# Patent

## Title
Cyclopropanecarboxamido-substituted aromatic compounds as anti-tumor agents

## Abstract

The present disclosure relates to novel cyclopropanecarboxamido-substituted aromatic compounds that inhibit protein kinases and their use in anti-tumor area. In particular, the disclosure relates to novel tyrosine-kinase inhibitors and Raf-kinase inhibitors as anti-tumor agents, their preparation, pharmaceutical composition, and their use in the treatment of cancer.

## Background
INTRODUCTION
This patent application is a continuation application of U.S. application Ser. No. 14/408,943, filed on Dec. 17, 2014, which is the national phase of PCT application No. PCT/CN2013/000731 filed on Jun. 24, 2013, which claims the benefit of priority from Chinese patent application No. 201210213116.4 filed on Jun. 26, 2012 and No. 201310172393.X filed on Apr. 16, 2013, the content of each of which is herein incorporated by reference in its entirety.


TECHNICAL FIELD
The present disclosure relates to novel cyclopropanecarboxamido-substituted aromatic compounds that inhibit protein kinases and their use in anti-tumor area. In particular, the disclosure relates to novel tyrosine-kinase inhibitors and Raf-kinase inhibitors as anti-tumor agents, their preparation, pharmaceutical composition, and their use in the treatment of cancer.
BACKGROUND OF THE DISCLOSURE
Protein kinases have become targets of intense drug discovery efforts for the past 10-15 years (Cohen, Nat. Rev. Drug Discov., 2002, 1(4), 309-315; Sebolt-Leopold, Nature Review Cancer, 2004, 4, 937-947). Small molecule inhibitors of Ras-Raf-Mek-Erk pathway have been the focus of many studies (Thompson, et al., Curr. Opin. Pharmacology, 2005, 5, 1-7; US patent Application 2003/0216446). Raf inhibitors have been proposed to be used in inhibiting cancer cell growth and in the treatment of cancers, including histiocytic lymphoma, lung adenocarcinoma, small-cell lung cancer, pancreatic cancer, and breast cancer. Recently, the approval of Zelboraf® (Vemurafenib by Plexxikon/Roche) for B-Raf mutated metastatic melanoma further demonstrates the attractiveness of the target.
Disruption of VEGFR signaling is a very attractive target in cancer therapy, as angiogenesis is a prerequisite for all solid tumor growth (Folkman, J. National Cancer Institute, 1990, 82, 4-6) and that the mature endothelium remains relatively quiescent (except for female reproductive system and wound healing), consequently, targeting of pro-angiogenesis pathway becomes a strategy being widely pursued to provide new therapeutics in cancer area.
WO2008/115263 to Curis relates to Raf kinase inhibitors containing zinc-binding and their use in the treatment of Raf related diseases and disorders such as cancer. The said derivatives may further act as HDAC inhibitors.
WO2008/033747 to Curis relates to the compositions, methods, and applications of a novel approach to selective inhibition of several cellular or molecular targets with a single small molecule. More specifically, the present disclosure relates to multi-functional small molecules wherein one functionality is capable of inhibiting histone deacetylases (HDAC) and the other functionality is capable of inhibiting a different cellular or molecular pathway involved in aberrant cell proliferation, differentiation or survival.
WO2009/042646 to Curis relates to the compositions, methods, and applications of a novel approach to selective inhibition of several cellular or molecular targets with a single small molecule. More specifically, the present disclosure relates to multi-functional small molecules capable of inhibiting a different cellular or molecular pathway involved in aberrant cell proliferation, differentiation or survival.
WO2002/32872 (EP1415987) to Eisai relates to nitrogeneous aromatic ring compounds as anti-cancer agents.
The present disclosure provides novel cyclopropanecarboxamido-substituted aromatic compounds which have improved anti-tumor activity and longer half-life in vivo over known, structurally-related compounds. These novel compounds are kinase inhibitors, preferably Raf inhibitors and VEGFR-2 (KDR/Flk-1) inhibitors. Since Raf is a key component of Ras-Raf-Mek-Erk pathway and VEGFR-2 is critical in angiogenesis, inhibitors of these kinases will be useful in pharmaceutical compositions for human and veterinary use where inhibition of Raf pathway or VEGFR-mediated angiogenesis is indicated, e.g., treatment of tumor or cancerous cell growth mediated by Raf and/or VEGFR-2 kinases. In particular, the compounds or the present disclosure are useful in the treatment of human and animal solid tumors.


DESCRIPTION OF THE DISCLOSURE
The objective of the present disclosure is to provide a series of amides of nitrogen-containing heteroaryls with anti-tumor activities.
In one aspect, the present disclosure novel compounds of formula (I), and pharmaceutically acceptable salts thereof




wherein:

    R1 is hydrogen, C1-6 alkyl, C1-6 haloalkyl, halo, or cyano;
    M is CH or N;
    L is O, NH, or N(CH3);
    A is CR5 or N;
    W is CR6 or N;
    R2, R5 and R6 are independently hydrogen, C1-6 alkyl, C1-6 haloalkyl, halo, C3-7 cycloalkyl or cyano;
    X, Y, and Z are independently CH or N;
    R3, R4 are independently hydrogen, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C2-6 alkenyl, hydroxyl-C1-6 alkyl, di-(C1-6 alkylamino)-C1-6 alkyl, amino, C1-6 alkylamino, C3-7 cycloalkylamino, di-(C1-6 alkyl)amino, amino-C1-6 alkylamino, C1-6 alkoxy-C1-6 alkylamino, C1-6 alkoxycarbonyl-C1-6 alkylamino, di-(C1-6 alkoxy-C1-6 alkyl)amino, aminocarbonyl, C1-6 alkylaminocarbonyl, di-(C1-6 alkyl)aminocarbonyl, C3-7 cycloalkylaminocarbonyl, C1-6 alkoxy, C3-7 cycloalkoxy, hydroxyl-C1-6 alkoxy, C1-6 haloalkoxy, amino-C1-6 alkyl, amino-C1-6 alkoxy, C1-6 alkylsulfonyl, C2-6 alkenylsulfonyl, C3-7 cycloalkylsulfonyl, heterocycle optionally substituted by B, aryl optionally substituted by B, heteroaryl optionally substituted by B, C1-6 alkylsulfonylamino, C2-6 alkenylsulfonylamino, C3-7 cycloalkylsulfonylamino, amido, C1-6 alkylcarbonylamino, C2-6 alkenylcarbonylamino, C3-7 cyclooalkylcarbonylamino, C1-6 alkoxycarbonylamino, C3-7 cycloalkoxycarbonylamino, ureido, C3-7 cycloalkyl, C3-7 halocycloalkyl, heterocyclyl-oxy, piperidinylamino, N-methyl-piperidinyl-4-carbonyl, piperazinyl-C1-6 alkyl, pyrrolylcarbonylamino, N-methyl-piperidinylcarbonylamino or heterocyclyl-C1-6 alkoxy; or
    R3 and R4 can form a 3 to 8-membered ring together with atoms in the aromatic ring to which they are attached; and
    B is hydrogen, C1-6 alkyl, C1-6 haloalkyl, halo, hydroxyl, aryl, amino, C1-6 alkylamino, C3-7 cycloalkylamino, di-(C1-6 alkyl)amino, cyano, or C3-7 cycloalkyl.


In one embodiment, the NHCONH and L substituents on the central aromatic ring are in a 1,3-configuration.
In another embodiment, the NHCONH and L substituents on the central aromatic ring are in a 1,4-configuration.
In another embodiment, L is O.
In another embodiment, L is NH or N(CH3).
In another embodiment, X, Y, and Z are all CH.
In another embodiment, one of X, Y, or Z is N and the other two are CH.
In another embodiment, M is CH.
In another embodiment, M is N.
In another embodiment, A is CR5.
In another embodiment, A is N.
In another embodiment, W is CR6.
In another embodiment, W is N.
In another embodiment, R1 is hydrogen or C1-3 alkyl.
In another embodiment, R2, R5 and R6 are independently hydrogen, C1-3 alkyl, C1-3 haloalkyl, halo, or cyano.
In another embodiment, R3 and R4 are independently hydrogen, halo, cyano, C1-6 alkyl, C1-6 haloalkyl, C2-6 alkenyl, hydroxyl-C1-6 alkyl, di-(C1-6 alkylamino)-C1-6 alkyl, amino, C1-6 alkylamino, C3-7 cycloalkylamino, di-(C1-6 alkyl)amino, amino-C1-6 alkylamino, C1-6 alkoxy-C1-6 alkylamino, C1-6 alkoxycarbonyl-C1-6 alkylamino, di-(C1-6 alkoxy-C1-6 alkyl)amino, aminocarbonyl, C1-6 alkylaminocarbonyl, di-(C1-6 alkyl)aminocarbonyl, C3-7 cycloalkylaminocarbonyl, C1-6 alkoxy, C3-7 cycloalkoxy, hydroxyl-C1-6 alkoxy, C1-6 haloalkoxy, amino-C1-6 alkyl, amino-C1-6 alkoxy, C1-6 alkylsulfonyl, C2-6 alkenylsulfonyl, C3-7 cycloalkylsulfonyl, heterocycle optionally substituted by B, aryl optionally substituted by B, heteroaryl optionally substituted by B, C1-6 alkylsulfonylamino, C2-6 alkenylsulfonylamino, C3-7 clcyoalkylsulfonylamino, amido, C1-6 alkylcarbonylamino, C2-6 alkenylcarbonylamino, C3-7 clcyoalkylcarbonylamino, C1-6 alkoxycarbonylamino, C3-7 cycloalkoxycarbonylamino, ureido, C3-7 cycloalkyl, C3-7 halocycloalkyl, heterocyclyl-oxy, piperidinylamino, N-methyl-piperidinyl-4-carbonyl, piperazinyl-C1-6 alkyl, pyrrolylcarbonylamino, N-methyl-piperidinylcarbonylamino or heterocyclyl-C1-6 alkoxy.
In another embodiment, one or both of R3 and R4 are independently a heterocyclyl-oxy group selected from pyrrolyloxy, piperidinyloxy, furanyloxy and azetidinyloxy.
In another embodiment, one or both of R3 and R4 are independently an optionally substituted aryl group selected from phenyl and naphthyl.
In another embodiment, one or both of R3 and R4 are independently an optionally substituted hereroaryl group selected from furyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzimidazolyl, indolyl, indazolyl, benzoisothiazolyl, benzoxazolyl, benzoisoxazolyl, quniolinyl and isoquinolinyl.
In another embodiment, the hereroaryl is pyridinyl, oxazolyl, or triazolyl.
In another embodiment, one or both of R3 and R4 are independently an optionally substituted heterocyclyl group selected from piperidinyl, piperazinyl, homopiperazinyl, azepinyl, azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, imidazolinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinuclidinyl, thiadiazolidinyl, benzothiazolidinyl, benzopyrrolidinyl, dihydrofuranyl, tetrahydrofuranyl, dihydropyranyl, tetrahydropyranyl, thiomorpholinyl, 1-oxo-thiomorpholinyl, 1,1-dioxothiomorpholinyl, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl and tetrahydroisoquinolinyl.
In another embodiment, the optionally substituted heterocyclyl is N(R7)piperazinyl, N(R7)piperidin-4-yl, pyrrolidinyl, 2-oxopyrrolidinyl, morpholinyl, 2-methylmorpholinyl, 2,6-dimethylmorpholinyl, 2-oxomorpholinyl, 3-(dimethylamino)pyrrolidinyl, 2-oxo-5-methyloxazolidin-3-yl, 3,3-difluoroazetidinyl, fluoropiperidinyl, hydroxylpiperidinyl, or 1,1,-dioxothiomorpholinyl, and R7 is hydrogen, C1-3 alkyl, C1-3 haloalkyl, C3-6 cycloalkyl, C1-5 acyl or C3-6 cycloalkylacyl.
In another embodiment, B is hydrogen, C1-3 alkyl, C1-3 haloalkyl, halo, hydroxyl, aryl, amino, C1-3 alkylamino, C3-5 cycloalkylamino, di-(C1-3 alkyl)amino, cyano, or C3-5 cycloalkyl.
In another embodiment, B is hydrogen, halo, hydroxyl, C1-3 alkyl, C1-3 haloalkyl, di-(C1-3 alkyl)amino.
In another embodiment, R3 and R4 are independently halo, cyano or C1-3 haloalkyl.
In another embodiment, R3 and R4 together form a 3 to 8-membered ring with the atoms in the aromatic ring to which they are attached.
Other embodiments of the compounds include Examples 1-242, and pharmaceutically acceptable salts thereof.
It is understood that the disclosure also contemplates all possible combinations of the embodiments listed above.
The compounds of the present disclosure have improved anti-tumor activity and longer half-life in vivo. For example, compared to the marketed drug sorafenib which is structurally somewhat similar to the compound in example 137 of this disclosure, the in vitro IC50 of compound 137 at B-Raf is four times more potent than that of sorafenib. In pre-clinical animal studies, sorafenib has a shorter half-life than many of the compounds in this disclosure; its in vivo anti-tumor activities are also weaker than that of many of the compounds in the present disclosure. Clinically, sorafenib is given twice daily with 200-400 mg each time. The preferred compounds in the present disclosure are expected to be given once daily with much lower dose.
A key difference between compounds in the present disclosure and those known in the art is the use of (substituted)cyclopropanecarboxamido group as the substituent on the aromatic rings. It is this (substituted)cyclopropanecarboxamido group that plays a key and irreplaceable role in providing the desired properties to compounds in the present disclosure. For example, when the cyclopropanecarboxamido group in example 22 is replaced by a cyclobutanecarboxamido group, the IC50 of the resulting compound is decreased by twenty folds in pERK assay using MDA-MB-231 cell line; whereas the replacement of cyclopropanecarboxamido group in example 7 with either cyclobutanecarboxamido group or propionamido group results in more than twenty fold loss of activity in pERK assay using MDA-MB-231 cell line. These examples further demonstrate the pivotal role that (substituted)cyclopropanecarboxamido group plays in the structures and properties of compounds in the present disclosure.
A second aspect of the disclosure relates to the processes for preparing the compounds of the disclosure and to the synthetic intermediates useful in such process as described below. Compounds of the present disclosure can be prepared in a number of conventional methods. Some suitable methods for preparing the compounds are described in the examples. Typically, compounds of formula (I) can be prepared with the methods below.
Because of the fixed (properly substituted) cyclopropanecarboxamide structural feature, it would be more efficient when a fragment containing this functional feature is allowed to couple with another fragment through urea bond formation to make the final products. There are many ways of forming ureas, including reaction of amines with isocynates; or reaction of amines with phenyl carbamates or other analogs. For example, Compounds in Formula (I) may be prepared through the following approach:
1) Amines with optionally substituted phenylisocynates or other arylisocynates




Wherein M, L, A, W, X, Y, Z and R1 to R4 are as defined above;
2) Amines with optionally substituted phenyl carbamates or other aryl carbamates




Wherein M, L, A, W, X, Y, Z, and R1 to R4 are as defined above.
Based on the same urea formation principle, the products can also be obtained through first forming the urea link and then installing the cyclopropanecarboxamide or substituted cyclopropanecarboxamide moiety, with P1 as the protective group (e.g., Boc-, etc.):




