Pyrazolyl compounds and methods of use thereof

Abstract

Provided are compounds having activity as chemotherapeutic agents. The compounds have the following structure (I): #imgabs0# or a pharmaceutically acceptable salt, stereoisomer, isotopic form or prodrug thereof, wherein R 1a , R 1b , R 1c , R 1d , L, A and B are as defined herein. Also provided are methods related to the preparation and use of such compounds, pharmaceutical compositions containing such compounds, and methods for treating cancer (e.g., blood cancer).

Description

[0001] background Technical Field

[0002] The present disclosure generally relates to novel compounds and methods for their preparation and use as therapeutic or prophylactic agents, such as for the treatment of cancer (eg, hematological cancer).

[0003] Description of Related Technology

[0004] Cancer is a group of diseases that involve abnormal cell growth with the potential to spread to various parts of the body. There are hundreds of types of cancer that affect humans. Millions of people have been diagnosed with cancer, and millions more are diagnosed every year. The most common types of cancer include lung cancer, breast cancer, prostate cancer, colorectal cancer, and more. Treatments for cancer include surgery, radiation therapy, chemotherapy, immunotherapy, hormone therapy, stem cell replacement therapy, and more. Treatment options can be invasive and have a variety of adverse side effects. Chemotherapy is a common type of cancer treatment, and while it can be effective, chemotherapy can have strong side effects.

[0005] Therefore, despite the progress that has been made in this field, there remains a need in the art for improved compounds and methods for treating cancer (e.g., blood cancers). The present disclosure satisfies this need and provides other related advantages.

[0006] Brief description of the invention

[0007] In short, the embodiments of the present disclosure provide compounds, including pharmaceutically acceptable salts, isotopic forms, stereoisomers and prodrugs thereof. Methods of using such compounds to treat various diseases or conditions such as cancer (e.g., blood cancer) are also provided.

[0008] In one embodiment, a compound having the following structure (I) is provided:

[0009]

[0010] or a pharmaceutically acceptable salt, stereoisomer, isotope form or prodrug thereof, wherein R 1a , R 1b , R 1c , R 1d , L, As defined herein. In various other embodiments, pharmaceutical compositions comprising one or more compounds of structure (I) and a pharmaceutically acceptable carrier are also provided.

[0011] In other embodiments, the disclosure provides methods for treating cancer (eg, hematological cancer), comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition comprising any one or more compounds of structure (I).

[0012] These and other aspects of the disclosure will become apparent upon reference to the following detailed description. DETAILED DESCRIPTION

[0013] In the following description, certain specific details are set forth in order to provide a thorough understanding of various embodiments of the present disclosure. However, it will be appreciated by those skilled in the art that the present disclosure may be practiced without these details.

[0014] Throughout this specification and claims, the word "comprise" and variations thereof (e.g., "comprises" and "comprising") are to be construed in an open, inclusive sense, i.e., as meaning "including, but not limited to," unless the context requires otherwise.

[0015] In the present description, any concentration range, percentage range, ratio range or integer range shall be understood to include the value of any integer within the range, and where appropriate, include its fraction (e.g., one tenth and one hundredth of an integer), unless otherwise specified. In addition, any numerical range described herein relating to any physical feature (e.g., polymer subunit, size or thickness) shall be understood to include any integer within the range, unless otherwise specified. As used herein, the terms "about" and "approximately" mean ±20%, ±10%, ±5% or ±1% of the range, value or structure shown, unless otherwise specified. It should be understood that the terms "a" and "an" as used herein refer to "one or more" of the listed components. The use of alternatives (e.g., "or") shall be understood to mean one, two, or any combination thereof in the alternatives.

[0016] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in relation to that embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrases "in one embodiment" or "in an embodiment" throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0017] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which the present disclosure belongs. As used in this specification and claims, the singular forms "a", "an", and "the" include plural references unless the context clearly indicates otherwise.

[0018] "Amino" refers to the -NH2 group.

[0019] "Carboxy" or "carboxyl" refers to a -CO2H group.

[0020] "Cyano" refers to a -CN group.

[0021] "Hydroxy" or "hydroxyl" refers to an -OH group.

[0022] "Oxo" refers to a =0 substituent.

[0023] "Alkyl" refers to a saturated straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms, having from one to twelve carbon atoms (C1-C 12 The term "alkyl" refers to an alkyl group, preferably one to eight carbon atoms (C1-C8 alkyl) or one to six carbon atoms (C1-C6 alkyl), and which is connected to the rest of the molecule by a single bond, such as methyl, ethyl, n-propyl, 1-methylethyl (isopropyl), n-butyl, n-pentyl, 1,1-dimethylethyl (tert-butyl), 3-methylhexyl, 2-methylhexyl, etc. Unless otherwise specifically stated in the specification, an alkyl group is optionally substituted.

[0024] "Alkenyl" refers to an unsaturated straight or branched hydrocarbon chain group consisting solely of carbon and hydrogen atoms and containing one or more carbon-carbon double bonds, having two to twelve carbon atoms (C2-C 12 The term "alkenyl" refers to an alkylene group, preferably a ...

[0025] "Alkynyl" refers to an unsaturated straight or branched hydrocarbon chain radical consisting solely of carbon and hydrogen atoms and containing one or more carbon-carbon triple bonds, having two to twelve carbon atoms (C2-C 12 Alkynyl), preferably two to eight carbon atoms (C2-C8 alkynyl) or two to six carbon atoms (C2-C6 alkynyl), and which is attached to the rest of the molecule by a single bond, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl, etc. Unless otherwise specifically stated in the specification, an alkynyl group is optionally substituted.

[0026] "Alkylidene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain consisting only of carbon and hydrogen, connecting the rest of the molecule to a radical, which is saturated or unsaturated (i.e., containing one or more double bonds or "alkenylene" and / or triple bonds or "alkynylene"), and has one to twelve carbon atoms, such as methylene, ethylene, propylene, n-butylene, vinylene, propenylene, n-butenylene, propynylene, n-butynylene, etc. The alkylene chain is connected to the rest of the molecule by a single bond or a double bond and to the radical by a single bond or a double bond. The point of attachment of the alkylene chain to the rest of the molecule and to the radical can be through one carbon or any two carbons within the chain. Unless otherwise specifically stated in this specification, the alkylene chain is optionally substituted.

[0027] "Alkoxy" refers to a group of the formula -OR a A group in which R a is an alkyl group as defined above containing from one to twelve carbon atoms. Unless stated otherwise specifically in the specification, an alkoxy group is optionally substituted.

[0028] "Alkylamino" refers to a group of the formula -NHR a or -NR a R a A group in which each R a is independently an alkyl group as defined above containing from one to twelve carbon atoms. Unless specifically stated otherwise, an alkylamino group is optionally substituted.

[0029] "Aminoalkyl" refers to an alkyl group containing at least one amino substituent. The amino substituent can be on a tertiary, secondary, or primary carbon. Unless specifically stated otherwise, an aminoalkyl group is optionally substituted.

[0030] "Alkylaminoalkyl" refers to an alkyl group containing at least one alkylamino substituent. The alkylamino substituent can be on a tertiary carbon, a secondary carbon, or a primary carbon. Unless otherwise specifically stated in the specification, an alkylaminoalkyl group is optionally substituted.

[0031] "Alkylsulfonyl" refers to a group of the formula -S(O)2R a A group in which R a is an alkyl group as defined above containing from one to twelve carbon atoms. Unless specifically stated otherwise, an alkylsulfonyl group is optionally substituted.

[0032] "Aromatic ring" refers to a cyclic planar portion of a molecule (i.e., a group) having a ring with a resonant bond, which exhibits increased stability relative to other connection arrangements with the same atomic group. Typically, an aromatic ring contains a set of covalently bonded coplanar atoms and contains a certain number of π-electrons (e.g., alternating double bonds and single bonds), the number of which is an even number but not a multiple of 4 (i.e., 4n+2 π-electrons, where n=0, 1, 2, 3, etc.). Aromatic rings include, but are not limited to, phenyl, naphthyl, imidazolyl, pyrrolyl, pyridyl, pyrimidyl, pyrazinyl, pyridone (pyridonyl), pyridazinyl, pyrimidone. Unless otherwise specifically stated in the specification, "aromatic ring" includes all groups that are optionally substituted.

[0033] "Aryl" refers to a carbocyclic ring system radical containing 6 to 18 carbon ring atoms and at least one aromatic ring. For the purposes of embodiments of the present invention, an aryl group is a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may include fused or bridged ring systems. Aryl groups include, but are not limited to, benzoacenaphthene, acenaphthylene, acephenanthren, anthracene, azulene, benzene, The aryl groups of fluoranthene, fluorene, unsymmetrical indacene, symmetrical indacene, indane, indene, naphthalene, phenalene, phenanthrene, pyrene and triphenylene. Unless otherwise specifically stated in the specification, the term "aryl" or the prefix "ar" (such as in "aryl") is intended to include optionally substituted aryl groups.

[0034] "Carbocyclic" or "carbocycle" refers to a ring system in which each of the ring atoms is carbon.

[0035] "Cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic carbocyclic group, which may include a fused or bridged ring system, having three to fifteen carbon atoms, preferably three to ten carbon atoms, and which is saturated or unsaturated and connected to the remainder of the molecule by a single bond. Monocyclic groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. Polycyclic groups include, for example, adamantyl, norbornyl, decahydronaphthyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, etc. Unless otherwise specifically stated in this specification, cycloalkyl groups are optionally substituted.

[0036] "Fused" refers to any ring structure described herein that is fused to an existing ring structure in a compound of the invention. When the fused ring is a heterocyclyl ring or a heteroaryl ring, any carbon atom on the existing ring structure that becomes part of the fused heterocyclyl ring or the fused heteroaryl ring may be replaced by a nitrogen atom.

[0037] "Halo" or "halogen" refers to bromo, chloro, fluoro or iodo.

[0038] "Haloalkyl" refers to an alkyl group as defined above substituted with one or more halogen groups as defined above, for example trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1,2-difluoroethyl, 3-bromo-2-fluoropropyl, 1,2-dibromoethyl, etc. "Perhaloalkyl" is an alkyl group as defined above in which each H atom is replaced by a halogen. Unless otherwise specifically stated in the specification, a haloalkyl group is optionally substituted.

[0039] "Haloalkoxy" refers to a group of the formula -OR a A group in which R a is a haloalkyl group as defined herein containing from one to twelve carbon atoms. Unless stated otherwise specifically in the specification, a haloalkoxy group is optionally substituted.

[0040] "Heteroalkylene" or "heteroalkylene chain" refers to a straight or branched divalent chain consisting of carbon, hydrogen and at least one heteroatom (e.g., oxygen, nitrogen, sulfur, phosphorus, etc.) within the chain bond, connecting the rest of the molecule to a radical, which is saturated or unsaturated (i.e., containing one or more double bonds and / or triple bonds). The heteroalkylene chain is connected to the rest of the molecule by a single bond or a double bond, and to the radical by a single bond or a double bond. The point of attachment of the heteroalkylene chain to the rest of the molecule and to the radical can be through one carbon, any two carbons, one heteroatom, or any two heteroatoms within the chain. Unless otherwise specifically stated in this specification, the heteroalkylene chain is optionally substituted.

[0041] "Heteroatom" or "heteroatom chain" refers to a straight or branched divalent chain that is composed of atoms other than carbon and hydrogen (e.g., oxygen, nitrogen, sulfur, phosphorus, etc.) in the chain bond, connecting the rest of the molecule to the group, which is saturated or unsaturated (i.e., containing one or more double bonds and / or triple bonds). The heteroatom chain is connected to the rest of the molecule by a single bond or a double bond, and is connected to the group by a single bond or a double bond. The point of connection of the heteroatom chain to the rest of the molecule and to the group can be through one heteroatom or any two heteroatoms in the chain. Unless otherwise specifically stated in this specification, the heteroatom chain is optionally substituted.

[0042] "Heterocyclyl" or "heterocyclic ring" refers to a stable 3- to 18-membered non-aromatic ring radical having from one to twelve ring carbon atoms (e.g., two to twelve) and from one to six ring heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specifically stated in the specification, a heterocyclyl group is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include fused, spirocyclic ("spiro-heterocyclyl"), and / or bridged ring systems; and the nitrogen, carbon, or sulfur atoms in the heterocyclyl group are optionally oxidized; the nitrogen atom is optionally quaternized; and the heterocyclyl group is partially or fully saturated. Examples of such heterocyclyl groups include, but are not limited to, dioxolanyl, thienyl [1,3] dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuranyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, 1,2,3,4-tetrahydroquinolinyl, and 1,1-dioxo-thiomorpholinyl. Unless otherwise specifically stated in the specification, heterocyclyl groups are optionally substituted.

[0043] As used herein, the term "substituted" means any of the above groups (e.g., alkyl, alkenyl, alkynyl, alkylene, alkoxy, alkylamino, aminoalkyl, alkylaminoalkyl, alkylsulfonyl, aryl, cycloalkyl, haloalkyl, haloalkoxy, heteroalkylene, and / or heterocyclyl) in which at least one hydrogen atom (e.g., 1, 2, 3, or all hydrogen atoms) is replaced by a bond to a non-hydrogen atom, such as, but not limited to, a halogen atom, such as F, Cl, Br, and I; a hydroxyl group, an alkoxy group, Oxygen atoms in groups such as thiol groups, alkylthio groups, sulfone groups, sulfonyl groups and sulfoxide groups; Nitrogen atoms in groups such as cyano groups, amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides and enamines; Silicon atoms in groups such as trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl and triarylsilyl; And other heteroatoms in various other groups. "Substituted" also means any of the above groups in which one or more hydrogen atoms are replaced by a higher valence bond (e.g., double bond or triple bond) with a heteroatom, such as: oxygen in groups of oxo, carbonyl, carboxyl and ester groups; And nitrogen in groups such as imines, oximes, hydrazones and nitriles. For example, "substituted" includes groups in which one or more hydrogen atoms are replaced by NR g R h 、-NR g C(=O)R h、-NR g C(=O)NR g R h 、-NR g C(=O)OR h 、-NR g S02R h 、-OC(=O)NR g R h 、-OR g 、-SR g 、-SOR g 、-SO2R g 、-OSO2R g 、-SO2OR g , =NSO2R g and -SO2NR g R h "Substituted" also means any of the above groups in which one or more hydrogen atoms are replaced by -C(=O)R g 、-C(=O)OR g 、-C(=O)NR g R h 、-CH2SO2R g 、-CH2SO2NR g R h In the foregoing, R g and R h The same or different, and independently hydrogen, optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkylamino, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted haloalkyl or optionally substituted heterocyclic radical. "Substituted" also means any of the above groups in which one or more hydrogen atoms are replaced by a key with amino, cyano, hydroxyl, oxo (=O), halo or optionally substituted alkyl, optionally substituted alkoxy, optionally substituted alkylamino, optionally substituted aryl, optionally substituted cycloalkyl, optionally substituted haloalkyl and / or optionally substituted heterocyclic radical groups. In addition, each of the above-mentioned substituents can also be optionally substituted by one or more of the above substituents.

[0044] It should be understood that for R 1a , R 1b , R 1c , R 1d , L, Each of the options is optionally substituted as described above, unless otherwise specifically stated, and provided that the substitution satisfies all valences. 1a , R 1b , R 1c , R 1d , L, Each alternative is optionally substituted unless otherwise specifically stated and provided that such substitution results in a stable molecule (eg, groups such as H and halogen are not optionally substituted).

[0045] The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound described herein sufficient to achieve the intended application, including but not limited to disease treatment as defined below. The therapeutically effective amount may vary depending on the intended therapeutic application (in vivo), or the individual and disease condition being treated, such as the weight and age of the individual, the severity of the disease condition, the mode of administration, etc., which can be readily determined by one of ordinary skill in the art. The term also applies to doses that will induce a specific response in target cells (e.g., a decrease in platelet adhesion and / or cell migration). The specific dose will vary depending on the specific compound selected, the dosing regimen followed, whether it is administered in combination with other compounds, the timing of administration, the tissue to which it is administered, and the physical delivery system that carries it.

[0046] As used herein, "treatment" or "treating" refers to a method of obtaining a beneficial or desired result about a disease, disorder, or medical condition, including but not limited to a therapeutic benefit and / or a preventive benefit. A therapeutic benefit means eliminating or improving the underlying disorder being treated. In addition, a therapeutic benefit is obtained by eliminating or improving one or more physiological symptoms associated with the underlying disorder such that an improvement is observed in the individual, although the individual may still suffer from the underlying disorder. In certain embodiments, in order to obtain a preventive benefit, the composition is administered to an individual at risk of developing a particular disease, or to an individual reporting one or more physiological symptoms of a disease, even though the disease may not have been diagnosed.

[0047] As used herein, the term "therapeutic effect" encompasses therapeutic benefits and / or prophylactic benefits as described above. Prophylactic effects include delaying or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, stopping or reversing the progression of a disease or condition, or any combination thereof.

[0048] As used herein, the terms "co-administered," "administered in conjunction with," and their grammatical equivalents encompass administration of two or more agents to animals, including humans, such that the agents and / or their metabolites are both present in the individual at the same time. Co-administration includes simultaneous administration in separate compositions, administration at different times in separate compositions, or administration in a composition in which both agents are present.

[0049] "Pharmaceutically acceptable salts" include both acid addition salts and base addition salts.

[0050] "Pharmaceutically acceptable acid addition salts" means those which retain the biological effectiveness and properties of the free bases, are not biologically or otherwise undesirable, and are formed with inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, as well as organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid Acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, etc.

[0051] "Pharmaceutically acceptable base addition salts" refer to those salts that retain the biological effectiveness and properties of the free acids and are not biologically or otherwise undesirable. These salts are prepared by the addition of inorganic or organic bases to the free acids. Salts derived from inorganic bases include, but are not limited to, sodium salts, potassium salts, lithium salts, ammonium salts, calcium salts, magnesium salts, iron salts, zinc salts, copper salts, manganese salts, aluminum salts, and the like. Preferred inorganic salts are ammonium salts, sodium salts, potassium salts, calcium salts, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines and basic ion exchange resins, such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, dimethylethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, phenethylbenzylamine, benzathine, ethylenediamine, glucosamine, methylglucosamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine.

[0052] In some embodiments, pharmaceutically acceptable salts include quaternary ammonium salts, such as quaternary ammonium alkyl halide salts (eg, methyl bromide).

[0053] The term "inhibitor" refers to a compound that has the ability to inhibit a biological function of a target protein, whether by inhibiting the activity or expression of the protein. Preferred biological activities inhibited by the inhibitor are related to the formation, growth or spread of a tumor.

[0054] "Anticancer", "anti-tumor agent" or "chemotherapeutic agent" refers to any agent useful for treating a neoplastic condition. One class of anti-cancer agents includes chemotherapeutic agents. "Chemotherapy" means administering one or more chemotherapeutic drugs and / or other agents to a cancer patient by various methods, including intravenous, oral, intramuscular, intraperitoneal, intravesical, subcutaneous, transdermal, buccal, or in the form of an inhalation or suppository.

[0055] The term "selective inhibition" or "selectively inhibit" refers to the ability of a biologically active agent to preferentially reduce target signaling activity over non-target signaling activity via direct or indirect interaction with the target.

[0056] "Subject" refers to an animal, such as a mammal, such as a human. The methods described herein can be used for human therapy and veterinary applications. In some embodiments, the subject is a mammal, and in some embodiments, the subject is a human.

[0057] "Mammal" includes humans, as well as domestic animals such as laboratory animals and household pets (e.g., cats, dogs, pigs, cows, sheep, goats, horses, rabbits), and non-domestic animals such as wild animals and the like.

[0058] "Radiation therapy" means exposing an individual to radiation emitters, such as alpha-particle emitting radionuclides (e.g., actinium and thorium radionuclides), low linear energy transfer (LET) radiation emitters (i.e., beta emitters), conversion electron emitters (e.g., strontium-89 and samarium-153-EDTMP), or high-energy radiation including, but not limited to, x-rays, gamma rays, and neutrons, using conventional methods and compositions known to practitioners.

[0059] The prodrugs of disclosed compounds are included in various embodiments. "Prodrug" means a compound that can be converted into a bioactive compound described herein (e.g., a compound of structure (I)) under physiological conditions or by solvolysis. Therefore, the term "prodrug" refers to a precursor of a pharmaceutically acceptable bioactive compound. In some aspects, the prodrug is inactive when applied to an individual, but is converted into an active compound in vivo, for example, by hydrolysis. Prodrug compounds generally provide advantages of solubility, tissue compatibility, or delayed release in mammalian organisms (see, e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier, Amsterdam)). Discussion of prodrugs is provided in Higuchi, T. et al. "Pro-drugs as Novel Delivery Systems," ACSSymposium Series, Vol. 14 and Bioreversible Carriers in Drug Design, Edward B. Roche, ed., American Pharmaceutical Association and Pergamon Press, 1987, both of which are incorporated herein by reference in their entirety. The term "prodrug" also means any covalently bonded carrier that releases the active compound in vivo when such prodrug is administered to a mammalian individual. Prodrugs of active compounds as described herein are typically prepared in the following manner, i.e., modifying the functional groups present in the active compound so that the modifier is cleaved into the parent active compound in conventional operations or in vivo. Prodrugs include compounds in which hydroxyl, amino or sulfhydryl groups are bonded to any group, and when the prodrug of the active compound is administered to a mammalian individual, it cleaves to form free hydroxyl, free amino or free sulfhydryl groups, respectively. Examples of prodrugs include, but are not limited to, acetate, formate and benzoate derivatives of hydroxyl functional groups in the active compound, or acetamide, formamide and benzamide derivatives of amine functional groups, and the like.

[0060] The term "in vivo" refers to events that occur within the body of a subject.

[0061] The embodiments disclosed herein are also intended to encompass all pharmaceutically acceptable compounds of structure (I) that are isotopically labeled by replacing one or more atoms with an atom having a different atomic mass or mass number (i.e., "isotopic forms" of compounds of structure (I)). Examples of isotopes that may be introduced into the disclosed compounds include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, chlorine, and iodine, e.g. 2 H. 3 H. 11 C. 13 C.14 C. 13 N 、15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 36 Cl, 123 I and 125 I. These radiolabeled compounds can be used to help determine or measure the effectiveness of a compound by characterizing, for example, the site or mode of action or binding affinity to a pharmacologically important site of action. Certain isotopically labeled compounds of structure (I) (e.g., compounds into which a radioactive isotope has been introduced) are useful in drug and / or substrate tissue distribution studies. The radioactive isotope tritium (i.e. 3 H) and carbon-14 (i.e. 14 C) is particularly suitable for this purpose due to its easy introduction and readily available detection means.

[0062] Using deuterium (i.e. 2 Heavier isotopic substitutions of H) may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances.

[0063] Use such as 11 C. 18 F. 15 O and 13 Positron emitting isotope substitution of N can be useful in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of structure (I) can generally be prepared by conventional techniques known to those skilled in the art or by methods analogous to those described in the Examples described below, using an appropriate isotopically labeled reagent in place of the unlabeled reagent previously used.

