1.0, about 5.0, about 10.0, about 15.0, about 20.0, about 25.0, about 50.0, about 75.0, about 100.0, about 150.0, about 200.0, about 250.0, about 300.0, about 400.0, about 500.0, about 600.0, about 750.0, about 800.0, about 900.0, and about 1,000.0 mg of the active ingredient, so that the dosage can be adjusted according to the symptoms of the patient being treated. The compound can be administered 1 to 4 times / day, such as once or twice / day. There may be periods without dosing, followed by another dosing regimen.

[0125] However, it is understood that the specific dose level and frequency for any particular patient can vary and depend on a variety of factors, including the activity of the specific compound used, the metabolic stability and duration of action of the compound, age, weight, general health condition, sex, diet, route and time of administration, excretion rate, drug combination, severity of the specific condition, and the host’s ongoing therapy.

[0126] The compounds of the present invention can be used alone or in combination with an effective amount of a therapeutic antibody (or a therapeutic fragment thereof), a chemotherapeutic agent, or an immunotoxic agent to treat tumors. Illustrative examples of chemotherapeutic agents that can be used for this purpose include doxorubicin, docetaxel, or paclitaxel. It should be further understood that the present invention includes combinations of the compounds of the present invention, including, but not limited to, vasculostatic agents, such as tyrosine, serine, or threonine kinase inhibitors, and any chemotherapeutic agent or therapeutic antibody.

[0127] The compounds of this invention have a series of advantages compared to known non-deuterated compounds in the prior art. These advantages include: First, the compounds and compositions employing the technical solutions of this invention provide a more advantageous therapeutic tool for treating JAK2-mediated diseases. Second, they improve the metabolism of the compounds in organisms, resulting in better pharmacokinetic parameters. In this case, dosage can be varied and long-acting formulations can be formed, improving applicability. Third, they increase the drug concentration of the compounds in animals, thereby improving drug efficacy. Fourth, they inhibit certain metabolites, improving the safety of the compounds. Example

[0128] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise stated, parts and percentages are parts by weight and weight percentages.

[0129] Typically, in the preparation process, the reactions are carried out in an inert solvent at room temperature to reflux temperature (e.g., 0°C to 100°C, preferably 0°C to 80°C). The reaction time is typically 0.1–60 hours, preferably 0.5–24 hours.

[0130] Example 1 N-(tert-butyl)-3-((5-methyl-2-((4-(2-(pyrrolid-1-yl)ethoxy-1,1,2,2-) Preparation of d4)phenyl)amino)pyrimidin-4-yl)amino)benzenesulfonamide (compound T-1).

[0131]

[0132] The synthesis is performed using the following route:

[0133]

[0134] Step 1: Synthesis of compound 3.

[0135] Compound 1 (10.0 g, 45.12 mmol) and THF (100 mL) were added sequentially to a 250 mL single-necked flask equipped with a magnetic stirrer. After stirring until dissolved, a THF solution (20 mL) of tert-butylamine (9.9 g, 135.37 mmol) was slowly added dropwise under an ice-water bath. After the addition was complete, the ice bath was removed, and the reaction mixture was stirred at room temperature under a nitrogen atmosphere for 1 hour. The organic solvent was removed by vacuum distillation, and the mixture was extracted with water (50 mL), followed by extraction with dichloromethane (50 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain 9.5 g of a white solid, with a yield of 81.6%. LC-MS (APCI): m / z = 259.1 (M+1) + . 1H NMR (500MHz, CDCl3) δppm: 8.75 (t, J=2.0Hz, 1H), 8.40 (dd, J=8.0Hz, J=2.0Hz, 1H), 8.24(dd,J=8.0Hz,J=2.0Hz,1H),7.73(t,J=8.0Hz,1H),4.99(s,1H),1.27(s,9H).

[0136] Step 2: Synthesis of compound 4.

[0137] Add 60 mL of ethanol / water mixture (2 / 1) and compound 3 (3.0 g, 11.66 mmol) to a 100 mL single-necked flask equipped with a magnetic stirrer and a condenser. Add reduced iron powder (6.51 g, 116.6 mmol) and ammonium chloride (3.12 g, 58.3 mmol) while stirring. Heat to 85 °C under nitrogen atmosphere and stir for 1 h.

[0138] Cool to room temperature, filter off insoluble solids, evaporate organic solvent under reduced pressure, add saturated NaHCO3 aqueous solution (5 mL), extract with ethyl acetate (50 mL x 3), combine organic phases, dry over anhydrous sodium sulfate, filter, and concentrate to give 2.55 g white solid, yield 95.79%. LC-MS (APCI): m / z = 229.1 (M+1) + .

[0139] Step 3: Synthesis of compound 6.

[0140] Compound 5 (1.0 g, 6.13 mmol) and a methanol / water mixture (15 mL, 1 / 1) were added sequentially to a 50 mL single-necked flask equipped with a magnetic stirrer at room temperature. After stirring until dissolved, compound 4 (1.26 g, 5.52 mmol) was added. The reaction mixture was heated to 45 °C under nitrogen and stirred overnight. Upon cooling to room temperature, a large amount of white solid precipitated. The mixture was filtered, and the filter cake was washed with methanol / water (3.4 mL / 4.0 mL), dried under vacuum at 50 °C, and yielded 1.31 g of white solid (60.17% yield). LC-MS (APCI): m / z = 354.1 (M+1) + . 1 H NMR (500MHz, DMSO-D6) δppm: 9.11 (s, 1H), 8.11-8.09 (m, 2H), 7.88-7.86 (m, 1H), 7.55-7.52 (m, 3H), 2.18 (s, 3H), 1.12 (s, 9H).

[0141] Step 4: Synthesis of compound 9.