Wherein M, L, A, W, X, Y, Z, and R1 to R4 are as defined above.
In a third aspect, the present disclosure provides pharmaceutical compositions comprising compounds of formula (I), or pharmaceutically acceptable salts thereof, together with a pharmaceutically acceptable carrier or excipient.
In a fourth aspect, the present disclosure provides a method of treating cancer using compounds of formula (I), or pharmaceutically acceptable salts thereof.
In a fifth aspect, the present disclosure provides use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, in the preparation of medicament for the treatment of cancer.
Compounds of this disclosure inhibit KDR and B-Raf kinases at the enzyme levels, they also inhibit Erk phosphorylation and PLC-PRF-5 cell proliferation at the cellular levels. These compounds can be used to treat hyperproliferative disorders such as cancers, especially colon cancer, breast cancer, lung cancer, prostate cancer, pancreatic cancer, gastrointestinal cancer, bladder cancer, ovarian cancer, melanoma, neuroblastoma, cervical cancer, kidney or renal cancer, leukemia and lymphoma. They are especially useful in treating or inhibiting solid tumors, for example, breast cancer, colon cancer, lung and prostate cancer. These compounds can be used as treatment for AML, ALL and GIST.
The compounds of the present disclosure are preferably formulated as pharmaceutical compositions administered by a variety of routes. Such pharmaceutical compositions and processes for preparing them are well known in the art. See, e.g., REMINGTON: THE SCIENCE AND PRACTICE OF PHARMACY (A. Gennaro, et al., eds, 19th ed. Mack Publishing Co., 1995)
Unless otherwise defined, the following definitions refer to the various terms used above and throughout the disclosure:
The term “halo” refers to fluoro-, chloro-, bromo- and iodo-; with fluoro-, chloro- and bromo-preferred.
The term “C1-6 alkyl”, alone or in combination with other groups, refers to monovalent, linear chain or branched chain alkyl groups containing from 1 to 6 carbon atoms. Exemplary C1-6 alkyl groups include but not limited to metheyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl and tert-butyl groups. More preferred are C1-4 alkyls.
The term “haloalkyl” refers to alkyl groups in which one or more hydrogen atoms are substituted with the same or different halogens. Exemplary haloalkyl groups include —CH2Cl, —CH2CF3, CH2CCl3, pan-fluoroalkyl (e.g., —CF3), etc.
The term “C3-7 cycloalkyl”, alone or in combination with other groups, refers to, unless otherwise defined, fully saturated hydrocarbon rings of 3 to 7 carbon atoms. For example, cyclopropyl, cyclobutyl, cyclohexyl.
The term “C3-6 cycloalkylacyl” also called “cycloalkanecarbonyl” refers to acyl groups in which the carbonyl is directly attached to saturated hydrocarbon rings of 3-6 carbon atoms. For example, cyclopropylacyl.
The term “C1-6 alkoxy” refers to, alone or in combination with other groups, R′—O—, where R′ is C1-6 alkyl.
The term “C2-6 alkenyl” refers to, alone or in combination with other groups, linear or branched-chain monovalent groups of 2 to 6 carbon atoms with at least one carbon-carbon double bond. Examples include vinyl, 2-propenyl.
The term “aryl” refers to, alone or in combination with other groups, a monovalent, mono- or bi-cyclic aromatic carbon ring system. The preferred aryls include, but are not limited to, phenyl, naphthyl, methylphenyl and dimethylphenyl.
The term “heterocyclyl” refers to, alone or in combination with other groups, 3 to 8-membered non-aromatic mono- or bi-cyclic heteroatom-containing ring systems where one or two heteroatoms may be selected from N, O, or S(O)n (n is an integral of 0 to 2).
Examples of “heterocyclyl” include, but are not limited to, optionally substituted piperidinyl, piperazinyl, homopiperazinyl, azepinyl, azetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, imidazolinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinuclidinyl, thiadiazolidinyl, benzothiazolidinyl, benzopyrrolidinyl, dihydrofuranyl, tetrahydrofuranyl, dihydropyranyl, tetrahydropyranyl, thiomorpholinyl, 1-oxo-thiomorpholinyl, 1,1-dioxothiomorpholinyl, dihydroquinolinyl, dihydroisoquinolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, etc.
The term “heteroaryl” refers to five- to six-membered monocyclic or nine to ten-membered bicyclic aromatic rings containing one, two or three heteroatoms selected from nitrogen, oxygen and/or sulfur atoms. Examples include, but are not limited to, furyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzimidazolyl, indolyl, indazolyl, benzoisothiazolyl, benzoxazolyl, benzoisoxazolyl, quniolinyl and isoquinolinyl. The preferred heteroaryls are pyridinyl, oxazolyl and triazolyl.
The term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgement, suitable for use in contact of the tissues of human and lower animals without undue toxicity, irritation, allergic reactions and the like. The salts are organic or inorganic salts of a compound of the disclosure which maintain the biological activities of it as drawn in Formula (I). The salts may be prepared from suitable, non-toxic organic or inorganic acids reacting with free base, or organic and inorganic bases reacting with acid group in the compounds of the present disclosure. Examples of acid addition salts include those salts derived from inorganic acids and organic acids. Examples of inorganic acid include, but are not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, animosulfonic acid, phosphoric acid, nitric acid; examples of organic acids include, but are not limited to, para-toluenesulfonic acid, salicylic acid, methanesulfonic acid, oxalic acid, succinic acid, citric acid, maleic acid, lactic acid, fumaric acid, etc. Examples of base addition salts include those derived from ammonium hydroxide, sodium hydroxide, potassium hydroxide, quaternary ammonium hydroxide such as tetramethylammonium hydroxide. Conversion of an acid or base compound into a salt is well known in the art to improve its physicochemical properties, chemical stability, moisture absorption property, liquidity and solubility.
“Pharmaceutically acceptable” carrier, excipient refers to those carriers and excipients which are compatible to the administration of the compounds to the subjects, and are non-toxic.
The “therapeutically effective amount” of a compound of this disclosure means an amount of the compound that effectively prevents or delays the progression of the disease, or attenuates, ameliorates some of the symptoms of the disease or extends the life of patients. Determination of therapeutically effective amount depends on a variety of factors well known in medical arts.
The therapeutically effective amount or dose may vary in a wide range, and can be determined by known arts in this field. The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the activity of specific compound being employed, route of administration, duration of treatment, and the age, body weight, general health, sex and diet of the patient. In general, the total daily dose of the compound for a 70 Kg adult, when administered orally or parenterally, may range from 10 mg to 10,000 mg, preferably from about 200 mg to 1,000 mg, albeit some evidences show higher dose level may be acceptable. Total daily dose of the compounds of this disclosure may be administered in a single dose or multiple doses, for parenteral administration, the total daily dose may be delivered through continuous infusion.
All compound names listed herein are generated with Symyx Draw 3.3 program from Accelrys.
Abbreviations used in this application are listed below:

    DCM: dichloromethane; PE: petroleum ether; THF: tetrahydrofuran; DMF: N,N,-dimethylformamide; DIEA: diisopropylethylamine; DMSO: dimethylsulfoxide; TEA: triethylamine; TFA: trifluoroacetic acid; MW: microwave; NMP: N-methyl-2-pyrrolidone; mCPBA: meta-chloroperoxybenzoic acid; BAST: bis-(2-methoxyethyl)aminosulfur trifluoride; EDCI: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride; TLC: thin-layer chromatography; EtOAc: ethyl acetate; MeOH: methanol; EtOH: ethanol;

MS data is obtained with ESI-MS (electrospray ionization mass spectrometry) method through a LC/MS system.



INTERMEDIATES
Preparation of Intermediate A




Step 1: Synthesis of N-(6-chloropyrimidin-4-yl)cyclopropanecarboxamide




Add slowly cyclopropanecarbonyl chloride (10.6 g, 102.3 mmol) to a mixture of 4-amino-6-chloro-pyrimidine (12 g, 91 mmol), pyridine (18 g, 227.5 mmol) in THF (150 mL) at 0° C. Stir the reaction at 60° C. for 4 hrs.
TLC (DCM:MeOH=20:1) shows that the reaction is complete. Cool the reaction to 0° C., add water (100 mL), extract with EtOAc (100 mL×2). Combine the organic layers, wash with diluted aqueous HCl (1M, 150 mL) and brine sequentially, dry over anhydrous Na2SO4. Evaporation under reduced pressure affords crude product (16 g, crude yield 88.8%). MS: (M+1): 198.1. Use crude product directly in next step without further purification.
Step 2: Synthesis of N-[6-(4-amino-3-methyl-phenoxy)pyrimidin-4-yl]cyclopropanecarboxamide




Mix the cyclopropanecarboxamide derivative obtained in Step 1 (400 mg, 2.03 mmol), 4-amino-3-methyl-phenol (272 mg, 2.23 mmol) and Cs2CO3 (1.3 g, 4.06 mmol) in DMF (10 mL). Stir the reaction at 150° C. under microwave conditions for 30 min.
TLC (EtOAc:PE=2:1) shows the reaction is complete. Cool the reaction to room temperature; add water (15 mL). Extract with EtOAc (15 mL×2). Wash the combined organic layers with brine, dry over anhydrous Na2SO4. Evaporation under reduced pressure affords crude product. Purification by chromatography (silica gel, EtOAc:petroleum ether=1:1) provides the title compound as a light-yellow solid (450 mg, 78.1%). MS: (M+1): 285.2.
Intermediates A2-A5 can be synthesized with similar method (Table A1).







TABLE A1







Intermediates A1-A5










Number
Starting material
Intermediate
MS [M + 1]+





A1








285.2


 


A2








305.1


 


A3








289.2


 


A4








305.1


 


A5








303.2













Step 1: Synthesis of N-(6-chloropyrimidin-4-yl)cyclopropanecarboxamide (see Step 1 for A1)




Step 2: Synthesis of N-[6-(3-bromo-5-methyl-phenoxy)pyrimidin-4-yl]cyclopropanecarboxamide




Add N-(6-chloropyrimidin-4-yl)cyclopropanecarboxamide (1 g, 5.07 mmol), 3-bromo-5-methyl-phenol (950 mg, 5.07 mmol), Cs2CO3 (2.14 g, 6.60 mmol) and DMF (10 mL) to a microwave reaction vessel (25 mL). Heat the reaction under microwave conditions at 150° C. for 1 hr.
Cool to room temperature, add water (50 mL), extract with EtOAc (100 mL×2), combine organic layers, dry over anhydrous Na2SO4. Concentration and purification by chromatography (silica gel, EtOAc:PE=1:3) afford the title compound (1.2 g, 67.9%). MS: (M+1): 348.1.
Step 3: Synthesis of N-[6-[3-(benzhydrylideneamino)-5-methyl-phenoxy]pyrimidin-4-yl]cyclopropanecarboxamide




Mix the cyclopropanecarboxamide derivative obtained in Step 2 (400 mg, 1.15 mmol), benzophenone imine (624 mg, 3.45 mmol), t-BuOK (257 mg, 2.3 mmol) and 1,4-dioxane in a 250 mL flask, add Xantphos (133 mg, 0.23 mmol), Pd(OAc)2 (26 mg, 0.115 mmol). Stir the reaction under N2 at 120° C. for 15 hrs.
TLC shows reaction is complete. Remove the solid through suction filtration. Concentration and purification by chromatography (silica gel, EtOAc:PE=1:3) afford the title compound (450 mg, 87.4%). MS: (M+1): 449.3.
Step 4: Synthesis of N-[6-(3-amino-5-methyl-phenoxy)pyrimidin-4-yl]cyclopropanecarboxamide




Add 1M HCl aqueous solution (10 mL) to a solution of the imine obtained in Step 3 (450 mg, 1.0 mmol) in THF (10 mL), stir the reaction at room temperature for 1 hr. Basicify with saturated NaHCO3 solution, extract with EtOAc (30 mL×2). Combine the organic layers, dry over anhydrous Na2SO4. Concentration and purification by chromatography (silica gel, EtOAc:PE=1:1) afford the title compound (230 mg, 80.8%). MS: (M+1): 285.2.
Intermediates A7-A8 can be synthesized with similar method (Table A2)







TABLE A2







Intermediates A6-A8










Number
Starting material
Intermediate
MS [M + 1]+





A6








285.2


 


A7








289.2


 


A8








296.2













Step 1: Synthesis of 4-chloro-6-(4-nitrophenoxy)pyrimidine




Dissolve NaOH (2.8 g, 71.9 mmol) in water (90 mL) at 0° C. Add para-nitrophenol (10 g, 71.9 mmol), and then a solution of 4,6-dichloropyrimidine (10.7 g, 71.9 mmol) in acetone (90 mL). Stir the reaction at 70° C. for 24 hrs. Cool the reaction, collect the solid, rinse the solid with the mixture of acetone-water (1:1, 20 mL), and dry the solid to afford the title compound (18 g, 77.8%). MS: (M+Na): 274.0.
Step 2: Synthesis of N-[6-(4-nitrophenoxy)pyrimidin-4-yl]cyclopropanecarboxamide




Mix 4-chloro-6-(4-nitrophenoxy)pyrimidine (10.6 g, 39.68 mmol), cyclopropanecarboxamide (8.7 g, 99.2 mmol) and 1,4-dioxane (150 mL). Then under N2, add tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 1.8 g, 1.98 mmol], (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 2.5 g, 3.97 mmol), Cs2CO3 (32 g, 99.5 mmol), and stir at 100° C. for 12 hrs. Cool to room temperature, filter, concentration and purification by chromatography (silica gel, DCM:EtOAc=1:5) afford the title compound (5.5 g, 43.6%). MS: (M+1): 301.
Step 3: Synthesis of N-[6-(4-aminophenoxy)pyrimidin-4-yl]cyclopropanecarboxamide




Dissolve N-[6-(4-nitrophenoxy)pyrimidin-4-yl]cyclopropanecarboxamide (2.2 g, 7.33 mmol) in DCM (50 mL), add Pd/C (10%, 340 mg), flush with H2, and stir at room temperature under H2 for 2 hrs. After the reaction is complete, flush with N2, and then filter, concentrate the filtrate, purify the crude product by chromatography (silica gel, EtOAc:PE=1:1) to provide the title compound (1.8 g, 91%). MS: (M+1): 271.
Intermediates A10-A16 can be synthesized by similar method (Table A3).







TABLE A3







Intermediates A9-A16










Number
Starting material
Intermediate
MS [M + 1]+













A9








271


 


A10








289.2


 


A11








289.2


 


A12








284.2


 


A13








286


 


A14








271.1


 


A15








289


 


A16








285.2













Step 1: Synthesis of 6-chloro-N-(3-fluoro-4-nitro-phenyl)pyrimidin-4-amine




Mix 4,6-dichloropyrimidine (3 g, 20.1 mmol), 3-fluoro-4-nitroaniline (2.4 g, 15.5 mmol), isopropanol (20 mL) and concentrated HCl (3 mL) in a 100 mL flask, heat at 120° C. under N2 for 4 hrs.
TLC (PE:EtOAc=3:1) shows the reaction is complete. Cool to room temperature, add water (20 mL), carefully adjust pH=8 with saturated NaHCO3 solution. Extract with EtOAc (100 mL×2), combine the organic layers; wash with brine (100 mL), dry over anhydrous Na2SO4. Filtration and concentration afford crude product. Add the mixture of EtOAc and PE (1:1, 100 mL), stir and filter to give the title compound (1.38 g, 67%). MS: (M+1): 269.1.
Step 2: Synthesis of 6-chloro-N-(3-fluoro-4-nitro-phenyl)-N-methyl-pyrimidin-4-amine




Add the compound obtained in Step 1 (700 mg, 2.6 mmol), NaH (60%, 208 mg, 5.2 mmol) to DMF (20 mL), stir for 1 hr. Cool to 0° C., add slowly iodomethane (554 mg, 3.9 mmol). Warm the reaction to 25° C. and stir for 2 hrs. Cool the reaction to 0° C., add water (20 mL), extract with EtOAc (15 mL×2), wash the combined organic layers with brine, dry over anhydrous Na2SO4. Filter and concentrate under reduced pressure to afford crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (350 mg, 47.6%). MS: (M+1): 283.1.
Step 3: Synthesis of N-[6-(3-fluoro-N-methyl-4-nitro-anilino)pyrimidin-4-yl]cyclopropanecarboxamide




Mix the compound obtained in Step 2 (350 mg, 1.24 mmol), cyclopropanecarboxamide (116 mg, 1.36 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 77 mg, 0.124 mmol), Cs2CO3 (1.01 g, 3.1 mmol) in 1,4-dioxane (30 mL), then under N2 atmosphere, add tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 113 mg, 0.124 mmol]. Stir the reaction at 126° C. under N2 for 16 hrs. Cool the reaction mixture, filter, and concentrate the filtrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (178 mg, 43.3%). MS: (M+1): 332.2.
Step 4: Synthesis of N-[6-(4-amino-3-fluoro-N-methyl-anilino)pyrimidin-4-yl]cyclopropanecarboxamide




Dissolve the compound obtained in Step 3 (180 mg, 0.543 mmol) in ethanol (20 mL), add iron powder (150 mg, 2.72 mmol) and saturated NH4Cl solution (10 mL). Stir the reaction at 75° C. for 2 hrs. TLC (DCM:MeOH=20:1) shows the reaction is complete. Cool the reaction to 0° C., add water (20 mL), extract with EtOAc (30 mL×2), combine the organic layers, wash with brine (30 mL), dry over anhydrous Na2SO4, filter and concentrate to give the crude product (150 mg). Use it directly without further purification. MS: (M+1): 302.2.




Step 1: Synthesis of 6-(6-chloropyrimidin-4-yl)oxypyridin-3-amine




Add 5-aminopyridin-2-ol (500 mg, 4.545 mmol), t-BuOK (555 mg, 4.545 mmol) in DMF (20 mL), stir at 25° C. for 30 min. Cool to 0° C., add slowly 4,6-dichloropyrimidine (614 mg, 4.13 mmol). Stir the reaction under N2 for 1 hr. TLC (EtOAc:PE=1:1) shows the reaction is complete. Cool the mixture to 0° C., add water (20 mL), extract with EtOAc (20 mL×2). Combine the organic layers; wash with brine, dry over anhydrous Na2SO4. Filter and concentrate under reduced pressure to give crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (650 mg, 70.8%). MS: (M+1): 223.1.
Step 2: Synthesis of tert-butyl N-[6-(6-chloropyrimidin-4-yl)oxy-3-pyridyl]carbamate




Add the compound obtained in Step 1 (650 mg, 2.93 mmol) and Boc2O (766 mg, 3.51 mmol) in t-BuOH (20 mL) and heat at 50° C. for 16 hrs. TLC (EtOAc:PE=1:1) shows the reaction is complete. Concentrate under reduced pressure to give crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (610 mg, 64.6%). MS: (M+1): 323.1.
Step 3: Synthesis of tert-butyl N-[6-[6-(cyclopropanecarbonylamino)pyrimidin-4-yl]oxy-3-pyridyl]carbamate




Mix the compound obtained in Step 2 (610 mg, 1.9 mmol), cyclopropanecarboxamide (193 mg, 2.27 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 118 mg, 0.19 mmol), Cs2CO3 (1.238 g, 3.8 mmol) and tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 122 mg, 0.133 mmol] in 1,4-dioxane (30 mL). Stir the reaction at 120° C. under N2 for 16 hrs. Cool the reaction, filter, concentrate the filtrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (500 mg, 70.8%). MS: (M+1): 372.2.
Step 4: Synthesis of N-[6-[(5-amino-2-pyridyl)oxy]pyrimidin-4-yl]cyclopropanecarboxamide




Add the compound obtained in Step 3 (500 mg, 1.9 mmol) to DCM (30 mL), cool to 0° C., add slowly trifluoroacetic acid (10 mL), then stir the reaction at 25° C. for 4 hrs. TLC (DCM:MeOH=20:1) shows the reaction is complete. Cool the reaction to 0° C., adjust to pH=8-9 with saturated NaHCO3 solution. Extract with DCM (30 mL×2), combine the organic layers; wash with brine (30 mL), dry over anhydrous Na2SO4. Filter and concentrate under reduced pressure to give crude product. Rinse the solid with ether, dry under reduced pressure to give the product (180 mg, 49.3%). MS: (M+1): 272.2.
Intermediate A19 can be synthesized with similar method (Table A4).