[0064] Certain embodiments are also intended to encompass metabolites in vivo of disclosed compounds. Such products can be produced by oxidation, reduction, hydrolysis, amidation, esterification, etc. (mainly due to enzymatic processes) of the compound, for example, administered. Therefore, embodiments include compounds produced by the following method, which includes administering a compound of the present disclosure to a mammal for a period of time sufficient to produce its metabolites. Such products are generally confirmed by administering a radiolabeled compound of the present disclosure of a detectable dose to an animal such as a rat, mouse, guinea pig, monkey, or to a human, allowing enough time for metabolism to occur, and separating its conversion products from urine, blood, or other biological samples.

[0065] "Stable compound" and "stable structure" are intended to indicate a compound that is sufficiently robust to survive isolation to a useful degree of purity from a reaction mixture, and formulation into an efficacious therapeutic agent.

[0066] Typically crystallization produces a solvate of the compound of the present invention. As used herein, the term "solvate" refers to an aggregate comprising one or more molecules of the compound of the present disclosure and one or more solvent molecules. In some embodiments, the solvent is water, in which case the solvate is a hydrate. Alternatively, in other embodiments, the solvent is an organic solvent. Therefore, the compound of the present disclosure can exist as a hydrate, including a monohydrate, a dihydrate, a hemihydrate, a sesquihydrate, a trihydrate, a tetrahydrate, etc., and corresponding solvated forms. In some aspects, the compound of the present disclosure is a true solvate, while in other cases, the compound of the present disclosure retains only adventitious water or a mixture of water plus some adventitious solvents.

[0067] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not occur. For example, "optionally substituted aryl" means that the aryl group may or may not be substituted, and that the description includes substituted aryl groups and aryl groups without substitution.

[0068] "Pharmaceutical composition" refers to a preparation of a compound of the present disclosure and a medium generally accepted in the art for delivering the biologically active compound to a mammal (eg, a human). Such a medium includes all pharmaceutically acceptable carriers, diluents or excipients.

[0069] "Pharmaceutically acceptable carrier, diluent or excipient" includes, but is not limited to, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that has been approved by the U.S. Food and Drug Administration as acceptable for use in humans or domestic animals.

[0070] The compounds of the present disclosure (i.e., compounds of structure (I) and embodiments thereof) or pharmaceutically acceptable salts thereof may contain one or more centers of geometric asymmetry and may therefore give rise to enantiomers, diastereomers and other stereoisomeric forms, which are defined as (R)- or (S)- in terms of absolute stereochemistry, or as (D)- or (L)- for amino acids. The embodiments therefore include all such possible isomers as well as racemic and optically pure forms thereof. Optically active (+) and (-), (R)- and (S)- or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques (e.g., chromatography and fractional crystallization). Conventional techniques for preparing / isolating individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). When the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included.

[0071] Embodiments of the present disclosure include all rotamer modes and conformationally restricted states of the compounds of the present invention. Also included are atropisomers, which are stereoisomers resulting from hindered rotation about a single bond, where energy differences due to steric strain or other contributing factors create sufficiently high barriers to rotation to allow separation of individual conformers. For example, certain compounds of the present disclosure may exist as a mixture of atropisomers, or in a purified or enriched form of one atropisomer.

[0072] "Stereoisomers" refer to compounds made up of the same atoms bonded by the same bonds but having different three-dimensional structures that are not interchangeable. The present disclosure encompasses various stereoisomers and mixtures thereof, and includes "enantiomers," which refer to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0073] "Tautomer" refers to a proton shift from one atom of a molecule to another atom of the same molecule. Embodiments therefore include tautomers of the disclosed compounds.

[0074] Chemical naming schemes and structural formulas used herein are revised forms of the IUPAC nomenclature system, which use ACD / named version 9.07 software programs and / or ChemDraw software naming programs (CambridgeSoft). For complex chemical names used herein, substituents are usually named before the group to which they are attached. For example, cyclopropylethyl includes an ethyl backbone with a cyclopropyl substituent. Except for description below, all bonds are indicated in this chemical structural formula, except for the assumed bonding of enough hydrogen atoms on some carbon atoms to complete all bonds of valence.

[0075] Compound

[0076] In one aspect, the present disclosure provides compounds for use as chemotherapeutic agents. In a specific embodiment, the present disclosure provides compounds having the following structure (I):

[0077]

[0078] or a pharmaceutically acceptable salt, isotopic form, stereoisomer or prodrug thereof, wherein:

[0079] is an optionally substituted cycloalkyl group or an optionally substituted heterocyclic group;

[0080] is optionally substituted aryl;

[0081] L is a direct bond or a heteroatom, heteroalkylene or alkylene linker; and

[0082] R 1a , R 1b , R 1c and R 1d Each is independently H, halogen, C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl.

[0083] In some more specific embodiments, R 1a , R 1b , R 1c and R 1d are each independently H or halogen. In some specific embodiments, R 1c and R 1d Each is H. In some embodiments, R 1a is a halogen, for example, R 1a In some more specific embodiments, R 1b is H. In some embodiments, R 1a and R 1b Each is H.

[0084] In some embodiments, L is a direct bond. In some other embodiments, L is a heteroalkylene linker, for example, a heteroalkylene linker having one of the following structures:

[0085]

[0086] Where R 2 is independently at each occurrence H or C1-C6 alkyl.

[0087] In some more specific embodiments, R 2 is independently at each occurrence H or methyl.

[0088] In some embodiments, the compound of structure (I) has one of the following structures (Ia), (Ib), (Ic), or (Id):

[0089]

[0090] in:

[0091] R 1a is H or halogen; and

[0092] R 2 It is H or methyl.

[0093] In certain embodiments, the compound has one of the following structures (Ia'), (Ia"), (Ib'), (Ib"), (Ic'), (Ic"), (Id'), or (Id"):

[0094]

[0095]

[0096] In certain embodiments of the compound of structure (I), is a cycloalkyl group. For example, in some specific embodiments, is cyclobutyl, cyclopentyl or cyclohexyl.

[0097] In other embodiments, is a heterocyclic group. For example, in some embodiments, It is piperidinyl.

[0098] As disclosed above, is optionally substituted. Thus, in some embodiments, In certain more specific embodiments, is substituted with at least one substituent selected from C1-C6 alkyl, hydroxyl, amino, alkylamino, aminoalkyl, alkylaminoalkyl or a combination thereof. In certain other embodiments, is unsubstituted. In some specific embodiments, Has one of the following structures:

[0099]

[0100] In some specific embodiments, Has one of the following structures:

[0101]

[0102] In certain embodiments, In some embodiments, In some embodiments, is substituted with at least one substituent selected from C1-C6 alkyl, halogen, haloalkyl, haloalkoxy, cyano, alkoxy, hydroxyl, alkylsulfonyl and heterocyclyl. In some more specific embodiments, is substituted with at least one substituent selected from methyl, fluoro, cyano, methoxy, hydroxy, trifluoromethoxy, ethylsulfonyl, morpholinyl and N-methylpiperazinyl. In certain other embodiments, is unsubstituted.

[0103] In some specific embodiments, Has one of the following structures:

[0104]

[0105]

[0106] In some embodiments, the compound of structure (I) is a pharmaceutically acceptable salt (e.g., a base addition salt or an acid addition salt), or exists as a substantially purified enantiomer or diastereomer. Therefore, some embodiments provide a pharmaceutically acceptable salt of a compound according to any of the foregoing embodiments. Other embodiments provide a substantially purified enantiomer or diastereomer according to any of the foregoing embodiments.

[0107] In various embodiments, the compound has one of the structures listed in Table 1 below. Prepared by the methods described in the Examples or by methods known in the art and characterized by mass spectrometry and / or 1 The exemplary compounds in Table 1 were analyzed by H NMR.

[0108] Table 1. Representative compounds of structure (I)

[0109]

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

[0116] It is understood that throughout this specification, combinations of substituents and / or variables in the depicted formulae are permissible only if such combinations result in stable compounds.

[0117] In addition, all compounds of the present disclosure that exist in free base or free acid form can be converted into their pharmaceutically acceptable salts by methods known to those skilled in the art with appropriate inorganic or organic bases or acids. The salts of compounds of the present disclosure can be converted into their free base or acid forms by standard techniques.

[0118] Pharmaceutical composition

[0119] Other embodiments relate to pharmaceutical compositions. The pharmaceutical composition comprises any one (or more) of the aforementioned compounds and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is formulated for oral administration. In other embodiments, the pharmaceutical composition is formulated for injection. In more embodiments, the pharmaceutical composition comprises a compound disclosed herein and an additional therapeutic agent. Non-limiting examples of such therapeutic agents are described herein below.

[0120] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ocular, pulmonary, transmucosal, transdermal, vaginal, aural, nasal, and topical administration. In addition, parenteral delivery includes, by way of example only, intramuscular, subcutaneous, intravenous, intramedullary injection, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic, and intranasal injection.

[0121] In certain embodiments, the compound as described herein is applied in a local manner rather than a systemic manner, for example, usually in a reservoir formulation or a sustained-release formulation via direct injection of the compound into an organ to achieve. In a specific embodiment, the long-acting preparation is applied by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. In addition, in other embodiments, the drug is delivered in a targeted drug delivery system, for example, in a liposome coated with an organ-specific antibody. In such embodiments, the liposome targets an organ and is selectively absorbed by the organ. In other embodiments, the compound as described herein is provided in a quick-release formulation form, in a delayed-release formulation form, or in an intermediate release formulation form. In other embodiments, the compound described herein is applied topically.

[0122] The compounds according to the present disclosure are effective over a wide dosage range. For example, in the treatment of adults, dosages of 0.01 to 1000 mg, 0.5 to 100 mg, 1 to 50 mg per day, and 5 to 40 mg per day are examples of dosages used in some embodiments. An exemplary dosage is 10 to 30 mg per day. The exact dosage will depend on the route of administration, the form of the compound being administered, the individual to be treated, the weight of the individual to be treated, and the preference and experience of the attending physician.

[0123] In some embodiments, the compounds of the present disclosure are administered in a single dose. Typically, such administration will be by injection, such as intravenous injection, so as to quickly introduce the agent. However, other routes are used as appropriate. A single dose of a compound of the present disclosure can also be used to treat acute conditions.

[0124] In some embodiments, the compounds of the present disclosure are administered in multiple doses. In some embodiments, administration is about once a day, twice, three times, four times, five times, six times or more than six times. In other embodiments, administration is about once a month, once every two weeks, once a week or once every other day. In another embodiment, the compounds of the present disclosure are administered together with other medicaments, about once a day to about 6 times a day. In another embodiment, the administration of the compounds of the present disclosure and medicaments continues to be less than about 7 days. In another embodiment, administration continues for more than about 6 days, 10 days, 14 days, 28 days, two months, six months or one year. In some cases, continuous administration is achieved and maintained as long as necessary.

[0125] Administration of the compounds of the present disclosure may be continued as long as necessary. In some embodiments, the compounds of the present disclosure are administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, the compounds of the present disclosure are administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, the compounds of the present disclosure are administered on an ongoing basis, for example, for the long-term effects of treatment.

[0126] In some embodiments, the compounds of the present disclosure are administered in divided doses. It is known in the art that due to the inter-individual variability of compound pharmacokinetics, personalization of dosing regimens is necessary for optimal therapy. The dosage of the compounds of the present disclosure can be found by routine experiments based on the present disclosure.

[0127] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions. In specific embodiments, the pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers comprising excipients and adjuvants that facilitate processing of the active compounds into pharmaceutically usable preparations. Suitable formulations depend on the selected route of administration. Any pharmaceutically acceptable techniques, carriers, and excipients are suitable for preparing the pharmaceutical compositions described herein: Remington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

[0128] Provided herein are pharmaceutical compositions comprising a compound of structure (I) and a pharmaceutically acceptable diluent, excipient or carrier. In certain embodiments, the compound is administered in the form of a pharmaceutical composition in which a compound of structure (I) is mixed with other active ingredients, such as in combination therapy. All combinations of the active agents described in the following combination therapy sections and throughout this disclosure are contemplated herein. In specific embodiments, the pharmaceutical composition comprises one or more compounds of structure (I).

[0129] As used herein, pharmaceutical composition refers to a mixture of a compound of structure (I) and other chemical components such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners and / or excipients. In certain embodiments, the pharmaceutical composition promotes the administration of the compound to an organism. In some embodiments, when practicing the treatment or use methods provided herein, the compound of structure (I) provided herein in a therapeutically effective amount is administered to a mammal suffering from a disease, disorder or medical condition to be treated in the form of a pharmaceutical composition. In a specific embodiment, the mammal is a human. In certain embodiments, the therapeutically effective amount varies depending on the severity of the disease, the age and relative health of the individual, the efficacy of the compound used, and other factors. The compounds described herein are used alone or in combination with one or more therapeutic agents as components of the mixture.

[0130] In one embodiment, one or more compounds of structure (I) are formulated into an aqueous solution. In specific embodiments, the aqueous solution is selected from (by way of example only) a physiologically compatible buffer, such as Hank's solution, Ringer's solution, or a saline buffer. In other embodiments, one or more compounds of structure (I) are formulated for transmucosal administration. In specific embodiments, transmucosal formulations include permeants suitable for permeation barriers. In other embodiments where the compounds described herein are formulated for other parenteral injections, appropriate formulations include aqueous or non-aqueous solutions. In specific embodiments, such solutions contain physiologically compatible buffers and / or excipients.

[0131] In another embodiment, the compounds described herein are formulated for oral administration. The compounds described herein are formulated by combining the active compound with, for example, a pharmaceutically acceptable carrier or excipient. In various embodiments, the compounds described herein are formulated into oral dosage forms, including, by way of example only, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions, and the like.

[0132] In certain embodiments, pharmaceutical preparations for oral use are obtained by mixing one or more solid excipients with one or more compounds described herein, optionally grinding the resulting mixture, and, if desired, processing the mixture of particles after adding suitable adjuvants to obtain tablets or dragee cores. In particular, suitable excipients are fillers such as sugars (including lactose, sucrose, mannitol or sorbitol); cellulose preparations such as, for example, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methylcellulose, microcrystalline cellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose; or other excipients such as polyvinyl pyrrolidone (PVP or povidone) or calcium phosphate. In a specific embodiment, a disintegrant is optionally added. Disintegrants include, by way of example only, cross-linked croscarmellose sodium, polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate.

[0133] In one embodiment, dosage forms such as dragee cores and tablets are provided with one or more applicable coatings. In a specific embodiment, concentrated sugar solution is used for coated dosage forms. Sugar solution optionally contains additional components, such as (for example only) gum arabic, talcum, polyvinyl pyrrolidone, carbopol gel (carbopol gel), polyethylene glycol and / or titanium dioxide, lacquer solution (lacquer solution) and suitable organic solvent or solvent mixture. For identification purposes, dye and / or pigment are also optionally added in coating. In addition, dye and / or pigment are optionally used to characterize the different combinations of active compound dosage.

[0134] In certain embodiments, a therapeutically effective amount of at least one of the compounds described herein is formulated into other oral dosage forms. Oral dosage forms include push-fit capsules made of gelatin, and soft sealed capsules made of gelatin and a plasticizer (e.g., glycerol or sorbitol). In specific embodiments, the push-fit capsules contain the active ingredient mixed with one or more fillers. Fillers include, by way of example only, lactose, binders (such as starches) and / or lubricants (such as talc or magnesium stearate) and optional stabilizers. In other embodiments, soft capsules contain one or more active compounds dissolved or suspended in a suitable liquid. Suitable liquids include, by way of example only, one or more fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers are optionally added.

[0135] In other embodiments, a therapeutically effective amount of at least one compound described herein is formulated for buccal or sublingual administration. Formulations suitable for buccal or sublingual administration include, by way of example only, tablets, lozenges, or gels. In other embodiments, the compounds described herein are formulated for parenteral injection, including formulations suitable for push injection or continuous infusion. In specific embodiments, the formulation for injection is presented in unit dosage form (e.g., ampoule) or in a multidose container. A preservative is optionally added to the injection formulation. In other embodiments, the pharmaceutical composition is formulated into a form suitable for parenteral injection, such as a sterile suspension, solution, or emulsion in an oily or aqueous solvent. Parenteral injection formulations optionally contain a formulation agent, such as a suspending agent, a stabilizer, and / or a dispersant. In specific embodiments, pharmaceutical formulations for parenteral administration include aqueous solutions of active compounds in water-soluble form. In additional embodiments, a suspension of an active compound (e.g., a compound of structure (I)) is prepared into a suitable oily injection suspension. Suitable lipophilic solvents or vehicles suitable for use in the pharmaceutical compositions described herein include, by way of example only, fatty oils (e.g., sesame oil) or synthetic fatty acid esters (e.g., ethyl oleate or triglycerides) or liposomes. In certain specific embodiments, aqueous injection suspensions contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension contains a suitable stabilizer or an agent that increases the solubility of the compound to allow the preparation of a highly concentrated solution. Alternatively, in other embodiments, the active ingredient is in powder form, which is constructed with a suitable solvent such as sterile pyrogen-free water before use.

[0136] In other embodiments, the compound of structure (I) is administered topically. The compounds described herein are formulated into various compositions for topical administration, such as solutions, suspensions, lotions, gels, pastes, medical sticks, balms, creams or ointments. Such pharmaceutical compositions optionally contain solubilizers, stabilizers, tonicity enhancers, buffers and preservatives.

[0137] In other embodiments, the compound of structure (I) is formulated for transdermal administration. In a specific embodiment, the transdermal preparation adopts a transdermal delivery device and a transdermal delivery patch, and can be a lipophilic emulsion or a buffered aqueous solution, dissolved and / or dispersed in a polymer or an adhesive. In various embodiments, such patches are constructed so that the agent is delivered continuously, pulsed or on demand. In additional embodiments, the transdermal delivery of the compound of structure (I) is achieved by means of an ion introduction patch, etc. In certain embodiments, a transdermal patch provides the controlled delivery of the compound of structure (I). In a specific embodiment, the absorption rate is slowed down by using a rate-controlling membrane or by trapping the compound in a polymer matrix or a gel. In an alternative embodiment, an absorption enhancer is used to increase absorption. An absorption enhancer or a carrier includes an absorbable drug-acceptable solvent that helps to pass through the skin. For example, in one embodiment, the transdermal device is in the form of a bandage, which includes a pad component, a drug storage layer containing a compound and an optional carrier, an optional rate-controlling barrier that delivers the compound to the skin of the host at a controlled and predetermined rate over an extended period of time, and a member for fixing the device to the skin.

[0138] In other embodiments, the compound of structure (I) is formulated for administration by inhalation. Various forms suitable for administration by inhalation include, but are not limited to, aerosols, sprays, or powders. Pharmaceutical compositions of any compound of structure (I) are conveniently delivered in the form of an aerosol spray presented from a pressurized bag or nebulizer using a suitable propellant (e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas). In a specific embodiment, the dosage unit of the pressurized aerosol is determined by providing a valve for delivering a quantitative amount. In certain embodiments, capsules and cartridges such as (for example only) gelatin for use in an inhaler or insufflator are formulated as a powder mixture containing the compound and a suitable powder base (e.g., lactose or starch).

[0139] In other embodiments, the compounds of structure (I) are formulated into rectal compositions such as enemas, rectal gels, rectal foams, rectal aerosols, suppositories, jelly suppositories or retention enemas containing conventional suppository bases such as cocoa butter or other glycerides and synthetic polymers such as polyvinylpyrrolidone, PEG, etc. In suppository forms, a low melting wax such as, but not limited to, a mixture of fatty acid glycerides, optionally in combination with cocoa butter, is first melted.

[0140] In certain embodiments, pharmaceutical compositions are prepared using one or more physiologically acceptable carriers in any conventional manner, the carrier comprising excipients and adjuvants that promote the processing of the active compound into pharmaceutically available preparations. Suitable preparations depend on the selected route of administration. When suitable, any pharmaceutically acceptable technology, carrier and excipient are optionally used. Pharmaceutical compositions comprising compounds of structure (I) are prepared in a conventional manner, such as (for example only) by means of conventional mixing, dissolving, granulating, making dragees, water-flying, emulsifying, encapsulating, encapsulating or compressing methods.

[0141] The pharmaceutical composition comprises at least one pharmaceutically acceptable carrier, diluent or excipient and at least one compound of structure (I) described herein as an active ingredient. The active ingredient is in the form of a free acid or free base, or in the form of a pharmaceutically acceptable salt. In addition, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also referred to as polymorphs) and active metabolites of these compounds having the same type of activity. All tautomers of the compounds described herein are included within the scope of the compounds presented herein. In addition, the compounds described herein encompass non-solvated forms and solvated forms with pharmaceutically acceptable solvents such as water, ethanol, etc. The solvated forms of the compounds presented herein should also be considered as disclosed herein. In addition, the pharmaceutical composition optionally comprises other medical or pharmaceutical agents, carriers, adjuvants, such as preservatives, stabilizers, wetting agents or emulsifiers, solubilizers, salts for regulating osmotic pressure, buffers and / or other therapeutically valuable substances.

[0142] Methods for preparing compositions comprising compounds described herein include formulating the compounds with one or more inert pharmaceutically acceptable excipients or carriers to form a solid, semisolid or liquid. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets and suppositories. Liquid compositions include solutions in which the compounds are dissolved, emulsions containing the compounds, or solutions containing liposomes, micelles or nanoparticles containing compounds as disclosed herein. Semisolid compositions include, but are not limited to, gels, suspensions and creams. The pharmaceutical compositions described herein may be in the form of liquid solutions or suspensions, solid forms suitable for forming solutions or suspensions in liquids prior to use, or emulsions. These compositions may also optionally contain small amounts of non-toxic auxiliary substances, such as wetting agents or emulsifiers, pH buffers, and the like.

[0143] In some embodiments, the pharmaceutical composition comprising at least one compound of structure (I) is illustratively in liquid form, wherein the medicament is present in solution, suspension or both. Typically, when the composition is administered in the form of a solution or suspension, the medicament of the first portion is present in solution and the medicament of the second portion is present in the form of particles suspended in a liquid matrix. In some embodiments, the liquid composition includes a gel formulation. In other embodiments, the liquid composition is aqueous.

[0144] In certain embodiments, available aqueous suspensions contain one or more polymers as suspending agents. Available polymers include water-soluble polymers and water-insoluble polymers, such as cellulose polymers, such as hydroxypropyl methylcellulose, and water-insoluble polymers such as cross-linked carboxyl-containing polymers. Certain pharmaceutical compositions described herein include mucoadhesive polymers, such as selected from carboxymethylcellulose, carbomer (acrylic acid polymer), poly (methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate and dextran.

[0145] Available pharmaceutical compositions also optionally include solubilizing agents to assist in the dissolution of the compound of structure (I). The term "solubilizing agent" generally includes agents that result in the formation of micellar solutions or true solutions of the medicament. Certain acceptable nonionic surfactants (e.g., polysorbate 80) can be used as solubilizing agents, and ophthalmologically acceptable ethylene glycol, polyethylene glycol (e.g., polyethylene glycol 400) and glycol ethers can also be used as solubilizing agents.

[0146] In addition, the pharmaceutical compositions that can be used optionally contain one or more pH adjusters or buffers, including acids such as acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and trishydroxymethylaminomethane; and buffers such as citrate / dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases, and buffers are included in amounts required to maintain the pH of the composition within an acceptable range.

[0147] In addition, useful compositions optionally also contain one or more salts in an amount required to bring the osmotic pressure of the composition into an acceptable range. Such salts include salts having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions; suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite and ammonium sulfate.