[0142] Compound 7 (300 mg, 2.16 mmol) and DMF (6 mL) were added sequentially to a 50 mL single-necked flask equipped with a magnetic stirrer. After stirring until dissolved, potassium carbonate (894 mg, 6.47 mmol) and compound 8 (827 mg, 4.31 mmol) were added. The reaction mixture was heated to 80 °C under a nitrogen atmosphere and stirred for 4 hours. After cooling to room temperature, 30 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with water (60 mL x 3), washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to give 450 mg of white solid, yield 83.4%. LC-MS (APCI): m / z = 250.0 and 252.0 (M+1). + .

[0143] Step 5: Synthesis of compound 10.

[0144] Compound 9 (450 mg, 1.8 mmol) and DMF (6 mL) were added sequentially to a 50 mL single-necked flask equipped with a magnetic stirrer. After stirring until dissolved, potassium carbonate (746 mg, 5.4 mmol) and pyrrolidine (256 mg, 3.6 mmol) were added. The reaction mixture was heated to 80 °C under a nitrogen atmosphere and stirred for 4 hours. After cooling to room temperature, 30 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with water (60 mL x 3), washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to obtain 250 mg of a yellow oil, yield 57.8%. LC-MS (APCI): m / z = 241 (M+1) + .

[0145] Step 6: Synthesis of compound 11.

[0146] Compound 10 (250 mg, 1.04 mmol) and methanol (10 mL) were added to a 50 mL single-necked flask equipped with a magnetic stirrer. The mixture was stirred until dissolved, then Pd / C (25 mg, 10%) was added. The mixture was evacuated under a hydrogen balloon and purged three times. The reaction was carried out overnight under a hydrogen atmosphere. The catalyst was filtered off, and the filter cake was washed with methanol (3 mL). The filtrates were combined and concentrated to give 220 mg of a yellow oil, with a yield of 98.6%. LC-MS (APCI): m / z = 211.1 (M+1) + .

[0147] Step 7: Synthesis of compound T-1.

[0148] Compound 6 (200 mg, 0.56 mmol), compound 11 (107 mg, 0.51 mmol), and ethylene glycol monomethyl ether (6 mL) were added to a 25 mL single-necked flask equipped with a magnetic stirrer and condenser. The mixture was stirred until dissolved, and a solution of isopropanol containing hydrogen chloride (1.41 mmol, 0.28 mL, 5 M) was added dropwise. The mixture was heated to 120 °C under a nitrogen atmosphere and stirred overnight. After cooling to room temperature, water (10 mL) and saturated sodium bicarbonate (5 mL) were added, and the mixture was extracted with dichloromethane (15 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was passed through a silica gel column to give 150 mg of a white solid, with a yield of 50.3%. LC-MS (APCI): m / z = 529.2 (M+1) + . 1 H NMR (500MHz, CDCl3) δppm: 8.11 (s, 1H), 7.92-7.89 (m, 2H), 7.57 (d, J = 8.0Hz, 1H), 7.41-7.38 (m, 3H), 6.91 (d, J = 8.0Hz ,1H),6.81(s,1H),6.44(s,1H),4.60(s,1H),4.13(t,J=6.0Hz,2H),2.94(t,J=6.0Hz,2H),2.13(s,3H),1.22(s,9H).

[0149] Example 2 N-(tert-butyl)-3-((5-(methyl-d3)-2-((4-(2-(pyrrolidone-1-yl)ethoxy)phenyl) Preparation of (amino)pyrimidin-4-yl-6-d)amino)benzenesulfonamide (compound T-2).

[0150]

[0151] The synthesis is performed using the following route:

[0152]

[0153] Step 1: Synthesis of compound 13.

[0154] Compound 12 (1.0 g, 7.93 mmol), Pd / C (200 mg, 10%), and heavy water (15 mL) were added to a 20 mL microwave tube containing a magnetic inlet. Hydrogen gas was bubbled through the tube for 2 minutes. The tube was then sealed and placed in a microwave reactor. The temperature was raised to 180 °C and the reaction was carried out for 1 hour. After cooling to room temperature, the catalyst was filtered off, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to give 700 mg of a white solid, with a yield of 67.8%. LC-MS (APCI): m / z = 131.1 (M+1) + . 1 H NMR (300MHz, DMSO-d6) δppm: 11.00 (s, 1H), 10.58 (s, 1H).

[0155] Step 2: Synthesis of compound 14.

[0156] Compound 13 (360 mg, 2.77 mmol) and phosphorus oxychloride (5 mL) were added sequentially to a 50 mL three-necked flask equipped with a magnetic stirrer. The mixture was heated to 110 °C under a nitrogen atmosphere and stirred overnight. Unreacted phosphorus oxychloride was removed by vacuum distillation. The residue was dissolved in ethyl acetate (40 mL), and saturated sodium bicarbonate solution (10 mL) was added. The mixture was stirred for 5 minutes, and the layers separated. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to give 400 mg of a white solid, with a yield of 86.5%. LC-MS (APCI): m / z = 168.1 (M+1) + .

[0157] Step 3: Synthesis of compound 15.

[0158] Compound 14 (400 mg, 2.39 mmol) and a 10 mL (1 / 1) mixture of MeOD / heavy water were added sequentially to a 50 mL single-necked flask equipped with a magnetic stirrer at room temperature. The mixture was stirred until dissolved, then compound 4 (382 mg, 1.68 mmol) was added. The reaction mixture was heated to 45 °C under nitrogen and stirred overnight. After cooling to room temperature, a large amount of white solid precipitated. The solid was filtered, and the filter cake was washed with MeOD / heavy water (2 mL / 2 mL), dried under vacuum at 50 °C, and yielded 400 mg of white solid (46.7%). LC-MS (APCI): m / z = 358.1 (M+1) + . 1 H NMR (400MHz, DMSO-D6)δ / ppm:9.11(s,1H),8.11-8.09(m,1H),7.88-7.86(m,1H),7.55-7.52(m,3H),1.12(s,9H).