TABLE A4







Intermediates A17-A19 










Number
Starting material
Intermediate
MS [M + 1]+





A17








302.2


 


A18








272.2


 


A19








286.2













Synthesis of phenyl
N-[4-[6-(cyclopropanecarbonylamino)pyrimidin-4-yl]oxy-2-methyl-phenyl]carbamate




Add N-[6-(4-amino-3-methyl-phenoxy)pyrimidin-4-yl]cyclopropanecarboxamide (Intermediate A1, 500 mg, 1.76 mmol), pyridine (280 mg, 3.52 mmol) to a mixed solvent of THF (5 mL) and DCM (5 mL). Cool to 0° C.; add slowly phenyl chloroformate (330 mg, 2.11 mmol). Stir the reaction at room temperature for 2 hrs. TLC (EtOAc:PE=1:1) shows the reaction is complete. Dilute with water, extract with DCM. Combine the organic layers, wash with 1M diluted HCl, brine sequentially, and dry over anhydrous Na2SO4. Concentration under reduced pressure affords the title compound (700 mg, 98.3%). MS: (M+1) 273.







TABLE A5







Intermediate A20










Number
Starting material
Intermediate
MS [M + 1]+





A20








273









Preparation of Intermediate B




Step 1: Synthesis of 4-[(3-methyl-5-nitro-2-pyridyl)oxy]pyridin-2-amine




Mix 2-aminopyridin-4-ol (3.3 g, 30 mmol), 2-chloro-3-methyl-5-nitropyridine (5.17 g, 30 mmol) in anhydrous DMF (50 mL), add K2CO3 (8.28 g, 60 mmol). Stir the reaction at 90° C. overnight. Concentrate under reduced pressure to give crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (3.3 g, 45%). MS: (M+1): 247.1.
Step 2: Synthesis of tert-butyl N-[4-[(3-methyl-5-nitro-2-pyridyl)oxy]-2-pyridyl]carbamate




Add 4-[(3-methyl-5-nitro-2-pyridyl)oxy]pyridin-2-amine (400 mg, 1.62 mmol), Boc2O (388 mg, 1.78 mmol) to t-BuOH (10 mL). Stir the reaction at 50° C. for 12 hrs. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the title compound (400 mg, 71%). MS: (M+1): 347.
Step 3: Synthesis of tert-butyl N-(cyclopropanecarbonyl)-N-[4-[(3-methyl-5-nitro-2-pyridyl)oxy]-2-pyridyl]carbamate




Add N-[4-[(3-methyl-5-nitro-2-pyridyl)oxy]-2-pyridyl]carbamate (300 mg, 0.87 mmol), Et3N (350 mg, 3.47 mmol) to anhydrous DCM (5 mL), cool to 0° C., add cyclopropanecarbonyl chloride (360 mg, 3.47 mmol) dropwise. Stir at room temperature for 2 hrs. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the title compound (350 mg, 97%). MS: (M+1): 415.
Step 4: Synthesis of N-[4-[(3-methyl-5-nitro-2-pyridyl)oxy]-2-pyridyl]cyclopropanecarboxamide




Add tert-butyl N-(cycl opropanecarbonyl)-N-[4-[(3-methyl-5-nitro-2-pyridyl)oxy]-2-pyridyl]carbamate (350 mg, 0.84 mmol) in DCM (10 mL), add trifluoroacetic acid (2 mL). Stir the reaction at room temperature for 2 hrs. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the title compound (260 mg, 98%). MS: (M+1): 315.
Step 5: Synthesis of N-[4-[(5-amino-3-methyl-2-pyridyl)oxy]-2-pyridyl]cyclopropanecarboxamide




Dissolve N-[4-[(3-methyl-5-nitro-2-pyridyl)oxy]-2-pyridyl]cyclopropanecarboxamide (269 mg, 0.83 mmol) in MeOH (5 mL) and DCM (5 mL). Add Pd/C (10%, 104 mg), flush with H2. Stir the reaction under H2 at room temperature for 2 hrs. After the reaction, flush with N2, filter the reaction mixture; concentrate the filtrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (70 mg, 30%). MS: (M+1): 285.
Intermediates B2-B9 can be Synthesized Using Similar Method (Table B1).







TABLE B1







Intermediates B1-B9











Starting




Number
material
Intermediate
MS [M + 1]+













B1








285.2


 


B2








285.2


 


B3








NA


 


B4








NA


 


B5








271.1


 


B6








270.2


 


B7








288


 


B8








284.2


 


B9








288













Step 1: Synthesis of 4-[(5-bromo-3-fluoro-2-pyridyl)oxy]pyridin-2-amine




Mix 2-aminopyridin-4-ol (1.13 g, 10.3 mmol), 5-bromo-2,3-difluoro-pyridine (2 g, 10.3 mmol) in DMF (10 mL). Stir the mixture at 120° C. for 3 hrs. TLC (EtOAc:PE=1:1) shows the reaction is complete. Filter, add water (50 mL) to the filtrate, extract with EtOAc (50 mL×3). Combine the organic layers, dry over anhydrous Na2SO4. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:4) affords the title compound (1.35 g, 46.3%). MS: (M+1): 284.
Step 2: Synthesis of tert-butyl N-[4-[(5-bromo-3-fluoro-2-pyridyl)oxy]-2-pyridyl]carbamate




Add the compound obtained in Step 1 (1.35 g, 4.7 mmol) and Boc2O (1.13 g, 5.2 mmol) in t-BuOH (10 mL). Stir the reaction at 75° C. for 2 hrs. TLC (EtOAc:PE=1:1) shows reaction is complete. Concentrate under reduced pressure to give the crude product (1.8 g) which is used without further purification.
Step 3: Synthesis of tert-butyl N-[4-[(5-bromo-3-fluoro-2-pyridyl)oxy]-2-pyridyl]-N-(cyclopropanecarbonyl)carbamate




Add the compound obtained in Step 2 (1.8 g, 4.7 mmol), Et3N (0.95 g, 9.4 mmol) to DCM (10 mL), then add cyclopropanecarbonyl chloride (0.95 g, 7.0 mmol). Stir the reaction at room temperature for 2.5 hrs. TLC (EtOAc:PE=1:1) shows the reaction is complete. Add water (10 mL) to quench the reaction. Extract with DCM (10 mL×3), combine the organic layers, dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product (2.14 g) which is used without further purification.
Step 4: Synthesis of N-[4-[(5-bromo-3-fluoro-2-pyridyl)oxy]-2-pyridyl]cyclopropanecarboxamide




Dissolve the crude product obtained in Step 3 (2.14 g, 4.7 mmol) in HCl solution in 1,4-dioxane (5 M, 10 mL). Stir the reaction at room temperature for 17 hrs. LC/MS shows reaction is complete. Remove the solvent under reduced pressure, suspend the solid in DCM and add carefully saturated NaHCO3 solution. Separate the organic layer. Extract the aqueous layer with DCM. Combine the organic layers, dry over anhydrous Na2SO4. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the title compound (0.9 g, 54% from step 2). MS: (M+1): 352.
Step 5: Synthesis of N-[4-[[5-(benzhydrylideneamino)-3-fluoro-2-pyridyl]oxy]-2-pyridyl]cyclopropanecarboxamide




Add the compound obtained in Step 4 (0.9 g, 2.5 mmol), benzophenone imine (0.59 g, 3.25 mmol), tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 200 mg, 0.25 mmol], (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 250 mg, 0.375 mmol), Cs2CO3 (1.6 g, 5.0 mmol) in 1,4-dioxane (10 mL). Stir the mixture under N2 at 100° C. for 16 hrs. Cool to room temperature, filter, and concentrate the filtrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the title compound (0.8 g, 69%). MS: (M+1): 453.
Step 6: Synthesis of N-[4-[(5-amino-3-fluoro-2-pyridyl)oxy]-2-pyridyl]cyclopropanecarboxamide




Dissolve the compound obtained in Step 5 (0.8 g, 1.77 mmol) in THF (10 mL), add water (1 mL) and 1N aqueous HCl (3 mL). Stir the reaction at room temperature for 2 hrs. TLC (EtOAc:PE=1:1) shows the reaction is complete. Extract with EtOAc. Wash the organic layer with saturated NaHCO3 solution and brine respectively. Collect the organic layer, dry over anhydrous Na2SO4. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=5:1) affords the title compound (0.4 g, 78%).




Step 1: Synthesis of N-(4-hydroxy-2-pyridyl)cyclopropanecarboxamide




Dissolve 2-amino-pyridin-4-ol (5 g, 45.4 mmol) in DCM (30 mL), add Et3N (13.8 g, 136 mmol) and then cyclopropanecarbonyl chloride (14.2 g, 136 mmol) dropwise while keeping the temperature below 10° C. After addition, stir the reaction at room temperature for 16 hrs. Remove the solvent under reduced pressure, dissolve the residue in THF (40 mL), adjust pH=12 with 1N NaOH solution. Stir the mixture for 3 hrs. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:MeOH=9:1) affords the title compound (2.6 g, 32%).
Step 2: Synthesis of 5-(benzhydrylideneamino)-2-fluoro-benzonitrile




Add 5-bromo-2-fluoro-benzonitrile (0.5 g, 2.5 mmol), benzophenone imine (0.543 g, 3 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 60 mg, 0.1 mmol), tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 50 mg, 0.05 mmol], and Cs2CO3 (1.6 g, 4.9 mmol) in 1,4-dioxane (20 mL). Stir the reaction under N2 at 110° C. for 16 hrs. Cool to room temperature, filter the solid, and concentrate the filtrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the title compound (0.78 g, 99%). MS: (M+1): 301.
Step 3: Synthesis of N-[4-(4-amino-2-cyano-phenoxy)-2-pyridyl]cyclopropanecarboxamide




Mix N-(4-hydroxy-2-pyridyl)cyclopropanecarboxamide (179 mg, 1 mmol), 5-(benzhydrylideneamino)-2-fluoro-benzonitrile (0.3 g, 1 mmol), Cs2CO3 (0.28 g, 2 mmol) and DMF (10 mL) in a microwave reaction vessel. Use microwave to heat the reaction at 150° C. for 30 min. TLC (EtOAc:PE=2:1) shows the reaction is complete. Cool the reaction to room temperature. Add water (15 mL), extract with EtOAc (15 mL×2). Combine the organic layers; wash with brine, dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude imine intermediate. Purification by chromatography (silica gel, EtOAc:PE=1:1) provides the imine intermediate (255 mg, 55.4%). MS: (M+1): 459.3.
Dissolve this imine intermediate in THF (10 mL), add 1M aqueous HCl (10 mL), stir for 1 hr. Adjust pH=7, extract with EtOAc (50 mL×3), combine the organic layers, wash with brine, dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=3:1) affords the title compound (124 mg, 78%). MS: (M+1): 295.2.




Step 1: Synthesis of 2-chloro-N-(2-fluoro-4-nitro-phenyl)pyridin-4-amine




Add 2-chloropyridin-4-amine (500 mg, 3.9 mmol), 1,2-difluoro-4-nitrobenzene (620 mg, 3.9 mmol), Cs2CO3 (3.2 g, 9.8 mmol) to DMF (15 mL). Stir the reaction at 110° C. for 16 hrs. Cool to room temperature, add water (100 mL), extract with EtOAc (15 mL×3). Combine the organic layers and wash with brine, dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:4) affords the title compound (280 mg, 27%). MS: (M+1): 268.1.
Step 2: Synthesis of N-[4-(2-fluoro-4-nitro-anilino)-2-pyridyl]cyclopropanecarboxamide




Mix 2-chloro-N-(2-fluoro-4-nitro-phenyl)pyridin-4-amine (280 mg, 1.05 mmol), cyclopropanecarboxamide (267 mg, 3.15 mmol), Cs2CO3 (863 mg, 2.63 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 65 mg, 1.05 mmol), tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 48 mg, 0.053 mmol] and 1,4-dioxane (6 mL) in a 50 mL round bottom flask, heat at 100° C. under N2 for 16 hrs. Cool the reaction to 25° C., add water (40 mL), extract with EtOAc (10 mL×2). Combine the organic layers; wash with brine (15 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, PE:EtOAc=7:3) affords the target compound (180 mg, 54%). MS: (M+1): 317.1.
Step 3: Synthesis of N-[4-(4-amino-2-fluoro-anilino)-2-pyridyl]cyclopropanecarboxamide




Dissolve the compound obtained in Step 2 (180 mg, 0.57 mmol) in methanol (6 mL), add wet Pd/C (10%, 70 mg), flush with H2, and stir the reaction at room temperature under H2 atmosphere for 16 hrs. After the reaction, remove H2, filter the reaction mixture and concentrate the filtrate to get the crude product (100 mg). Use the crude product directly without further purification. MS: (M+1): 287.2.




Step 1: Synthesis of 2-chloro-N-(3-fluoro-4-nitro-phenyl)-N-methyl-pyridin-4-amine




Add 2-chloro-N-(3-fluoro-4-nitro-phenyl)pyridin-4-amine (300 mg, 1.12 mmol), methyl iodide (174 mg, 1.23 mmol) and K2CO3 (201 mg, 1.45 mmol) in DMF (6 mL). Stir the reaction at 70° C. for 3 hrs. TLC (EtOAc:PE=1:1) shows the reaction is complete, cool the reaction to 0° C., add water, extract with EtOAc. Combine the organic layers. Wash with brine, dry over anhydrous Na2SO4. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (190 mg, 45%). MS: (M+1): 282.2.
Step 2: Synthesis of N-[4-(3-fluoro-4-nitro-anilino)-2-pyridyl]cyclopropanecarboxamide




Add 2-chloro-N-(3-fluoro-4-nitro-phenyl)-N-methyl-pyridin-4-amine (190 mg, 0.68 mmol), cyclopropanecarboxamide (85 mg, 2.03 mmol), Cs2CO3 (440 mg, 1.35 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 43 mg, 0.05 mmol) and tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 42 mg, 0.07 mmol] in 1,4-dioxane (6 mL) under N2. Heat the reaction at 125° C. for 16 hrs. TLC (EtOAc:PE=2:1) shows the reaction is complete. Cool to room temperature, filter, and concentrate the filtrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=2:1) affords the target compound (160 mg, 71.7%). MS: (M+1): 331.
Step 3: Synthesis of N-[4-(4-amino-3-fluoro-anilino)-2-pyridyl]cyclopropanecarboxamide




Dissolve the compound obtained in Step 2 (160 mg, 0.48 mmol) in methanol (15 mL) and dichloromethane (15 mL). Add Pd/C (10%, 32 mg), flush with H2. Stir the reaction at room temperature for 3.5 hrs under H2. LC/MS shows the reaction is complete. Remove the H2, filter the reaction mixture, and concentrate the filtrate under reduced pressure to give the crude product (120 mg). MS: (M+1): 301.




Step 1: Synthesis of N-[4-(2-fluoro-4-nitro-phenoxy)-2-pyridyl]-1-methyl-cyclopropanecarboxamide




Add oxalyl chloride (760 mg, 6.0 mmol) slowly to a solution of 1-methylcyclopropanecarboxylic acid (500 mg, 5.0 mmol) in DCM (10 mL) at 0° C., add a few drops of DMF. Stir the reaction at 0° C. for 30 min, and then stir at room temperature for 3 hrs. Remove the volatiles under reduced pressure to give the crude 1-methylcyclopropanecarbonyl chloride.
Add the solution of the above 1-methylcyclopropanecarbonyl chloride in DCM slowly to a solution of 4-(2-fluoro-4-nitro-phenoxy)pyridin-2-amine (500 mg, 2.0 mmol) and Et3N (505 mg, 5.0 mmol) in DCM (15 mL) at 0° C. Stir the reaction at room temperature for 15 hrs. TLC (EtOAc:PE=1:1) shows the reaction is complete. Concentrate the mixture to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the target compound (160 mg, 24%). MS: (M+1): 332.2.
Step 2: Synthesis of N-[4-(4-amino-2-fluoro-phenoxy)-2-pyridyl]-1-methyl-cyclopropanecarboxamide




Dissolve N-[4-(2-fluoro-4-nitro-phenoxy)-2-pyridyl]-1-methyl-cyclopropanecarboxamide (160 mg, 0.48 mmol) in DCM (30 mL), add Pd/C (10%, 55 mg), flush with H2. Stir the reaction at room temperature under H2 atmosphere for 2 hrs. Filter the reaction mixture; concentrate the filtrate under reduced pressure to give the crude product (100 mg) which is used without further purification. MS: (M+1): 302.2.