[0148] Other useful pharmaceutical compositions optionally contain one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as phenylmercuric nitrate and thimerosal; stabilized chlorine dioxide; and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide and cetylpyridinium chloride.

[0149] Other useful compositions include one or more surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, such as polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkyl ethers and alkylphenyl ethers, such as octoxynol 10, octoxynol 40.

[0150] Other useful compositions may contain one or more antioxidants to enhance chemical stability as desired. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.

[0151] In certain embodiments, aqueous suspension compositions are packaged in single-dose non-reclosable containers. Alternatively, multiple-dose reclosable containers are used, in which case a preservative is generally included in the composition.

[0152] In alternative embodiments, other delivery systems for hydrophobic pharmaceutical compounds are used. Liposomes and emulsions are examples of delivery vehicles or carriers available herein. In certain embodiments, organic solvents such as N-methylpyrrolidone are also used. In additional embodiments, sustained release systems are used to deliver compounds as described herein, such as semipermeable matrices of solid hydrophobic polymers containing therapeutic agents. Various sustained release materials are available herein. In some embodiments, sustained release capsules release compounds for weeks to more than 100 days. Depending on the chemical properties and biological stability of the therapeutic agent, other strategies for protein stabilization are adopted.

[0153] In certain embodiments, the formulations described herein include one or more antioxidants, metal chelators, thiol-containing compounds, and / or other general stabilizers. Examples of such stabilizers include, but are not limited to: (a) about 0.5% to about 2% w / v glycerol; (b) about 0.1% to about 1% w / v methionine; (c) about 0.1% to about 2% w / v monothioglycerol; (d) about 1 mM to about 10 mM EDTA; (e) about 0.01% to about 2% w / v ascorbic acid; (f) 0.003% to about 0.02% w / v polysorbate 80; (g) 0.001% to about 0.05% w / v polysorbate 20; (h) arginine; (i) heparin; (j) dextran sulfate; (k) cyclodextrin; (l) pentosan polysulfate and other heparinoids; (m) divalent cations, such as magnesium and zinc; or (n) combinations thereof.

[0154] In some embodiments, the concentration of the compound of structure (I) provided in the pharmaceutical composition of the present disclosure is less than 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 0.10%, 0.11%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.30%, 0.31%, 0.32%, 0.33%, 0.34%, 0.36%, 0.37%, 0.38%, 0.39%, 0.40%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.50%, 0.51%, 0.52%, 0.53%, 0.54%, 0.55%, 0.56%, 0.57%, 0.58%, 0.59%, 0.60%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.70%, 0.71%, 0.72 %. %, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002% or 0.0001% w / w, w / v or v / v.

[0155] In some embodiments, the concentration of the compound of structure (I) provided in the pharmaceutical composition of the present disclosure is greater than 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 19.75%, 19.50%, 19.25%, 19%, 18.75%, 18.50%, 18.25%, 18%, 17.75%, 17.50%, 17.25%, 17%, 16.75%, 16.50%, 16.25%, 16%, 15.75%, 15.50%, , 15.25%, 15%, 14.75%, 14.50%, 14.25%, 14%, 13.75%, 13.50%, 13.25%, 13%, 12.75%, 12.50%, 12.25%, 12%, 11.75%, 11.50%, 11.25%, 11%, 10.75%, 10.50%, 10.25%, 10%, 9.75%, 9.50%, 9.25%, 9%, 8.75%, 8.50%, 8.25%, 8%, 7.75 %, 7.50%, 7.25%, 7%, 6.75%, 6.50%, 6.25%, 6%, 5.75%, 5.50%, 5.25%, 5%, 4.75%, 4.50%, 4.25%, 4%, 3.75%, 3.50%, 3.25%, 3%, 2.75%, 2.50%, 2.25%, 2%, 1.75%, 1.50%, 125%, 1%, 0.5%, 0.4%, 0.3%, 0.2%, 0.1%, 0.09%, 0.08%, 0.07% , 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002% or 0.0001% w / w, w / v or v / v.

[0156] In some embodiments, the concentration of the compound of structure (I) provided in the pharmaceutical composition of the present disclosure is about 0.0001% to about 50%, about 0.001% to about 40%, about 0.01% to about 30%, about 0.02% to about 29%, about 0.03% to about 28%, about 0.04% to about 27%, about 0.05% to about 26%, about 0.06% to about 25%, about 0.07% to about 24%, about 0.08% to about 26%, about 0.09% to about 27%, about 0.10% to about 28%, about 0.11% to about 29%, about 0.12% to about 29%, about 0.13% to about 29%, about 0.14% to about 29%, about 0.15% to about 29%, about 0.16% to about 29%, about 0.17% to about 29%, about 0.18% to about 29%, about 0.19% to about 29%, about 0.20% to about 29%, about 0.21% to about 29%, about 0.22% to about 29%, about 0.23% to about 29%, about 0.24% to about 29%, about 0.25% to about 29%, about 0.26% to about 29%, about 0.27% to about 29%, about 0. About 0.08% to about 23%, about 0.09% to about 22%, about 0.1% to about 21%, about 0.2% to about 20%, about 0.3% to about 19%, about 0.4% to about 18%, about 0.5% to about 17%, about 0.6% to about 16%, about 0.7% to about 15%, about 0.8% to about 14%, about 0.9% to about 12%, about 1% to about 10% w / w, w / v or v / v.

[0157] In some embodiments, the concentration of the compound of structure (I) provided in the pharmaceutical compositions of the present disclosure is about 0.001% to about 10%, about 0.01% to about 5%, about 0.02% to about 4.5%, about 0.03% to about 4%, about 0.04% to about 3.5%, about 0.05% to about 3%, about 0.06% to about 2.5%, about 0.07% to about 2%, about 0.08% to about 1.5%, about 0.09% to about 1%, about 0.1% to about 0.9% w / w, w / v, or v / v.

[0158] In some embodiments, the amount of the compound of structure (I) provided in the pharmaceutical composition of the present disclosure is equal to or less than 10g, 9.5g, 9.0g, 8.5g, 8.0g, 7.5g, 7.0g, 6.5g, 6.0g, 5.5g, 5.0g, 4.5g, 4.0g, 3.5g, 3.0g, 2.5g, 2.0g, 1.5g, 1.0g, 0.95g, 0.9g, 0.85g, 0.8g, 0.75g, 0.7g, 0.65g, 0.6g, 0.55g, 0.5g, 0.45g, 0.4g, 0.35g, 0.3g, 0.25g, 0.2g, 0.15g, 0.1g, 0.09g, 0.08g, 0.07g, 0.06g, 0.05g, 0.04g, 0.03g, 0.02g, 0.01g, 0.009g, 0.008g, 0.007g, 0.006g, 0.005g, 0.004g, 0.003g, 0.002g, 0.001g, 0.0009g, 0.0008g, 0.0007g, 0.0006g, 0.0005g, 0.0004g, 0.0003g, 0.0002g or 0.0001g.

[0159] In some embodiments, the amount of the compound of structure (I) provided in the pharmaceutical composition of the present disclosure is greater than 0.0001 g, 0.0002 g, 0.0003 g, 0.0004 g, 0.0005 g, 0.0006 g, 0.0007 g, 0.0008 g, 0.0009 g, 0.001 g, 0.0015 g, 0.002 g, 0.0 025g, 0.003g, 0.0035g, 0.004g, 0.0045g, 0.005g, 0.0055g, 0.006g, 0.0065g, 0. 007g, 0.0075g, 0.008g, 0.0085g, 0.009g, 0.0095g, 0.01g, 0.015g, 0.02g, 0.025g ,0.03g,0.035g,0.04g,0.045g,0.05g,0.055g,0.06g,0.065g,0.07g,0.075g,0 .08g, 0.085g, 0.09g, 0.095g, 0.1g,, 0.15g, 0.2g,, 0.25g, 0.3g,, 0.35g, 0.4g,, 0 .45g, 0.5g, 0.55g, 0.6g,, 0.65g, 0.7g, 0.75g, 0.8g, 0.85g, 0.9g, 0.95g, 1g, 1.5 g, 2g, 2.5, 3g, 3.5, 4g, 4.5g, 5g, 5.5g, 6g, 6.5g, 7g, 7.5g, 8g, 8.5g, 9g, 9.5g or 10g.

[0160] In some embodiments, the amount of the compound of structure (I) provided in the pharmaceutical composition of the present disclosure is 0.0001-10 g, 0.0005-9 g, 0.001-8 g, 0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g.

[0161] Kits / Products

[0162] Also provided are kits and articles for the therapeutic applications described herein. In some embodiments, such kits comprise carriers, packaging, or containers spaced apart to accommodate one or more containers such as vials, tubes, etc., each of which contains one of the individual elements used in the methods described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The containers are formed of a variety of materials such as glass or plastic.

[0163] The articles provided herein contain packaging materials. Packaging materials for packaging pharmaceutical products include, for example, packaging materials found in U.S. Patent No. 5,323,907, U.S. Patent No. 5,052,558, and U.S. Patent No. 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packages, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging materials suitable for selected formulations and intended modes of administration and treatment. For example, the container contains one or more compounds described herein, optionally in a composition or in combination with other medicaments disclosed herein. The container optionally has a sterile access port (e.g., the container is an intravenous solution bag or a vial with a stopper that can be pierced by a hypodermic needle). Such kits optionally include a compound and an identification description or label or instructions related to its use in the methods described herein.

[0164] For example, the kit typically includes one or more additional containers, each of which has one or more of the various materials (e.g., reagents, optionally in concentrated form, and / or devices) required to use the compounds described herein from a commercial and user perspective. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes; carriers, packaging, containers, vials and / or tubes, labels listing the contents and / or instructions for use, and package inserts with instructions for use. A set of instructions is also typically included. The label is optionally located on or associated with the container. For example, when the letters, numbers or other characters forming the label are attached, stenciled or etched into the container itself, the label is located on the container; when the label is present in a receiver or carrier that also supports the container, the label is associated with the container, for example as a package insert. In addition, the label is used to indicate the contents to be used for a specific therapeutic application. In addition, the label indicates the contents such as instructions for use in the methods described herein. In certain embodiments, the pharmaceutical composition is present in a package or dispenser device containing one or more unit dosage forms containing the compounds provided herein. The package, for example, contains metal foil or plastic foil, such as a blister pack. Alternatively, the package or dispenser device is accompanied by instructions for administration. Alternatively, the package or dispenser is accompanied by a notice associated with the container in a form specified by a governmental agency that regulates the manufacture, use, or sale of pharmaceuticals, which reflects the agency's approval of the drug form for human or veterinary administration. Such notices are, for example, labels approved by the U.S. Food and Drug Administration for prescription drugs, or approved product inserts. In some embodiments, a composition containing a compound provided herein formulated in a compatible pharmaceutical carrier is prepared and placed in an appropriate container and labeled for treatment of a specified condition.

[0165] Treatment and medication methods

[0166] Embodiments of the present disclosure provide methods for treating cancer. One embodiment provides a method for treating cancer, the method comprising administering to an individual in need thereof an effective amount of a pharmaceutical composition as disclosed in any embodiment herein.

[0167] In some embodiments, the cancer is a blood cancer. For example, in some embodiments, the cancer is a leukemia (e.g., acute myeloid leukemia). In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is breast cancer (e.g., triple negative breast cancer). In some embodiments, the cancer is lung cancer (e.g., non-small cell lung cancer). In some specific embodiments, the cancer is brain cancer (e.g., glioblastoma).

[0168] Other therapeutic agents that can be combined with the compounds of the present disclosure are found in "The Pharmacological Basis of Therapeutics" by Goodman and Gilman, Tenth Edition, Eds. Hardman, Limbird and Gilman, or the Physician's Desk Reference, both of which are incorporated herein by reference in their entirety.

[0169] The compounds described herein may be used in combination with the agents disclosed herein or other suitable agents depending on the condition being treated. Therefore, in some embodiments, one or more compounds of the present disclosure will be co-administered with other agents as described above. When used for combined therapy, the compounds described herein are administered simultaneously or separately with the second agent. This combined administration may include administering the two agents simultaneously in the same dosage form, administering them simultaneously in a separate dosage form, and administering them separately. That is, the compounds described herein and any of the agents described above may be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds of the present disclosure and any of the agents described above may be administered simultaneously, wherein the two agents are present in separate preparations. In other alternatives, the compounds of the present disclosure may be administered after any of the agents described above, or vice versa. In some embodiments of the separate administration scheme, the compounds of the present disclosure and any of the agents described above are administered a few minutes apart, or a few hours apart, or a few days apart.

[0170] Compounds disclosed herein include 1H-pyrazole-pyrimidine compounds as inhibitors of MELK (maternal embryonic leucine zipper kinase), its mutant MELK (T460M), FLT3 (FMS-like tyrosine kinase, cluster of differentiation antigen 135 (CD135), fetal liver kinase 2 (Flk2)) and its mutants (FLT3 mutant forms: FLT3 (D835Y), FLT3 (F594R595insR), FLT3 (F594R595insREY), FLT3 (ITD), FLT3 (ITD)-NPOS, FLT3 (ITD)-W51 and FLT3 (R595_E596insEY) kinase). Pharmaceutical compositions containing compounds of structure (I) and methods of using the compounds or compositions to treat various types of diseases or conditions mediated by MELK, FLT3 (e.g., disease states associated with abnormal cell growth, such as cancer). Furthermore, these compounds are important therapeutic agents for the treatment of AML, breast cancer, TNBC, GBM, lung cancer, and prostate cancer.

[0171] Preparation method

[0172] Compounds of structure (I) can be prepared according to methods known in the art and according to methods disclosed herein. In general, starting components can be obtained from sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, or synthesized according to sources known to those skilled in the art (see, e.g., Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th Edition (Wiley, December 2000)) or prepared as described herein.

[0173] General Reaction Scheme 1 ("Method A")

[0174]

[0175] Embodiments of compounds of structure (I) can be prepared according to General Reaction Scheme 1 ("Method A"), wherein R 1a , R 1b , R 1c , R 1d , L, As defined herein with respect to structure (I). Compounds of structure A1 are purchased or prepared according to methods known in the art. Conversion of hydrazine hydrochloride A1 to desired product A2 can be carried out using appropriately selected reagents and solvents (e.g., 1,1,3,3-tetramethoxypropane in ethanol) under appropriate reaction conditions (e.g., at 90° C. for 16 hours). Product A2 can then be converted to add reactive moiety X. The overall reaction scheme can be considered, based on compatibility with other synthetic steps (e.g., with Y 1 and / or Y 2 The reactive moiety X is selected (and alternatively, modified if necessary) based on the complementary reactivity of the moiety. X may include, but is not limited to, a halide, a pseudohalide (e.g., a triflate), a boronic acid, or a boric acid ester. The reaction conditions for adding the reactive moiety X are well known in the art and may be based on the desired X to be added (e.g., N-iodosuccinimide in acetic acid; or pinacol borane, triethylamine, S-phos, and bis(acetonitrile)dichloropalladium(II); or bis(pinacol)diboron, a base, and [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II)). Next, under suitable conditions (e.g., Suzuki coupling conditions, such as a base and [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) in dioxane / H2O solution with heating), A3 is coupled to A4 (which may also be purchased or prepared according to methods known in the art, having a reactive group Y 1 and Y2 (e.g., halides such as chlorine)) to obtain the desired product A5. After the coupling step is completed, A5 can be optionally converted as needed to remove Y 2 (eg, using palladium on carbon, a base, methanol and H2), and the protecting groups can be removed as necessary to give the desired product, ie, a compound of structure (I).

[0176] General Reaction Scheme 2 ("Method B")

[0177]

[0178] Embodiments of compounds of structure (I) can be prepared according to General Reaction Scheme 2 ("Method B"), wherein R 1a , R 1b , R 1c , R 1d , L, As defined herein for structure (I). Compounds of structures B1 and B2 are purchased or prepared according to methods known in the art. 1 , X 2 , Y 1 and Y 2 is a reactive group, for example, a reactive group with complementary reactivity to form a covalent bond (e.g., a halide and a boronate to form a carbon-carbon bond or an amine and a halide to form a secondary amine via a Suzuki cross-coupling reaction). Using appropriate conditions (e.g., wherein Y 1 is chlorine and X 1 is a boronate ester, and using a base and tetrakis(triphenylphosphine)palladium) to react compound B1 with compound B2 to give compound B3. Then under suitable conditions (e.g., wherein X 2 is an amine and Y 2 is chlorine, and B3 is reacted with B4 (including protected forms thereof) using Cs2CO3 and chlorine (2-dicyclohexyl-phosphino-2',4',6'-triisopropyl-1,1'-biphenyl) [2-(2'-amino-1,1'-biphenyl)]-palladium (II)) to give compounds of structure (I).

[0179] Alternatively, B3 can be connected with or its protected form (which is similar to but having a suitable reaction group Y 2 The coupled reactive moiety (e.g., when Y 2 is a halogen and When containing secondary amines)) reaction. B3 and Reaction under appropriate conditions (e.g., using Cs2CO3 and chloro(2-dicyclohexyl-phosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]-palladium(II)) produces compounds of structure (I) (e.g., wherein L is a direct bond).

[0180] General Reaction Scheme 3 ("Method C")

[0181]

[0182] Embodiments of compounds of structure (I) can be prepared according to General Reaction Scheme 3 ("Method C"), wherein R 1a , R 1b , R 1c , R 1d , L, As defined herein for structure (I). Compounds of structures C1 and C2 are purchased or prepared according to methods known in the art. 1 , X 2 , Y 1 , Y 2 and Y 3 X is a reactive group, for example, a reactive group having complementary reactivity to form a covalent bond (e.g., -OH, halogen / boronate). Compound C1 is reacted with Compound C2 using appropriate conditions (e.g., DIAD, triphenylphosphine) to obtain Compound C3. Then, under appropriate conditions (e.g., wherein X 1 is pinacol borane and Y 1 is chlorine, and C3 is reacted with C4 (including protected forms thereof) using Na2CO3 and PdCl2(dppf). The product of the reaction between C3 and C4 can be deprotected (e.g., using hydrochloric acid in ether) to give a compound of structure (I). Alternatively, prior to the deprotection step (e.g., using hydrochloric acid in ether), an additional reaction can be performed to modify Y 3 (For example, conversion from chlorine to hydrogen using palladium on carbon and H2 and a base).

[0183] General Reaction Scheme 4 ("Method D")

[0184]

[0185] Embodiments of compounds of structure (I) can be prepared according to General Reaction Scheme 4 ("Method D"), wherein R 1a , R 1b , R 1c , R 1d , L, As defined herein for structure (I). Compounds of structures D1 and D2 are purchased or prepared according to methods known in the art. 1 , X 2 , Y 1 and Y 2 is a reactive group, for example, a reactive group with complementary reactivity to form a covalent bond (e.g., a halogen to react with the secondary amine of D1). Compound D1 is reacted with compound D2 using appropriate conditions (e.g., CuI, K2CO3) to give compound D3. D3 is then derivatized to add reactive groups (e.g., using Br2 and acetic acid), which can be further modified as desired (e.g., using bis(pinacolato)diboron, PdCl2(dppf) and potassium acetate). Then under appropriate conditions (e.g., wherein X 2 is pinacol borane and Y 1 is chlorine, and using Na2CO3 and PdCl2(dppf), D4 is reacted with D5 (including protected forms thereof). The reaction product between D4 and D5 can be deprotected (e.g., using hydrochloric acid in diethyl ether) to obtain a compound of structure (I). Alternatively, prior to the deprotection step (e.g., using hydrochloric acid in diethyl ether), an additional reaction can be performed to modify Y 2 (For example, conversion from chlorine to hydrogen using palladium on carbon and H2 and a base).

[0186] General Reaction Scheme 5 ("Method E")

[0187]

[0188] Embodiments of compounds of structure (I) can be prepared according to General Reaction Scheme 5 ("Method E"), wherein R 1a , R 1b , R 1c , R 1d , L, As defined herein for structure (I). Compounds of structures E1 and E2 are purchased or prepared according to methods known in the art. 1 , X 2 , Y 1 and Y 2 is a reactive group, for example, a reactive group having complementary reactivity to form a covalent bond (e.g., X 1 and Y 1 can be borate ester and halogen, respectively, and X 2 (may be halogen to react with the secondary amine of E3). Compound E1 is reacted with compound E2 using appropriate conditions (e.g., bis(pinacolato)diboron, PdCl2(dppf) and sodium carbonate) to give compound E3. Then, under appropriate conditions (e.g., wherein X 2is boronic acid, and using copper acetate and base), E3 and E4 (including protected forms thereof) are reacted. The reaction product between E3 and E4 can be deprotected (for example, using hydrochloric acid in diethyl ether) to obtain a compound of structure (I). Alternatively, before performing the deprotection step (for example, using hydrochloric acid in diethyl ether), an additional reaction can be performed to modify Y 2 (For example, conversion from chlorine to hydrogen using palladium on carbon and H2 and a base).

[0189] The above reaction scheme can be modified as needed to prepare other compounds of structure (I) by adding, removing or modifying substituents. For example, substituents or reactive moieties can be added, removed or modified to achieve the desired or its protected form.

[0190] It should be noted that various alternative strategies for preparing compounds of structure (I) are available to those of ordinary skill in the art. For example, other compounds of structure (I) can be prepared using appropriate starting materials according to similar methods. It will also be appreciated by those skilled in the art that in the method for preparing the compounds described herein, the functional groups of the intermediate compounds may need to be protected by suitable protecting groups. Such functional groups include, but are not limited to, hydroxyl, amino, sulfhydryl and carboxylic acid. Suitable protecting groups for hydroxyl include trialkylsilyl or diarylalkylsilyl (e.g., tert-butyldimethylsilyl, tert-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, etc. Suitable protecting groups for amino include tert-butyloxycarbonyl ("BOC"), benzyloxycarbonyl, etc. Suitable protecting groups for sulfhydryl include -C(O)-R" (wherein R" is alkyl, aryl or arylalkyl), p-methoxybenzyl, trityl, etc. Suitable protecting groups for carboxylic acids include alkyl esters, aryl esters or arylalkyl esters. Protecting groups are optionally added or removed according to standard techniques known to those skilled in the art and standard techniques described herein. The use of protecting groups is described in detail in Green, TW and PGM Wutz, Protective Groups in Organic Synthesis (1999), 3rd edition, Wiley. As will be appreciated by those skilled in the art, the protecting group may also be a polymer resin, such as Wang resin, Rink resin or 2-chlorotrityl-chloride resin.

[0191] It will also be appreciated by those skilled in the art that, although such protected derivatives of compounds of the present disclosure may not themselves have pharmacological activity, they may be administered to mammals and thereafter metabolized in vivo to form compounds of the present disclosure that have pharmacological activity. Such derivatives may therefore be described as "prodrugs". Prodrugs of compounds of the present disclosure are all included within the scope of embodiments of the present invention.

[0192] The examples and preparations provided below further illustrate and exemplify the compounds of the present disclosure and methods for preparing such compounds. It should be understood that the scope of the present disclosure is not limited in any way to the scope of the following examples and preparations. In the following examples, as well as throughout the specification and claims, unless otherwise stated, molecules with a single stereocenter exist as a racemic mixture. Unless otherwise stated, those molecules with two or more stereocenters exist as a racemic mixture of diastereomers. Single enantiomers / diastereomers can be obtained by methods known to those skilled in the art.