[0159] Step 4: Synthesis of compound 17.

[0160] Compound 7 (1.0 g, 7.19 mmol), compound 16 (1.58 g, 9.35 mmol), and DMF (20 mL) were added sequentially to a 50 mL single-necked flask equipped with a magnetic stirrer. Cs₂CO₃ (cesium carbonate, 7.0 g, 21.6 mmol) was added while stirring. The reaction mixture was heated to 100 °C under a nitrogen atmosphere and stirred overnight. After cooling to room temperature, 30 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with water (60 mL x 3), washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and separated by silica gel column chromatography to give 1.2 g of a white solid, yield 70.4%. LC-MS (APCI): m / z = 237.1 (M+1) +.1H NMR (300MHz, CDCl3) δppm: 8.20 (d, J = 9.0Hz, 2H), 6.98 (d, J = 9.0Hz, 2H), 4.21 ( t,J=6.0Hz,2H),2.96(t,J=6.0Hz,2H),2.67-2.64(m,4H),1.88-1.82(m,4H).

[0161] Step 5: Synthesis of compound 18.

[0162] Compound 17 (250 mg, 1.04 mmol) and methanol (10 mL) were added to a 50 mL single-necked flask equipped with a magnetic stirrer. The mixture was stirred until dissolved, then Pd / C (25 mg, 10%) was added. The mixture was evacuated under a hydrogen balloon and purged three times. The reaction was carried out overnight under a hydrogen atmosphere. The catalyst was filtered off, and the filter cake was washed with methanol (3 mL). The filtrates were combined and concentrated to give 220 mg of a yellow oil, with a yield of 98.6%. LC-MS (APCI): m / z = 207.1 (M+1) + .

[0163] Step 6: Synthesis of compound T-2.

[0164] Compound 15 (200 mg, 0.56 mmol), compound 18 (107 mg, 0.51 mmol), and ethylene glycol monomethyl ether (6 mL) were added to a 25 mL single-necked flask equipped with a magnetic stirrer and condenser. The mixture was stirred until dissolved, and a solution of isopropanol containing hydrogen chloride (1.41 mmol, 0.28 mL, 5 M) was added dropwise. The mixture was heated to 120 °C under a nitrogen atmosphere and stirred overnight. After cooling to room temperature, water (10 mL) and saturated sodium bicarbonate (5 mL) were added, and the mixture was extracted with dichloromethane (15 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was passed through a silica gel column to give 150 mg of a white solid, with a yield of 50.3%. LC-MS (APCI): m / z = 529.2 (M+1) + . 1 H NMR(500MHz, CDCl3)δ / ppm:8.11(s,1H),7.90(dd,J=8.0Hz,J=1.0Hz,1H),7.57(d,J=8.0Hz,1H),7.41-7.38(m,3H),6.91(d,J=8.0Hz,1H) ,6.81(s,1H),6.44(s,1H),4.75(s,1H),4.13(t,J=6.0Hz,2H),2.94(t,J=6.0Hz,2H),2.69-2.64(m,4H),1.85-1.81(m,4H),1.22(s,9H).

[0165] Example 3 3-((5-methyl-2-((4-(2-(pyrrolidone-1-yl)ethoxy)phenyl)amino)pyrimidin-4-yl) Preparation of amino)-N-(2-methylpropane-2-yl-1,1,1,3,3,3-d6)benzenesulfonamide (compound T-3).

[0166]

[0167] The synthesis is performed using the following route:

[0168]

[0169] Step 1: Synthesis of compound 20.

[0170] Compound 19 (4.0 g, 62.39 mmol) and diethyl ether (40 mL) were added sequentially to a 100 mL three-necked flask equipped with a magnetic stirrer and a condenser. The mixture was cooled to -10 °C under a nitrogen atmosphere, and methyl magnesium bromide (20.80 mL, 62.39 mmol, 3 M) was slowly added dropwise while maintaining the temperature. After the addition was complete, the mixture was slowly heated to room temperature and then refluxed for 2 hours. The mixture was cooled to room temperature, and the reaction was quenched by adding saturated NH4Cl aqueous solution (10 mL) dropwise under an ice bath. The mixture was stirred for 10 minutes, and the layers were separated. The aqueous layer was extracted with diethyl ether (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and carefully distilled under reduced pressure at room temperature to obtain 1.7 g of a colorless liquid, with a yield of 33.99%. This was then directly added to the next step.

[0171] Step 2: Synthesis of compound 21.

[0172] Under an ice-water bath, concentrated sulfuric acid (10 g) was slowly added dropwise to water (10 g), controlling the temperature to not exceed 5°C. Sodium azide (1.52 g, 23.33 mmol) was slowly added in portions, stirred until dissolved, and compound 20 (1.7 g, 21.21 mmol) was added. The reaction was stirred overnight at room temperature under a nitrogen atmosphere. Diethyl ether (20 mL) was added, and the mixture was allowed to stand to separate the layers. The upper organic phase was separated, and the lower phase was extracted again with diethyl ether (20 mL). The organic phases were combined, washed with saturated NaHCO3 (5 mL), dried over anhydrous sodium sulfate, and the solvent was carefully evaporated at room temperature to give 1.2 g of a yellow liquid, with a yield of 53.80%. This was directly added to the next step.

[0173] Step 3: Synthesis of compound 22.

[0174] Compound 21 (1.2 g, 11.41 mmol) and methanol (20 mL) were added to a 50 mL single-necked flask equipped with a magnetic stirrer. After stirring until dissolved, Pd / C (120 mg, 10%) was added. The mixture was then evacuated under a hydrogen balloon and purged three times. The reaction was carried out overnight under a hydrogen atmosphere. The catalyst was filtered off, and isopropanol hydrochloride solution (5 M) was slowly added dropwise to adjust the pH to approximately 2. The mixture was stirred for 10 minutes, and the solvent was removed under reduced pressure to give 1.04 g of a yellow solid, with a yield of 79.23%. This was then directly added to the next step.