Step 1: Synthesis of tert-butyl N-(4-bromo-2-pyridyl)carbamate




Dissolve 2-amino-4-bromopyridine (1.0 g, 5.8 mmol), di-tert-butyl dicarbonate (Boc2O, 1.4 g, 6.4 mmol) in t-BuOH (15 mL), heat the mixture at 50° C. overnight. Cool to room temperature, pour to water (50 mL), extract with EtOAc (15 mL×3). Combine the organic layers, dry over anhydrous Na2SO4; concentrate under reduced pressure to afford the crude product (1.24 g) which is used without further purification. MS: (M+1): 219.0.
Step 2: Synthesis of tert-butyl N-(4-bromo-2-pyridyl)-N-(cyclopropanecarbonyl)carbamate




Add cyclopropanecarbonyl chloride (1.42 g, 13.68 mmol) slowly to a solution of tert-butyl N-(4-bromo-2-pyridyl)carbamate (1.24 g, 4.6 mmol), Et3N (1.38 g, 13.68 mmol) in DCM (15 mL) at 0° C. After addition, stir the reaction at room temperature for 2 hrs. Pour the reaction mixture to water (50 mL), extract with EtOAc (15 mL×3), combine the organic layers and wash with brine (100 mL). Dry over anhydrous Na2SO4; concentrate under reduced pressure to give the crude product (1.55 g) which is used without further purification. MS: (M+1): 241.0
Step 3: Synthesis of N-(4-bromo-2-pyridyl)cyclopropanecarboxamide




Dissolve tert-butyl N-(4-bromo-2-pyridyl)-N-(cyclopropanecarbonyl)carbamate (900 mg, 2.8 mmol) in DCM (8 mL), add slowly trifluoroacetic acid (4 mL). Stir the reaction at room temperature for 3 hrs. Pour the reaction mixture to water (50 mL), adjust to pH=7 with saturated NaHCO3 solution. Extract with EtOAc (15 mL×3), combine the organic layers; wash with brine (100 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product (670 mg) which is used without further purification. MS: (M+1): 243.1.
Step 4: Synthesis of N-[4-(3-fluoro-4-nitro-anilino)-2-pyridyl]cyclopropanecarboxamide




Under N2, mix N-(4-bromo-2-pyridyl)cyclopropanecarboxamide (460 mg, 1.9 mmol), 3-fluoro-4-nitroaniline (359 mg, 2.3 mmol), palladium(II) acetate [Pd(OAc)2, 43.2 mg, 0.19 mmol], 2-dicyclohexylphosphino-2′,6′-dimethoxybiphenyl (S-Phos, 158 mg, 0.38 mmol), Cs2CO3 (1.2 g, 3.8 mmol) and anhydrous toluene (10 mL). Stir the mixture at 120° C. under N2 for 16 hrs. Cool to room temperature, pour the reaction mixture to water (50 mL), extract with EtOAc (15 mL×3), combine the organic layers and dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, PE:EtOAc=54:46) affords the target compound (160 mg, 22%). MS: (M+1): 317.2.
Step 5: Synthesis of N-[4-(4-amino-3-fluoro-anilino)-2-pyridyl]cyclopropanecarboxamide




Dissolve N-[4-(3-fluoro-4-nitro-anilino)-2-pyridyl]cyclopropanecarboxamide (160 mg, 0.51 mmol) in methanol (10 mL), add wet Pd/C (10%, 60 mg), flush with H2 and then stir the reaction under H2 atmosphere at room temperature for 6 hrs. Remove H2, filter the mixture. Concentrate the filtrate under reduced pressure to give the crude product (140 mg) which is used directly without further purification. MS: (M+1): 287.2.




Step 1: Synthesis of tert-butyl N-[4-[(2-chloro-4-pyridyl)oxy]-2-fluoro-phenyl]carbamate




Dissolve tert-butyl N-(2-fluoro-4-hydroxy-phenyl)carbamate (10.47 g, 46.1 mmol) in N-methyl-2-pyrrolidone (NMP, 100 mL), add t-BuOK (5.67 g, 50.5 mmol) and stir at 0° C. for 30 min. Add 2-chloro-4-fluoropyridine (5.5 g, 41.9 mmol), then stir at 70° C. under N2 for 12 hrs. Quench the reaction with water, extract with EtOAc (300 mL×3), combine the organic layers, wash with brine (300 mL×2), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:4) affords the title compound (11.23 g, 79.5%). MS: (M+1): 339.
Step 2: Synthesis of tert-butyl N-[4-[[2-(cyclopropanecarbonylamino)-4-pyridyl]oxy]-2-fluoro-phenyl]carbamate




Dissolve tert-butyl N-[4-[(2-chloro-4-pyridyl)oxy]-2-fluoro-phenyl]carbamate (13 g, 14.7 mmol), cyclopropanecarboxamide (8.1 g, 36.7 mmol) in 1,4-dioxane (240 mL). Then add tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3, 1.75 g, 0.73 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 2.38 g, 1.47 mmol) and Cs2CO3 (24.8 g, 29.4 mmol) under N2. Stir the reaction at 100° C. for 12 hrs. Cool to room temperature, filter, and concentrate the filtrate to give the crude product. Purification by chromatography (silica gel, DCM:EtOAc=1:5) affords the title compound (12.14 g, 83.2%). MS: (M+1): 388.
Step 3: Synthesis of N-[4-(4-amino-3-fluoro-phenoxy)-2-pyridyl]cyclopropanecarboxamide




Dissolve tert-butyl N-[4-[[2-(cyclopropanecarbonylamino)-4-pyridyl]oxy]-2-fluoro-phenyl]carbamate (12.14 g, 31.28 mmol) in DCM (280 mL), add trifluoroacetic acid (35 mL). Stir the reaction at room temperature for 5 hrs. Remove the volatiles under reduced pressure. Add EtOAc (300 mL), washed with saturated NaHCO3 solution (300 mL×2), and brine (300 mL×2). Dry the organic layer with anhydrous Na2SO4. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (7 g, 78.4%). MS: (M+1): 288.




Step 1: Synthesis of 2-chloro-4-(4-methyl-3-nitro-phenoxy)pyridine




Add 4-methyl-3-nitrophenol (6.95 g, 45.46 mmol) and t-BuOK (5.94 g, 53.03 mmol) to N-methylpyrrolidin-2-one (100 mL), stir for 30 min, then add 2-chloro-4-fluoro-pyridine (5 g, 37.88 mmol) and stir the reaction at 70° C. overnight. TLC (PE:EtOAc=3:1) shows the reaction is complete. Add water (250 mL) to the reaction mixture, extract with EtOAc (100 mL×4), combine the organic layers, wash with water, brine respectively and dry over Na2SO4. Concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:4) affords the title compound (5.8 g, 58%). MS: (M+1): 265.1.
Step 2: Synthesis of N-[4-(4-methyl-3-nitro-phenoxy)-2-pyridyl]cyclopropanecarboxamide




Add the compound obtained in Step 1 (5.0 g, 18.87 mmol), cyclopropanecarboxamide (4.81 g, 56.61 mmol) to 1,4-dioxane (100 mL). Then under N2, add tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 0.86 g, 0.94 mmol], (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 1.17 g, 1.89 mmol), Cs2CO3 (12.27 g, 37.74 mmol). Stir the reaction under N2 at 100° C. overnight. TLC (PE:EtOAc=2:1) shows the reaction is complete. Cool the reaction to room temperature, filter, and concentrate the filtrate to get the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:4) affords the title compound (3.0 g, 51%). MS: (M+1): 314.2.
Step 3: Synthesis of N-[4-(3-amino-4-methyl-phenoxy)-2-pyridyl]cyclopropanecarboxamide




Dissolve the above nitro compound (3.0 g, 9.6 mmol) in methanol (150 mL), add Pd/C (10%, 0.9 g) to the solution, flush with H2 and stir under H2 atmosphere at room temperature overnight. TLC (PE:EtOAc=2:1) shows reaction is complete. Filter and concentrate the filtrate under reduced pressure to give a crude product. Purification by chromatography (silica gel, EtOAc:PE=1:2) affords the title compound (1.90 g, 70%). MS: (M+1): 284.2.
Intermediate B18 can be synthesized with similar method (Table B2).







TABLE B2







Intermediates B10-B18










Number
Starting material
Intermediate
MS [M + 1]+













B10








289.1


 


B11








295.2


 


B12








287.2


 


B13








301


 


B14








302.2


 


B15








287.2


 


B16








288


 


B17








284.2


 


B18








284.2













Step 1: Synthesis of phenyl N-[4-[[2-(cyclopropanecarbonylamino)-4-pyridyl]oxy]-2-fluoro-phenyl]carbamate




Mix N-[4-(4-amino-3-fluoro-phenoxy)-2-pyridyl]cyclopropanecarboxamide (500 mg, 1.7 mmol), DCM (8 mL) and pyridine (345 mg) and cool to 0-5° C. Add a solution of phenyl chloroformate (350 mg, 2.2 mmol) in DCM (1 mL). Stir the reaction at room temperature for 1.5 hrs. TLC (EtOAc:PE=1:1) shows the reaction is complete. Quench the reaction with water (5 mL). Wash the organic layer with 1N HCl solution, saturated NaHCO3 solution and brine respectively. Dry the organic layer with anhydrous Na2SO4. Filter and concentrate to give the crude product (710 mg) which is used without further purification. MS: (M+1): 408.
Intermediates B20-21 can be synthesized with similar method (Table B3).







TABLE B3







Intermediates B19-B21













MS


Number
Starting material
Intermediate
[M + 1]+





B19








408


 


B20








408


 


B21








NA









Preparation of Intermediate C




Step 1: Synthesis of tert-butyl 4-(4-nitro-2-(trifluoromethyl)phenyl)-5,6-dihydropyridine-1 (21)-carboxylate




Add 2-bromo-5-nitrobenzotrifluoride (250 mg, 0.926 mmol) to dioxane (3 mL), then add tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate (207 mg, 0.926 mmol), tetrakis(triphenylphosphine)palladium [Pd(PPh3)4, 35 mg, 0.03 mmol] and saturated sodium bicarbonate solution (1 mL) under nitrogen atmosphere. Stir the reaction mixture overnight at 120° C. TLC (PE:EtOAc=5:1) shows the reaction is complete. Partition the mixture between EtOAc and water, dry the organic layer over sodium sulfate, filter and concentrate the filtrate under reduced pressure. Purify the residue by flash chromatography (silica gel, PE:EtOAc=5:1) to afford the desired product (289 mg, 84%).
Step 2: Synthesis of tert-butyl 4-(4-amino-2-(trifluoromethyl)phenyl)piperidine-1-carboxylate




Stir the mixture of tert-butyl 4-(4-nitro-2-(trifluoromethyl)phenyl)-5,6-dihydropyridine-1(2H)-carboxylate (289 mg, 0.776 mmol), methanol (50 mL) and Pd/C (10%, 25 mg) under hydrogen atmosphere for 30 hrs. Remove H2, filter the mixture and concentrate the filtrate under reduced pressure. Purify the residue with flash chromatography (silica gel, PE:EtOAc=5:1) to afford the desired product (151 mg, 84%).
Step 3: Synthesis of 4-(1-methylpiperidin-4-yl)-3-(trifluoromethyl)aniline




Add LiAlH4 (66.5 mg, 1.75 mmol) at 0° C. to the mixture of tert-butyl 4-(4-amino-2-(trifluoromethyl)phenyl)piperidine-1-carboxylat (151 mg, 0.438 mmol) in THF (3 mL), stir the reaction mixture overnight at room temperature. Quench the reaction with 15% sodium hydroxide solution, filter and concentrate the filtrate under reduced pressure. Purify the residue by flash chromatography (silica gel, DCM:MeOH=15:1) to afford the desired product (70 mg, 56%). MS: (M+1): 259.1.




Step 1: Synthesis of 1-(methylsulfonyl)-3-nitro-5-(trifluoromethyl)benzene




Stir the mixture of 1-bromo-3-nitro-5-(trifluoromethyl)benzene (6 g, 22.2 mmol), sodium methanesulfinate (2.8 g, 26.7 mmol), cuprous iodide (0.5 g, 2.22 mmol), L(−)-proline (0.5 g, 4.44 mmol) and sodium hydroxide (0.088 g, 4.44 mmol) in DMSO (20 mL) at 100° C. for 15 hrs under nitrogen atmosphere. Add water, extract the mixture with ethyl acetate (200 mL×3), combine the organic layers, dry over anhydrous sodium sulfate, and concentrate the mixture under reduced pressure. Purify the residue by flash chromatography to afford the desired product (2 g, 33.3%).
Step 2: Synthesis of 3-(methylsulfonyl)-5-(trifluoromethyl)aniline




Stir the mixture of 1-(methylsulfonyl)-3-nitro-5-(trifluoromethyl)benzene (2 g, 7.43 mmol), Pd/C (10%, 500 mg) in methanol (100 mL) under hydrogen atmosphere at room temperature for 15 hrs. Filter off the solid and concentrate the filtrate under reduced pressure. Purify the residue with flash chromatography (silica gel, EtOAc:PE=1:5) to afford the desired product (1.3 g, 73%). MS: (M+23): 262.0.




Step 1: Synthesis of 1-methyl-3-(3-nitro-5-(trifluoromethyl)phenoxy)pyrrolidine




Add 3-nitro-5-(trifluoromethyl)phenol (1.6 g, 7.7 mmol), 1-methylpyrrolidin-3-ol (940 mg, 9.3 mmol), triphenylphosphine (3.4 g, 11.6 mmol) in THF (20 mL), stir on ice bath, then add diethyl azodicarboxylate (DEAD, 2.0 g, 11.6 mmol). Stir the mixture at ambient temperature for 15 hrs. Add water (100 mL), then extract the mixture with ethyl acetate (100 mL×3), combine the organic layers and dry over anhydrous sodium sulfate. Filter and concentrate the filtrate to get the crude product. Purify by flash chromatography (silica gel, DCM:MeOH=20:1) to afford the target product (1.5 g, 67%). MS: (M+1): 291.1.
Step 2: Synthesis of 3-(1-methylpyrrolidin-3-yloxy)-5-(trifluoromethyl)aniline




Add Pd/C (10%, 400 mg) to the solution of 1-methyl-3-(3-nitro-5-(trifluoromethyl) phenoxy)pyrrolidine (1.54 g, 5.31 mmol) in methanol (10 mL). Stir the mixture under hydrogen atmosphere at room temperature for 15 hrs. Filter off the solid and concentrate the filtrate to give crude product. Purify by flash chromatography (silica gel, EtOAc:PE=1:1) to afford the target compound (900 mg, 65%). MS: (M+1): 261.1.
Intermediate C4 can be prepared with similar method (Table C1).







Step 1: Synthesis of tert-butyl 4-(methoxy(methyl)carbamoyl)piperidine-1-carboxylate




Add N,O-dimethylhydroxylamine hydrochloride (2.52 g, 26 mmol), 1-hydroxybenzotriazole (HOBt) (2.8 g, 20.8 mmol), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI.HCl) (4 g, 20.8 mmol) and 4-methylmorpholine (5.28 g, 52 mmol) at 0° C. to the solution of N-Boc-piperidine-4-carboxylic acid (4 g, 17.4 mmol) in DMF (50 mL), stir the mixture overnight at room temperature. TLC (PE:EtOAc=1:1) shows the reaction is complete. Partition between ethyl acetate and water, collect the organic layer and wash with brine, dry over anhydrous sodium sulfate, filter and concentrate the filtrate under reduced pressure to give the crude product (4.5 g, 95%) which is used in next step without further purification.
Step 2: Synthesis of tert-butyl-4-(3-bromo-5-(trifluoromethyl)benzoyl)piperidine-1-carboxylate




Add n-butyllithium (2.5 N, 1.44 mL) to the solution of 1,3-dibromo-5-(trifluoromethyl)benzene (1 g, 3.28 mmol) in THF (15 mL) at −78° C. and stir the mixture at −78° C. for 1 hr. Then add tert-butyl 4-(methoxy(methyl)carbamoyl) piperidine-1-carboxylate (1.4 g, 4.92 mmol) obtained in Step 1 to the mixture and continue stirring. TLC (PE:EtOAc=5:1) shows the reaction is complete. Quench the reaction mixture with saturated ammonium chloride solution. Partition between EtOAc and water, separate the organic layer, and wash with brine, dry over anhydrous sodium sulfate, concentrate to give the crude product. Purify by flash chromatography (silica gel, DCM:MeOH=20:1) to afford the product (387 mg, 27%). MS: (M+1): 436.
Step 3: Synthesis of (3-bromo-5-(trifluoromethyl)phenyl)(piperidin-4-yl)methanone




Add trifluoroacetic acid in dichloromethane (20%, 5 mL) to the solution of tert-butyl-4-(3-bromo-5-(trifluoromethyl)benzoyl)piperidine-1-carboxylate (387 mg, 0.88 mmol) in dichloromethane (5 mL) at 0° C., stir the mixture. TLC (PE:EtOAc=5:1) shows the reaction is complete. Concentrate under reduced pressure to dryness to afford the crude product (400 mg, 100%) which is used directly in next step. MS: (M+1): 336.0.
Step 4: Synthesis of (3-bromo-5-(trifluoromethyl)phenyl)(1-methylpiperidin-4-yl)methanone




Stir the mixture of (3-bromo-5-(trifluoromethyl)phenyl)(piperidin-4-yl)methanone (400 mg, 0.96 mmol), formic acid (1 mL) and formaldehyde (1 mL) at 100° C. overnight. Upon the reaction completion, concentrate under reduced pressure to provide the crude product (250 mg, 77%) which is used in next step without further purification.
Step 5: Synthesis of (3-(diphenylmethyleneamino)-5-(trifluoromethyl)phenyl)(1-methylpiperidin-4-yl)methanone




Add benzophenone imine (195 mg, 1.07 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 37 mg, 0.059 mmol), tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 25 mg, 0.027 mmol] and cesium carbonate (585 mg, 1.79 mmol) to the solution of (3-bromo-5-(trifluoromethyl)phenyl)(1-methylpiperidin-4-yl)methanone (251 mg, 0.717 mmol) in dioxane (5 mL), then stir the reaction mixture overnight at 100° C. Upon reaction completion, filter the mixture and concentrate the filtrate under reduced pressure. Purify the residue with flash chromatography (silica gel, PE:EtOAc=1:1) to afford the product (210 mg, 65%). MS: (M+1): 451.2.
Step 6: Synthesis of (3-amino-5-(trifluoromethyl)phenyl)(1-methylpiperidin-4-yl)methanone




Dissolve (3-(diphenylmethyleneamino)-5-(trifluoromethyl)phenyl)(1-methylpiperidin-4-yl)methanone (210 mg, 0.47 mmol) in THF (2 mL) in an ice-bath, add hydrochloric acid (4N, 0.5 mL), then stir it at room temperature for one hour. TLC (PE:EtOAc=1:1) shows the reaction is complete. Partition the mixture between EtOAc and water. Collect the aqueous layer, adjust pH=9 with sodium hydroxide solution, extract with ethyl acetate three times, combine the organic layers, dry over anhydrous sodium sulfate, concentrate to yield the crude product (78 mg, 59%). MS: (M+1): 287.1.