[0193] Example

[0194] The following examples are provided for illustrative purposes.Methods for preparing compounds of structure (I) are known in the art or can be obtained by one of ordinary skill in the art.

[0195] The in vivo efficacy of compounds of exemplary structure (I) in established human AML, breast, GBM xenograft growth inhibition experiments was performed together with multi-kinase, targeted kinase reference agents and standard chemotherapeutic agents / drugs. Effective and selective MELK, FLT3 inhibitors were orally administered in an in vivo dose escalation experiment starting at 10, 20, 40 and 60 mg / Kg after subcutaneous injection of AML, breast and GBM cancer cells into mice 4 to 6 weeks.

[0196] Example 1

[0197] General reaction: Exemplary preparation of A2 (Method A)

[0198]

[0199] Method 1A - A solution of A1 (1 eq) and 1,1,3,3-tetramethoxypropane (1.1 eq) in ethanol (10 volumes) was stirred at 90°C for 16 hours. The reaction mass was cooled to ambient temperature and concentrated in vacuo to remove ethanol (a small amount of product distillation was observed while distilling the solvent). The residue thus obtained was diluted with water and extracted with dichloromethane (2x). The combined organic layers were washed with saturated brine solution, dried over anhydrous sodium sulfate, filtered and concentrated to give the crude desired product A2.

[0200] Method 1B - The crude product obtained above was purified by silica gel column chromatography to give Compound A2.

[0201] Example 2

[0202] General reaction: Exemplary preparation of A2' (Method A)

[0203]

[0204] Method 2A - N-iodosuccinimide (1.4 eq) was added to a stirred solution of A2 (1 eq) in acetic acid (10 volumes) at ambient temperature. The reaction mixture was stirred at 130°C for 0.5 hours. The reaction mass was cooled to ambient temperature, poured into ice water and extracted with ethyl acetate (2x). The combined organic layers were washed with water, saturated brine solution, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography to give compound A2'.

[0205] Method 2B - N-iodosuccinimide (1.4 eq) was added to a stirred solution of A2 (1 eq) in acetic acid (10 volumes) at ambient temperature. The reaction mixture was stirred at 120°C for 0.5 hours. The reaction mass was cooled to ambient temperature, poured into ice water and extracted with dichloromethane (2x). The combined organic layers were washed with water, saturated brine solution, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give compound A2'.

[0206] Method 2C - N-iodosuccinimide (1 eq) was added to a stirred solution of A2 (1 eq) in acetic acid (10 volumes) at ambient temperature. The reaction mixture was stirred at ambient temperature for 16 hours. The reaction mass was poured into ice water and extracted with dichloromethane (2x). The combined organic layers were washed with water, saturated brine solution, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give compound A2'.

[0207] Method 2D - N-iodosuccinimide (1.4 eq) was added to a stirred solution of A2 (1 eq) in acetic acid (10 vol) at ambient temperature. The reaction mixture was stirred at 130°C for 0.5 h. The reaction mass was cooled to ambient temperature, poured into ice water and a solid precipitated. The obtained solid was filtered, washed with water and dried under vacuum to give compound A2'.

[0208] Example 3

[0209] General reaction: Exemplary preparation of A3 (Method A)

[0210]

[0211] Method 3A - Under nitrogen atmosphere, pinacol borane (1.2 eq.), triethylamine (2.5 eq.), S-Phos (0.09 eq.) and bis(acetonitrile) dichloropalladium (II) (0.03 eq.) were added to a stirred solution of A2' (1 eq.) in toluene (10 volumes) at ambient temperature. The reaction mixture was stirred at 90 °C for 16 hours. The reaction mass was cooled to ambient temperature, quenched with saturated NH4Cl solution and extracted with dichloromethane (2×). The combined organic layers were washed with saturated brine solution, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography to give compound A3.

[0212] Method 3B - Bis(pinacolato)diboron (2 eq.), potassium acetate (3 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) or PdCl2(dppf) (1:1) (0.1 eq.) were added to a stirred solution of A2' (1 eq.) in DMSO (10 vols) at ambient temperature under nitrogen atmosphere. The reaction mixture was stirred at 80°C for 16 hours. The reaction mass was cooled to ambient temperature, diluted with water and extracted with dichloromethane (2x). The combined organic layers were washed with saturated brine solution, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography to afford compound A3.

[0213] Example 4

[0214] General reaction: Exemplary preparation of A5 (Method A)

[0215]

[0216] Method 4A - A3 (1.1 eq.), Na2CO3 (2 eq.) and [1,1'-bis(diphenylphosphino)ferrocene]-dichloropalladium(II) (1:1) (0.1 eq.) were added to a stirred solution of A4 (1 eq.) in dioxane (9 vol.) and water (1 vol.) at ambient temperature under nitrogen atmosphere. The reaction mixture was stirred at 90°C for 8 hours. The reaction mass was cooled to ambient temperature and concentrated in vacuo. The crude product was purified by silica gel column chromatography to afford compound A5.

[0217] Example 5

[0218] General reaction: Exemplary preparation of A5' (Method A)

[0219]

[0220] Method 5A - Palladium on carbon (10%) (10 wt%) and NaHCO (2 eq) were added to a stirred solution of A5 (1 eq) in methanol (10 vol) at ambient temperature under nitrogen atmosphere. The reaction mixture was stirred under hydrogen atmosphere (balloon) at ambient temperature for 16 hours. The reaction mass was filtered through a celite bed, washed with methanol and the filtrate was concentrated in vacuo. The crude product was purified by silica gel column chromatography to give compound A5'.

[0221] Example 6

[0222] General reaction: Exemplary preparation of compounds of structure (I) (Method A)

[0223]

[0224] Method 6A (Deprotection) - 2M hydrochloric acid in diethyl ether was added to a stirred solution of A5' in dichloromethane at 0°C. The reaction mixture was stirred at ambient temperature for 16 hours. The reaction mass was concentrated in vacuo and the residue was lyophilized to give a compound of structure (I).

[0225] Method 6B (Deprotection) - 2M hydrochloric acid in diethyl ether was added to a stirred solution of A5' in dichloromethane at 0°C. The reaction mixture was stirred at ambient temperature for 1 hour. The reaction mass was filtered, the solid was washed with diethyl ether and dried under vacuum to give a compound of structure (I).

[0226] Example 7

[0227] General reaction: Exemplary preparation of B3 (Method B)

[0228] Method 7A - Under nitrogen atmosphere, B1 (1.1 eq.), K2CO3 (2 eq.) and tetrakis(triphenylphosphine)palladium (0.05 eq.) were added to a stirred solution of B2 (1 eq.) in dimethoxyethane (DME; 8 vol.) and water (2 vol.) at ambient temperature. The reaction mixture was stirred at 90°C for 16 hours. The reaction mass was cooled to ambient temperature and concentrated in vacuo. The crude product was purified by silica gel column chromatography to afford compound B3.

[0229] Example 8

[0230] General reaction: Exemplary preparation of (protected) compounds of structure (I) (Method B)

[0231] Method 8A - Under nitrogen atmosphere, to a stirred solution of B3 (1 eq.) in toluene (15 vols) were added B4 (1.5 eq.), Cs2CO3 (3 eq.) and chloro(2-dicyclohexyl-phosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]-palladium(II) (0.1 eq.) at ambient temperature. The reaction mixture was stirred at 130°C for 40 hours. The reaction mass was cooled to ambient temperature and concentrated in vacuo. The crude product was purified by silica gel column chromatography to afford the compound of formula (I) in protected form.

[0232] Example 9

[0233] General reaction: Exemplary deprotection of compounds of structure (I)

[0234] Method 9A (deprotection) - A 2M solution of hydrochloric acid in ether was added to a stirred solution of the protected intermediate in dichloromethane at 0° C. The reaction mixture was stirred at ambient temperature for 3 hours. The reaction mass was concentrated in vacuo and the residue was purified by preparative HPLC to give a compound of structure (I).

[0235] Method 9B (Deprotection) - A 2M solution of hydrochloric acid in diethyl ether was added to a stirred solution of the protected intermediate in dichloromethane at 0° C. The reaction mixture was stirred at ambient temperature for 3 hours. The reaction mass was concentrated in vacuo and the residue was lyophilized to give a compound of structure (I).

[0236] Example 10

[0237] General procedure for Boc protection of amino acids (Method 10A)

[0238] Method 10A: To an ice-cold solution of an acetone / water mixture of R-amino acid and KCO was added di-tert-butyl dicarbonate (boc-anhydride), and the reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete (TLC), acetone was removed in vacuo at a temperature not exceeding 50°C, and the water layer was washed with hexane to remove any excess boc-anhydride. The water layer was then acidified with 1.5N HCl (pH-5), and the solid formed was collected with a Buchner funnel, the filter cake was thoroughly washed with water and dried to give the title compound as an off-white solid.

[0239] Embodiment 11

[0240] General procedure for the reduction of Boc amino acids to alcohols (Method 11A)

[0241] Method 11A: A solution of RN-Boc amino acid and triethylamine (3.0 eq.) in dichloromethane (10 vol.) obtained according to Example 10 was cooled to -40°C, and isobutyl chloroformate (1.1 eq.) was added dropwise. The reaction mixture was warmed to room temperature and stirred for an additional hour. After completion (TLC), the reaction mixture was neutralized with 1.5N HCl, and the crude product was extracted with dichloromethane. The organic phase was washed with water and brine, dried over Na2SO4, and concentrated in vacuo to give a crude mixed anhydride of a pale yellow gum. Sodium borohydride was added to the crude residue dissolved in THF (10 vol.) at -30°C. Methanol was slowly added through an addition funnel at the same temperature, during which time slow bubbling began, and the reaction mixture was warmed to room temperature and continued to stir for an additional hour. The reaction mixture was neutralized with 1.5N HCl, and the crude product was extracted with ethyl acetate. The organic layer was washed with water and brine, dried over Na2SO4, concentrated, and purified to give the title compound.

[0242] Example 12

[0243] Synthesis of compound intermediate 12

[0244]

[0245] Pd(dppf)Cl2 was added to a solution of 4,6-dichloro-5-methoxypyrimidine (3 g, 0.1675 mol), 1-(2-fluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (4.20 g, 0.01843 mol) and sodium carbonate (2.66 g, 0.02513 mol) in dioxane:water. The dichloromethane complex (50 mg, 0.0006 mol) and the reaction mixture were heated at 90 °C for 16 hours. After the reaction was complete, the crude product was filtered through a diatomaceous earth bed, and the filtrate was diluted with ethyl acetate and washed thoroughly with water. The crude material obtained was purified by column chromatography (230-400 silica gel (10% EA / hexane as eluent)) to give the product as a brown solid.

[0246] Yield: 40% (2 g).

[0247] Appearance: brown solid.

[0248] LC / MS: Calculated 304.71; Found m / z [M+H] + 305.1.

[0249] To a solution of 4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazole-4-yl)-5-methoxypyrimidine (6g, 00197mol) in THF: methanol (50mL) sodium bicarbonate (3.31g, 0.0394mol) and 10% Pd / C (600mg) was added. The resulting mixture was stirred at RT under a hydrogen atmosphere for 16 hours. After the reaction was complete, the crude mixture was filtered through a diatomaceous earth bed and washed with methanol: ethyl acetate. The organic layer was concentrated in vacuo, and the crude material was purified by column chromatography (230-400 silica gel (50% EA / hexane as eluent)) to obtain the product as an off-white solid.

[0250] Yield: 75% (4 g).

[0251] Appearance: off-white solid.

[0252] LC / MS: Calculated 270.27; Found m / z [M+H] + 271.1.

[0253] To a solution of 4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)-5-methoxypyrimidine (4 g, 0.01481 mol) in dichloroethane (DCE; 100 mL) was added aluminum chloride (7.8 g, 0.02962 mol) in two portions at 0°C. The reaction mixture was stirred at 0°C for 10 minutes and then at 60°C for 16 hours. Upon completion, the reaction mixture was cooled to 0°C and quenched with 1M aqueous HCl (80 mL), followed by the slow addition of methanol (8 mL) with vigorous stirring. The mixture was poured into water (50 mL), the DCE layer was separated, and the aqueous phase was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered and concentrated. The crude material obtained was purified by column chromatography (230-400 silica gel (5% methanol / dichloromethane as eluent)) to give a colorless, gummy product.

[0254] Yield: 63% (2.4 g).

[0255] Appearance: colorless colloid.

[0256] LC / MS: Calculated 256.24; Found m / z [M+H] + 257.1.

[0257] Example 13

[0258] Synthesis of compound intermediate 13

[0259]

[0260] To a solution of 1-fluoro-2-iodobenzene (19.5 g, 87.8 mmol) in DMSO (40 mL) was added pyrazole (4 g, 58.8 mmol), followed by CuI (0.52 g, 2.73 mmol), L-proline (0.67 g, 5.217 mmol) and potassium carbonate (4.4 g, 31.88 mmol). The reaction mixture was heated to 100 ° C for 48 hours. The reaction mixture was cooled to room temperature, filtered through a celite bed, and the filtrate was extracted with dichloromethane. The organic layer was concentrated and the residue was purified by flash column chromatography to give 1- (2-fluorophenyl) -1H-pyrazole (3 g, 33% yield).

[0261] LC / MS APCI: Calculated 162.06 Found m / z [M+H] + 163.2.

[0262] 1 H-NMR 400MHz, DMSO-d6: δ8.21(d,J=5.60Hz,1H),7.81(t,J=8.00Hz,1H),7.80-7.78(m,1H),7.50-7.34(m,3H),6.58(d,J=4.00Hz,1H).

[0263] At 0°C, a solution of bromine (3 g, 18.5 mmol) in acetic acid (10 mL) was added to a solution of 1-(2-fluorophenyl)-1H-pyrazole in acetic acid (5 mL). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was cooled to room temperature, poured into ice and H2O in a 100 mL beaker, and an excess of saturated NaHCO3 aqueous solution was added until all acetic acid was quenched. Ethyl acetate (50 mL) was added and the layers were separated. The aqueous layer was extracted twice with ethyl acetate, and the combined organics were dried over Na2SO4 and concentrated in vacuo to give a crude solid, which was purified by flash column chromatography (SiO2, 1:1 hexane-ethyl acetate) to give 4-bromo-1-(2-fluorophenyl)-1H-pyrazole (2 g, 46% yield).

[0264] LC / MS APCI: Calculated Measured m / z [M+H] + 241.2.

[0265] 1 H-NMR 400MHz, DMSO-d6: δ8.48(s,1H),7.94(s,1H),7.64(t,J=8.00Hz,1H),7.52-7.47(m,2H),7.39-7.36(m,1H).

[0266] 4-Bromo-1-(2-fluorophenyl)-1H-pyrazole (3.0 g, 12.4 mmol), potassium acetate (2.43 g, 24.7 mmol), bis(pinacol)diboron (3.77 g, 14.9 mmol) and Pd(dppf)Cl2 (0.9 g, 1.23 mmol) were suspended in DMSO (4 mL) and 1,4-dioxane (15 mL). The system was exchanged with N2. The mixture was stirred at 100 ° C for 16 hours. The reaction mixture was diluted with water (60 mL). The reaction mixture was extracted with dichloromethane (50 mL×3). The combined organic layer was washed with saturated NaCl aqueous solution (15 mL), dried over anhydrous Na2SO4, and concentrated in vacuo. The residue was purified by silica gel column chromatography with 20:1 hexanes / ethyl acetate to give 1-(2-fluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (2.0 g, 58% yield).

[0267] LC / MS APCI: Calculated 288.14 Found m / z [M+H] + 289.1.

[0268] 1 H-NMR 400MHz, DMSO-d6: δ8.35(s,1H),7.91(s,1H),7.80(t,J=8.00Hz,1H),7.51-7.44(m,2H),7.39-7.34(m,1H),1.25(s,12H).

[0269] Embodiment 14

[0270] Synthesis of Compound I-1

[0271]

[0272] Using Method 1A, 4.0 g of (2,6-difluorophenyl)hydrazine hydrochloride was converted to 1-(2,6-difluorophenyl)-1H-pyrazole.

[0273] Yield (96%): 3.8 g.

[0274] Appearance: Light brown liquid.

[0275] LC / MS: Calculated 180.16; Found m / z [M+H] + 180.9.

[0276] LC / MS purity: 68.32%.

[0277]

[0278] Using Method 2A, 3.8 g of 1-(2,6-difluorophenyl)-1H-pyrazole were converted to 1-(2,6-difluorophenyl)-4-iodo-1H-pyrazole.

[0279] Yield (62%): 4.0 g.

[0280] Appearance: Light brown liquid.

[0281] LC / MS: Calculated 306.05; Found m / z [M+H] + 307.0.

[0282] LC / MS purity: 97.53%.

[0283]

[0284] Using Method 3A, 4.0 g of 1-(2,6-difluorophenyl)-4-iodo-1H-pyrazole was converted to 1-(2,6-difluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole.

[0285] Yield (63%): 2.5 g.

[0286] Appearance: light brown solid.

[0287] LC / MS: Calculated 306.12; Found m / z [M+H] + 307.3.

[0288] LC / MS purity: 75.14%.

[0289]

[0290] Using Method 4A, 447 mg of 1-(2,6-difluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was converted to tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0291] Yield (43%): 300 mg.

[0292] Appearance: white solid.

[0293] LC / MS: Calculated 519.98; Found m / z [M+H] + 520.2.

[0294] LC / MS purity: 92.50%.

[0295]

[0296] Using Method 5A, 300 mg of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to tert-butyl ((1s,4s)-4-(((4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0297] Yield (54%): 150 mg.

[0298] Appearance: white solid.

[0299] LC / MS: Calculated 485.54; Found m / z [M+H] + 486.3.

[0300] LC / MS purity: 99.91%.

[0301]

[0302] Using Method 6A, 150 mg of tert-butyl ((1s,4s)-4-(((4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to (1s,4s)-4-(((4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)-cyclohexane-1-amine hydrochloride.

[0303] Yield (100%): 130 mg.

[0304] Appearance: off-white solid.

[0305] LC / MS: Calculated 421.88; Found m / z [M+H] + 386.1.

[0306] HPLC purity: 99.05%.

[0307] Embodiment 15

[0308] Preparation of Compound I-2

[0309]

[0310] Using Method 1A (Example 1), 4.5 g of (2-(trifluoromethyl)phenyl)hydrazine hydrochloride was converted to 1-(2-(trifluoromethyl)phenyl)-1H-pyrazole.

[0311] Yield (90%): 4.0 g.

[0312] Appearance: Light brown liquid.

[0313] LC / MS: Calculated 212.18; Found m / z [M+H] + 213.1.

[0314] LC / MS purity: 87.78%.

[0315]

[0316] Using Method 2B, 4 g of 1-(2-(trifluoromethyl)phenyl)-1H-pyrazole was converted to 4-iodo-1-(2-(trifluoromethyl)phenyl)-1H-pyrazole.

[0317] Yield (79%): 4.8 g.

[0318] Appearance: light brown solid.

[0319] LC / MS: Calculated value 338.07; Found value m / z [M+H] + 339.0.

[0320] LC / MS purity: 98.74%.

[0321]

[0322] Using Method 3A, 4.8 g of 4-iodo-1-(2-(trifluoromethyl)phenyl)-1H-pyrazole was converted to 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2-(trifluoromethyl)-phenyl)-1H-pyrazole.

[0323] Yield (63%): 3 g.

[0324] Appearance: Light brown colloidal solid.

[0325] LC / MS: Calculated value 338.14; Found value m / z [M+H] + 339.1.

[0326] LC / MS purity: 82.42%.

[0327]

[0328] Using Method 4A, 493 mg of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2-(trifluoromethyl)phenyl)-1H-pyrazole was converted to tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)-carbamate.

[0329] Yield (34%): 250 mg.

[0330] Appearance: white solid.

[0331] LC / MS: calculated value 552.00; found value m / z [M+H] + 552.2.

[0332] LC / MS purity: 91.55%.

[0333]

[0334] Using Method 5A, 250 mg of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to tert-butyl ((1s,4s)-4-(((4-(1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)-pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0335] Yield (85%): 200 mg.

[0336] Appearance: white solid.

[0337] LC / MS: Calculated 517.55; Found m / z [M+H] + 518.1.

[0338] LC / MS purity: 99.75%.

[0339]

[0340] Using Method 6A, 200 mg of tert-butyl ((1s,4s)-4-(((4-(1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to ((1s,4s)-4-(((4-(1-(2-(trifluoromethyl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexan-1-amine hydrochloride.

[0341] Yield (100%): 180 mg.

[0342] Appearance: light brown solid.

[0343] LC / MS: Calculated 453.89; Found m / z [M+H] + 418.1.

[0344] HPLC purity: 99.40%.

[0345] Example 16

[0346] Synthesis of Compound I-3

[0347]

[0348] Using Method 1A, 2 g of (2-(trifluoromethoxy)phenyl)hydrazine hydrochloride was converted to 1-(2-(trifluoromethoxy)phenyl)-1H-pyrazole.

[0349] Yield (100%): 2 g.

[0350] Appearance: Light brown liquid.

[0351] LC / MS: Calculated 228.17; Found m / z [M+H] + 229.1.

[0352] LC / MS purity: 94.19%.

[0353]

[0354] Using Method 2C, 1.0 g of 1-(2-(trifluoromethoxy)phenyl)-1H-pyrazole was converted to 4-iodo-1-(2-(trifluoromethoxy)phenyl)-1H-pyrazole.

[0355] Yield (72%): 1.1 g.

[0356] Appearance: Light brown liquid.

[0357] LC / MS: Calculated value 354.07; Found value m / z [M+H] + 355.0.

[0358] LC / MS purity: 58.62%.

[0359]

[0360] Using Method 3B, 1.1 g of 4-iodo-1-(2-(trifluoromethoxy)phenyl)-1H-pyrazole was converted to 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2-(trifluoromethoxy)-phenyl)-1H-pyrazole.

[0361] Yield (54%): 600 mg.

[0362] Appearance: light brown solid.

[0363] LC / MS: Calculated 354.14; Found m / z [M+H] + 355.1.

[0364] LC / MS purity: 87.77%.

[0365]

[0366] Using Method 4A, 600 mg of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2-(trifluoromethoxy)phenyl)-1H-pyrazole was converted to tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2-(trifluoromethoxy)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)-carbamate.

[0367] Yield (33%): 300 mg.

[0368] Appearance: white solid.

[0369] LC / MS: Calculated 567.99; Found m / z [M+H] + 568.2.

[0370] LC / MS purity: 98.97%.

[0371]

[0372] Using Method 5A, 300 mg of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2-(trifluoromethoxy)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to tert-butyl ((1s,4s)-4-(((4-(1-(2-(trifluoromethoxy)phenyl)-1H-pyrazol-4-yl)-pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0373] Yield (81%): 230 mg.

[0374] Appearance: white solid.

[0375] LC / MS: Calculated 533.55; Found m / z [M+H] + 534.3.

[0376] LC / MS purity: 99.40%.

[0377]

[0378] Using Method 6A, 230 mg of tert-butyl ((1s,4s)-4-(((4-(1-(2-(trifluoromethoxy)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to (1s,4s)-4-(((4-(1-(2-(trifluoromethoxy)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)-oxy)methyl)cyclohexan-1-amine hydrochloride.

[0379] Yield (99%): 200 mg.