[0175] Step 4: Synthesis of compound 23.

[0176] Compound 1 (2.0 g, 9.02 mmol) and THF (30 mL) were added sequentially to a 50 mL single-necked flask equipped with a magnetic stirrer. After stirring until dissolved, compound 22 (1.04 g, 9.02 mmol) and triethylamine (3.65 g, 36.08 mmol) were slowly added dropwise under an ice-water bath. After the addition was complete, the ice bath was removed, and the reaction was stirred at room temperature for 1 hour under a nitrogen atmosphere. The organic solvent was removed by vacuum distillation, water (50 mL) was added, and the mixture was extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give 700 mg of a white solid, yield 29.34%. LC-MS (APCI): m / z = 265.1 (M+1) + . 1 H NMR (500MHz, CDCl3) δ8.75 (t, J=2.0Hz, 1H), 8.40 (dd, J=8.0Hz, J=2.0Hz, 1H), 8. 24(dd,J=8.0Hz,J=2.0Hz,1H),7.73(t,J=8.0Hz,1H),4.99(s,1H),1.27(s,3H).

[0177] Step 5: Synthesis of compound 24.

[0178] To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser, add 15 mL of ethanol / water mixture (2 / 1) and compound 23 (300 mg, 1.17 mmol). While stirring, add reduced iron powder (651 mg, 11.67 mmol) and ammonium chloride (0.31 mg, 5.83 mmol). Heat to 85 °C under nitrogen and maintain the temperature with stirring for 1 h. Cool to room temperature, filter to remove insoluble solids, evaporate organic solvent under reduced pressure, add saturated NaHCO3 aqueous solution (5 mL), extract with ethyl acetate (20 mL x 3), combine organic phases, dry to anhydrous sodium sulfate, filter, and concentrate to give 250 mg of white solid, yield 95.79%. LC-MS (APCI): m / z = 235.1 (M+1) + .

[0179] Step 6: Synthesis of compound 25.

[0180] Compound 24 (200 mg, 1.23 mmol) and a methanol / water mixture (15 mL, 1 / 1) were added sequentially to a 50 mL single-necked flask equipped with a magnetic stirrer at room temperature. The mixture was stirred until dissolved, and then compound 5 (250 mg, 1.10 mmol) was added. The reaction mixture was heated to 45 °C under nitrogen and stirred overnight. After cooling to room temperature, a large amount of white solid precipitated. The solid was filtered, and the filter cake was washed with methanol / water (3.4 mL / 4.0 mL), dried under vacuum at 50 °C, and yielded 230 mg of white solid (52.83%). LC-MS (APCI): m / z = 361.1 (M+1) + .1H NMR (500MHz, DMSO-d6) δ9.11(s,1H),8.11-8.09(m,2H),7.88-7.86(m,1H),7.55-7.52(m,3H),2.18(s,3H),1.12(s,3H).

[0181] Step 7: Synthesis of compound T-2.

[0182] Compound 25 (200 mg, 0.56 mmol), compound 18 (107 mg, 0.51 mmol), and ethylene glycol monomethyl ether (6 mL) were added to a 25 mL single-necked flask equipped with a magnetic stirrer and condenser. The mixture was stirred until dissolved, and isopropanol hydrochloride solution (1.41 mmol, 0.28 mL, 5 M) was added dropwise. The mixture was heated to 120 °C under a nitrogen atmosphere and stirred overnight. After cooling to room temperature, water (10 mL) and saturated sodium bicarbonate (5 mL) were added. The mixture was extracted with dichloromethane (15 mL x 3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was passed through a silica gel column to give 150 mg of a white solid, with a yield of 50.3%. LC-MS (APCI): m / z = 531.2 (M+1) + . 1 H NMR (300MHz, CDCl3) δ8.11(s,1H),7.92-7.89(m,2H),7.57(d,J=8.1Hz,1H),7.41-7.38(m,3H),6.91(d,J=8.1Hz,1H),6.81(s,1H),6. 44(s,1H),4.60(s,1H),4.13(t,J=6.0Hz,2H),2.94(t,J=6.0Hz,2H),2.69-2.64(m,4H),2.13(s,3H),1.85-1.81(m,4H),1.22(s,3H).

[0183] Example 4 3-((5-methyl-2-((4-(2-(pyrrolidone-1-yl)ethoxy)phenyl)amino)pyrimidin-4-yl) Preparation of amino)-N-(2-(methyl-d3)propane-2-yl-1,1,1,3,3,3-d6)benzenesulfonamide (compound T-4).

[0184]

[0185] The synthesis is performed using the following route:

[0186]

[0187] Step 1: Synthesis of compound 28.

[0188] Magnesium powder (1.80 g, 74.87 mmol) was added to a 100 mL three-necked flask equipped with a magnetic stirrer and a condenser. The flask was evacuated and purged with nitrogen three times. Diethyl ether (30 mL) and CD3I (10.0 g, 68.96 mmol) were added under nitrogen atmosphere. After the addition was complete, the mixture was refluxed for 2 hours. The mixture was cooled to -10 °C, and a solution of compound 19 (4.0 g, 62.39 mmol) in diethyl ether (10 mL) was added dropwise. After the addition was complete, the mixture was slowly heated to room temperature and then refluxed for 2 hours. The mixture was cooled to room temperature, and the reaction was quenched by adding saturated NH4Cl aqueous solution (10 mL) dropwise under ice bath conditions. The mixture was stirred for 10 minutes, and the layers were separated. The aqueous layer was extracted with diethyl ether (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and carefully distilled under reduced pressure at room temperature to obtain 1.7 g of a colorless liquid (33.99% yield). This was then directly added to the next step.