Step 1: Synthesis of 1-(3-bromo-5-(trifluoromethyl) phenylamino)propan-2-ol




Add propylene oxide (36 mg, 0.624 mmol) and lithium perchlorate (56 mg, 0.458 mmol) to the solution of 3-bromo-5-(trifluoromethyl)aniline (100 mg, 0.416 mmol) in acetonitrile (5 mL), stir the resulting mixture overnight at ambient temperature. Upon completion of the reaction, concentrate the mixture under reduced pressure. Purify the residue by flash chromatography (silica gel, PE:EtOAc=5:1) to afford the product (136 mg, 100%).
Step 2: Synthesis of 3-(3-bromo-5-(trifluoromethyl)phenyl)-5-methyloxazolidin-2-one




Add triethylamine (339 mg, 3.35 mmol) and triphosgene (250 mg, 0.838 mmol) to the solution of 1-(3-bromo-5-(trifluoromethyl)phenylamino)propan-2-ol (500 mg, 1.68 mmol) in dichloromethane (8 mL), stir the resulting mixture overnight at ambient temperature. Partition the reaction mixture between dichloromethane and water, separate the organic layer, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to give the crude product. Purify with flash chromatography (silica gel, PE:EtOAc=1:1) to afford the product (530 mg, 97%). MS: (M+1): 324.0.
Step 3: Synthesis of 3-(3-(diphenylmethyleneamino)-5-trifluoromethyl)phenyl)-5-methyloxazolidin-2-one




Add benzophenone imine (3.56 g, 2 mmol), (±)-2,2-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 43 mg, 0.069 mmol), tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 90 mg, 0.098 mmol] and cesium carbonate (858 mg, 2.63 mmol) to the solution of 3-(3-bromo-5-(trifluoromethyl)phenyl)-5-methyloxazolidin-2-one (430 mg, 1.32 mmol) in dioxane (5 mL). Stir the mixture under nitrogen atmosphere at 100° C. overnight. TLC (PE:EtOAc=1:1) shows the reaction is complete. Filter the mixture and concentrate the filtrate to yield the crude product. Purify by flash chromatography (silica gel, PE:EtOAc=1:1) to afford the product (500 mg, 89%).
Step 4: Synthesis of 3-(3-amino-5-(trifluoromethyl)phenyl)-5-methyloxazolidin-2-one




Add hydrochloric acid (4 N, 1 mL) to the solution of the above imine (310 mg, 0.731 mmol) in THF (5 mL) in an ice-bath. Then stir the reaction for half hour at ambient temperature. TLC (PE:EtOAc=1:1) shows the reaction is complete. Partition the mixture between water and ethyl acetate, collect the aqueous layer, adjust pH=9 with sodium hydroxide solution, extract the aqueous layer with ethyl acetate three times, combine the organic layers, dry over anhydrous sodium sulfate, concentrate under reduced pressure to get the crude product (170 mg, 89%). MS: (M+1): 261.0.




Step 1: Synthesis of methyl 3-nitro-5-(trifluoromethyl)benzoate




Add thionyl chloride (2 mL) dropwise to the solution of 3-nitro-5-(trifluoromethyl) benzoic acid (1.5 g, 6.4 mmol) in methanol (20 mL), reflux for 5 hrs. TLC (PE:EtOAc=1:1) shows the reaction is complete. Concentrate under reduced pressure to give the crude product (1.8 g, 100%) which is used in next step without further purification.
Step 2: Synthesis of 3-amino-5-(trifluoromethyl)benzohydrazide




Mix methyl 3-nitro-5-(trifluoromethyl)benzoate (1.8 g, 12 mmol), ethanol (6 mL) and 80% (w/w) hydrazine (10 mL), reflux overnight. Concentrate the mixture, partition the resulting residue between ethyl acetate and water, collect the organic layer, wash with sodium bicarbonate, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to give the crude product (1.23 g, 77%) which is used in next step without further purification.
Step 3: Synthesis of 5-(3-amino-5-(trifluoromethyl)phenyl)-1,3,4-oxadiazol-2-amine




Add sodium bicarbonate (270 mg, 3.21 mmol) and cyanogen bromide (187 mg, 1.76 mmol) to the solution of 3-amino-5-(trifluoromethyl)benzohydrazide (400 mg, 1.6 mmol) in ethanol (5 mL), reflux for 5 hrs. TLC (PE:EtOAc=1:1) shows the reaction is complete. Concentrate the mixture, partition the residue between EtOAc and water, collect the organic layer, wash with saturated sodium bicarbonate solution, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to afford the crude product. Recrystallization in ethyl acetate yields the pure product (280 mg, 63%).




Step 1: Synthesis of 2-methoxy-N-(3-nitro-5-(trifluoromethyl)phenyl)acetamide




Stir the mixture of 3-nitro-5-(trifluoromethyl)aniline (500 mg, 2.4 mmol), 2-methoxyacetic acid (220 mg, 2.4 mmol), 1-hydroxybenzotriazole (HOBt, 324 mg, 2.4 mmol), N-methyl morpholine (NMM, 480 mg, 4.8 mmol), 1-ethyl-3-(3-dimethyllaminopropyl)carbodiimide hydrochloride (EDCI.HCl, 920 mg, 4.8 mmol) in DMF (8 mL) at 80° C. for 16 hrs. Cool the reaction mixture to room temperature, add water (100 mL), extract with ethyl acetate (30 mL×2), combine the organic layers, wash with brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to give the crude product. Purify by flash chromatography (silica gel, EtOAc:PE=15:85) to yield the target compound (260 mg, 39%).
Step 2: Synthesis of N-(3-amino-5-(trifluoromethyl)phenyl)-2-methoxyacetamide




Add Pd/C (10%, 30 mg) to the solution of 2-methoxy-N-(3-nitro-5-(trifluoromethyl) phenyl)acetamide (100 mg, 0.36 mmol) in methanol (6 mL), stir the mixture under hydrogen atmosphere at room temperature for 16 hrs. Filter off the solid, concentrate the filtrate under reduced pressure to yield the crude product (89 mg, 100%) which is used without further purification.
Intermediate C9 can be synthesized with similar method (Table C1).







TABLE C1







Intermediates C1-C9










Number
Starting material
Intermediate
MS [M + 1]+





C1








259.2


 


C2








[M + 23] 262.0


 


C3








261.1


 


C4








248.1


 


C5








287.1


 


C6








NA


 


C7








NA


 


C8








249.1


 


C9








219.1













Step 1: Synthesis of 1-(3-nitro-5-(trifluoromethyl)phenyl)ethanol




Add sodium borohydride (95 mg, 2.44 mmol) to a solution of 1-(3-nitro-5-(trifluoromethyl)phenyl)ethanone (570 mg, 2.44 mmol) in methanol (5 mL), stir the mixture for 15 min at ambient temperature. Quench the reaction with water, adjust pH=7 with 1N hydrochloric acid, extract with ethyl acetate (50 mL×3), combine the organic layers, wash with brine, dry over sodium sulfate. Concentrate under reduced pressure to provide the crude product. Purify by flash chromatography (silica gel, EtOAc:PE=1:10) to afford the title compound (520 mg, 91.2%).
Step 2: Synthesis of 1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl 4-methylbenzenesulfonate




Add p-toluenesulfonyl chloride (1.7 g, 8.9 mmol) dropwise at 0° C. to the mixture of 1-(3-nitro-5-(trifluoromethyl)phenyl)ethanol (520 mg, 2.23 mmol) and triethylamine (1.4 g, 13.4 mmol) in dichloromethane (20 mL), stir the resulting mixture for 12 hrs at ambient temperature. Concentrate the mixture under reduced pressure to get a residue, purify the residue by chromatography (silica gel, EtOAc:PE=1:10) to afford the title compound (700 mg, 80.6%).
Step 3: Synthesis of tert-butyl 4-(1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl)piperazine-1-carboxylate




Stir the mixture of 1-(3-nitro-5-(trifluoromethyl)phenyl)ethyl 4-methylbenzenesulfonate (350 mg, 0.90 mmol), 1-piperazinecarboxylic acid tert-butyl ester (335 mg, 1.8 mmol) and diisopropylethylamine (232 mg, 1.8 mmol) in THF (10 mL) for 12 hrs at 80° C. Concentrate the reaction mixture to obtain the crude product, purify by flash chromatography (silica gel, EtOAc:PE=1:10) to afford the title compound (280 mg, 77.1%). MS: (M+1): 404.2.
Step 4: Synthesis of tert-butyl 4-(1-(3-amino-5-(trifluoromethyl)phenyl)ethyl) piperazine-1-carboxylate




Add Pd/C (10%, 75 mg) to the solution of tert-butyl 4-(1-(3-nitro-5-(trifluoromethyl)-phenyl)ethyl)piperazine-1-carboxylate (280 mg, 0.69 mmol) in methanol (20 mL), stir the resulting mixture under hydrogen atmosphere for 2 hrs at room temperature. Filter and concentrate the filtrate to afford the crude product. Purify by flash chromatography (silica gel, EtOAc:PE-1:1) to afford the title compound (165 mg, 63.7%). MS: (M+1): 374.1.
Intermediates C11-C12 can be synthesized with similar method (Table C2).







TABLE C2







Intermediates C10-C12













MS 


Num-


[M +


ber
Starting material
Intermediate
1]+





C10








374.2


 


C11








288.2


 


C12








NA













Step 1: Synthesis of 4-fluoro-1-(3-nitro-5-(trifluoromethyl)phenyl)piperidine




Mix 1-bromo-3-nitro-5-(trifluoromethyl)benzene (700 mg, 2.59 mmol), 4-fluoropiperidine hydrochloride (544 mg, 3.89 mmol), cesium carbonate (2.5 g, 7.77 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 48 mg, 0.077 mmol), tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 47 mg, 0.052 mmol] in dioxane (10 mL), Stir the mixture under nitrogen atmosphere at 100° C. for 16 hrs. Cool the reaction mixture to room temperature, dilute it with water (10 mL), extract with ethyl acetate (10 mL×2), combine the organic layers, wash with brine (15 mL) and dry over anhydrous sodium sulfate. Concentrate the filtrate to get the crude product. Purify by flash chromatography (silica gel, 100% PE) to yield the target compound (480 mg, 63%).
Step 2: Synthesis of 3-(4-fluoropiperidin-1-yl)-5-(trifluoromethyl)aniline




Add Pd/C (10%, 70 mg) to the solution of 4-fluoro-1-(3-nitro-5-(trifluoromethyl)phenyl)piperidine (300 mg, 1.0 mmol) in methanol (10 mL), stir the mixture under hydrogen atmosphere for 16 hrs at room temperature. Filter and concentrate the filtrate to get the crude product (270 mg, 100%) which is used directly without further purification.
Intermediates C14-20 can be synthesized with similar method (Table C3).







TABLE C3







Intermediates C13-C20










Number
Starting material
Intermediate
MS [M + 1]+





C13








263.1


 


C14








[M + 23] 382.2


 


C15








275.2


 


C16








274.2


 


C17








219.2


 


C18








NA


 


C19








NA


 


C20








NA













Step 1: Synthesis of 1-(chloromethyl)-3-nitro-5-(trifluoromethyl)benzene




Add slowly tosyl chloride (1.1 g, 6.02 mmol) to the mixture of (3-nitro-5-(trifluoromethyl)phenyl)methanol (1.1 g, 5.18 mmol) and triethylamine (1.62 mL) in dichloromethane (20 mL) at 0° C., stir the resulting mixture for 15 hrs at room temperature. Concentrate the mixture to obtain the crude product, purify by flash chromatography (silica gel, EtOAc:PE=1:15) to afford the title compound (655 mg, 55%).
Step 2: Synthesis of 1-methyl-4-(3-nitro-5-(trifluoromethyl)benzyl)piperazine




Stir the mixture of 1-(chloromethyl)-3-nitro-5-(trifluoromethyl)benzene (640 mg, 2.68 mmol) and N-methylpiperazine (536 mg, 5.36 mmol) in THF (8 mL) for 2 hrs at 60° C. TLC (EtOAc:PE=1:1) shows the reaction is complete. Concentrate the reaction mixture under reduced pressure to obtain the crude product (812 mg, 100%). MS: (M+1): 303.1.
Step 3: Synthesis of 3-((4-methylpiperazin-1-yl)methyl)-5-(trifluoromethyl)aniline




Add Pd/C (10%, 200 mg) to the solution of 1-methyl-4-(3-nitro-5-(trifluoromethyl)benzyl)piperazine (812 mg, 2.68 mmol) in methanol (15 mL), flush with H2, stir under hydrogen atmosphere for 16 hrs at room temperature. Filter and concentrate the filtrate to give the crude product (732 mg, 100%). MS: (M+1): 273.0.




Step 1: Synthesis of N-(3-amino-5-(trifluoromethyl)phenyl)-4-chlorobutanamide




Mix 5-(trifluoromethyl)benzene-1,3-diamine (300 mg, 1.70 mmol), 4-chlorobutanoic acid (230 mg, 1.87 mmol), 1-ethyl-3-(3-dimethyllaminopropyl)carbodiimide hydrochloride (EDCI•HCl, 650 mg, 3.40 mmol), 1-hydroxybenzotriazole (HOBt, 230 mg, 1.70 mmol) and diisopropylethylamine (450 mg, 3.40 mmol) in dichloromethane (10 mL), stir overnight at room temperature. Concentrate the reaction mixture under reduced pressure to obtain the crude product. Purify by flash chromatography (silica gel, PE:EtOAc=1:10) to yield the product (250 mg, 52%). MS: (M+1): 281.2.
Step 2: Synthesis of 1-(3-amino-5-(trifluoromethyl)phenyl)pyrrolidin-2-one




Add sodium hydride (60%, 65 mg, 1.64 mmol) portionwise to the solution of N-(3-amino-5-(trifluoromethyl)phenyl)-4-chlorobutanamide (230 mg, 0.82 mmol) in THF (5 mL), stir the resulting mixture for 2 hrs at ambient temperature. Quench the reaction with water, extract the mixture with ethyl acetate, wash the organic layer with water and brine sequentially, and dry over anhydrous sodium sulfate, concentrate to give brownish oil. Purify by flash chromatography (silica gel, PE:EtOAc=1:1) to yield white solid as the title compound (180 mg, 90%). MS: (M+1): 245.1.




Step 1: Synthesis of 2-fluoroethyl 4-methylbenzenesulfonate




Add toluenesulfonyl chloride (1.8 g, 9.4 mmol) to the mixture of 2-fluoroethanol (500 mg, 7.8 mmol) and triethylamine (1.6 g, 15.6 mmol) in dichloromethane (30 mL) at 0-5° C. Stir the reaction at ambient temperature for 5 hrs. Remove the volatiles under reduced pressure to yield the crude product. Purify by flash chromatography (silica gel, PE:EtOAc=2:1) to afford a white solid as the title compound (1.3 g, 77%).
Step 2: Synthesis of 1-(2-fluoroethyl)-4-(3-nitro-5-(trifluoromethyl)phenyl)piperazine




Mix (3-nitro-5-(trifluoromethyl)phenyl)piperazine (200 mg, 0.73 mmol), 2-fluoroethyl 4-methylbenzenesulfonate (320 mg, 1.46 mmol) and 4-dimethylaminopyridine (50 mg, 0.37 mmol) in THF (15 mL), stir at 90° C. overnight. TLC (EtOAc:PE=1:1) shows the reaction is complete. Concentrate under reduced pressure to give a residue, purify the residue by flash chromatography (silica gel, PE:EtOAc=1:1) to yield a white solid as the target compound (120 mg, 51%).
Step 3: Synthesis of 3-(4-(2-fluoroethyl)piperazin-1-yl)-5-(trifluoromethyl)aniline




Stir the mixture of 1-(2-fluoroethyl)-4-(3-nitro-5-(trifluoromethyl)phenyl)piperazine (120 mg, 0.50 mol) and Pd/C (10%, 30 mg) in methanol (5 mL) under hydrogen atmosphere for 2 hrs at room temperature. Filter and concentrate the filtrate under reduced pressure to afford the white solid as the target compound (90 mg, 84%).
Intermediates C24-C25 can be synthesized with similar method (Table C4).