[0380] Appearance: off-white solid.

[0381] LC / MS: Calculated 469.89; Found m / z [M+H] + 434.1.

[0382] HPLC purity: 95.79%.

[0383] Embodiment 17

[0384] Synthesis of Compound I-4

[0385]

[0386] 1H-pyrazole (1.12 g, 0.0165 mol, 1 eq.) and KCO (2.73 g, 0.0198, 1.2 eq.) were added to a stirred solution of 2-fluorobenzonitrile (2 g, 0.165 mol, 1 eq.) in DMF (30 mL) at ambient temperature. The reaction mixture was stirred at 120 °C for 16 hours. The reaction mass was concentrated in vacuo and the residue was diluted with water and extracted with ethyl acetate (2 x 75 mL). The combined organic layers were washed with water (100 mL), saturated brine solution (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give 2-(1H-pyrazol-1-yl)benzonitrile.

[0387] Yield (89%): 2.5 g.

[0388] Appearance: Light brown liquid.

[0389] LC / MS: calculated value 169.19; found value m / z [M+H] + 170.1.

[0390] LC / MS purity: 99.15%.

[0391]

[0392] Using Method 2C, 2.5 g of 2-(1H-pyrazol-1-yl)benzonitrile was converted to 2-(4-iodo-1H-pyrazol-1-yl)benzonitrile.

[0393] Yield (69%): 3 g.

[0394] Appearance: light brown solid.

[0395] LC / MS: Calculated 295.08; Found m / z [M+H] + 295.9.

[0396] LC / MS purity: 92.10%.

[0397]

[0398] Using Method 3B, 3 g of 2-(4-iodo-1H-pyrazol-1-yl)benzonitrile was converted to 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)benzonitrile.

[0399] Yield (50%): 1.5 g.

[0400] Appearance: light brown solid.

[0401] LC / MS: Calculated 295.15; Found m / z [M+H] + 296.2.

[0402] LC / MS purity: 57.56%.

[0403]

[0404] 431 mg of 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazol-1-yl)benzonitrile was converted to tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2-cyanophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate using Method 4A.

[0405] Yield (44%): 300 mg.

[0406] Appearance: white solid.

[0407] LC / MS: calculated value 509.01; found value m / z [M+H] + 509.2.

[0408] LC / MS purity: 98.91%.

[0409]

[0410] Using Method 5A, 150 mg of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2-cyanophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to tert-butyl ((1s,4s)-4-(((4-(1-(2-cyanophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)-cyclohexyl)carbamate.

[0411] Yield (72%): 100 mg.

[0412] Appearance: white solid.

[0413] LC / MS: Calculated 474.57; Found m / z [M+H] + 475.2.

[0414] LC / MS purity: 98.63%.

[0415]

[0416] Using Method 6B, 80 mg of tert-butyl ((1s,4s)-4-(((4-(1-(2-cyanophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to 2-(4-(5-(((1s,4s)-4-aminocyclohexyl)methoxy)pyrimidin-4-yl)-1H-pyrazol-1-yl)benzonitrile hydrochloride.

[0417] Yield (65%): 45 mg.

[0418] Appearance: off-white solid.

[0419] LC / MS: Calculated 410.91; Found m / z [M+H] + 375.2.

[0420] HPLC purity: 97.00%.

[0421] Embodiment 18

[0422] Synthesis of Compound I-5

[0423]

[0424] Using Method 1B, 4.5 g of (2-methoxyphenyl)hydrazine hydrochloride was converted to 1-(2-methoxyphenyl)-1H-pyrazole.

[0425] Yield (40%): 2.2 g.

[0426] Appearance: Light brown liquid.

[0427] LC / MS: Calculated 174.20; Found m / z [M+H] + 175.1.

[0428] LC / MS purity: 98.56%.

[0429]

[0430] Using Method 2C, 2.0 g of 1-(2-methoxyphenyl)-1H-pyrazole was converted to 4-iodo-1-(2-methoxyphenyl)-1H-pyrazole.

[0431] Yield (73%): 2.5 g.

[0432] Appearance: Light brown liquid.

[0433] LC / MS: Calculated 300.10; Found m / z [M+H] + 301.1.

[0434] LC / MS purity: 90.58%.

[0435]

[0436] Using Method 3B, 1 g of 4-iodo-1-(2-methoxyphenyl)-1H-pyrazole was converted to 1-(2-methoxyphenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole.

[0437] Yield (60%): 600 mg.

[0438] Appearance: white solid.

[0439] LC / MS: Calculated 300.17; Found m / z [M+H] + 301.1.

[0440] LC / MS purity: 91.95%.

[0441]

[0442] Using Method 4A, 600 mg of 1-(2-methoxyphenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was converted to tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0443] Yield (34%): 350 mg.

[0444] Appearance: white solid.

[0445] LC / MS: calculated value 514.02; found value m / z [M+H] + 515.1.

[0446] LC / MS purity: 89.35%.

[0447]

[0448] Using Method 5A, 300 mg of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to tert-butyl ((1s,4s)-4-(((4-(1-(2-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0449] Yield (79%): 220 mg.

[0450] Appearance: white solid.

[0451] LC / MS: Calculated 479.58; Found m / z [M+H] + 480.3.

[0452] LC / MS purity – 99.66%.

[0453]

[0454] Using Method 6A, 220 mg of tert-butyl ((1s,4s)-4-(((4-(1-(2-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to (1s,4s)-4-(((4-(1-(2-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)-cyclohexane-1-amine hydrochloride.

[0455] Yield (100%): 200 mg.

[0456] Appearance: light yellow solid.

[0457] LC / MS: Calculated 415.92; Found m / z [M+H] + 380.1.

[0458] HPLC purity: 99.60%.

[0459] Embodiment 19

[0460] Synthesis of Compound I-6

[0461]

[0462] Using Method 10A, 2 g of ((1S,3R)-3-aminocyclopentyl)(11-oxyalkyl)methanone was converted to tert-butyl ((1R,3S)-3-((11-oxyalkyl)carbonyl)cyclopentyl)carbamate.

[0463] Yield (75%): 1.6 g.

[0464] Appearance: off-white solid.

[0465] LC / MS: calcd. 228; found m / z 174 [M+H] -56.

[0466] LC / MS purity: 99%.

[0467]

[0468] Using Method 11A, 2.0 g of tert-butyl ((1R,3S)-3-((11-oxoalkyl)carbonyl)cyclopentyl)carbamate was converted to tert-butyl ((1R,3S)-3-(hydroxymethyl)cyclopentyl)carbamate.

[0469] Yield: (2.7 g, 75% yield).

[0470] Appearance: off-white solid.

[0471] LC / MS: calcd. 215.29; found m / z 160; [M+H-56].

[0472]

[0473] Using a modified Mitsunobu reaction for the synthesis of compound 1-8, tert-butyl ((1R,3S)-3-(hydroxymethyl)cyclopentyl)carbamate (180 mg) was converted to tert-butyl ((1R,3S)-3-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-yl)oxy)methyl)cyclopentyl)carbamate.

[0474] Yield: (800 mg) crude product impure with triphenylphosphine.

[0475] Appearance: Off-white solid

[0476] LC / MS: Calculated 453.52; Found m / z [M+H] + 454.2.

[0477]

[0478] Using Method 6A, tert-butyl ((1R,3S)-3-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclopentyl)carbamate (800 mg crude product containing TPPO) was converted to (1R,3S)-3-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclopentan-1-amine hydrochloride.

[0479] Yield: (200 mg).

[0480] Appearance: white solid.

[0481] LC / MS: Calculated 389.86; Found m / z [M+H] + 354.1.

[0482] HPLC purity: 95.31%.

[0483] Embodiment 20

[0484] Synthesis of Compound I-7

[0485]

[0486] To a stirred solution of (1R,3S)-3-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclopentane-1-amine (170 mg, 0.4810 mmol) in methanol were added paraformaldehyde (300 mg), NaCNBH3 (200 mg) and AcOH (catalyst). The resulting mixture was stirred at room temperature for 16 hours. After the reaction was complete, the mixture was filtered through a celite bed, the celite was washed with methanol and the combined organic layers were concentrated in vacuo. The crude material obtained was purified by preparative HPLC to give a gummy substance, which was dissolved in water, basified with saturated sodium bicarbonate solution and extracted with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered and concentrated to give a colorless gummy substance.

[0487] Yield: 26.2% (48 mg).

[0488] Appearance: colorless colloid.

[0489] LC / MS: Calculated 381.46; Found m / z [M+H] + 382.1.

[0490] HPLC purity: 94.24%.

[0491] 1H-NMR (400MHz, DMSO-d6): δ8.86 (d, J = 2.48Hz, 1H), 8.78 (t, J = 0.92Hz, 1H), 4. 64(d,J=1.72Hz,1H),8.50(d,J=1.88Hz,1H),7.93(t,J=8.00Hz,1H),7.48-7. 54(m,2H),7.40-7.44(m,1H),4.21(d,J=6.72Hz,2H),2.42-2.51(m,1H),2.07 -2.12(m,7H),1.80-1.89(m,3H),1.48-1.58(m,2H),1.25(d,J=11.20Hz,1H).

[0492] Embodiment 21

[0493] Synthesis of Compound I-8

[0494]

[0495] To an ice-cold solution of (1R, 3S)-3-(methoxycarbonyl)cyclopentane-1-carboxylic acid (2g, 11.6mmol), dimethylamine.HCl (1.22g, 15.12mmol) and triethylamine (3.5g, 35mmol) in dichloromethane (20mL) was added propylphosphonic anhydride (T3P, 5.5g, 17.4mmol), and the mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with water, and the crude product was extracted with dichloromethane. The combined extracts were concentrated and purified by column chromatography to obtain (1S, 3R)-3-(dimethylcarbamoyl)cyclopentane-1-methyl formate as an off-white solid.

[0496] Yield: (1.6 g, 70% yield).

[0497] LC / MS: calcd. 199.15; found m / z 200; [M+H].

[0498]

[0499] To the ice-cold solution of THF (20mL) of (1S, 3R)-3-(dimethylcarbamoyl)cyclopentane-1-methyl formate (1.5g, 7.5mmol), LAH (7.5mL, 2M in THF, 15mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with ethyl acetate / ice, and the crude product was extracted with ethyl acetate. The combined extracts were concentrated and purified by column chromatography to obtain ((1S, 3R)-3-((dimethylamino)methyl)cyclopentyl)methanol as a light brown liquid.

[0500] Yield: (0.7 g, 45% yield).

[0501] LC / MS: calcd. 157.26; found m / z 158.26 [M+H].

[0502]

[0503] To a solution of 4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-ol (100 mg, 0.3902 mmol) in toluene (3 mL) at 0°C, ((1R, 3S)-3-((dimethylamino)methyl)cyclopentyl)methanol (67.50 mg, 0.4292 mmol) and (cyanomethylene)tri-n-butylphosphine (CMBP, modified Mitsunobu reaction, 117.46 mg, 0.4866 mmol) were added and the mixture was stirred at 100°C for 18 hours. The organic layer was concentrated in vacuo. The crude material obtained was purified by preparative HPLC.

[0504] Yield: 23.2% (36 mg).

[0505] Appearance: colorless colloid.

[0506] LC / MS: Calculated 395.48; Found m / z [M+H] + 396.3.

[0507] HPLC purity: 99.45%.

[0508] Embodiment 22

[0509] Preparation of Compound I-9

[0510]

[0511] Paraformaldehyde (75 mg) and Na(OAc)3BH (259 mg, 1.224 mmol, 6 equiv) were added to a stirred solution of (1s, 4s)-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-amine (75 mg, 0.204 mmol, 1 equiv) in AcOH (2 mL) and ethanol (2 mL) at ambient temperature. The reaction mixture was stirred at 100 °C for 48 hours. The reaction mass was cooled to ambient temperature and concentrated in vacuo. The residue thus obtained was diluted with water (20 mL) and extracted with dichloromethane (2×30 mL). The combined organic layers were washed with saturated brine solution, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by preparative HPLC to give compound I-9.

[0512] Yield (18%): 15 mg.

[0513] Appearance: Off-white solid

[0514] LC / MS: Calculated 395.48; Found m / z [M+H] + 396.1.

[0515] HPLC purity: 99.03%.

[0516] Embodiment 23

[0517] Synthesis of Compound I-10

[0518]

[0519] Using Method 8A, 300 mg of 5-chloro-4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidine was converted to tert-butyl (1-(4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)piperidin-4-yl)carbamate.

[0520] Yield (7%): 30 mg.

[0521] Appearance: white solid.

[0522] LC / MS: Calculated 438.51; Found m / z [M+H] + 439.2.

[0523] LC / MS purity: 59.64%.

[0524]

[0525] Using Method 9B, 30 mg of tert-butyl (1-(4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)piperidin-4-yl)carbamate was converted to 1-(4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)piperidin-4-amine hydrochloride.

[0526] Yield (100%): 25 mg.

[0527] Appearance: light yellow solid.

[0528] LC / MS: Calculated 374.85; Found m / z [M+H] + 339.0.

[0529] HPLC purity: 99.61%.

[0530] Embodiment 24

[0531] Synthesis of Compound I-11

[0532]

[0533] Using Method 7A, 1.0 g of 4,5-dichloropyrimidine was converted to 5-chloro-4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidine.

[0534] Yield (43%): 0.8 g.

[0535] Appearance: white solid.

[0536] LC / MS: Calculated 274.68; Found m / z [M+H] + 275.0.

[0537] LC / MS purity: 99.14%.

[0538]

[0539] Using Method 8A, 200 mg of 5-chloro-4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidine was converted to tert-butyl ((1s,4s)-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)-pyrimidin-5-yl)amino)methyl)cyclohexyl)carbamate.

[0540] Yield (29%): 100 mg.

[0541] Appearance: Light brown colloidal solid.

[0542] LC / MS: Calculated 466.56; Found m / z [M+H] + 467.3.

[0543] LC / MS purity: 91.69%.

[0544]

[0545] Using Method 9A, 100 mg of tert-butyl (((1s,4s)-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)amino)methyl)cyclohexyl)carbamate was converted to N-(((1s,4s)-4-aminocyclohexyl)methyl)-4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-amine trifluoroacetate.

[0546] Yield (45%): 50 mg.

[0547] Appearance: light yellow solid.

[0548] LC / MS: Calculated 480.47; Found m / z [M+H]+ 367.2.

[0549] HPLC purity: 99.28%.

[0550] Embodiment 25

[0551] Synthesis of Compound I-12

[0552]

[0553] Using Method 7A, 500 mg of 4,5-dichloropyrimidine was converted to 5-chloro-4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidine.

[0554] Yield (51%): 500 mg.

[0555] Appearance: white solid.

[0556] LC / MS: Calculated 292.67; Found m / z [M+H] + 293.0.

[0557] LC / MS purity: 99.19%.

[0558]

[0559] Using Method 8A, 300 mg of 5-chloro-4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidine was converted to tert-butyl ((1s,4s)-4-(((4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)amino)methyl)cyclohexyl)carbamate.

[0560] Yield (8%): ​​40 mg.

[0561] Appearance: light brown solid.

[0562] LC / MS: Calculated 484.55; Found m / z [M+H] + 485.3.

[0563] LC / MS purity: 75.46%.

[0564]

[0565] Using Method 9A, 40 mg of tert-butyl ((1s,4s)-4-(((4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)amino)methyl)cyclohexyl)carbamate was converted to N-(((1s,4s)-4-aminocyclohexyl)methyl)-4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)-pyrimidin-5-amine trifluoroacetate.

[0566] Yield (60%): 25 mg.

[0567] Appearance: light yellow solid.

[0568] LC / MS: Calculated 498.2; Found m / z [M+H] + 385.0 (without trifluoroacetate ion).

[0569] HPLC purity: 97.70%.

[0570] Embodiment 26

[0571] Synthesis of Compound I-13

[0572]

[0573] Using Method 11A, 0.22 g of (1S,2R)-2-((tert-butoxycarbonyl)amino)cyclobutane-1-carboxylic acid was converted to tert-butyl ((1R,2S)-2-(hydroxymethyl)cyclobutyl)carbamate.

[0574] Yield (14%): 30 mg. It was used as such in the next step without any purification.

[0575]

[0576] Using Method 11A, 0.03 g of tert-butyl ((1R,2S)-2-(hydroxymethyl)cyclobutyl)carbamate was converted to tert-butyl ((1R,2S)-2-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)-oxy)methyl)cyclobutyl)carbamate.

[0577] Yield (38%): 25 mg. It was used as such in the next step without any purification.

[0578]

[0579] Using Method 6A, 0.03 g of tert-butyl ((1R,2S)-2-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclobutyl)carbamate was converted to (1R,2S)-2-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)-cyclobutane-1-amine (HCl salt).

[0580] Yield (78%): 15 mg.

[0581] Appearance: off-white solid.

[0582] LC / MS: calcd. 339.37; found m / z 340 [M+H].

[0583] Embodiment 27

[0584] Preparation of Compound I-14

[0585]

[0586] Using Method 1B, 3.0 g of (2-fluoro-6-methylphenyl)hydrazine hydrochloride was converted to 1-(2-fluoro-6-methylphenyl)-1H-pyrazole.

[0587] Yield (46%): 1.4 g.

[0588] Appearance: Light brown liquid.

[0589] LC / MS: Calculated value 176.19; Found value m / z [M+H] + 177.1.

[0590] LC / MS purity: 97.71%.

[0591]

[0592] Using Method 2C, 1.4 g of 1-(2-fluoro-6-methylphenyl)-1H-pyrazole was converted to 1-(2-fluoro-6-methylphenyl)-4-iodo-1H-pyrazole.

[0593] Yield (67%): 1.6 g.

[0594] Appearance: Light brown liquid.

[0595] LC / MS: Calculated 302.09; Found m / z [M+H] + 303.0.

[0596] LC / MS purity: 89.86%.

[0597]

[0598] Using Method 3B, 1.6 g of 1-(2-fluoro-6-methylphenyl)-4-iodo-1H-pyrazole was converted to 1-(2-fluoro-6-methylphenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole.

[0599] Yield (62%): 1 g.

[0600] Appearance: Light brown liquid.

[0601] LC / MS: Calculated 302.16; Found m / z [M+H] + 303.2.

[0602] LC / MS purity: 95.28%.

[0603]

[0604] Using Method 4A, 250 mg of 1-(2-fluoro-6-methylphenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was converted to ((1s,4s)-4-(((4-chloro-6-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)-cyclohexyl)-carbamic acid tert-butyl ester.

[0605] Yield (35%): 150 mg.

[0606] Appearance: white solid.

[0607] LC / MS: calculated value 516.01; found value m / z [M+H] + 516.1.

[0608] LC / MS purity: 97.87%.

[0609]

[0610] Using Method 5A, 150 mg of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to tert-butyl ((1s,4s)-4-(((4-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)-pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0611] Yield (72%): 100 mg.

[0612] Appearance: white solid.

[0613] LC / MS: Calculated 481.57; Found m / z [M+H] + 482.2.

[0614] LC / MS purity: 99.89%.

[0615]

[0616] Using Method 6A, 100 mg of tert-butyl ((1s,4s)-4-(((4-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to (1s,4s)-4-(((4-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)-cyclohexane-1-amine hydrochloride.

[0617] Yield (97%): 85 mg.

[0618] Appearance: off-white solid.

[0619] LC / MS: Calculated 417.91; Found m / z [M+H] + 382.2.

[0620] HPLC purity: 98.05%.

[0621] Embodiment 28

[0622] Preparation of Compound I-15

[0623]

[0624] Acetone (0.2 mL) was added to a stirred solution of (1s, 4s)-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-amine (75 mg, 0.204 mmol, 1 eq) in methanol (2 mL) at ambient temperature. The reaction mixture was concentrated in vacuo. To the residue dissolved in methanol was added Na(OAc)3BH (216.36 mg, 1.02 mmol, 5 eq) at 0°C. The reaction mixture was stirred at ambient temperature for 4 hours. The reaction mass was concentrated in vacuo. The residue thus obtained was diluted with water (20 mL) and extracted with dichloromethane (2×30 mL). The combined organic layers were washed with saturated brine solution, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by preparative HPLC to give compound I-15.

[0625] Yield (18%): 15 mg.

[0626] Appearance: off-white solid.

[0627] LC / MS: Calculated 523.53; Found m / z [M+H] + 410.2.

[0628] HPLC purity: 96.02%.

[0629] Embodiment 29

[0630] Synthesis of Compound I-16

[0631]

[0632] 4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-ol (100 mg) was converted to 4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)-5-((1-isopropylpiperidin-4-yl)methoxy)pyrimidine using a modified Mitsunobu reaction for the synthesis of compound 1-8.

[0633] Yield (39%): 60 mg.

[0634] Appearance: colorless colloid.

[0635] LC / MS: Calculated 395.48; Found m / z [M+H] + 396.21.

[0636] HPLC purity: 98.59%.

[0637] Embodiment 30

[0638] Synthesis of Compound I-17

[0639]

[0640] To the ice-cold solution of (1s, 4s)-4-((tert-butoxycarbonyl) amino) cyclohexane-1-formic acid (5g, 20mmol), N, O-dimethylhydroxylamine HCl (2.52g, 26mmol) and triethylamine (6.2g, 60mmol) in dichloromethane (50mL), add propylphosphonic anhydride (T3P, 6.7g, 30mmol), and the reaction mixture is stirred at room temperature for 16 hours. The reaction mixture is diluted with water, and the crude product is extracted with dichloromethane. The combined extract is concentrated and purified by column chromatography to obtain the tert-butyl ((1s, 4s)-4-(methoxy (methyl) carbamoyl) cyclohexyl) carbamate of off-white solid.

[0641] Yield (74%): 4.2 g.

[0642] LC / MS: calcd. 286.37; found m / z 287 [M+H].

[0643]

[0644] Under nitrogen atmosphere, MeMgBr (20mL, 2M in THF, 40mmol) is added to the ice-cold solution of THF (30mL) of ((1s, 4s)-4-(methoxy (methyl) carbamoyl) cyclohexyl) tert-butyl carbamate (3.0g, 10.4mmol), and the reaction mixture is stirred at room temperature for 4 hours. The reaction mixture is quenched with saturated ammonium chloride, and the crude product is extracted with ethyl acetate. The combined extracts are concentrated and purified to obtain tert-butyl ((1s, 4s)-4-acetylcyclohexyl) carbamate as an off-white solid.

[0645] Yield (68%): 1.7 g.

[0646] LC / MS: calcd. 241.33; found m / z 142.1 [M+H]-100.

[0647]

[0648] Sodium borohydride (0.057g, 1.5mmol) is added to the ice-cold solution of methanol (5mL) of ((1s, 4s)-4-acetylcyclohexyl) tert-butyl carbamate (0.25g, 1.0mmol), and the reaction mixture is stirred at room temperature for 3 hours. The reaction mixture is concentrated, diluted with water, and the crude product is extracted with dichloromethane. The combined extract is concentrated and purified by column chromatography to obtain ((1R, 4s)-4-((S)-1-hydroxyethyl) cyclohexyl) tert-butyl carbamate of transparent jelly.

[0649] Yield (60%): 0.15 g.

[0650] LC / MS: calcd. 243.35; found m / z 142.1 [M+H]-100.