[0189] Step 2: Synthesis of compound 29.

[0190] Under an ice-water bath, concentrated sulfuric acid (10 g) was slowly added dropwise to water (10 g), controlling the temperature to not exceed 5°C. Sodium azide (1.52 g, 23.33 mmol) was slowly added in portions, stirred until dissolved, and compound 28 (1.7 g, 21.21 mmol) was added. The reaction was stirred overnight at room temperature under a nitrogen atmosphere. Diethyl ether (20 mL) was added, and the mixture was allowed to stand to separate the layers. The upper organic phase was separated, and the lower phase was extracted again with diethyl ether (20 mL). The organic phases were combined, washed with saturated NaHCO3 (5 mL), dried over anhydrous sodium sulfate, and the solvent was carefully evaporated at room temperature to give 1.2 g of a yellow liquid, with a yield of 53.80%. This was directly added to the next step.

[0191] Step 3: Synthesis of compound 30.

[0192] Compound 29 (1.2 g, 11.41 mmol) and methanol (20 mL) were added to a 50 mL single-necked flask equipped with a magnetic stirrer. After stirring until dissolved, Pd / C (120 mg, 10%) was added. The mixture was then evacuated under a hydrogen balloon and purged three times. The reaction was carried out overnight under a hydrogen atmosphere. The catalyst was filtered off, and isopropanol hydrochloride solution (5 M) was slowly added dropwise to adjust the pH to approximately 2. The mixture was stirred for 10 minutes, and the solvent was removed under reduced pressure to give 1.04 g of a yellow solid, with a yield of 79.23%. This was then directly added to the next step.

[0193] Step 4: Synthesis of compound 31.

[0194] Compound 1 (2.0 g, 9.02 mmol) and THF (30 mL) were added sequentially to a 50 mL single-necked flask equipped with a magnetic stirrer. After stirring until dissolved, compound 30 (1.04 g, 9.02 mmol) and triethylamine (3.65 g, 36.08 mmol) were slowly added dropwise under an ice-water bath. After the addition was complete, the ice bath was removed, and the reaction was stirred at room temperature for 1 hour under a nitrogen atmosphere. The organic solvent was removed by vacuum distillation, water (50 mL) was added, and the mixture was extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to give 700 mg of a white solid, yield 29.34%. LC-MS (APCI): m / z = 268.1 (M+1) + . 1 H NMR (500MHz, CDCl3) δ8.75(t,J=2.0Hz,1H),8.40(dd,J=8.0Hz,J=2.0Hz,1H),8.24(dd,J=8.0Hz,J=2.0Hz,1H),7.73(t,J=8.0Hz,1H),4.99(s,1H).

[0195] Step 5: Synthesis of compound 32.

[0196] To a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser, add 15 mL of ethanol / water mixture (2 / 1) and compound 31 (300 mg, 1.17 mmol). While stirring, add reduced iron powder (651 mg, 11.67 mmol) and ammonium chloride (0.31 mg, 5.83 mmol). Heat to 85 °C under nitrogen and maintain the temperature with stirring for 1 h. Cool to room temperature, filter to remove insoluble solids, evaporate organic solvent under reduced pressure, add saturated NaHCO3 aqueous solution (5 mL), extract with ethyl acetate (20 mL x 3), combine organic phases, dry to anhydrous sodium sulfate, filter, and concentrate to give 250 mg of white solid, yield 95.79%. LC-MS (APCI): m / z = 238.1 (M+1) + .

[0197] Step 6: Synthesis of compound 33.

[0198] Compound 32 (200 mg, 1.23 mmol) and a methanol / water mixture (15 mL, 1 / 1) were added sequentially to a 50 mL single-necked flask equipped with a magnetic stirrer at room temperature. The mixture was stirred until dissolved, and then compound 5 (250 mg, 1.10 mmol) was added. The reaction mixture was heated to 45 °C under nitrogen and stirred overnight. After cooling to room temperature, a large amount of white solid precipitated. The solid was filtered, and the filter cake was washed with methanol / water (3.4 mL / 4.0 mL), dried under vacuum at 50 °C, and yielded 230 mg of white solid (52.83%). LC-MS (APCI): m / z = 364.1 (M+1)+ .1H NMR(500MHz,DMSO-d6)δ9.11(s,1H),8.11-8.09(m,2H),7.88-7.86(m,1H),7.55-7.52(m,3H),2.18(s,3H).

[0199] Step 7: Synthesis of compound T-4.

[0200] Compound 33 (200 mg, 0.56 mmol), compound 18 (107 mg, 0.51 mmol), and ethylene glycol monomethyl ether (6 mL) were added to a 25 mL single-necked flask equipped with a magnetic stirrer and condenser. The mixture was stirred until dissolved, and isopropanol hydrochloride solution (1.41 mmol, 0.28 mL, 5 M) was added dropwise. The mixture was heated to 120 °C under a nitrogen atmosphere and stirred overnight. After cooling to room temperature, water (10 mL) and saturated sodium bicarbonate (5 mL) were added, and the mixture was extracted with dichloromethane (15 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was passed through a silica gel column to give 150 mg of a white solid, with a yield of 50.3%. LC-MS (APCI): m / z = 534.2 (M+1) + . 1 H NMR (300MHz, CDCl3) δ8.11(s,1H),7.92-7.89(m,2H),7.57(d,J=8.1Hz,1H),7.41-7.38(m,3H),6.91(d,J=8.1Hz,1H),6.81(s, 1H),6.44(s,1H),4.60(s,1H),4.13(t,J=6.0Hz,2H),2.94(t,J=6.0Hz,2H),2.69-2.64(m,4H),2.13(s,3H),1.85-1.81(m,4H).