TABLE C4







Intermediates C21-C25










Number
Starting material
Intermediate
MS [M + 1]+





C21








274.2


 


C22








245.1


 


C23








NA


 


C24








NA


 


C25








NA













Step 1: Synthesis of 3-bromo-5-(1,1-difluoroethyl) pyridine




Heat the mixture of 1-(5-bromopyridin-3-yl)ethanone (500 mg, 2.5 mmol), bis-(2-methoxyethyl)aminosulfur trifluoride (BAST, 2.2 g, 10 mmol) in dichloromethane (8 mL) in a sealed tube at 60° C. overnight. TLC (PE:EtOAc=1:1) shows the reaction is complete. Concentrate the mixture, purify the residue by flash chromatography (silica gel, PE:EtOAc=1:1) to afford the title compound (200 mg, 36%). MS: (M+1): 222/224.
Step 2: Synthesis of 5-(1,1-difluoroethyl)-N-(diphenylmethylene)pyridin-3-amine




Add benzophenone imine (215 mg, 1.19 mmol), cesium carbonate (640 mg, 1.96 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 98 mg, 0.16 mmol), tris(dibenzylideneacetone)dipalladium [Pd2(dba)3, 90 mg, 0.098 mmol] to the solution of 3-bromo-5-(1,1-difluoroethyl)pyridine (200 mg, 0.9 mmol) in dioxane (10 mL), stir the resulting mixture under nitrogen atmosphere at 100° C. for 16 hrs. Cool the reaction mixture to room temperature, filter off the solid, concentrate the filtrate under reduced pressure, purify by flash chromatography (silica gel, EtOAc:PE=2:1) to yield the title compound (296 mg, 92%). MS: (M+1): 323.
Step 3: Synthesis of 5-(1,1-difluoroethyl) pyridin-3-amine




Add 5-(1,1-difluoroethyl)-N-(diphenylmethylene)pyridin-3-amine (296 mg, 0.92 mmol), water (1 mL) and hydrochloric acid (1N, 4 mL) to THF (10 mL), stir at room temperature for 3 hrs. TLC (PE:EtOAc=2:1) shows the reaction is complete. Extract the mixture with ethyl acetate, wash the organic layer with saturated sodium bicarbonate solution and brine sequentially, and dry over anhydrous sodium sulfate. Concentrate under reduced pressure to give the crude product. Purification by flash chromatography (silica gel, EtOAc:PE=1:1) yields the title compound (140 mg, 96.5%). MS: (M+1): 159.2.




Synthesis of 3-cyclopropylaniline
Stir the mixture of 3-bromoaniline (400 mg, 2.35 mmol), cyclopropylboronic acid (240 mg, 2.79 mmol), tetrakis(triphenylphosphine)palladium(0) (140 mg, 0.12 mmol), cesium carbonate (1.5 g, 4.60 mmol) in dioxane (5 mL) and water (0.3 mL) under nitrogen atmosphere for 17 hrs at 100° C. Cool the reaction to room temperature, filter and concentrate the filtrate under reduced pressure. Purify by flash chromatography (silica gel, EtOAc:PE=3:7) to afford the title compound (120 mg, 38.8%). MS: (M+1): 134.0.




Step 1: Synthesis of 5-bromo-2-fluoroaniline




Mix 4-bromo-1-fluoro-2-nitrobenzene (2 g, 9.17 mmol) and stannous chloride dihydrate (8 g, 35 mmol) in ethanol (20 mL), heat the mixture to reflux for 4 hrs. Cool the reaction to room temperature, dilute with water and adjust pH=12 with sodium hydroxide solution, filter off the solid. Partition the filtrate in water and dichloromethane, collect the organic layer, dry over anhydrous sodium sulfate, and concentrate under reduced pressure to give crude product (800 mg, 46.5%). MS: (M+1): 190/192.
Step 2: Synthesis of 5-cyclopropyl-2-fluoroaniline




Stir the mixture of 5-bromo-2-fluoroaniline (800 mg, 4.26 mmol), cyclopropylboronic acid (434 mg, 5.05 mmol), tetrakis(triphenylphosphine)palladium(0) (242 mg, 0.21 mmol), cesium carbonate (2.7 g, 8.28 mmol) in dioxane (5 mL) and water (0.3 mL) under N2 atmosphere at 100° C. for 17 hrs. TLC (EtOAc:PE=1:3) shows the reaction is complete. Filter off the solid, concentrate the filtrate. Purify by flash chromatography (silica gel, EtOAc:PE=3:7) to afford the title compound (350 mg, 55%). MS: (M+1): 152.1.




Step 1: Synthesis of 3,3-difluoro-5-nitroindolin-2-one




Add 5-nitro-indoline-2,3-dione (5 g, 15.7 mmol) and bis-(2-methoxyethyl)aminosulfur trifluoride (20 mL) in dichloromethane (100 mL), stir at room temperature for 12 hrs. Quench the reaction with methanol (10 mL), pour the mixture into ice water (50 mL), extract with dichloromethane, combine the organic layers, wash with brine, dry over anhydrous sodium sulfate, concentrate under reduced pressure, purify by flash chromatography (silica gel, EtOAc:PE=1:4) to afford the title compound (3.5 g, 62.8%).
Step 2: Synthesis of 3-fluoro-5-nitro-1H-indole




Add borane-tetrahydrofuran complex (1M, 15 mL) dropwise to the solution of 3,3-difluoro-5-nitroindolin-2-one (650 mg, 3 mmol) in THF (10 mL) at 0° C. After addition, stir the reaction at 70° C. for 5 hrs. Concentrate under reduced pressure, add ethyl acetate to the residue, wash the organic layer with saturated sodium bicarbonate solution twice and then with brine twice, dry over anhydrous sodium sulfate. Concentrate under reduced pressure to obtain the crude product. Purify by flash chromatography (silica gel, EtOAc:PE=1:3) to yield the title compound (500 mg, 91.5%).
Step 3: Synthesis of 3-fluoro-1H-indol-5-amine




Add zinc powder (387 mg, 5 mmol) portionwise to a solution of 3-fluoro-5-nitro-1H-indole (360 mg, 2 mmol) in methanol (10 mL). Then add dropwise 15.5% ammonium chloride solution (10 mL, 20 mmol), stir the reaction at ambient temperature for 5 hrs. Filter, dilute the filtrate with water (50 mL), extract with ethyl acetate (50 mL×2), wash with brine (50 mL×2), dry over anhydrous sodium sulfate. Concentrate under reduced pressure to give the crude product. Purify by flash chromatography (silica gel, EtOAc:PE=1:3) to yield the title compound (150 mg, 50%). MS: (M+1): 151.2.




Step 1: Synthesis of methyl 2-(2-nitrophenyl)acetate




Add sulfonyl chloride (3 mL) dropwise to a solution of 2-nitrophenylacetic acid (7.0 g, 38.6 mmol) in methanol (20 mL) at 0° C., reflux the resulting mixture overnight. TLC (PE:EtOAc=3:1) shows the reaction is complete. Concentrate the reaction mixture under reduced pressure, add water, extract with ethyl acetate twice, and wash the combined organic layers with brine, dry over anhydrous sodium sulfate. Concentrate under reduced pressure to yield the crude product (7.5 g, 94.7%) which is used directly without further purification. MS: (M+1): 196.1.
Step 2: Synthesis of methyl 2-(2-aminophenyl)acetate




Add Pd/C (10%, 800 mg) to a solution of methyl 2-(2-nitrophenyl)acetate (7.5 g, 38.5 mmol) in methanol (50 mL) under N2, then flush with H2, stir the reaction under H2 atmosphere at room temperature overnight. Remove the hydrogen; filter off the solid, concentrate the filtrate under reduced pressure to obtain the crude product (4.8 g, 76.2%) which is used directly without further purification. MS: (M+1):166.
Step 3: Synthesis of indolin-2-one




Stir the mixture of methyl 2-(2-aminophenyl)acetate (4.8 g, 29.1 mmol) and potassium carbonate (8.0 g, 58.2 mmol) in DMF (15 mL) at room temperature overnight. TLC (PE:EtOAc=2:1) shows the reaction is complete. Concentrate the reaction mixture under reduced pressure to give the crude product. Purify by chromatography (silica gel, EtOAc:PE=1:2) to afford the title compound (4.0 g, 95%). MS: (M+1): 134.2.
Step 4: Synthesis of 1-methylindolin-2-one




Add diethyl azodicarboxylate (DEAD, 8.0 g, 44.9 mmol) dropwise to a solution of indolin-2-one (4.0 g, 30.0 mmol), methanol (1.4 g, 44.9 mmol) and triphenylphosphine (12.0 g, 44.9 mmol) in THF (40 mL) at 0° C., stir the reaction at room temperature overnight. Concentrate under reduced pressure, purify the resulting residue with flash chromatography (silica gel, EtOAc:PE=1:3) to afford the title compound (2.6 g, 59.1%). MS: (M+1): 148.2.
Step 5: Synthesis of 1′-methylspiro[cyclopropane-1,3′-indolin]-2′-one




Add sodium hydride (60%, 1.4 g, 35.4 mmol) portionwise to the solution of 1-methylindolin-2-one (2.6 g, 17.7 mmol) in DMF (10 mL) at 0° C., stir for 30 min. Then add 1,2-dibromoethane (3.31 g, 17.7 mmol) to the mixture, stir the resulting mixture at room temperature overnight. Pour the reaction mixture to ice water, extract with ethyl acetate (100 mL×2), and wash the combined organic layers with brine (50 mL), dry over anhydrous sodium sulfate. Concentrate under reduced pressure, purify the residue by chromatography (silica gel, EtOAc:PE=1:3) to provide the title compound (1.5 g, 50.0%). MS: (M+1): 174.
Step 6: Synthesis of 5′-bromo-1′-methylspiro[cyclopropane-1,3′-indolin]-2′-one




Add bromine (1.4 g, 8.7 mmol) dropwise to the mixture of 1′-methylspiro[cyclopropane-1,3′-indolin]-2′-one (1.5 g, 8.7 mmol), sodium acetate (0.7 g, 8.7 mmol) and acetic acid (0.52 g, 8.7 mmol) in dichloromethane (15 mL) at 0° C. After addition, stir the resulting mixture at room temperature overnight. Pour the reaction mixture to the cold sodium thiosulfate solution, extract with ethyl acetate (100 mL×2), combine the organic layers, and wash with brine (20 mL), dry over anhydrous sodium sulfate. Concentrate the solution under reduced pressure to give the crude product, purify by flash chromatography (silica gel, EtOAc:PE=1:5) to afford the title compound (1.0 g, 47.6%). MS: (M+1): 254.
Step 7: Synthesis of 5′-amino-1′-methylspiro[cyclopropane-1,3′-indolin]-2′-one




Add 5′-bromo-1′-methylspiro[cyclopropane-1,3′-indolin]-2′-one (1.0 g, 4.0 mmol), benzophenone imine (1.4 g, 8.0 mmol), cesium carbonate (3.2 g, 10.0 mmol), (±)-2,2′-bis(diphenylphosphino)-1,1′-binaphthalene (BINAP, 500 mg, 0.80 mmol), tris(dibenzylideneacetone)dipalladium(0) [Pd2(dba)3, 500 mg, 0.55 mmol] to dioxane (20 mL), stir the reaction under nitrogen atmosphere at 110° C. overnight. Cool to room temperature; filter off the solid, concentrate the filtrate under reduced pressure to give a residue. Add hydrochloric acid (1N, 2 mL) and THF (15 mL) to the residue; stir the mixture for 30 mins at room temperature. TLC (PE:EtOAc=1:1) shows the reaction is almost complete. Concentrate the mixture under reduce pressure to give the crude product, purify by flash chromatography (silica gel, EtOAc:PE=1:1) to afford the title compound as a yellow solid (0.7 g, 93.3%). MS: (M+1): 189.1.
Data of intermediate C26-30 are summarized in Table C5 below.







TABLE C5







Intermediates C26-C30










Num-


MS


ber
Starting material
Intermediate
[M + 1]+





C26








159.1


 


C27








134.1


 


C28








152.1


 


C29








151.1


 


C30








189.1













Step 1: Synthesis of tert-butyl 4-(3-nitro-5-(trifluoromethyl)phenyl)-5,6-dihydropyridine-1(2H)-carboxylate




Mix 1-bromo-3-nitro-5-(trifluoromethyl)benzene (200 mg, 0.65 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5,6-dihydropyridine-1(2H)-carboxylate (175 mg, 0.65 mmol), tetrakis(triphenylphosphine)palladium(0) (35 mg, 0.03 mmol) in dioxane and saturated sodium bicarbonate solution (3:1, 10 mL), stir the mixture at 120° C. for 1.5 hrs. Cool to room temperature; concentrate under reduced pressure to give the crude product. Purify by flash chromatography (silica gel, EtOAc:PE=20:1) to afford the product as a yellow solid (310 mg, 82.8%). MS: (M+23): 395.2.
Step 2: Synthesis of tert-butyl 4-[3-amino-5-(trifluoromethyl)phenyl]piperidine-1-carboxylate




Add tert-butyl 4-(3-nitro-5-(trifluoromethyl)phenyl)-5,6-dihydropyridine-1(2H)-carboxylate (300 mg, 1.87 mmol) and Pd/C (10%, 50 mg) in ethanol (10 mL), flush with H2, stir under hydrogen atmosphere for 48 hrs at room temperature, filter and concentrate the filtrate to yield a white solid (220 mg, 73.9%). MS: (M+23): 367.
Step 3: Synthesis of 3-(1-methyl-4-piperidyl)-5-(trifluoromethyl)aniline




Add lithium aluminum hydride (120 mg, 3.2 mmol) to a solution of the compound prepared in Step 2 (220 mg, 0.64 mmol) in THF (20 mL), stir the mixture at 90° C. for 12 hrs, TLC (MeOH:DCM=1:1) shows the reaction is complete, quench the reaction with water (2 mL), extract with ethyl acetate (20 mL), wash the organic layer with water and brine sequentially, dry over anhydrous sodium sulfate. Concentrate under reduced pressure to give the crude product, purify by flash chromatography (silica gel, MeOH:DCM=1:10) to afford the product as a yellow oil (110 mg, 48.3%).




Step 1: Synthesis of 4-(3-nitro-5-(trifluoromethyl) phenyl)-1,2,3,6-tetrahydropyridine




Dissolve tert-butyl 4-(3-nitro-5-(trifluoromethyl)phenyl)-5,6-dihydropyridine-1(2H)-carboxylate (300 mg, 0.81 mmol) in dichloromethane (10 mL), add trifluoroacetic acid (3 mL), stir at room temperature overnight. Adjust pH=8 with saturated NaHCO3 solution, extract with ethyl acetate, wash the organic extract with water and brine sequentially, dry over anhydrous sodium sulfate. Concentrate under reduced pressure to provide yellow oil (220 mg, 100%) which is used directly in next step.
Step 2: Synthesis of 1-cyclopropyl-4-(3-nitro-5-(trifluoromethyl)phenyl)-1,2,3,6-tetrahydropyridine




Stir the mixture of 4-(3-nitro-5-(trifluoromethyl)phenyl)-1,2,3,6-tetrahydropyridine (150 mg, 0.55 mmol), (1-ethoxycyclopropoxy)trimethylsilane (700 mg, 2.4 mmol), sodium cyanoborohydride (200 mg, 3.3 mmol) and acetic acid (240 mg, 4.0 mmol) in ethanol (10 mL) at 60° C. overnight. Concentrate the mixture to get a residue, partition between EtOAc and water, collect the organic layer, wash with water and brine sequentially, and dry over anhydrous sodium sulfate. Concentrate under reduced pressure to give the product as a yellow solid (135 mg, 79%). MS: (M+1): 313.1.
Step 3: Synthesis of 3-(1-cyclopropylpiperidin-4-yl)-5-(trifluoromethyl)aniline




Stir the mixture of Pd/C (10%, 30 mg), 1-cyclopropyl-4-(3-nitro-5-(trifluoromethyl)phenyl)-1,2,3,6-tetrahydropyridine (120 mg, 0.32 mmol) in methanol (10 mL) under hydrogen atmosphere at room temperature for 18 hrs. Remove hydrogen atmosphere, filter off the solid, concentrate the filtrate to yield the product as a white solid (85 mg, 94%). MS: (M+1): 285.1.
Data of intermediates C31-C32 are summarized in Table C6 below.







TABLE C6







Intermediates C31-C32










Number
Starting material
Intermediate
MS [M + 1]+





C31








NA


 


C32








285.2









Data of other commercially available intermediates C33-C37 are summarized in Table C7 below.







TABLE C7







Other intermediates C33-C37











Number
Intermediate
MS [M + 1]+







C33




NA



 



C34




NA



 



C35




NA



 



C36




NA



 



C37




176.1










Preparation of Intermediate D




Step 1: Synthesis of phenyl N-[4-chloro-3-(trifluoromethyl)phenyl]carbamate




Add 4-chloro-3-(trifluoromethyl)aniline (1.0 g, 5.1 mmol) and pyridine (1.0 g, 12.8 mmol) in DCM (10 mL), stir well and cool to 0° C. Add phenyl chloroformate (1.04 g, 6.6 mmol) at 0° C. After addition, stir the reaction at room temperature for 1 hr. TLC (EtOAc:PE=1:2) shows the reaction is complete. Wash the organic layer with 1M HCl solution (20 mL) and brine (20 mL) respectively. Dry over anhydrous Na2SO4; concentrate under reduced pressure to give the target compound (1.16 g) which is used without further purification. MS: (M+1): 316.1.
Intermediates D2-D16 can be synthesized with similar method (Table D1).