[0651]

[0652] 4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-ol (80 mg) was converted to tert-butyl ((1S,4s)-4-((R)-1-((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)ethyl)cyclohexyl)carbamate using a modified Mitsunobu reaction for compound 1-8.

[0653] Yield (39%): 60 mg.

[0654] Appearance: colorless colloid.

[0655] LC / MS: Calculated 481.57; Found m / z [M+H] + 482.2.

[0656]

[0657] Using Method 6A, tert-butyl ((1S,4s)-4-((R)-1-((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)ethyl)cyclohexyl)carbamate (70 mg) was converted to (1S,4s)-4-((R)-1-((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)ethyl)-cyclohexan-1-amine.

[0658] Yield (95%): 70 mg.

[0659] Appearance: colorless colloid.

[0660] LC / MS: Calculated 381.46; Found m / z [M+H] + 382.3.

[0661] HPLC purity: 94.10%.

[0662] Embodiment 31

[0663] Synthesis of Compound I-18

[0664]

[0665] Using Method 10A, 2.0 g of (1S,3R)-3-aminocyclohexane-1-carboxylic acid was converted to (1S,3R)-3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid.

[0666] Yield (75%): 2.5 g.

[0667] LC / MS: calcd. 243.30; found m / z 144.1 [M+H]-100.

[0668]

[0669] Using Method 11A, 1.0 g of (1S,3R)-3-((tert-butoxycarbonyl)amino)cyclohexane-1-carboxylic acid was converted to tert-butyl ((1R,3S)-3-(hydroxymethyl)cyclohexyl)carbamate.

[0670] Yield (70% yield): 0.6 g

[0671] LC / MS: calcd. 229.32; found m / z 130.12 [M+H]-100.

[0672]

[0673] Using a modification of the Mitsunobu reaction for the preparation of compound 1-8, tert-butyl ((1R,3S)-3-(hydroxymethyl)cyclohexyl)carbamate (400 mg) was converted to tert-butyl ((1R,3S)-3-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0674] Yield (61%): 400 mg.

[0675] Appearance: colorless colloid.

[0676] LC / MS: Calculated 376.28; Found m / z [M+H] + 322.1.

[0677]

[0678] Using Method 4A, tert-butyl ((1R,3S)-3-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (250 mg) was converted to tert-butyl ((1R,3S)-3-(((4-chloro-6-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0679] Yield (40%): 200 mg.

[0680] Appearance: brown solid.

[0681] LC / MS: calculated value 516.01; found value m / z [M+H] + 516.5.

[0682]

[0683] Using Method 5A, tert-butyl ((1R,3S)-3-(((4-chloro-6-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (200 mg) was converted to tert-butyl ((1R,3S)-3-(((4-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0684] Yield (64%): 120 mg.

[0685] Appearance: colorless colloid.

[0686] LC / MS: Calculated 481.57; Found m / z [M+H] + 482.2.

[0687]

[0688] Using Method 6A, tert-butyl ((1R,3S)-3-(((4-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (200 mg) was converted to (1R,3S)-3-(((4-(1-(2-fluoro-6-methylphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)-oxy)-methyl)-cyclohexane-1-amine hydrochloride.

[0689] Yield (64%): 120 mg.

[0690] Appearance: white solid.

[0691] LC / MS: Calculated 381.46; Found m / z [M+H] + 382.2.

[0692] HPLC purity: 98.27%.

[0693] Embodiment 32

[0694] Synthesis of Compound I-19

[0695]

[0696] Using the modified Mitsunobu method for synthesizing compound 1-8, 150 mg of 4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-ol was converted into tert-butyl ((1s,4s)-4-(2-((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)ethyl)cyclohexyl)carbamate.

[0697] Yield (32%): 90 mg.

[0698] Appearance: Brown liquid.

[0699] LC / MS: Calculated 481.57; Found m / z [M+H] + 482.3.

[0700]

[0701] Using Method 6A, 90 mg of tert-butyl ((1s,4s)-4-(2-((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)ethyl)cyclohexyl)carbamate was converted to (1s,4s)-4-(2-((4-(1-(2-fluoro-phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)ethyl)cyclohexan-1-amine.

[0702] Appearance: colorless colloid.

[0703] LC / MS: Calculated 381.46; Found m / z [M+H] + 382.2.

[0704] HPLC purity: 97.02%.

[0705] Embodiment 33

[0706] Synthesis of Compound I-20

[0707]

[0708] Using Method 1A, 3 g of (3,5-difluorophenyl)hydrazine hydrochloride was converted to 1-(3,5-difluorophenyl)-1H-pyrazole.

[0709] Yield (97%): 2.8 g.

[0710] Appearance: Light brown liquid.

[0711] LC / MS: Calculated 180.16; Found m / z [M+H] + 181.1.

[0712] LC / MS purity: 98.27%.

[0713]

[0714] Using Method 2D, 2.8 g of 1-(3,5-difluorophenyl)-1H-pyrazole was converted to 1-(3,5-difluorophenyl)-4-iodo-1H-pyrazole.

[0715] Yield (76%): 3.5 g.

[0716] Appearance: light brown solid.

[0717] LC / MS: Calculated 306.05; Found m / z [M+H] + 306.9.

[0718] LC / MS purity: 98.32%.

[0719]

[0720] Using Method 3A, 3 g of 1-(3,5-difluorophenyl)-4-iodo-1H-pyrazole were converted to 1-(3,5-difluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole.

[0721] Yield (67%): 2 g.

[0722] Appearance: white solid.

[0723] LC / MS: Calculated 306.12; Found m / z [M+H] + 307.0.

[0724] LC / MS purity: 89.93%.

[0725]

[0726] Using Method 4A, 447 mg of 1-(3,5-difluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was converted to tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(3,5-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0727] Yield (36%): 250 mg.

[0728] Appearance: white solid.

[0729] LC / MS: Calculated 519.98; Found m / z [M+H] + 464.1 (Boc fragment mass).

[0730] LC / MS purity: 97.43%.

[0731]

[0732] Using Method 5A, 250 mg of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(3,5-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to tert-butyl ((1s,4s)-4-(((4-(1-(3,5-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0733] Yield (76%): 180 mg.

[0734] Appearance: white solid.

[0735] LC / MS: Calculated 485.54; Found m / z [M+H] + 486.1.

[0736] LC / MS purity: 99.49%.

[0737]

[0738] Using Method 6A, 180 mg of tert-butyl ((1s,4s)-4-(((4-(1-(3,5-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)-methyl)cyclohexyl)carbamate was converted to (1s,4s)-4-(((4-(1-(3,5-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)-methyl)-cyclohexane-1-amine hydrochloride.

[0739] Yield (96%): 150 mg.

[0740] Appearance: off-white solid.

[0741] LC / MS: Calculated 421.88; Found m / z [M+H] + 386.1.

[0742] HPLC purity: 97.62%.

[0743] Embodiment 34

[0744] Preparation of Compound I-21

[0745]

[0746] BBr3 (1M in dichloromethane) (4 mL, 4.019 mmol, 10 equiv) was added to a stirred solution of tert-butyl ((1s, 4s)-4-(((4-(1-(3-fluoro-4-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)-methyl)-cyclohexyl)carbamate (200 mg, 0.401 mmol, 1 equiv) in dichloromethane (5 mL) at -78 °C. The reaction mixture was stirred at ambient temperature for 16 hours. The reaction mass was cooled to 0 °C, quenched with methanol and concentrated in vacuo. The crude product was purified by preparative HPLC to give compound 4-(4-(5-(((1s, 4s)-4-aminocyclohexyl)-methoxy)pyrimidin-4-yl)-1H-pyrazol-1-yl)-2-fluorophenol trifluoroacetic acid.

[0747] Yield (72%): 145 mg.

[0748] Appearance: light yellow solid.

[0749] LC / MS: Calculated 497.45; Found m / z [M+H] + 384.2.

[0750] HPLC purity: 99.66%.

[0751] Embodiment 35

[0752] Preparation of Compound I-22

[0753]

[0754] Using Method 1A, 2 g of (3-fluoro-4-methoxyphenyl)hydrazine hydrochloride was converted to 1-(3-fluoro-4-methoxyphenyl)-1H-pyrazole.

[0755] Yield (100%): 2 g.

[0756] Appearance: Light brown liquid.

[0757] LC / MS: Calculated value 192.19; Found value m / z [M+H] + 193.1.

[0758] LC / MS purity: 84.96%.

[0759]

[0760] Using Method 2C, 2.0 g of 1-(3-fluoro-4-methoxyphenyl)-1H-pyrazole was converted to 1-(3-fluoro-4-methoxyphenyl)-4-iodo-1H-pyrazole.

[0761] Yield (67%): 2.2 g.

[0762] Appearance: light brown solid.

[0763] LC / MS: Calculated value 318.09; Found value m / z [M+H] + 318.9.

[0764] LC / MS purity: 76.57%.

[0765]

[0766] Using Method 3B, 2.2 g of 1-(3-fluoro-4-methoxyphenyl)-4-iodo-1H-pyrazole was converted to 1-(3-fluoro-4-methoxyphenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole.

[0767] Yield (59%): 1.3 g.

[0768] Appearance: white solid.

[0769] LC / MS: Calculated value 318.16; Found value m / z [M+H] + 319.2.

[0770] LC / MS purity: 49.77%.

[0771]

[0772] Using Method 4A, 557 mg of 1-(3-fluoro-4-methoxyphenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole was converted to tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(3-fluoro-4-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)-carbamate.

[0773] Yield (47%): 400 mg.

[0774] Appearance: white solid.

[0775] LC / MS: calculated value 532.01; found value m / z [M+H] + 532.2.

[0776] LC / MS purity: 99.13%.

[0777]

[0778] Using Method 5A, 400 mg of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(3-fluoro-4-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to tert-butyl ((1s,4s)-4-(((4-(1-(3-fluoro-4-methoxyphenyl)-1H-pyrazol-4-yl)-pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate.

[0779] Yield (80%): 300 mg.

[0780] Appearance: white solid.

[0781] LC / MS: Calculated 497.57; Found m / z [M+H] + 498.1.

[0782] LC / MS purity: 98.75%.

[0783]

[0784] Using Method 6A, 100 mg of tert-butyl ((1s,4s)-4-(((4-(1-(3-fluoro-4-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate was converted to (1s,4s)-4-(((4-(1-(3-fluoro-4-methoxyphenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)-cyclohexane-1-amine hydrochloride.

[0785] Yield (97%): 85 mg.

[0786] Appearance: light yellow solid.

[0787] LC / MS: Calculated 433.91; Found m / z [M+H] + 398.1.

[0788] HPLC purity: 99.13%.

[0789] Embodiment 36

[0790] Synthesis of Compound I-23 and Compound I-27

[0791]

[0792] At 0 ° C, aluminum chloride (7.5 g, 56.4 mmol) was added to a solution of DCE (50 mL) of 4,6-dichloro-5-methoxypyrimidine (5.0 g, 28.24 mmol) in two portions. The reaction mixture was stirred at 0 ° C for 10 minutes and then at 50 ° C for 4 hours. The mixture was cooled to 0 ° C, and HCl aqueous solution (1 M, 20 mL) was slowly added, followed by methanol (20 mL) while stirring vigorously. The mixture was poured into water and extracted with ether. The combined organic layers were washed with brine, dried over magnesium sulfate, filtered and concentrated. The residue was concentrated in vacuo to give 4,6-dichloropyrimidine-5-ol as a beige solid.

[0793] LC / MS APCI: Calculated 163.95; Found m / z [M+H] + 165.1.

[0794] 1 H NMR (DMSO-d6): δ (ppm) 11.67 (s, br, 1H), 8.38 (s, 1H).

[0795] Triphenylphosphine (3.17 g, 12 mmol) and 4,6-dichloropyrimidine-5-ol (1.0 g, 6 mmol) were added to a solution of tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (1.95 g, 9.06 mmol) in THF (10 mL). Diisopropyl azodicarboxylate (2.4 mL, 12 mmol) was added to the mixture at 0 ° C., and the mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 40: 1 as eluent) to give tert-butyl 4-(((4,6-dichloropyrimidine-5-yl) oxy) methyl) piperidine-1-carboxylate (0.6 g, 28% yield) as a colorless oil.

[0796] LC / MS APCI: Calculated 361.10; Found m / z [M+H] + 362.2.

[0797] To 1,4-dioxane (5 mL) was added tert-butyl 4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.6 g, 1.6 mmol), 1-(2-fluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.47 g, 1.65 mmol), Pd(dppf)Cl (0.12 g, 0.16 mmol) and NaCO (0.21 g, 1.98 mmol in 2 mL water) under N2 atmosphere. The contents were heated and maintained at 110°C overnight. After the reaction was complete, the reaction mixture was cooled to room temperature. Dioxane was removed in vacuo, and the resulting aqueous solution was extracted with EtOAc. The combined organic phases were washed three times with water and saturated brine, and dried over Na2SO4. After removing the solvent under reduced pressure, the residue was purified by flash column chromatography to give the pure product tert-butyl 4-(((4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.4 g, 50% yield).

[0798] LC / MS APCI: Calculated 487.18 Found m / z [M+H] + 488.2.

[0799] 1H-NMR 400MHz, DMSO-d6: δ8.88(s,1H),8.75(s,1H),8.46(s,1H),7.94-7.89(m,1H),7.58-7.48(m,2H),7.44-7.40(m,1H),4.79- 4.77(m,2H),4.04-3.98(m,2H),3.92-3.90(m,2H),2.81-2.77(m,2H),2.14-2.10(m,1H),1.85-1.81(m,2H),1.07(s,9H).

[0800] To a solution of tert-butyl 4-(((4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.3 g, 0.61 mmol) in methanol was added sodium bicarbonate (0.11 g, 1.23 mmol) and 30 mg of 10% Pd / carbon. The flask was equipped with a hydrogen-filled balloon fitted to a three-way stopcock. After several evacuation / hydrogen flush cycles, the reaction mixture was stirred under 1 atm of hydrogen for 16 hours. The mixture was filtered through a pad of celite and the pad was rinsed with copious amounts of methanol. The volatiles were removed in vacuo and the crude solid was purified by flash column chromatography to give tert-butyl 4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.15 g, 56% yield).

[0801] LC / MS APCI: Calculated 453.22 Found m / z [M+H] + 454.3.

[0802] 1 H-NMR 400MHz, DMSO-d6: δ8.87(s,1H),8.79(s,1H),8.65(s,1H),8.49(s,1H),7.95-7.92(m,1H),7.57-7.49(m,2H),7.44-7.40(m,1H),4 .79-4.60(m,2H),4.21(d,J=6.40Hz,2H),4.04-4.01(m,2H),2.15-2.12(m,1H),1.84-1.81(m,2H),1.18-1.16(m,2H),1.09(s,9H).

[0803] To a solution of tert-butyl 4-(((4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.05 g, 0.1 mmol) in dichloromethane was added 4M HCl in diethyl ether (2.0 mL), and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated in vacuo and triturated with diethyl ether to give 4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)-5-(piperidin-4-ylmethoxy)pyrimidine (Compound I-27; 18 mg, 46% yield).

[0804] LC / MS APCI: Calculated 387.13 Found m / z [M+H] + 388.0.

[0805] 1 H-NMR 400MHz, DMSO-d6: δ8.87(s,1H),8.77(s,1H),8.46(s,1H),7.92(t,J=8.00Hz,1H),7.58-7.49(m,2H),7.45-7.41(m,1 H),4.78-4.75(m,2H),4.02-3.98(m,2H),3.92-3.90(m,2H),2.81-2.77(m,2H),2.14-2.12(m,1H),1.86-1.83(m,2H).

[0806] To a solution of tert-butyl 4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.15 g, 0.33 mmol) in dichloromethane was added 4M HCl in diethyl ether (2.0 mL), and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated in vacuo and triturated with diethyl ether to give 4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)-5-(piperidin-4-ylmethoxy)pyrimidine (Compound I-23; 40 mg, 37% yield).

[0807] LC / MS APCI: Calculated 353.17 Found m / z [M+H] + 354.2.

[0808] 1H-NMR 400MHz, DMSO-d6: δ8.89(s,1H),8.74(bs,1H),8.61(s,1H),8.44(s,1H),8.36(s,1H),7.90(t,J=8.00Hz,1H),7.56-7.51(m,2H),7 .44(t,J=8.00Hz,1H),3.29-3.26(m,2H),3.18(s,2H),2.84-2.81(m,2H),1.96-1.93(m,1H),1.93-1.90(m,2H),1.42-1.39(m,2H).

[0809] Embodiment 37

[0810] Synthesis of Compound I-24

[0811]

[0812] Under N2 atmosphere, 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (1.39 g, 4.8 mol), 4,5-dichloropyrimidine (0.718 g, 4.85 mmol), Pd(dppf)Cl2 (0.35 g, 0.47 mmol) and Na2CO3 (0.617 g, 5.82 mmol in 2 mL water) were added to 1,4-dioxane (10 mL). The contents were heated and maintained at 110°C overnight. After the reaction was complete, the reaction mixture was cooled to room temperature, dioxane was removed under reduced pressure, and the resulting aqueous solution was extracted with EtOAc. The combined organic phase was washed three times with water and saturated brine, and dried over Na2SO4. After the solvent was removed in vacuo, the residue was purified by flash chromatography to give the pure product 5-chloro-4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidine (0.2 g, 54% yield).

[0813] LC / MS APCI: Calculated 180.02 Found m / z [M+H] + 181.0.

[0814] Under N2 atmosphere, 5-chloro-4-(1-(2-fluorophenyl)-1H-pyrazole-4-yl)pyrimidine (0.15g, 0.54mmol), 4-(aminomethyl)piperidine-1-tert-butyl formate (0.116g, 1.08mmol), Xphos second generation catalyst (42mg, 0.053mmol) and Cs2CO3 (0.354g, 5.82mmol) are added to toluene (5mL). Content is heated in microwave reactor and kept at 130 ℃ for 1 hour. After the reaction is completed, the reaction mixture is cooled to room temperature, and toluene is removed in vacuo. Residue is extracted with EtOAc. The organic phase merged is washed three times with water and saturated brine, and dried over Na2SO4. After removing the solvent in vacuo, the residue was purified by flash chromatography to give the pure product tert-butyl 4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)amino)methyl)piperidine-1-carboxylate (40 mg, 18% yield).

[0815] LC / MS APCI: Calculated 452.23 Found m / z [M+H] + 453.0.

[0816] To a solution of tert-butyl 4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)amino)methyl)piperidine-1-carboxylate 5 (40 mg, 0.088 mmol) in dichloromethane was added 4M HCl in ether (2.0 mL), and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated in vacuo and purified by preparative HPLC to give 4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)-N-(piperidin-4-ylmethyl)pyrimidin-5-amine (Compound I-24; 17 mg, 54% yield).

[0817] LC / MS APCI: Calculated 352.18 Found m / z [M+H] + 353.2.

[0818] 1H-NMR 400MHz, DMSO-d6: δ8.89(s,1H),8.86(bs,1H),8.71(bs,1),8.61(s,1H),8.44(s,1H),8.36(s,1H),7.91(t,J=8.00Hz,1H),7.56-7.51(m,2H ),7.44(t,J=8.00Hz,1H),3.39-3.26(m,2H),3.19-3.17(m,2H),2.84- 2.81(m,2H),1.95-1.93(m,1H),1.93-1.90(m,2H),1.42-1.39(m,2H).

[0819] Embodiment 38

[0820] Synthesis of Compound I-25

[0821]

[0822] To a solution of tert-butyl (cis-4-(hydroxymethyl)cyclohexyl)carbamate (0.55 g, 2.40 mmol) in THF (15 mL) was added triphenylphosphine (0.96 g, 3.6 mmol) and 4,6-dichloropyrimidin-5-ol (0.4 g, 2.4 mmol). At 0° C., diisopropyl azodicarboxylate (0.98 g, 4.85 mmol) was added to the mixture, and the mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 40: 1 as eluent) to give tert-butyl (cis-4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (0.3 g, 38% yield) as a colorless oil.

[0823] LC / MS APCI: Calculated 375.11; Found m / z [M-99 (boc)] + 276.2.

[0824] 1 H-NMR 400MHz, DMSO-d6: δ8.68(s,1H),6.75(d,J=7.60Hz,1H),3.94(d,J=6.00Hz,2H),3.14 -3.12(m,1H),1.89-1.79(m,4H),1.73-1.71(m,1H),1.38(s,9H),1.19-1.15(m,4H).

[0825] Under N2 atmosphere, tert-butyl (cis-4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (0.15 g, 0.52 mmol), 1-(2-fluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.19 g, 0.52 mmol), Pd(dppf)Cl2 (38 mg, 0.6 mmol) and Na2CO3 (66 mg, 0.56 mmol in 2 mL water) were added to 1,4-dioxane (5 mL) under N2 atmosphere. The contents were heated and maintained at 110°C overnight. After the reaction was complete, the reaction mixture was cooled to room temperature, dioxane was removed in vacuo, and the resulting aqueous solution was extracted with EtOAc. The combined organic phase was washed three times with water and saturated brine and dried over Na2SO4. After the solvent was removed in vacuo, the residue was purified by flash chromatography to give the pure product (cis-tert-butyl 4-(((4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (0.1 g, 50% yield).

[0826] LC / MS APCI: Calculated 501.19 Found m / z [M+H] + 502.2.

[0827] 1 H-NMR 400MHz, DMSO-d6: δ8.88(s,1H),8.76(s,1H),8.46(s,1H),7.92(t,J=8.00Hz,1H),7.59-7.48(m,2H),7.44-7.41(m,1H) ,6.77(bd,J=8.00Hz,1H),3.86(d,J=6.40Hz,2H),3.25-3.15(m,1H),1.93-1.82(m,5H),1.38(s,9H),1.26-1.17(m,4H).

[0828] To a methanol solution of (cis-4-(((4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamic acid tert-butyl ester (0.1 g, 0.19 mmol) was added sodium bicarbonate (35 mg, 0.38 mmol) and 30 mg of 10% Pd / carbon. The flask was equipped with a hydrogen-filled balloon fitted to a three-way stopcock. After several evacuation / hydrogen flushing cycles, the reaction mixture was stirred under 1 atm of hydrogen for 16 hours. The mixture was filtered through a diatomaceous earth pad, and the pad was rinsed with a large amount of methanol. The volatiles were removed in vacuo, and the crude solid was purified by flash column chromatography to give (cis-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamic acid tert-butyl ester (60 mg, 64% yield).

[0829] LC / MS APCI: Calculated 467.23 Found m / z [M+H] + 468.2.

[0830] To a solution of tert-butyl (cis-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (60 mg, 0.13 mmol) in dichloromethane was added 4M HCl in diethyl ether (2.0 mL), and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated in vacuo and triturated with diethyl ether to give cis-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-amine (Compound I-25; 22 mg, 46% yield).

[0831] LC / MS APCI: Calculated 367.18 Found m / z [M+H] + 368.1.

[0832] 1 H-NMR 400MHz, DMSO-d6: δ8.87(d,J=2.80Hz,1H),8.79(s,1H),8.68(s,1H),8.49(s,1H),7.95-7.92(m,1H),7.90(bs,2H),7 .55-7.47(m,2H),7.43-7.39(m,1H),4.21(d,J=7.20Hz,2H),3.27-3.25(m,1H),2.18-2.16(m,1H),1.73-1.71(m,8H).