[0201] Example 5 N-(tert-butyl)-3-((5-methyl-2-((4-(2-(pyrrolid-1-yl-d8)ethoxy)phenyl)amino Preparation of 4-pyrimidinylaminobenzenesulfonamide (compound T-5).

[0202]

[0203] The synthesis is performed using the following route:

[0204]

[0205] Step 1: Synthesis of compound 36.

[0206] NaH (595 mg, 14.88 mmol, 60%) was added sequentially to a 50 mL three-necked flask equipped with a magnetic stirrer. The flask was evacuated and purged with nitrogen three times. Anhydrous THF (20 mL) was added under nitrogen atmosphere. Then, a solution of 2-ethanolamine (909 mg, 14.88 mmol) in anhydrous THF (2 mL) was added dropwise under an ice-water bath. The mixture was stirred for 10 minutes. Then, a solution of compound 34 (2.0 g, 14.17 mmol) in anhydrous THF (3 mL) was added dropwise. After the addition was complete, the ice bath was removed, and the mixture was stirred at room temperature for 2 hours. The reaction was quenched with ethyl acetate (50 mL) saturated with water. The mixture was filtered, dried over anhydrous sodium sulfate, filtered again, concentrated, and passed through a silica gel column to give 1.6 g of a yellow oil, yield 61.69%. LC-MS (APCI): m / z = 183.1 (M+1) + . 1 H NMR (300MHz, CDCl3) δ8.21(d,J=9.3Hz,2H), 6.97(d,J=9.3Hz,2H), 4.09(t,J=5.1Hz,2H), 3.15(t,J=5.1Hz,2H).

[0207] Step 2: Synthesis of compound 38.

[0208] Under ice-water bath conditions, aqueous hydrobromic acid solution (7.57 g, 93.57 mmol, 48%) and concentrated sulfuric acid (3.67 g) were added to a 50 mL single-necked flask equipped with a magnetic stirrer and a condenser. The mixture was stirred until homogeneous, and compound 37 (3.0 g, 37.43 mmol) was added dropwise. After the addition was complete, the reaction mixture was heated to 90 °C and stirred for 2 hours. The mixture was cooled to room temperature, extracted with ethyl acetate (20 mL x 2), and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to give 1.5 g of anhydrous oil, yield 17.89%.

[0209] Step 3: Synthesis of compound 39.

[0210] Compound 36 (420 mg, 2.31 mmol), compound 38 (640 mg, 2.86 mmol), and acetonitrile (15 mL) were added to a 50 mL single-necked flask equipped with a magnetic stirrer and condenser. The mixture was stirred until dissolved. Potassium iodide (76 mg, 0.46 mmol) and potassium carbonate (382 mg, 2.77 mmol) were added. The reaction mixture was heated to reflux under a nitrogen atmosphere and reacted overnight. After cooling to room temperature, the solvent was removed by vacuum distillation. Water (15 mL) and ethyl acetate (20 mL) were added. The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and passed through a silica gel column to give 250 mg of a yellow oil, in 44.2% yield. LC-MS (APCI): m / z = 245.1 (M+1) + .1 H NMR (300MHz, CDCl3) δ 8.20 (d, J = 9.0 Hz, 2H), 6.98 (d, J = 9.0 Hz, 2H), 4.21 (t, J = 6.0 Hz, 2H), 2.96 (t, J = 6.0 Hz, 2H).

[0211] Step 4: Synthesis of compound 40.

[0212] Compound 39 (250 mg, 1.04 mmol) and methanol (10 mL) were added to a 50 mL single-necked flask equipped with a magnetic stirrer. The mixture was stirred until dissolved, then Pd / C (25 mg, 10%) was added. The mixture was evacuated under a hydrogen balloon and purged three times. The reaction was carried out overnight under a hydrogen atmosphere. The catalyst was filtered off, and the filter cake was washed with methanol (3 mL). The filtrates were combined and concentrated to give 220 mg of a yellow oil, with a yield of 98.6%. LC-MS (APCI): m / z = 215.1 (M+1) + .

[0213] Step 5: Synthesis of compound T-5.

[0214] Compound 40 (200 mg, 0.56 mmol), compound 6 (107 mg, 0.51 mmol), and ethylene glycol monomethyl ether (6 mL) were added to a 25 mL single-necked flask equipped with a magnetic stirrer and condenser. The mixture was stirred until dissolved, and isopropanol hydrochloride solution (1.41 mmol, 0.28 mL, 5 M) was added dropwise. The mixture was heated to 120 °C under a nitrogen atmosphere and stirred overnight. After cooling to room temperature, water (10 mL) and saturated sodium bicarbonate (5 mL) were added, and the mixture was extracted with dichloromethane (15 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was passed through a silica gel column to give 150 mg of a white solid, with a yield of 50.3%. LC-MS (APCI): m / z = 533.2 (M+1) + . 1 H NMR (500MHz, CDCl3) δ8.11(s,1H),7.92-7.89(m,2H),7.57(d,J=8.0Hz,1H),7.41-7.38(m,3H),6.91(d,J=8.0Hz,1 H),6.81(s,1H),6.44(s,1H),4.60(s,1H),4.13(t,J=5.5Hz,2H),2.94(t,J=5.5Hz,2H),2.13(s,3H),1.22(s,9H).

[0215] Bioactivity testing.

[0216] (1) Kinase inhibition

[0217] Reagents and consumables:

[0218] JAK2, JAK2 / V617F, ATP (Sigma, catalog number A7699-1G), DMSO (Sigma, catalog number D2650), 96-well plate (Corning, catalog number 3365), 384-well plate (Greiner, catalog number 784076), HTRF Kinase TK kit (Cisbio), 5x Kinase Buffer A (Life Technologies, catalog number PV3186), Kinase Tracer 199 (Life Technologies, catalog number PV5830). Eu-anti-GST antibody (Life Technologies, catalog number PV5594).