TABLE D1







Intermediates D1-D16










Number
Starting material
Intermediate
MS [M + 1]+





D1








316.1


 


D2








300.1


 


D3








NA


 


D4








296.1


 


D5








NA


 


D6








NA


 


D7








NA


 


D8








NA


 


D9








NA


 


D10








282.1


 


D11








283.2


 


D12








316.1


 


D13








296.2


 


D14








359.1


 


D15








360.1


 


D16








NA













Step 1: Synthesis of 1-methyl-N-[3-nitro-5-(trifluoromethyl)phenyl]piperidin-4-amine




Add 1-bromo-3-nitro-5-(trifluoromethyl)benzene (3 g, 11.11 mmol), Cs2CO3 (9.5 g, 27.78 mmol), 1-methylpiperidin-4-amine (1.9 g, 16.67 mmol) to 1,4-dioxane (25 mL), then under N2, add Pd(OAc)2 (50 mg, 0.22 mmol) and BINAP (207 mg, 0.33 mmol). Stir the reaction at 120° C. under N2 overnight. TLC (DCM:MeOH=10:1) shows the reaction is complete. Cool to room temperature and filter off the solid. Concentrate the filtrate to get the crude product. Purification by chromatography (silica gel, DCM:MeOH=10:1) affords the title compound (3.44 g, 99%). MS: (M+1): 304.
Step 2: Synthesis of N1-(1-methyl-4-piperidyl)-5-(trifluoromethyl)benzene-1,3-diamine




Dissolve the compound obtained in last step (3.44 g, 11.3 mmol) in methanol (30 mL), add Pd/C (10%, 0.6 g), flush with H2. Stir the reaction at 30° C. under H2 atmosphere overnight. TLC (DCM:MeOH=10:1) shows the reaction is complete. Filter, and concentrate the filtrate to get the crude product (2.9 g) which is used without further purification. MS: (M+1): 294.1.
Step 3: Synthesis of phenyl N-[3-[(1-methyl-4-piperidyl)amino]-5-(trifluoromethyl)phenyl]carbamate




Dissolve the compound obtained in Step 2 (2.9 g, 10.61 mmol) in THF (30 mL), cool to −30° C., then add pyridine (1.68 g, 21.22 mmol) and phenyl chloroformate (1.83 g, 11.67 mmol). Stir at room temperature for 3 hrs. TLC (DCM:MeOH=10:1) shows the reaction is complete. Pour the solution to a mixture of EtOAc and water, separate the organic layer, and wash with brine, dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product (4.3 g). MS: (M+1): 394.1.
Intermediates D18-D20 can be synthesized with similar method (Table D2).







TABLE D2







Intermediates D17-D20










Number
Starting material
Intermediate
MS [M + 1]+





D17








394.1


 


D18








373.1


 


D19








NA


 


D20








381.1













Step 1: Synthesis of tert-butyl 4-[3-nitro-5-(trifluoromethyl)phenyl]pyrazole-1-carboxylate




Add 1-bromo-3-nitro-5-(trifluoromethyl)benzene (0.165 g, 0.61 mmol), tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole-1-carboxylate (0.18 g, 0.61 mmol), sodium acetate (0.1 g, 1.2 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium (II) (22 mg, 0.03 mmol) to 1,4-dioxane (15 mL) and water (2 mL). Stir the mixture at 100° C. under N2 for 6 hrs. TLC (EtOAc:PE=1:1) shows that the reaction is complete. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (0.17 g, 80.9%). MS: (M+1): 358.2.
Step 2: Synthesis of tert-butyl 4-[3-amino-5-(trifluoromethyl)phenyl]pyrazole-1-carboxylate




Add the compound obtained in Step 1 (0.17 g, 0.48 mmol) to methanol (15 mL), then add Pd/C (10%) under N2. Flush with H2, stir the reaction under H2 atmosphere at room temperature for 2 hrs. Filter off the solid, concentrate the filtrate to give the crude product (0.08 g) which is used directly without further purification. MS: (M+1): 328.1.
Step 3: Synthesis of tert-butyl 4-[3-(phenoxycarbonylamino)-5-(trifluoromethyl)phenyl]pyrazole-1-carboxylate




Mix the compound obtained in Step 2 (0.08 g, 0.24 mmol), pyridine (0.018 g, 0.24 mmol) and DCM (5 mL), add slowly phenyl chloroformate (0.042 g, 0.26 mmol) at 0° C. Stir the reaction at room temperature for one hour. Remove the solvent under reduced pressure to give a crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (65 mg, 78.3%).
Intermediates D22 can be prepared with similar method (Table D3).







TABLE D3







Intermediates D21-D22










Number
Starting material
Intermediate
MS [M + 1]+





D21








NA


 


D22








[M + 23] 487.2













Step 1: Synthesis of N-[3-nitro-5-(trifluoromethyl)phenyl]prop-2-enamide




Dissolve 3-nitro-5-(trifluoromethyl)aniline (150 mg, 0.73 mmol), triethylamine (220 mg, 2.18 mmol) in DCM (8 mL), cool to 0° C. at ice bath, add slowly acryloyl chloride (198 mg, 2.18 mmol). After addition, stir the reaction at room temperature for 2 hrs. Pour the mixture into water (30 mL), extract with EtOAc (10 mL×3), combine the organic layers; wash with brine (50 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product (190 mg) which is used without further purification.
Step 2: Synthesis of N-[3-amino-5-(trifluoromethyl)phenyl]prop-2-enamide




Dissolve the product obtained in Step 1 (190 mg, 0.73 mmol) in ethanol (8 mL), add iron powder (204 mg, 3.65 mmol), concentrated HCl (0.43 mL). Stir the reaction at room temperature for 2 hrs. After the reaction is complete, adjust pH to neutral with sodium hydroxide solution. Filter off the solid, mix the filtrate with ice water (50 mL), extract with EtOAc (15 mL×3). Combine the organic layers, wash with brine (100 mL), dry over anhydrous Na2SO4, and concentrate under reduced pressure to give the crude product (168 mg) which is used without further purification.
Step 3: Synthesis of phenyl N-[3-(prop-2-enoylamino)-5-(trifluoromethyl)phenyl]carbamate




Dissolve the product obtained in Step 2 (168 mg, 0.73 mmol) in DCM (6 mL), add pyridine (115 mg, 1.46 mmol) and then cool to 0° C. on ice bath. Add slowly phenyl chloroformate (125 mg, 0.8 mmol), stir at 0-5° C. for 2 hrs. Pour the mixture into water (50 mL), adjust pH to neutral with 1M HCl and extract with EtOAc (15 mL×3). Combine the organic layers, wash with brine (100 mL) and dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product (240 mg) which is used without further purification.
Intermediate D24 can be synthesized with similar method (Table D4).







TABLE D4







Intermediates D23-D24










Number
Starting material
Intermediate
MS [M + 1]+





D23








351.1


 


D24








[M + 23] 487.2













Step 1: Synthesis of tert-butyl 3-[3-nitro-5-(trifluoromethyl)phenoxy]azetidine-1-carboxylate




Dissolve 3-nitro-5-(trifluoromethyl)phenol (500 mg, 2.4 mmol), tert-butyl 3-hydroxyazetidine-1-carboxylate (500 mg, 2.9 mmol), PPh3 (9.5 g, 3.6 mmol) in THF (8 mL) and cool to 0° C. on ice bath. Add slowly diethyl azodicarboxylate (630 mg, 3.6 mmol). Stir the mixture at 35° C. for 16 hrs. Pour the mixture into water (50 mL), adjust pH to neutral with saturated NaHCO3 solution. Extract with EtOAc (15 mL×3), combine the organic layers; wash with brine (100 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=15:85) affords the target compound (760 mg, 87%).
Step 2: Synthesis of tert-butyl 3-[3-amino-5-(trifluoromethyl)phenoxy]azetidine-1-carboxylate




Dissolve the compound obtained in Step 1 (250 mg, 0.69 mmol) in methanol (10 mL), add wet Pd/C (10%, 70 mg), flush the system with H2 and stir the reaction under H2 atmosphere for 16 hrs at room temperature. After the reaction is complete, remove H2, flush with N2, filter and concentrate the filtrate to give the crude product (230 mg) which is used without further purification.
Step 3: Synthesis of tert-butyl 3-[3-(phenoxycarbonylamino)-5-(trifluoromethyl)phenoxy]azetidine-1-carboxylate




Dissolve the product obtained in Step 2 (230 mg, 0.39 mmol) in THF (10 mL), add pyridine (230 mg, 0.69 mmol) and cool to 0° C. on ice bath. Add slowly phenyl chloroformate (118 mg, 0.76 mmol), stir the reaction at 0-5° C. for 30 mins. Pour the reaction mixture into water (50 mL), adjust pH to neutral with 1M HCl solution. Extract with EtOAc (15 mL×3), combine the organic layers, wash with brine (100 mL) and dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product (312 mg) which is used without further purification.
Intermediates D26-D29 can be synthesized with similar method (Table D5).







TABLE D5







Intermediates D25-D29










Number
Starting material
Intermediate
MS [M + 1]+





D25








[M + 23] 475.2


 


D26








395.2


 


D27








NA


 


D28








312.1


 


D29








NA













Step 1: Synthesis of 1-tert-butoxycarbonylpiperidine-4-carboxylic acid




Mix piperidine-4-carboxylic acid (0.5 g, 3.9 mmol) and Boc2O (1 g, 4.6 mmol) in t-BuOH (10 mL) and stir at 70° C. for 4 hrs. TLC (PE:EtOAc=1:1) shows the reaction is complete. Remove the volatiles under reduced pressure to get white solid as the crude product (1.1 g) which is used without further purification.
Step 2: Synthesis of tert-butyl 4-[[3-amino-5-(trifluoromethyl)phenyl]carbamoyl]piperidine-1-carboxylate




Mix the compound obtained in Step 1 (1.1 g, crude), HOBt (1.6 g, 12 mmol), 5-(trifluoromethyl)benzene-1,3-diamine (845 mg, 4.8 mmol) in DCM (16 mL), add diisopropylethylamine (1.55 g, 12 mmol), 1-ethyl-3-(3-dimethyllaminopropyl)carbodiimide hydrochloride (2.3 g, 12 mmol) and stir at room temperature for 17 hrs. TLC (PE:EtOAc=1:1) shows the reaction is complete. Remove the volatiles under reduced pressure, purify the residue by chromatography (silica gel, EOAc:PE=4:6) to give the title compound (600 mg, 40% two-step yield).
Step 3: Synthesis of tert-butyl 4-[[3-(phenoxycarbonylamino)-5-(trifluoromethyl)phenyl]carbamoyl]piperidine-1-carboxylate




Mix the compound obtained in Step 2 (600 mg, 1.55 mmol), pyridine (306 mg, 3.9 mmol) in DCM (10 mL), cool to 0-5° C. Add the solution of phenyl chloroformate (315 mg, 2 mmol) in DCM (1 mL). Stir the reaction at room temperature for 1.5 hrs. TLC (PE:EtOAc=1:1) shows the reaction is complete. Quench the reaction with water (5 mL), wash the organic layer with 1N HCl solution, saturated NaHCO3 solution and brine sequentially, dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (400 mg, 51%). MS: (M+1): 508.1.
Intermediate D31 can be synthesized with similar method (Table D6).







TABLE D6







Intermediates D30-D31










Number
Starting material
Intermediate
MS [M + 1]+













D30








[M + 23] 530.2


 


D31








NA













Step 1: Synthesis of N-[3-nitro-5-(trifluoromethyl)phenyl]methanesulfonamide




Dissolve 3-nitro-5-(trifluoromethyl)aniline (300 mg, 1.45 mmol) in DCM (5 mL), add triethylamine (294 mg, 2.9 mmol) and then methylsulfonyl chloride (182 mg, 1.6 mmol). Stir the reaction at room temperature for 17 hrs. TLC (EtOAc:PE=1:3) shows the reaction is complete. Concentrate under reduced pressure to afford the crude product. Purification by chromatography (silica gel, EtOAc:PE=3:7) affords the title compound (150 mg, 36%).
Step 2: Synthesis of N-[3-amino-5-(trifluoromethyl)phenyl]methanesulfonamide




Dissolve the compound obtained in Step 1 (150 mg) in DCM (5 mL), add Pd/C (20%, 50 mg). Flush the system with H2 and then stir the reaction under H2 atmosphere at room temperature for 2 hrs. TLC (EtOAc:PE=1:2) shows the reaction is complete. Remove the H2, filter and concentrate the filtrate to give the product (110 mg, 82%). MS: (M+1): 255.1.
Step 3: Synthesis of phenyl N-[3-(methanesulfonamido)-5-trifluoromethyl)phenyl]carbamate




Use similar procedure used in the preparation of D1.





Step 1: Synthesis of 4-[3-nitro-5-(trifluoromethyl)phenyl]thiomorpholine




Add 1-bromo-3-nitro-5-(trifluoromethyl)benzene (500 mg, 2.9 mmol), thiomorpholine (455 mg, 4.4 mmol), Cs2CO3 (2.86 g, 8.8 mmol), BINAP (55 mg, 0.09 mmol) and Pd2(dba)3 (54 mg, 0.06 mmol) under N2 to 1,4-dioxane (10 mL), stir the reaction under N2 at 100° C. for 16 hrs. TLC (100% PE) shows the reaction is complete. Cool the reaction to room temperature, filter and concentrate the filtrate under reduced pressure to get crude product. Purification by chromatography (silica gel, 100% PE) affords the target compound (475 mg, 55%). MS: (M+1): 293.1.
Step 2: Synthesis of 4-[3-nitro-5-(trifluoromethyl)phenyl]-1,4-thiazinane 1,1-dioxide




Dissolve the compound obtained in Step 1 (370 mg, 1.3 mmol) in DCM (30 mL), add mCPBA (655 mg, 3.8 mmol), stir the mixture at room temperature for 1 hr. Add water (20 mL), extract with DCM (20 mL×2), combine the organic layers, wash sequentially with saturated Na2SO3 solution (50 mL), saturated NaHCO3 solution, brine (50 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (400 mg, 97.5%). MS: (M+1): 325.2.
Step 3: Synthesis of 3-(1,1-dioxo-1,4-thiazinan-4-yl)-5-(trifluoromethyl)aniline




Dissolve the compound obtained in Step 2 (510 mg, 1.57 mmol) in DCM (30 mL), add Pd/C (10%, 100 mg) under N2 atmosphere, then flush the system with H2 and stir the reaction under H2 atmosphere at room temperature for 15 hrs. Remove the H2, filter the reaction mixture, and concentrate the filtrate to give the crude product (390 mg) which is used directly without further purification. MS: (M+1): 295.1.
Step 4: Synthesis of phenyl N-[3-(1,1-dioxo-1,4-thiazinan-4-yl)-5-(trifluoromethyl)phenyl]carbamate




Mix the compound obtained in Step 3 (390 mg, 1.33 mmol) and pyridine (262 mg, 3.31 mmol) in DCM (10 mL), cool to 0° C., add slowly phenyl chloroformate (270 mg, 1.72 mmol). After addition, stir the reaction at room temperature for 1 hr. TLC (EtOAc:PE=1:1) shows the reaction is complete. Wash the reaction mixture sequentially with 1M HCl solution (20 mL), brine (20 mL), and dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the target product (600 mg) which is used directly without further purification. MS: (M+1): 415.1.




Step 1: Synthesis of 1-(chloromethyl)-3-nitro-5-(trifluoromethyl)benzene




Dissolve [3-nitro-5-(trifluoromethyl)phenyl]methanol (1.1 g, 5.18 mmol) and triethylamine (1.62 mL) in DCM (20 mL), cool to 0° C., add slowly p-toluenesulfonyl chloride (1.1 g, 6.02 mmol). After the addition, stir the reaction at room temperature for 15 hrs. TLC (EtOAc:PE=1:15) shows the reaction is complete. Concentrate under reduced pressure to get the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:15) affords the title compound (655 mg, 55%).
Step 2: Synthesis of 1-methyl-4-[[3-nitro-5-(trifluoromethyl)phenyl]methyl]piperazine




Mix the compound obtained in Step 1 (640 mg, 2.68 mmol) and 1-methylpiperazine (536 mg, 5.36 mmol) in THF (8 mL). Stir the reaction at 60° C. for 2 hrs. TLC (EtOAc:PE=1:10 shows the reaction is complete. Concentrate under reduced pressure to get the title compound (812 mg, 100%). MS: (M+1): 303.1.
Step 3: Synthesis of 3-[(4-methylpiperazin-1-yl)methyl]-5-(trifluoromethyl)aniline




Dissolve the nitro compound obtained in Step 2 (812 mg, 2.68 mmol) in methanol (15 mL), add Pd/C (10%, 200 mg) under N2 atmosphere. Flush the system with H2 and stir the action under H2 atmosphere at room temperature for 16 hrs. Remove the H2, filter off the solid, concentrate the filtrate to get the crude product (732 mg, 100%). MS: (M+1): 273.1.
Step 4: Synthesis of phenyl N-[3-[(4-methylpiperazin-1-yl)methyl]-5-(trifluoromethyl)phenyl]carbamate




Dissolve the amine obtained in Step 3 (732 mg, 2.68 mmol) and pyridine (423 mg, 3.52 mmol) in DCM (10 mL), cool to 0° C., add slowly phenyl chloroformate (460 mg, 2.9 mmol). After addition, stir the reaction at room temperature for 2 hrs. TLC (EtOAc:PE=4:1) shows the reaction is complete. Add water, separate the organic layer, and extract the aqueous layer with DCM. Combine the organic layers, wash sequentially with 1M HCl solution, brine, dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=4:1) affords the target compound (380 mg, 36%). MS: (M+1): 393.




Step 1: Synthesis of 1-methyl-N-[4-nitro-2-(trifluoromethyl)phenyl]piperidin-4-amine




Dissolve 1-chloro-4-nitro-2-(trifluoromethyl)benzene (1.0 g, 4.44 mmol) in DMSO (5 mL), add 1-methylpiperidin-4-amine (0.56 g, 4.89 mmol) and DIEA (1.72 g, 13.3 mmol) and stir the reaction at 80° C. overnight. TLC (DCM:MeOH=10:1) shows the reaction is complete. Add water, extract with EtOAc (10 mL×3), combine the organic layers, wash with water and brine sequentially, and dry over anhydrous Na2SO4. Concentrate under reduced pressure to get the crude product. Purification by chromatography (silica gel, MeOH:DCM=1:15) affords the title compound (587 mg, 44%). MS: (M+1): 304.2.
Step 2: Synthesis of N1-(1-methyl-4-piperidyl)-2-(trifluoromethyl)benzene-1,4-diamine




Dissolve the nitro compound obtained in Step 1 (587 mg, 1.94 mmol) in methanol (10 mL), add Pd/C (10%, 176 mg). Flush the system with H2. Stir the reaction under H2 atmosphere at room temperature overnight. TLC (DCM:MeOH=5:1) shows the reaction is complete. Filter off the solid, concentrate the filtrate under reduced pressure to give a crude residue. Purification by chromatography (silica gel, MeOH:DCM=1:10) affords the product (300 mg, 57%). MS: (M+1) 274.2.
Step 3
Use the same procedure used for D1 to get the product: MS: (M+1): 394.2.