[0833] Embodiment 39

[0834] Synthesis of Compound I-26

[0835]

[0836] Triphenylphosphine (1.43 g, 5.45 mmol) and 4,6-dichloropyrimidin-5-ol (0.68 g, 3.65 mmol) were added to a solution of tert-butyl (trans-4-(hydroxymethyl)cyclohexyl)carbamate (0.84 g, 3.65 mmol) in THF (15 mL). Diisopropyl azodicarboxylate (1.47 g, 7.27 mmol) was added to the mixture at 0°C, and the mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo, and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 40: 1 as eluent) to give tert-butyl (trans-4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (0.45 g, 33% yield) as a colorless oil.

[0837] LC / MS APCI: Calculated 375.11; Found m / z [M-99 (boc)] + 276.1.

[0838] 1 H-NMR 400MHz, DMSO-d6: δ8.68(s,1H),6.75(d,J=7.60Hz,1H),3.94(d,J=6.00Hz,2H),3.16 -3.14(m,1H),1.89-1.79(m,4H),1.73-1.71(m,1H),1.38(s,9H),1.19-1.16(m,4H).

[0839] Under N2 atmosphere, tert-butyl (trans-4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (0.26 g, 0.69 mmol), 1-(2-fluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.2 g, 0.69 mmol), Pd(dppf)Cl2 (50 mg, 0.069 mmol) and Na2CO3 (88 mg, 0.83 mmol in 2 mL water) were added to 1,4-dioxane (5 mL) under N2 atmosphere. The contents were heated and maintained at 110°C overnight. After the reaction was complete, the reaction mixture was cooled to room temperature, dioxane was removed in vacuo, and the resulting aqueous solution was extracted with EtOAc. The combined organic phase was washed three times with water and saturated brine and dried over Na2SO4. After the solvent was removed in vacuo, the residue was purified by flash chromatography to give the pure product (trans-tert-butyl 4-(((4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (0.15 g, 56% yield).

[0840] LC / MS APCI: Calculated 501.19 Found m / z [M+H] + 502.2.

[0841] To a methanol solution of (trans-4-(((4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamic acid tert-butyl ester (200 mg, 0.39 mmol) was added sodium bicarbonate (71 mg, 0.78 mmol) and 50 mg of 10% Pd / carbon. The flask was equipped with a hydrogen-filled balloon fitted to a three-way stopcock. After several evacuation / hydrogen flushing cycles, the reaction mixture was stirred under 1 atm of hydrogen for 16 hours. The mixture was filtered through a diatomaceous earth pad, and the pad was rinsed with a large amount of methanol. The volatiles were removed in vacuo, and the crude solid was purified by flash column chromatography to give (trans-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamic acid tert-butyl ester (150 mg, 55% yield). The crude reaction mixture was used in the next step without further analysis.

[0842] To a solution of tert-butyl (trans-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (150 mg, 0.32 mmol) in dichloromethane was added 4M HCl in diethyl ether (2.0 mL), and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated in vacuo and triturated with diethyl ether to give trans-4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-amine (Compound I-26; 80 mg, 72% yield for two steps).

[0843] LC / MS APCI: Calculated 367.18 Found m / z [M+H] + 368.1.

[0844] 1 H-NMR 400MHz, DMSO-d6: δ8.89(s,1H),8.80(s,1H),8.65(s,1H),8.51(s,1H),8.30-8.02(m,2H),7.95(t,J=8.00Hz,1H),7.54-7.49( m,2H),7.45-7.41(m,1H),4.16(d,J=5.60Hz,2H),3.02-3.00(m,1H),2.05-1.95(m,4H),1.43-1.40(m,1H),1.29-1.20(m,4H).

[0845] Embodiment 40

[0846] Synthesis of Compound I-28

[0847]

[0848] Under N2 atmosphere, 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.4 g, 0.20 mmol), 4,5-dichloropyrimidine (0.305 g, 0.20 mmol), Pd(dppf)Cl2 (0.15 g, 0.02 mmol) and Na2CO3 (0.26 g, 0.24 mmol) were added to 1,4-dioxane (5 mL). The contents were heated and maintained at 110°C overnight. After the reaction was complete, the reaction mixture was cooled to room temperature, dioxane was removed in vacuo, and the resulting aqueous solution was extracted with EtOAc. The combined organic phases were washed three times with water and saturated brine, and dried over Na2SO4. After the solvent was removed in vacuo, the residue was purified by flash chromatography to give the pure product 5-chloro-4-(1H-pyrazol-4-yl)pyrimidine (0.2 g, 54% yield).

[0849] LC / MS APCI: Found value m / z [M+H] + 181.0.

[0850] To a solution of 1-iodobenzene (0.35 g, 23.3 mmol) in DMSO (40 mL) was added 5-chloro-4-(1H-pyrazol-4-yl)pyrimidine (0.47 g, 19.4 mmol), followed by CuI (0.037 g, 1.94 mmol), L-proline (0.022 g, 1.94 mmol) and potassium carbonate (0.536 g, 3.88 mmol). The reaction mixture was heated to 100 °C for 48 hours. The reaction mixture was cooled to room temperature, filtered through a celite bed, and the filtrate was extracted with dichloromethane. The organic layer was concentrated and the residue was purified by flash column chromatography to give 5-chloro-4-(1-phenyl-1H-pyrazol-4-yl)pyrimidine (0.20 g, 45% yield).

[0851] LC / MS APCI: Calculated 256.05 Found m / z [M+H] + 257.1.

[0852] At 0 ° C, sodium hydride (0.185 g, 0.96 mmol) was added to a solution of tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (0.138 g, 0.64 mmol) in DMSO (2 mL), and the reaction was stirred at the same temperature for 10 minutes. At 0 ° C, 5-chloro-4-(1-phenyl-1H-pyrazole-4-yl) pyrimidine was added dropwise to the reaction mixture, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was quenched with ice and extracted with ethyl acetate. The combined organic layer was washed with a saturated NaCl aqueous solution (15 mL), dried over anhydrous Na2SO4, and concentrated in vacuo. The residue was used in the next step as is without further purification.

[0853] To a solution of tert-butyl 4-(((4-(1-phenyl-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate in dichloromethane was added 4M HCl in diethyl ether (2.0 mL), and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated in vacuo and purified by preparative HPLC to give 4-(1-phenyl-1H-pyrazol-4-yl)-5-(piperidin-4-ylmethoxy)pyrimidine (Compound I-28; 40 mg, 20% yield for two steps).

[0854] LC / MS APCI: Calculated 335.17 Found m / z [M+H] + 336.1.

[0855] 1 H-NMR 400MHz, DMSO-d6: δ9.09(s,1H),8.88(bs,1H),8.80(d,J=8.00Hz,1H),8.67(d,J=14.40Hz,1H),8 .45(d,J=6.40Hz,1H),7.96(d,J=8.00Hz,2H),7.56(t,J=8.00Hz,2H),7.40(t,J=7.20Hz,1H),4. 25(d,J=6.00Hz,2H),3.43-3.39(m,1H),3.35-3.31(m,1H),3.00-2.95(m,1H),2.83-2.80(m,1H) ,2.33-2.29(m,1H),2.00-1.97(m,1H),1.75-1.00(m,1H),1.56-1.53(m,1H),1.36-1.33(m,1H).

[0856] Embodiment 41

[0857] Synthesis of Compound I-29 and Compound I-30

[0858]

[0859] Pyrazole (2.0 g, 29.41 mmol) was added to a solution of iodobenzene (7.19 g, 35.2 mmol) in DMSO (40 mL), followed by CuI (0.55 g, 2.89 mmol), L-proline (0.337 g, 2.93 mmol) and potassium carbonate (8.1 g, 58.7 mmol). The reaction mixture was heated to 100 ° C for 48 hours. The reaction mixture was cooled to room temperature, filtered through a diatomaceous earth bed, and the filtrate was extracted with dichloromethane. The organic layer was concentrated and the residue was purified by flash column chromatography to give 1-phenyl-1H-pyrazole (3 g, 71% yield).

[0860] LC / MS APCI: Calculated 144.07 Found m / z [M+H] + 145.1.

[0861] 1 H-NMR 400MHz, DMSO-d6: δ8.50(d,J=2.00Hz,1H),7.85(d,J=8.40Hz,2H),7.75(s, 1H), 7.49 (t, J = 7.20Hz, 2H), 7.30 (t, J = 7.60Hz, 1H), 6.54 (t, J = 1.60Hz, 1H).

[0862] Under argon atmosphere, to a solution of 1-phenyl-1H-pyrazole (0.5 g, 3.47 mmol) in trifluoromethylbenzene, N-fluorobenzenesulfonimide (NFSI, 3.28 g, 10.4 mmol) was added, followed by palladium acetate (0.077 g, 0.34 mmol), and the reaction mixture was heated at 100 ° C for 16 hours. The reaction mixture was filtered through celite and concentrated in vacuo. The crude residue was purified by flash column chromatography to give 1- (2,6-difluorophenyl) -1H-pyrazole (0.4 g, 64% yield).

[0863] LC / MS APCI: Calculated 180.05 Found m / z [M+H] + 181.1.

[0864] 1 H-NMR 400MHz, DMSO-d6: δ8.11(d,J=2.00Hz,1H),7.82(d,J=5.20Hz,1H),7.64-7.57(m,1H),7.40-7.34(m,2H),6.57(t,J=2.00Hz,1H).

[0865] To a solution of 1-(2,6-difluorophenyl)-1H-pyrazole (1.5 g, 8.33 mmol) in acetic acid (5 mL) was added a solution of bromine (0.64 g, 4.02 mmol) in acetic acid (5 mL) at 0°C. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was cooled to room temperature, poured into ice and H2O in a 100 mL beaker, and an excess of saturated aqueous NaHCO3 was added until all the acetic acid had been quenched. Ethyl acetate (50 mL) was added and the layers were separated. The aqueous layer was extracted twice with ethyl acetate, and the combined extracts were dried over Na2SO4 and concentrated in vacuo to give a crude solid, which was purified by flash column chromatography (SiO2, 1:1 hexane-ethyl acetate) to give 4-bromo-1-(2,6-difluorophenyl)-1H-pyrazole (1 g, 46% yield).

[0866] LC / MS APCI: Calculated 257.96 Found m / z [M+H] + 258.9.

[0867] 1 H-NMR 400MHz, DMSO-d6: δ8.46(s,1H),7.96(s,1H),7.67-7.61(m,1H),7.43-7.38(m,2H).

[0868] A mixture of 4-bromo-1-(2,6-difluorophenyl)-1H-pyrazole (1.6 g, 6.1 mmol), potassium acetate (1.21 g, 12.34 mmol), bis(pinacol)diboron (1.87 g, 7.39 mmol) and Pd(dppf)Cl2 (0.45 g, 0.615 mmol) was suspended in DMSO (4 mL) and 1,4-dioxane (15 mL). The system was purged with N2. The mixture was stirred at 100°C for 16 hours, cooled, and diluted with water (60 mL). The resulting mixture was extracted with dichloromethane (50 mL×3), and the combined organic layers were washed with saturated aqueous NaCl solution (15 mL), dried over anhydrous Na2SO4, and concentrated in vacuo. The residue was purified by silica gel column chromatography eluting with 20:1 hexanes / ethyl acetate to give 1-(2,6-difluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.6 g, 33% yield).

[0869] LC / MS APCI: Calculated 306.14 Found m / z [M+H] + 307.0.

[0870] Under N2 atmosphere, tert-butyl 4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.6 g, 1.6 mmol), 1-(2,6-difluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.50 g, 1.65 mmol), Pd(dppf)Cl2 (0.12 g, 0.16 mmol) and Na2CO3 (0.21 g, 1.98 mmol in 2 mL water) were added to 1,4-dioxane (5 mL) under N2 atmosphere. The contents were heated and maintained at 110°C overnight. After the reaction was complete, the reaction mixture was cooled to room temperature, dioxane was removed in vacuo, and the resulting aqueous solution was extracted with EtOAc. The combined organic phase was washed three times with water and saturated brine and dried over Na2SO4. After the solvent was removed in vacuo, the residue was purified by flash chromatography to give the pure product tert-butyl 4-(((4-chloro-6-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.4 g, 40% yield).

[0871] LC / MS APCI: Calculated 505.17 Found m / z [M+H] + 506.1.

[0872] 1H-NMR 400MHz, DMSO-d6: δ8.88(s,1H),8.84(s,1H),8.45(s,1H),7.67-7.63(m,1H),7.46-7.38(m,2H),4.80-4.78(m,2 H),4.04-4.00(m,2H),3.96-3.92(m,2H),2.32-2.29(m,1H),1.99-1.96(m,2H),1.62-1.57(m,2H),1.17(s,9H).

[0873] To a solution of tert-butyl 4-(((4-chloro-6-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.2 g, 0.39 mmol) in methanol was added sodium bicarbonate (0.071 g, 0.788 mmol) and 25 mg of 10% Pd / carbon. The flask was equipped with a hydrogen-filled balloon fitted to a three-way stopcock. After several evacuation / hydrogen flushing cycles, the reaction mixture was stirred under 1 atm of hydrogen for 16 hours. The mixture was filtered through a pad of celite and the pad was rinsed with copious amounts of methanol. The volatiles were removed in vacuo to give tert-butyl 4-(((4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.10 g, 54% yield) as a crude solid which was used in the next step without further purification.

[0874] To a solution of tert-butyl 4-(((4-chloro-6-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.1 g, 0.19 mmol) in dichloromethane was added 4M HCl in diethyl ether (2.0 mL), and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated in vacuo and purified by preparative HPLC to give 4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)-5-(piperidin-4-ylmethoxy)pyrimidine (Compound I-29; 12 mg, 15% yield).

[0875] LC / MS APCI: Calculated 405.12 Found m / z [M+H] + 406.0.

[0876] 1H-NMR 400MHz, DMSO-d6: δ8.86(s,1H),8.79(s,1H),8.64(s,1H),8.46(s,1H),7.70-7.65(m,1H),7.46-7.42(m,2H),3.92 (d,J=6.00Hz,2H),2.94-2.91(m,2H),2.51-2.47(m,2H),2.03-1.99(m,1H),1.57-1.48(m,2H),1.24-1.16(m,2H).

[0877] To a solution of tert-butyl 4-(((4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)piperidine-1-carboxylate (0.10 g, 0.19 mmol) in dichloromethane was added 4M HCl in diethyl ether (2.0 mL), and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was concentrated in vacuo and purified by preparative HPLC to give 4-(1-(2,6-difluorophenyl)-1H-pyrazol-4-yl)-5-(piperidin-4-ylmethoxy)pyrimidine (Compound I-30; 29.4 mg, 37% yield).

[0878] LC / MS APCI: Calculated 371.16 Found m / z [M+H] + 372.1.

[0879] 1 H-NMR 400MHz, DMSO-d6: δ8.81(s,1H),8.80(s,1H),8.68(s,1H),8.57(bs,1H),8.51(s,1H),7.68-7.64(m,1H),7.45-7.41(m,2H ), 4.23 (d, J = 6.00Hz, 2H), 3.32-3.29 (m, 2H), 2.93-2.88 (m, 2H), 2.28-2.26 (m, 1H), 1.97-1.94 (m, 2H), 1.58-1.52 (m, 2H).

[0880] Embodiment 42

[0881] Synthesis of Compound I-31

[0882]

[0883] To a solution of 4-(hydroxymethyl)cyclohexane-1-ol (0.237 g, 1.82 mmol) in THF (15 mL) were added triphenylphosphine (0..718 g, 2.74 mmol) and 4,6-dichloropyrimidin-5-ol (0.3 g, 1.82 mmol). Di-tert-butyl azodicarboxylate (0.63 g, 2.74 mmol) was added to the mixture at 0°C, and the mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated in vacuo. The residue was purified by silica gel column chromatography (hexane: ethyl acetate = 9: 1 as eluent) to give 4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexane-1-ol (0.15 g, 30% yield).

[0884] LC / MS APCI: Calculated 276.04; Found m / z [M+H)] + 277.0.

[0885] 4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexane-1-ol (0.10 g, 0.36 mmol), 1-(2-fluorophenyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.104 g, 0.36 mmol), Pd(dppf)Cl (25 mg, 0.034 mmol) and NaCO (46 mg, 0.43 mmol in 1 mL water) were added to 1,4-dioxane (5 mL) under N2 atmosphere. The contents were heated and maintained at 110°C overnight. After the reaction was complete, the reaction mixture was cooled to room temperature, dioxane was removed in vacuo, and the resulting aqueous solution was extracted with EtOAc. The combined organic phases were washed three times with water and saturated brine, and dried over Na2SO4. After the solvent was removed in vacuo, the residue was purified by flash chromatography to give the pure product 4-(((4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexan-1-ol (60 mg, 41% yield).

[0886] LC / MS APCI: Calculated 402.13 Found m / z [M+H] + 403.2.

[0887] To a solution of 4-(((4-chloro-6-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-ol (60 mg, 0.15 mmol) in methanol was added sodium bicarbonate (26 mg, 0.28 mmol) and 20 mg of 10% Pd / carbon. The flask was equipped with a hydrogen-filled balloon fitted to a three-way stopcock. After several evacuation / hydrogen flush cycles, the reaction mixture was stirred under 1 atm of hydrogen for 16 hours. The mixture was filtered through a pad of celite, and the pad was rinsed with copious amounts of methanol. The volatiles were removed in vacuo, and the crude solid was purified by flash column chromatography to give 4-(((4-(1-(2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-ol (Compound I-31; 15.0 mg, 28% yield).

[0888] LC / MS APCI: Calculated 368.16 Found m / z [M+H] + 369.0.

[0889] 1 H-NMR 400MHz, MeOD: δ8.92-8.89(m,1H),8.73(d,J=8.80Hz,1H),8.53-7.98(m,2H),7.98-7.91(m,1H),7.48-7.37(m,3H),4.64-4.62( m,1H),4.15(d,J=14.00Hz,2H),3.58-3.56(m,1H),2.36-2.34(m,1H),2.08-2.05(m,2H),1.68-1.65(m,2H),1.40-1.37(m,4H).

[0890] Embodiment 43

[0891] Synthesis of Compound I-32

[0892]

[0893] To a solution of tert-butyl ((1s,4s)-4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (1 g, 2.68 mmol) in 1,4-dioxane (30 mL) was added 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.51 g, 2.68 mmol) and 2M aqueous Na2CO3 (2.68 mL, 2 equiv). The resulting reaction mixture was purged with nitrogen for 20 min, and then Pd(dppf)Cl2 (195 mg, 0.268 mmol) was added. The reaction mixture was heated to 100°C in a sealed tube for 12 h. The reaction mixture was cooled, diluted with water and extracted with ethyl acetate. The combined organic phases were washed with water and brine solution and dried over Na2SO4. After removing the solvent in vacuo, the crude compound was purified by silica gel column chromatography to obtain tert-butyl ((1s,4s)-4-(((4-chloro-6-(1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (300 mg, 27.6%) as a gray-brown solid.

[0894] LC / MS: Calculated 407.17 Found m / z [MH] + 406.2.

[0895] To a solution of tert-butyl ((1s, 4s)-4-(((4-chloro-6-(1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (250mg, 0.61mmol) and (4-(4-methylpiperazine-1-yl)phenyl)boronic acid (270mg, 1.22mmol) in dichloromethane (20mL) was added Cu(OAc)2(445mg, 2.45mmol), molecular sieve powder (400mg) and piperidine (145mg, 1.83mmol). The reaction mixture was stirred at room temperature for 48 hours. The mixture was filtered through a bed of celite and concentrated in vacuo. The crude compound was purified by preparative HPLC to afford tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(4-(4-methylpiperazin-1-yl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (110 mg, 30.8%).

[0896] LC / MS: Calculated 581.29 Found m / z [M+H] + 582.3.

[0897] To a solution of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(4-(4-methylpiperazin-1-yl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (110 mg, 0.19 mmol) in methanol (10 mL) was added triethylamine (76 mg, 0.76 mmol) followed by 10% Pd / carbon (220 mg) and stirred under a hydrogen atmosphere for 2 hours. The reaction mixture was concentrated in vacuo and purified by silica gel column chromatography to give tert-butyl ((1s,4s)-4-(((4-(1-(4-(4-methylpiperazin-1-yl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (60 mg, 58%).

[0898] LC / MS: Calculated 547.33 Found m / z [M+H] + 548.3.

[0899] To a solution of tert-butyl ((1s,4s)-4-(((4-(1-(4-(4-methylpiperazin-1-yl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (60 mg, 0.11 mmol) in 3 mL of dioxane was added 4M HCl in dioxane (3.0 mL) and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated in vacuo and triturated with diethyl ether to give (1s,4s)-4-(((4-(1-(4-(4-methylpiperazin-1-yl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-amine hydrochloride (Compound I-32; 50 mg, 94.5%) as a yellow solid.

[0900] LC / MS: Calculated 447.27 Found m / z [M+H] + 448.2.

[0901] HPLC (purity): 97.6%.

[0902] 1H-NMR (400MHz, DMSO-d6): δ10.81(brs,1H),8.95(s,1H),8.78(s,1H),8.67(s,1H),8.38(s,1H),8.02(brs,3H),7.79(d,J=9.2Hz,2H),7.16(d,J= 9.2Hz,2H),4.22(d,J=7.2Hz,2H),3.90(d,J=9.2Hz,2H),3.38(m,1H),3. 25-3.17(m,4H),2.83&2.81(s,3H),2.32-2.20(m,1H),1.73-1.70(m,8H).

[0903] Embodiment 44

[0904] Synthesis of Compound I-33

[0905]

[0906] To a solution of tert-butyl ((1s, 4s)-4-(((4-chloro-6-(1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (250mg, 0.61mmol) and (4-morpholinophenyl)boronic acid (253mg, 1.22mmol) in dichloromethane (20mL), Cu(OAc)2(445mg, 2.45mmol), molecular sieve powder (400mg) and piperidine (145mg, 1.83mmol) were added. The reaction mixture was stirred at room temperature for 48 hours. The mixture was filtered through a bed of diatomaceous earth and concentrated in vacuo. The crude compound was purified by preparative HPLC to afford tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(4-morpholinophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (130 mg, 37%).

[0907] LC / MS: Calculated 568.26 Found m / z [M+H] + 569.3.

[0908] To a solution of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(4-morpholinophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (130 mg, 0.23 mmol) in methanol (10 mL) was added triethylamine (92 mg, 0.91 mmol) followed by 10% Pd / carbon (260 mg) and the mixture was stirred under a hydrogen atmosphere for 2 hours. The reaction mixture was concentrated in vacuo and purified by silica gel column chromatography to give tert-butyl ((1s,4s)-4-(((4-(1-(4-morpholinophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (55 mg, 45%).

[0909] LC / MS: Calculated 534.3 Found m / z [M+H] + 535.2.

[0910] To a solution of tert-butyl ((1s,4s)-4-(((4-(1-(4-morpholinophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (55 mg, 0.10 mmol) in 3 mL of dioxane was added 4M HCl in dioxane (3.0 mL), and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated in vacuo and triturated with diethyl ether to give (1s,4s)-4-(((4-(1-(4-morpholinophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-amine hydrochloride (Compound I-33, 32 mg, 66%) as a yellow solid.

[0911] LC / MS: Calculated 434.24 Found m / z [M+H] + 435.2.