[0219] Specific experimental methods:

[0220] Compound preparation: Dissolve the test compound in DMSO to prepare a 20 mM stock solution. Then, dilute it 3-fold in DMSO at equal serial rates, for a total of ten dilutions. Before adding the drug, dilute it 10-fold with buffer solution.

[0221] JAK2 and JAK2[V617F] kinase assay: In 5x kinase buffer A, JAK2 or JAK2[V617F] kinase was mixed with pre-diluted compounds of different concentrations for 10 minutes, with double replicates for each concentration. The corresponding substrate and ATP were added, and the reaction was incubated at room temperature for 20 minutes (positive and negative controls were provided: negative control was a blank control, and positive control was erlotinib). After the reaction, the assay reagent (from the HTRF Kinase TK kit) was added, and the mixture was incubated at room temperature for 30 minutes. The enzyme activity was measured using an Evnvision microplate reader in the presence of the compounds of this invention at various concentrations, and the inhibitory activity of different concentrations of the compounds on the enzyme activity was calculated. Then, the inhibitory activity of the enzyme activity at different concentrations of the compounds was fitted using a four-parameter equation and Graphpad 5.0 software to calculate the IC50. 50 value.

[0222] In the kinase inhibition experiments described above, the compounds of the present invention and the undeuterated compound Fedratinib were tested. It was found that the compounds of the present invention exhibited stronger or comparable activity against JAK2 kinase and JAK2 / V617F kinase. The results of the inhibitory effects of the compounds in representative examples on the kinases are summarized in Table 1 below.

[0223] Table 1

[0224] Examples of compounds JAK2 IC 50 (nM) <![CDATA[JAK2 / V617F IC 50 (nM)]]> Fedratinib 3.08 4.05 T-1 3.16 4.06 T-2 2.89 3.69 T-3 3.06 3.78 T-4 3.24 4.05 T-5 3.47 4.55

[0225] (2) Cytotoxicity test

[0226] The inhibitory effects of the compounds in the examples on the activity of BaF3-EpoR-JAK2 and BaF3-EpoR-JAK2 / V617F cells were investigated.

[0227] Consumables and reagents: RPMI-1640 medium (GIBCO, catalog number A10491-01), fetal bovine serum (GIBCO, catalog number 10099-141), antibiotics (GIBCO, catalog number 10010-031), IL-3 (CST, catalog number 8923SF), phosphate-buffered saline (PBS) (GIBCO, catalog number 10010-0312), penicillin-streptomycin (GIBCO, catalog number 15140-122);

[0228] Cell lines: BaF3-EpoR-JAK2 cells (Pharmaron), BaF3-EpoR-JAK2 / V617F cells (Pharmaron), CellTiter-Glo4 live cell assay kit (Promega, catalog number G7572), 96-well black-walled transparent flat-bottomed cell culture plates (Corning, catalog number 3340).

[0229] Experimental Methods: 1. Cell Plate Preparation: BaF3-EpoR-JAK2 cells and BaF3-EpoR-JAK2 / V617F cells were seeded in 96-well plates. 1 ng / ml IL-3 was added to the Ba / F3 cells. The cells were incubated overnight in a CO2 incubator. 2. Dissolving the Test Compounds in DMSO and Performing 3-fold serial dilutions (9 compound concentrations) for replicates. 3. Cell Treatment with Compounds: The compounds were transferred to the cells at an initial concentration of 10 μM. The cells were incubated in a CO2 incubator for 3 days. 4. Detection: CellTiter-Glo reagent was added to the cells and incubated at room temperature for 30 minutes to stabilize the luminescence signal. Readings were performed using a PerkinElmer Envision multilabel analyzer.

[0230] In the above-mentioned cytotoxicity experiments, the compounds of the present invention and the undeuterated compound Fedratinib were tested. It was found that the compounds of the present invention exhibited stronger or comparable activity against BaF3-EpoR-JAK2 cells and BaF3-EpoR-JAK2 / V617F cells. The results of the inhibitory effects of the compounds of representative examples on the in vitro proliferation of cancer cells are summarized in Table 2 below.

[0231] Table 2

[0232]

[0233] (3) Evaluation of metabolic stability

[0234] Microsomal assay: Human liver microsomes: 0.5 mg / mL, Xenotech; Rat liver microsomes: 0.5 mg / mL, Xenotech; Mouse liver microsomes: 0.5 mg / mL, Xenotech; Coenzymes (NADPH / NADH): 1 mM, Sigma LifeScience; Magnesium chloride: 5 mM, 100 mM phosphate buffer (pH 7.4).

[0235] Preparation of stock solution: Accurately weigh a certain amount of the powder of the compound in the example and dissolve it in DMSO to 5mM.

[0236] Preparation of phosphate buffer (100mM, pH 7.4): Mix 150mL of 0.5M potassium dihydrogen phosphate and 700mL of 0.5M dipotassium hydrogen phosphate solution, then adjust the pH of the mixture to 7.4 with 0.5M dipotassium hydrogen phosphate solution. Before use, dilute with ultrapure water 5 times and add magnesium chloride to obtain phosphate buffer (100mM), which contains 100mM potassium phosphate, 3.3mM magnesium chloride, and pH 7.4.

[0237] Prepare the NADPH regeneration system solution (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U / mL G-6-PD, and 3.3 mM magnesium chloride) and place it on wet ice before use.