[0912] HPLC (purity): 93.66%

[0913] 1 H-NMR (400MHz, DMSO-d6): δ8.94(s,1H),8.79(s,1H),8.68(s,1H),8.38(s,1H),7.97(brs,3H),7.76(d,J=9.2Hz,2H),7.11(d, J=9.2Hz,2H),4.22(d,J=7.2Hz,2H),3.78(t,J=4.8Hz,4H),3.26(m,1H),3.19(t,J=4.8Hz,4H),2.23(m,1H),1.73-1.70(m,8H).

[0914] Embodiment 45

[0915] Synthesis of Compound I-34

[0916]

[0917] To a solution of 4-fluorobenzylthiophenol (5 g, 39 mmol) and iodoethane (3.78 mL, 46.8 mmol) in tetrahydrofuran (50 mL) was added triethylamine (5.44 mL, 39 mmol). The reaction mixture was stirred at ambient temperature for 2 hours and then filtered. The filtrate was concentrated, triturated with pentane and dried in vacuo to give ethyl (4-fluorophenyl) sulfane (2.8 g, 46%). The reaction mixture was used in the next step without further analysis.

[0918] A solution of ethyl (4-fluorophenyl) sulfane (2.5 g, 16 mmol) in dichloromethane (50 mL) was treated with 3-chloroperoxybenzoic acid (8.65 g, 35.2 mmol). The reaction mixture was stirred at ambient temperature for 6 hours and then filtered. The filtrate was washed with saturated aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography to obtain 1- (ethylsulfonyl) -4-fluorobenzene (2.2 g, 73%).

[0919] 1 H-NMR (400MHz, CDCl3): δ7.97-7.93 (m, 2H), 7.30-7.25 (m, 2H), 3.14 (q, J = 10.0Hz, 2H), 1.28 (t, J = 10.0Hz, 3H).

[0920] To a solution of 1-(ethylsulfonyl)-4-fluorobenzene (2.2 g, 11.7 mmol) and 4-bromo-1H-pyrazole (1.7 g, 11.7 mmol) in DMF (20 mL) was added CsCO (11.4 g, 35.1 mmol), and the reaction was heated at 80 ° C for 3 hours. The reaction mixture was cooled, treated with water and extracted with ethyl acetate (2×50 mL). The combined organic layers were washed with brine solution, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography to obtain 4-bromo-1-(4-(ethylsulfonyl)phenyl)-1H-pyrazole (1.4 g, 38%).

[0921] LC / MS: Calculated 315.97 Found m / z [M+H] + 317.0.

[0922] To a solution of 4-bromo-1-(4-(ethylsulfonyl)phenyl)-1H-pyrazole (1.2 g, 3.8 mmol) and (n-Bu)6Sn2 (6.6 g, 11.4 mmol) in xylene (10 mL) was added Pd(PPh3)4 (439 mg, 0.38 mmol) and heated at 130°C for 48 hours. The reaction mixture was filtered, evaporated and purified by neutral alumina column to give 1-(4-(ethylsulfonyl)phenyl)-4-(tributylstannyl)-1H-pyrazole (250 mg, 12.5%) which was used directly in the next step.

[0923] To a degassed solution of tert-butyl ((1s,4s)-4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (160 mg, 0.43 mmol) and 1-(4-(ethylsulfonyl)phenyl)-4-(tributylstannyl)-1H-pyrazole (247 mg, 0.47 mmol) in dioxane (10 mL) was added PdCl2(PPh3)2 (30 mg, 0.043 mmol) and the reaction mixture was heated at 110° C. for 16 h. The reaction mixture was filtered through a bed of celite and concentrated in vacuo. The crude compound was purified by silica gel column chromatography to obtain tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(4-(ethylsulfonyl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate 7 (120 mg, 49%).

[0924] LC / MS: Calculated 575.20 Found m / z [M+H] + 476.0 (corresponding to the mass of the lost BOC group)

[0925] To a solution of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(4-(ethylsulfonyl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate 7 (120 mg, 0.21 mmol) in methanol (10 mL) was added triethylamine (84 mg, 0.83 mmol), followed by 10% Pd / carbon (240 mg), and the mixture was stirred under a hydrogen atmosphere for 2 hours. The reaction mixture was concentrated in vacuo and purified by silica gel column chromatography to give tert-butyl ((1s,4s)-4-(((4-(1-(4-(ethylsulfonyl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (32 mg, 28%).

[0926] LC / MS: Calculated 541.24 Found m / z [M+H] + 442.2 (corresponding to the mass lost from the BOC group).

[0927] To a solution of tert-butyl ((1s,4s)-4-(((4-(1-(4-(ethylsulfonyl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (32 mg, 0.06 mmol) in 2 mL of dioxane was added 4M HCl in dioxane (2.0 mL), and the reaction mixture was stirred at room temperature for 8 hours. The reaction mixture was concentrated in vacuo and triturated with diethyl ether to give (1s,4s)-4-(((4-(1-(4-(ethylsulfonyl)phenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-amine hydrochloride (Compound I-34; 20 mg, 71%) as an off-white solid.

[0928] LC / MS: Calculated 441.18 Found m / z [M+H] + 442.1.

[0929] HPLC (purity): 95.19%.

[0930] 1 H-NMR (400MHz, DMSO-d6): δ9.24(s,1H),8.83(s,1H),8.75(s,1H),8.51(s,1H),8.23(d,J=8.8Hz,2H),8.04(d,J=8.8Hz,2H), 7.91(brs,3H),4.24(d,J=7.2Hz,2H),3.36(q,7.2Hz,2H),3.25(m,1H),2.24(m,1H),1.73-1.70(m,8H),1.13(t,J=7.2Hz,3H).

[0931] Embodiment 46

[0932] Synthesis of Compound I-35

[0933]

[0934] To a solution of 3,4-difluorobenzylthiophenol (3 g, 20.5 mmol) and iodoethane (2.0 mL, 24.6 mmol) in tetrahydrofuran (30 mL) was added triethylamine (2.86 mL, 20.5 mmol). The reaction mixture was stirred at ambient temperature for 2 hours and then filtered. The filtrate was concentrated, triturated with pentane and dried in vacuo to give (3,4-difluorophenyl)(ethyl)sulfane (2.6 g, 73%).

[0935] GC-MS: Calculated 174.03 Found m / z 174.1.

[0936] A solution of (3,4-difluorophenyl)(ethyl)sulfane (2.6 g, 14.9 mmol) in dichloromethane (50 mL) was treated with 3-chloroperbenzoic acid (8.1 g, 32.8 mmol). The reaction mixture was stirred at ambient temperature for 6 hours and then filtered. The filtrate was washed with saturated aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography to obtain 4-(ethylsulfonyl)-1,2-difluorobenzene (3.0 g, 97.4%).

[0937] GC-MS: Calculated 206.02 Found m / z 206.0.

[0938] To a solution of 4-(ethylsulfonyl)-1,2-difluorobenzene (2.5 g, 12.1 mmol) and 4-bromo-1H-pyrazole (1.77 g, 12.1 mmol) in DMF (20 mL) was added CsCO (11.8 g, 36.3 mmol), and the reaction was heated at 80 ° C for 3 hours. The reaction mixture was cooled, treated with water and extracted with ethyl acetate (2×50 mL). The combined organic layers were washed with saline solution, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography to obtain 4-bromo-1-(4-(ethylsulfonyl)-2-fluorophenyl)-1H-pyrazole (2.3 g, 57%).

[0939] LC / MS: Calculated 331.96 Found m / z [M+H] + 333.0.

[0940] To a solution of 4-bromo-1-(4-(ethylsulfonyl)-2-fluorophenyl)-1H-pyrazole (500 mg, 1.5 mmol) and (n-Bu)6Sn2 (2.62 g, 4.5 mmol) in xylene (10 mL) was added Pd(PPh3)4 (173 mg, 0.15 mmol) and heated at 130°C for 24 hours. The reaction mixture was filtered, evaporated and purified by silica gel column chromatography to give 1-(4-(ethylsulfonyl)-2-fluorophenyl)-4-(tributylstannyl)-1H-pyrazole (200 mg, 24.5%).

[0941] LC / MS: Calculated 544.16 Found m / z [M+H] + 545.2.

[0942] To a solution of tert-butyl ((1s,4s)-4-(((4,6-dichloropyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (125 mg, 0.33 mmol) and 1-(4-(ethylsulfonyl)-2-fluorophenyl)-4-(tributylstannyl)-1H-pyrazole (200 mg, 0.37 mmol) in dioxane (10 mL) was added PdCl2(PPh3)2 (23.1 mg, 0.033 mmol) and the reaction mixture was heated at 110° C. for 16 h. The reaction mixture was filtered through a bed of celite and concentrated in vacuo. The crude compound was purified by silica gel column chromatography to obtain tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(4-(ethylsulfonyl)-2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate 7 (150 mg, 76%).

[0943] LC / MS: Calculated 593.19 Found m / z [M+H] + 494.1 (corresponding to the mass lost from the BOC group).

[0944] To a solution of tert-butyl ((1s,4s)-4-(((4-chloro-6-(1-(4-(ethylsulfonyl)-2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate 7 (130 mg, 0.22 mmol) in methanol (10 mL) was added triethylamine (89 mg, 0.88 mmol) followed by 10% Pd / carbon (260 mg) and the mixture was stirred under a hydrogen atmosphere for 2 hours. The reaction mixture was concentrated in vacuo and purified by silica gel column chromatography to give tert-butyl ((1s,4s)-4-(((4-(1-(4-(ethylsulfonyl)-2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (70 mg, 57%).

[0945] LC / MS: Calculated 559.23 Found m / z [M+H] + 460.1 (corresponding to the mass lost from the BOC group).

[0946] To a solution of tert-butyl ((1s,4s)-4-(((4-(1-(4-(ethylsulfonyl)-2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexyl)carbamate (70 mg, 0.125 mmol) in 4 mL of dioxane was added 4M HCl in dioxane (4.0 mL), and the reaction mixture was stirred at room temperature for 3 hours. The reaction mixture was concentrated in vacuo and triturated with diethyl ether to give (1s,4s)-4-(((4-(1-(4-(ethylsulfonyl)-2-fluorophenyl)-1H-pyrazol-4-yl)pyrimidin-5-yl)oxy)methyl)cyclohexane-1-amine hydrochloride (Compound I-35; 60 mg, 96%) as an off-white solid.

[0947] LC / MS: Calculated 459.17 Found m / z [M+H] + 460.1.

[0948] HPLC (purity): 98.57%.

[0949] 1 H-NMR (400MHz, DMSO-d6): δ9.01(d,J=2.8Hz,1H),8.83(s,1H),8.72(s,1H),8.59(s,1H),8.27(t,J=8.0Hz,1H),8.08(dd,J=1.6Hz,11.2Hz,1H),8.0 2(brs,3H),7.92(d,J=8.1Hz,1H),4.23(d,J=7.2Hz,2H),3.34(q,J=7.2Hz ,2H),3.29(m,1H),2.17(m,1H),1.75-1.72(m,8H),1.16(t,J=7.2Hz,3H).

[0950] Embodiment 47

[0951] Formation of Exemplary Pharmaceutically Acceptable Salts

[0952] The hydrochloride salt of the foregoing compound was prepared by dissolving the compound in a minimum amount of ethanol and adding 20% ​​ethanolic HCl solution dropwise while the mixture was stirred for 1 hour, then adding diethyl ether. The precipitated off-white solid hydrochloride salt was isolated by filtration, washed with diethyl ether and dried.

[0953] In addition, salts can be prepared by reacting the free acid or free base form of a compound of the present disclosure with a stoichiometric amount of an appropriate base or acid in water or an organic solvent, or in a mixture of the two; generally, non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile are preferred. A list of suitable salts is found in Remington's Pharmaceutical Sciences. 18th edition, Mack Publishing Company, Easton, PA, 1990, p. 1445.

[0954] Embodiment 48

[0955] Inhibition assay

[0956] Experimental Procedure:

[0957] Kinase assays were performed at room temperature. Compounds were quantified using acoustic technology at a dose of 10 IC 50 The pattern was added to the enzyme / substrate mixture and the compound was pre-incubated for 20 minutes. Five concentrations of ATP were then added to initiate the reaction. Activity was monitored every 5-15 minutes for time course studies. The ATP, substrate, and compound concentrations tested were as follows:

[0958] ATP competitive evaluation and K of MELK and FLT3 i Determination:

[0959] ATP concentrations were tested with 10, 30, 75, 150 and 300 μM ATP and the substrate concentration was kept constant at 20 μM. For test compounds, 10 doses of IC were tested in 3-fold serial dilutions starting from 100 μM. 50 The time points were measured at 0, 5, 10, 15, 20, 30, 45, 60, 75, 90, 105 and 120 minutes.

[0960] Design experiments to obtain the K of ATP in the presence of different concentrations of inhibitors m value; ideally including IC 25 、IC 50 and IC 75 However, if the compound is an ATP-competitive inhibitor, then the IC 50 The value will shift. Therefore, a 10 dose IC 50 The mode was set to ensure that the optimal concentration range was covered. A 20-minute preincubation of the compound and enzyme was performed to ensure that the compound and enzyme were bound and balanced. The reaction was monitored every 5-15 minutes to obtain a progress curve that varied with time. At each time point, the radioisotope signal ( 33The slope of the linear part of the progress curve was obtained by linear regression using Microsoft Excel. The slope (or speed; μM / min) was then plotted against ATP concentration in Michaelis-Menten diagrams and subsequent Lineweaver-Burk diagrams (double reciprocal diagrams) using GraphPad Prism software. The results were further analyzed using the global fit of GraFit software. The inhibition of MELK and FLT3 (ITD) by the test compound was not time-dependent and was competitive relative to ATP.

[0961] Embodiment 49

[0962] General Protein Kinase Assay Method for Selected Kinases

[0963] In vitro analysis of the MELK, MELK(T460M) kinome was performed at Reaction Biology using the "HotSpot" assay platform. In vitro analysis of FLT3, FLT3 mutant forms (FLT3(D835Y), FLT3(F594R595insR), FLT3(F594R595insREY), FLT3(ITD), FLT3(ITD)-NPOS, FLT3(ITD)-W51, and FLT3(R595_E596insEY) kinome) was performed in the same manner.

[0964] Briefly, the specific kinase / substrate pair and the required cofactors were prepared in reaction buffer: 20 mM Hepes (pH 7.5), 10 mM MgCl2, 1 mM EGTA, 0.02% Brij35, 0.02 mg / mL BSA, 0.1 mM Na3VO4, 2 mM DTT, 1% DMSO. The compounds were added to the reaction and ATP (Sigma) and 33 The mixture of P ATP (PerkinElmer) to a final concentration of 10 μM. The reaction was carried out at 25°C for 120 minutes, and then the reaction was spotted onto P81 ion exchange filter paper (Whatman). Unbound phosphate was removed by washing the filter thoroughly with 0.75% phosphoric acid. After subtracting the background from the control reaction containing inactive enzyme, the kinase activity data are expressed as the percentage of the remaining kinase activity in the test sample compared to the vehicle (dimethyl sulfoxide) reaction. IC was obtained using Prism (GraphPad software) 50 Values ​​and curve fitting.

[0965] Table 2. Compound IC for MELK and FLT3 50List of values ​​(nM)

[0966]

[0967]

[0968] + indicates IC greater than 1,000 nM 50 value

[0969] ++ indicates IC between 200 and 1,000 nM 50 value

[0970] +++ indicates IC up to 200nM 50 value

[0971] - means no data

[0972] Embodiment 50

[0973] Cancer cell efficacy research

[0974] The compound of test structure (I) is to the influence of cancer cell lines from different cancers, to determine the ability of compound to influence the growth of cell lines from different tissue sources. This study is carried out on a wider range of disease groups and multiple hematological cell lines on the Oncolead GmbH cancer cell line test system. Cell lines are purchased directly from ATCC, NCI, CLS and DSMZ cell line collections by Oncolead. Master libraries and working aliquots are prepared from these cell lines, and the cell experience for research is less than 20 generations. In order to ensure that there is no potential contamination or wrong allocation, all cell lines are tested by STR analysis. Confirm that all cell lines used in the study are free of mycoplasma and SMRV contamination. In addition, all studies include a group of 6 reference agents-paclitaxel, 5-FU, SN-38 (active metabolite of irinotecan), sorafenib, geldanamycin and doxorubicin-to ensure the reproducible response of cell lines and provide a control of day-to-day experimental changes. 82 cell lines are tested in parallel. The test concentration range represents 6 data points of 10 times dilution. Each experiment was performed in duplicate, arranged in different microtiter plates and using independent compound dilutions. The experimental setup allowed minimizing systematic errors and accounting for intra-experimental variations.

[0975] The most sensitive and resistant cell lines are displayed by using a box plot or by selecting the 8 most and least sensitive cell lines using the z-score for each agent. This also applies to the cell lines where the activity of the test compound against the panel of cancer cell lines is plotted against the Z-score sensitivity and resistance readout. In this analytical assay, the specific AML cancer cell line MV4-11 was sensitive and a few blood cancer cell lines were observed to be sensitive to the test compounds.

[0976] Embodiment 51

[0977] In vivo models of AML

[0978] To evaluate the antitumor efficacy of compounds of structure (I) in MV4-11 (ATCC CRL-9591) human biphenotypic B myelomonocytic leukemia (lymphoblastic) cell line xenografts in female SCID-Beige mice, MV4-11 (ATCC CRL-9591 cell line obtained from ATCC) was propagated in cell culture RPMI + 10% fetal bovine serum.

[0979] Experimental design:

[0980] MV4-11 cells were harvested at logarithmic growth and implanted subcutaneously (5 × 10 6 Tumor growth was monitored daily and when tumors reached 100-120 mm 3 At , mice were randomly divided into 3 groups of 8 mice each and administered compounds using a positive control (paritinib) and a negative control (vehicle only) and a vehicle containing 10% PG, 25% WFI of Solutol and 65% PBS. All groups were orally administered once daily at a dose volume of 10 mL / Kg.

[0981] Table 3. Dosing parameters used in the in vivo xenograft model

[0982]

[0983] When the tumor reaches 2000mm 3 Animals were sacrificed when the tumor volume decreased to 10% of the body weight or when ulceration and necrosis occurred. After sacrifice, tumor tissues were dissected and snap-frozen in liquid nitrogen, and the samples were stored at -80°C until shipped to the funder. Tumor volume was measured by digital vernier caliper using the following formula:

[0984] (minimum diameter 2 )×maximum diameter / 2

[0985] Body weight was monitored daily and tumor volume was measured three times per week. Plasma samples were collected at 1, 2, 4 and 6 hours for bioanalysis and PK / PD correlation studies were performed before study termination. Four animals from the second group of test compound treated animals were retained without sacrifice for observation after completion of the 33-day dosing.

[0986] All U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications (e.g., US62 / 751,405 filed on October 26, 2018) mentioned in this specification or the attached application data sheets are incorporated herein by reference in their entirety to the extent not inconsistent with this specification.

[0987] From the foregoing it will be appreciated that although specific embodiments of the present disclosure have been described herein for purposes of illustration, various modifications may be made without departing from the spirit and scope of the present invention. Accordingly, the present disclosure is not to be limited, except as by the appended claims.

Claims

1. A compound having the following structure (I): or a pharmaceutically acceptable salt or stereoisomer thereof, wherein: is cyclobutyl, cyclopentyl, cyclohexyl or piperidinyl, wherein the cyclobutyl, cyclopentyl, cyclohexyl and piperidinyl are each optionally substituted with at least one substituent selected from C1-C6 alkyl, hydroxyl, amino, C1-C6 alkylamino, aminoC1-C6 alkyl, C1-C6 alkylaminoC1-C6 alkyl or a combination thereof; is phenyl, wherein the phenyl is optionally substituted with at least one substituent selected from C1-C6 alkyl, halogen, halogenated C1-C6 alkyl, halogenated C1-C6 alkoxy, cyano, C1-C6 alkoxy, hydroxy, C1-C6 alkylsulfonyl, morpholinyl and N-methylpiperazinyl; L is a direct bond or a heteroalkylene linker having one of the following structures: Where R 2 is independently H or C1-C6 alkyl at each occurrence; R 1a is H, halogen or C1-C6 alkyl; and R 1b , R 1c and R 1d Each is independently H or C1-C6 alkyl.

2. The compound of claim 1, wherein R 1a is H or halogen, and R 1b , R 1c and R 1d Each is independently H or C1-C6 alkyl.

3. The compound according to any one of claims 1 to 2, wherein R 1c and R 1d Each is H.

4. The compound according to any one of claims 1 to 2, wherein R 1a It's a halogen.

5. The compound of claim 4, wherein the halogen is chlorine.

6. The compound according to any one of claims 1 to 2, wherein R 1b It's H.

7. The compound according to any one of claims 1 to 2, wherein R 1a and R 1b Each is H.

8. The compound of any one of claims 1 to 2, wherein L is the direct bond.

9. The compound of any one of claims 1 to 2, wherein L is the heteroalkylene linker.

10. The compound of claim 1, wherein R 2 is independently at each occurrence H or methyl.

11. The compound of any one of claims 1 to 2, wherein the compound has one of the following structures (Ia), (Ib), (Ic) or (Id): in: R 1a is H or halogen; and R 2 It is H or methyl.

12. The compound of claim 11, wherein the compound has one of the following structures (Ia'), (Ia"), (Ib'), (Ib"), (Ic'), (Ic"), (Id') or (Id"):

13. The compound according to any one of claims 1 to 2, wherein is replaced.

14. The compound according to any one of claims 1 to 2, wherein is unsubstituted.

15. The compound according to any one of claims 1 to 2, wherein Has one of the following structures:

16. The compound of claim 15, wherein Has one of the following structures:

17. A compound according to any one of claims 1 to 2, wherein is replaced.

18. A compound according to any one of claims 1 to 2, wherein The alkylene group is substituted with at least one substituent selected from the group consisting of methyl, fluoro, cyano, methoxy, hydroxy, trifluoromethoxy, ethylsulfonyl, morpholinyl and N-methylpiperazinyl.

19. The compound according to any one of claims 1 to 2, wherein is unsubstituted.

20. The compound of any one of claims 1 to 2, wherein Has one of the following structures:

21. The compound of claim 1, having one of the following structures:

22. A pharmaceutically acceptable salt of a compound according to any one of claims 1 to 21.

23. The compound of any one of claims 1 to 22 which is a purified enantiomer or diastereomer.

24. A pharmaceutical composition comprising a compound according to any one of claims 1 to 23 and a pharmaceutically acceptable carrier.

25. Use of the pharmaceutical composition according to claim 24 in the preparation of a medicament for treating cancer.

26. The use according to claim 25, wherein the cancer is a blood cancer.

27. The use of claim 26, wherein the cancer is leukemia.

28. The use of claim 27, wherein the cancer is acute myeloid leukemia.

29. The use of claim 25, wherein the cancer is a solid tumor.

30. The use of claim 25, wherein the cancer is breast cancer.

31. The use of claim 25, wherein the cancer is triple-negative breast cancer.

32. The use of claim 25, wherein the cancer is lung cancer.

33. The use of claim 25, wherein the cancer is non-small cell lung cancer.

34. The use of claim 25, wherein the cancer is brain cancer.

35. The use of claim 25, wherein the cancer is glioblastoma.

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