[0238] Preparation of stop solution: An acetonitrile solution containing 50 ng / mL propranolol hydrochloride and 200 ng / mL tolbutamide (internal standard). Add 25057.5 μL of phosphate buffer (pH 7.4) to a 50 mL centrifuge tube, and add 812.5 μL of human liver microsomes, rat liver microsomes, and mouse liver microsomes respectively. Mix well to obtain a liver microsome dilution with a protein concentration of 0.625 mg / mL. Sample incubation: Dilute the stock solutions of the corresponding compounds to 0.25 mM with an aqueous solution containing 70% acetonitrile to prepare working solutions. Add 398 μL of each of the human liver microsome, rat liver microsome, or mouse liver microsome dilutions to a 96-well incubation plate (N=2), and add 2 μL of 0.25 mM working solution to each well. Mix well.

[0239] Determination of metabolic stability: 300 μL of pre-chilled stop solution was added to each well of a 96-well deep-well plate and placed on ice to serve as the stop plate. The 96-well incubation plate and NADPH regeneration system were placed in a 37°C water bath and incubated at 100 rpm for 5 min. 80 μL of incubation solution was taken from each well of the incubation plate and added to the stop plate, mixed well, and 20 μL of NADPH regeneration system solution was added to serve as the 0 min sample. Another 80 μL of NADPH regeneration system solution was added to each well of the incubation plate to start the reaction and begin timing. The reaction concentration of the corresponding compound was 1 μM, and the protein concentration was 0.5 mg / mL. At 10, 30, and 90 min of reaction, 100 μL of reaction solution was taken and added to the stop plate, and the reaction was terminated by vortexing for 3 min. The stop plate was centrifuged at 5000 × g at 4°C for 10 min. Take 100 μL of supernatant into a 96-well plate pre-filled with 100 μL of distilled water, mix well, and analyze the sample using LC-MS / MS.

[0240] Data Analysis: The peak areas of the corresponding compounds and internal standards were detected using an LC-MS / MS system, and the peak area ratio of the compounds to the internal standard was calculated. The slope was obtained by plotting the natural logarithm of the percentage of the remaining compound against time, and t was calculated using the following formula. 1 / 2 and CL int V / M is equal to 1 / protein concentration.

[0241]

[0242] In the metabolic stability evaluation experiments described above, the compounds of the present invention and the undeuterated compound Fedratinib were tested. It was found that the compounds of the present invention have a longer half-life and a shorter clearance rate, exhibiting superior metabolic stability. The metabolic stability results of the representative example compounds are summarized in Table 3.

[0243] Table 3

[0244]

[0245]

[0246] (4) Rat pharmacokinetics experiment

[0247] Six male Sprague-Dawley rats, 7-8 weeks old and weighing approximately 210g, were divided into two groups of three. Each group received a single dose of the compound via intravenous or oral administration (3 mg / kg intravenously; 10 mg / kg orally), and the pharmacokinetic differences were compared.

[0248] Rats were fed a standard diet and given water. Fasting began 16 hours prior to the experiment. The drug was dissolved in PEG400 and dimethyl sulfoxide. Blood was collected from the orbital rim at 0.083, 0.25, 0.5, 1, 2, 4, 6, 8, 12, and 24 hours post-drug administration.

[0249] Rats were briefly anesthetized after inhaling ether, and 300 μL of blood was collected from the orbital cavity in a test tube. The test tube contained 30 μL of 1% heparin solution. Before use, the test tube was dried overnight at 60°C. After the last blood sample was collected, the rats were euthanized under ether anesthesia.

[0250] After blood sample collection, the test tube was immediately and gently inverted at least five times to ensure thorough mixing before being placed on ice. The blood sample was centrifuged at 5000 rpm for 5 minutes at 4°C to separate the plasma from the red blood cells. 100 μL of plasma was pipetted into a clean plastic centrifuge tube, labeled with the compound name and time point. The plasma was stored at -80°C before analysis. The concentration of the compound of the present invention in the plasma was determined by LC-MS / MS. Pharmacokinetic parameters were calculated based on the plasma drug concentrations of each animal at different time points.

[0251] In the above-mentioned rat pharmacokinetic experiments, the compounds of the present invention and the undeuterated compound Fedratinib were tested, and it was found that the compounds of the present invention have better oral bioavailability. The oral bioavailability results of the representative example compounds are summarized in Table 4.

[0252] Table 4

[0253] Examples of compounds Fedratinib T-5 Oral bioavailability F (%) 7.24 7.48

[0254] The above description, in conjunction with specific preferred embodiments, provides a further detailed explanation of the present invention. It should not be construed that the specific implementation of the present invention is limited to these descriptions. For those skilled in the art, various simple deductions or substitutions can be made without departing from the concept of the present invention, and all such modifications and substitutions should be considered within the scope of protection of the present invention.

Claims

1. Compounds of formula (II): Equation (II) in, R 1 R 2 R 3 and R 4 It is hydrogen; R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 It is hydrogen; Y 5 It is hydrogen and X 4 It is CH3; X 1 X 2 and X 3 Selected from CH3 or CD3; The condition is that the above-mentioned compound contains at least three deuterium atoms; Or its pharmaceutically acceptable salt.

2. The compound according to claim 1, wherein the compound is selected from the group consisting of: Formula (11), or a pharmaceutically acceptable salt thereof.

3. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-2.

4. Use of the compound of any one of claims 1-2 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 3 in the preparation of a medicament for treating at least part of a disease mediated by JAK2.

5. The use according to claim 4, wherein the disease is myeloproliferative disorder, polycythemia vera, idiopathic thrombocythemia, cancer, eye disease, inflammation, any disease or viral infection related to angiogenesis.

6. The use according to claim 5, wherein the disease is polycythemia vera.

7. The use according to claim 5, wherein the disease is idiopathic thrombocytosis.

8. The use according to claim 5, wherein the disease is myelofibrosis with myeloid metaplasia.

9. The use according to claim 5, wherein the disease is proliferative diabetic retinopathy.

10. The use according to claim 5, wherein the disease is selected from cardiovascular disease or chronic myeloid leukemia.

11. The use according to claim 5, wherein the disease is selected from psoriasis.

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