Step 1: Synthesis of 1,1-diphenyl-N-[5-(trifluoromethyl)-3-pyridyl]methanimine




Add 3-bromo-5-(trifluoromethyl)pyridine (600 mg, 2.67 mmol), benzophenone imine (701 mg, 4.0 mmol), Cs2CO3 (2.17 g, 6.68 mmol), BINAP (25 mg, 0.040 mmol), Pd2(dba)3 (24 mg, 0.027 mmol) in 1,4-dioxane (10 mL), stir at 110° C. for 16 hrs. Cool to room temperature, add water (100 mL), extract with EtOAc (15 mL×3), wash the combined organic layers with brine, dry over anhydrous Na2SO4, concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=6:94) affords the target compound (850 mg, 98%).
Step 2: Synthesis of 5-(trifluoromethyl)pyridin-3-amine




Dissolve the imine obtained in Step 1 (850 mg, 2.6 mmol) in THF (10 mL), add water (2 mL), 1N HCl solution (5 mL), stir the reaction at room temperature for 2 hrs. After the reaction, extract with EtOAc, wash the organic layer with saturated NaHCO3 solution and brine sequentially, dry over anhydrous Na2SO4, and concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the target compound (380 mg, 90%).
Step 3: Synthesis of phenyl N-[5-(trifluoromethyl)-3-pyridyl]carbamate




Dissolve the amine obtained in Step 2 (380 mg, 2.35 mmol), pyridine (371 mg, 4.7 mmol) in DCM (8 mL), cool to 0° C. in ice bath. Add slowly phenyl chloroformate (403 mg, 2.35 mmol). After addition, stir the reaction at 0-5° C. for 2 hrs. Pour the mixture into water (50 mL), adjust pH to neutral with 1M HCl solution, extract with EtOAc (15 mL×3).
Combine the organic layers; wash with brine (100 mL), dry over anhydrous Na2SO4, and concentrate under reduced pressure to get the crude product. Purification by chromatography (silica gel, EtOAc:PE=15:85) affords the target compound (390 mg, 59%).




Step 1: Synthesis of 6-(trifluoromethyl)pyrimidin-4-amine




Add 6-(trifluoromethyl)pyrimidin-4-ol (1.5 g, 9.15 mmol) and oxalyl chloride (2.3 mL, 22.9 mmol) to EtOAc (15 mL), then add 5 drops of DMF. Heat the mixture to reflux for 2 hrs. After reaction, partition between EtOAc and brine, separate the organic layer; dry the organic layer over anhydrous Na2SO4. Transfer the organic solution to a sealed tube, add a solution of NH3 in methanol (7M, 10 mL), seal the tube, heat at 70° C. for 15 hrs. Cool to room temperature; concentrate under reduced pressure to get the crude product. Purification by chromatography (silica gel, EtOAc:PE=2:1) affords the target compound (920 mg, 61.3%).
Step 2: Synthesis of phenyl N-[6-(trifluoromethyl)pyrimidin-4-yl]carbamate




Add the amine obtained in Step 1 (550 mg, 3.37 mmol), Et3N (680 mg, 6.74 mmol) to THF (10 mL), cool to 0° C. on ice bath. Add slowly a solution of phenyl chloroformate (789 mg, 5.06 mmol) in THF. After addition, stir the reaction on ice bath for 20 hrs. Remove the volatiles under reduced pressure. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the target compound (160 mg, 17%). MS: (M+1): 284.1.
Data on intermediates D32-37 are summarized in Table D7.







TABLE D7







Intermediates D31-D37










Number
Starting material
Intermediate
MS [M + 1]+





D32








NA


 


D33








415.1


 


D34








394.2


 


D35








394.2


 


D36








283.1


 


D37








284.1













Step 1: Synthesis of 4-[3-nitro-5-(trifluoromethyl)phenyl]morpholine




Dissolve 1-bromo-3-nitro-5-(trifluoromethyl)benzene (270 mg, 1 mmol) and DIEA (258 mg, 2 mmol) in DMF (5 mL), add morpholine (174 mg, 2 mmol). Stir the reaction at 120° C. overnight. Cool to room temperature; remove the volatiles under reduced pressure to give a crude residue. Purification of the residue by chromatography (silica gel, EtOAc:PE=1:5) affords the product (138 mg, 50%).
Step 2: Synthesis of 4-[3-nitro-5-(trifluoromethyl)phenyl]morpholin-3-one




Dissolve the nitro compound obtained in Step 1 (276 mg, 1 mmol) and Bu3BnNCl (1.67 g, 6 mmol) in DCM (20 mL). Add KMnO4 portionwise (948 mg, 6 mmol). Stir the mixture at 70° C. overnight. Cool to room temperature; remove the volatiles under reduced pressure to give a residue. Purification of the residue by chromatography (silica gel, EtOAc:PE=1:5) affords the product (64 mg, 22%).
Step 3: Synthesis of 4-[3-amino-5-(trifluoromethyl)phenyl]morpholin-3-one




Dissolve the nitro compound obtained in Step 2 (290 mg, 1 mmol) in methanol (5 mL), add Pd/C (10%, 29 mg), flush with H2, and then stir at room temperature under 1 atm H2 atmosphere for 12 hrs. After the reaction, remove the H2, filter off the solid, concentrate the filtrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the product (221 mg, 85%). MS: (M+1): 261.
Step 4: Synthesis of phenyl N-[3-(3-oxomorpholin-4-yl)-5-(trifluoromethyl)phenyl]carbamate




Dissolve the amine obtained in Step 3 (260 mg, 1 mmol) and pyridine (79 mg, 1 mmol) in DCM (2 mL), cool to 0° C. Add phenyl chloroformate (156 mg, 1 mmol), then stir at room temperature for 2 hrs. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:5) affords the product (300 mg, 79%). MS: (M+1): 381.1.




Step 1: Synthesis of 1-[6-chloro-4-(trifluoromethyl)-2-pyridyl]-4-methyl-piperazine




Dissolve 2,6-dichloro-4-(trifluoromethyl)pyridine (215 mg, 1 mmol) and DIEA (129 mg, 1 mmol) in DMF (2 mL), add 1-methyl piperazine (100 mg, 1 mmol). Stir the reaction at 120° C. overnight. Cool to room temperature; concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, MeOH:DCM=1:10) affords the product (253 mg, 91%). MS: (M+1): 280.
Step 2: Synthesis of N-[6-(4-methylpiperazin-1-yl)-4-(trifluoromethyl)-2-pyridyl]-1,1-diphenyl-methanimine




Dissolve the compound obtained in Step 1 (279 mg, 1 mmol) in 1,4-dioxane (4 mL), add benzophenone imine (181 mg, 1 mmol), DIEA (181 mg, 1 mmol), Cs2CO3 (650 mg, 2 mmol), Pd2(dba)3 (18 mg, 0.02 mmol) and BINAP (18 mg, 0.03 mmol). Stir the reaction at 100° C. overnight. Cool to room temperature; filter off the solid, concentrate the filtrate under reduced pressure to give a crude product. Purification by chromatography (silica gel, MeOH:DCM=1:10) affords the product (200 mg, 91%). MS: (M+1): 425.
Step 3: Synthesis of 6-(4-methylpiperazin-1-yl)-4-(trifluoromethyl)pyridin-2-amine




Suspend the imine obtained in Step 2 (424 mg, 1 mmol) in methanol and water (1:1, 2 mL), add aqueous HCl (1N, 2 mL), stir the reaction at room temperature for 1 hr. After the reaction, extract with EtOAc, discard the organic layer. Adjust the aqueous layer with NaOH solution to pH=10, extract with EtOAc, wash the organic layer with water, brine sequentially, and dry over anhydrous Na2SO4. Concentrate under reduced pressure to get the product (200 mg, 77%). MS: (M+1): 261.
Step 4: Synthesis of phenyl N-[6-(4-methylpiperazin-1-yl)-4-(trifluoromethyl)-2-pyridyl]carbamate




Dissolve the amine obtained in Step 3 (260 mg, 1 mmol) and pyridine (79 mg, 1 mmol) in DCM (2 mL), cool to 0° C. and add dropwise phenyl chloroformate (156 mg, 1 mmol). After addition, stir the reaction at room temperature for 2 hrs. Concentrate under reduced pressure to give a crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the product (300 mg, 79%). MS: (M+1): 381.
Intermediates D38-D41 can be synthesized with similar method (Table D8).







TABLE D8







Intermediates D38-D41










Number
Starting material
Intermediate
MS [M + 1]+













D38








381.1


 


D39








381


 


D40








395.1


 


D41








368.1













Step 1: Synthesis of phenyl N-[3-(trifluoromethoxy)phenyl]carbamate




Add 3-(trifluoromethoxy)aniline (1.0 g, 5.65 mmol), pyridine (0.89 g, 11.3 mmol) to DCM (20 mL) and cool to 0° C. Add dropwise phenyl chloroformate (1.15 g, 7.34 mmol). After addition, stir for 1 hr. TLC (PE:EtOAc=3:1) shows the reaction is complete. Add water to the mixture, extract with DCM (10 mL×3), combine the organic layer, wash with 1N HCl solution, brine sequentially, and dry over anhydrous Na2SO4. Concentrate under reduced pressure to get the product (1.50 g, 89%).
Intermediates D42-D46 can be synthesized with similar method (Table D9).







TABLE D9







Intermediates D42-D46










Number
Starting material
Intermediate
MS [M + 1]+





D42








298.1


 


D43








316.1


 


D44








NA


 


D45








332.1


 


D46








NA













Step 1: Synthesis of 2-chloro-4-(1,1-difluoroethyl)pyridine




Mix 1-(2-chloro-4-pyridyl)ethanone (2 g, 12.90 mmol), DCM (2 mL) and bis-(2-methoxyethyl)aminosulfur trifluoride (BAST, 11.7 g, 51.6 mmol) in an unsealed tube, add 1 drop of methanol. Seal the tube and heat at 60° C. for 15 hrs. Cool to room temperature, add DCM to dilute the mixture, basicify with NaOH solution, pour into water (100 mL), extract with DCM (100 mL×2), combine the organic layers, dry over anhydrous Na2SO4. Concentrate under reduced pressure to get a crude residue. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the target compound (1.5 g, 65.7%).
Step 2: Synthesis of N-[4-(1,1-difluoroethyl)-2-pyridyl]-1,1-diphenyl-methanimine




Add the compound obtained in Step 1 (1.21 g, 6.81 mmol), benzophenone imine (1.83 g, 10.22 mmol), Cs2CO3 (4.43 g, 13.6 mmol) to 1,4-dioxane (15 mL), then under N2, add BINAP (635 mg, 1.02 mmol), Pd2(dba)3 (623 mg, 0.68 mmol), stir the reaction at 120° C. under N2 atmosphere for 15 hrs. Cool to room temperature; filter off the solid, concentrate the filtrate under reduced pressure to give a crude residue. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the target compound (1.68 g, 76.3%). MS: (M+1): 323.2.
Step 3: Synthesis of 4-(1,1-difluoroethyl)pyridin-2-amine




Dissolve the imine obtained in Step 2 (1.68 g, 5.22 mmol) in THF (10 mL), add 1M HCl solution (10 mL), stir the reaction at room temperature for 1 hr. Use saturated NaHCO3 solution to adjust pH>7. Extract with EtOAc (50 mL×2), combine the organic layers, dry over anhydrous Na2SO4, concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the target compound (624 mg, 75.7%). MS: (M+1): 159.1.
Step 4: Synthesis of phenyl N-[4-(1,1-difluoroethyl)-2-pyridyl]carbamate




Add the amine obtained in Step 3 (624 mg, 3.95 mmol), pyridine (780 mg, 9.85 mmol) in DCM (15 mL), cool to 0° C. on ice bath. Add slowly a solution of phenyl chloroformate (678 mg, 4.34 mmol) in DCM. After addition, stir on ice bath for 1 hr. Concentrate the mixture to get a crude residue. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the target compound (780 mg, 71%). MS: (M+1): 279.1.




Step 1: 1-(3-fluoro-5-nitro-phenyl)ethanone




Mix 1-fluoro-3-iodo-5-nitro-benzene (1 g, 3.75 mmol), tributyl(1-ethoxyvinyl)stannane (1.5 g, 4.15 mmol) and Pd(dppf)Cl2 (78 mg, 0.1 mmol) in toluene (5 mL) under N2. Stir at 100° C. under N2 atmosphere for 3 hrs. Cool to room temperature, add 1N HCl solution (5 mL) and stir for 20 min. Load directly onto silica gel column and purify by chromatography (EtOAc:PE=3:7) to give the title compound (660 mg, 96%).
Step 2: Synthesis of 1-(1,1-difluoroethyl-3-fluoro-5-nitro-benzene




In an unsealed tube, add the compound obtained in Step 1 (660 mg, 3.6 mmol), bis-(2-methoxyethyl)aminosulfur trifluoride (BAST, 3.2 g, 14.5 mmol) and DCM (8 mL), seal the tube, heat at 60° C. with stirring overnight. Cool the reaction mixture, purify by chromatography (silica gel, 100% PE) to give the title compound (500 mg, 67.6%).
Step 3: Synthesis of 3-(1,1-difluoroethyl)-5-fluoro-aniline




Add the nitro compound obtained in Step 2 (500 mg, 2.44 mmol) and SnCl2.2H2O (2.6 g, 11.5 mmol) in ethanol (20 mL), heat to reflux for 4 hrs. TLC (100% PE) shows the reaction is complete. Cool to room temperature, basicify to pH=10-12 with NaOH solution to get a suspension. Filter off the white solid, partition the filtrate between DCM and water. Collect the organic layer, dry over anhydrous Na2SO4, concentrate to get the crude product. Purification by chromatography (silica gel, PE:EtOAc=70:30) affords the title compound (350 mg, 82%). MS: (M+1): 176.
Step 4: Synthesis of phenyl N-[3-(1,1-difluoroethyl)-5-fluoro-phenyl]carbamate




Dissolve the amine obtained in Step 3 (350 mg, 2.0 mmol) in DCM (8 mL), add pyridine (395 mg, 5 mmol), cool to 0-5° C. on ice bath. Add dropwise a solution of phenyl chloroformate (406.7 mg, 2.6 mmol) in DCM (1 mL). Stir the reaction at room temperature for 1.5 hrs. Quench the reaction with water (5 mL), collect the organic layer, wash the organic layer with 1N HCl solution, then saturated NaHCO3 solution and brine sequentially. Dry the organic layer over anhydrous Na2SO4; concentrate under reduced pressure to give the crude product (400 mg, 80% pure) which is used without further purification. MS: (M+1): 296.




Step 1: Synthesis of 1-(1,1-difluoroethyl)-3-nitro-benzene




Dissolve 1-(3-nitrophenyl)ethanone (1 g, 6.1 mmol) in DCM (10 mL) in an unsealed tube, add dropwise bis-(2-methoxyethyl)aminosulfur trifluoride (BAST, 5.4 g, 24.2 mmol) and one drop of methanol, seal the tube and heat at 60° C. for 16 hrs. Cool to room temperature, pour the mixture to ice water (50 mL), neutralize with saturated NaHCO3 solution. Extract with EtOAc (15 mL×3), combine the organic layers; wash with brine (100 mL), dry over anhydrous Na2SO4, concentrate to give a crude residue. Purification by chromatography (silica gel, PE:EtOAc=99:1) affords the target compound (970 mg, 85%).
Step 2: Synthesis of 3-(1,1-difluoroethyl)aniline




Dissolve the nitro compound obtained in Step 1 (870 mg, 4.8 mmol) in ethanol (10 mL), add iron powder (1.3 g, 23.3 mmol), concentrated aqueous HCl (37%, 2.7 mL), stir the reaction at room temperature for 2 hrs. After the reaction, carefully neutralize the mixture with NaOH solution. Filter off the solid, pour the filtrate to ice water (50 mL), extract with EtOAc (15 mL×3), combine the organic layers, wash with brine (100 mL) and dry over anhydrous Na2SO4. Concentrate under reduced pressure to get the crude product (760 mg) which is used without further purification.
Step 3: Synthesis of phenyl N-[3-(1,1-difluoroethyl)phenyl]carbamate




Dissolve the amine obtained in Step 2 (760 mg, 4.8 mmol) and pyridine (765 mg, 9.7 mmol) in DCM (15 mL) and cool to 0-5° C. on ice bath. Add dropwise phenyl chloroformate (831 mg, 5.3 mmol). After addition, stir the mixture at 0-5° C. for 2 hrs. Pour the mixture to water (50 mL), adjust pH to neutral with 1M HCl solution. Extract with EtOAc (15 mL×3), combine the organic layers and wash with brine (100 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product (1.15 g) which is used without further purification.
Intermediates D50-D52 can be synthesized with similar method (Table D10).







TABLE D10







Intermediates D47-D52










Number
Starting material
Intermediate
MS [M + 1]+





D47








279.1


 


D48








296.2


 


D49








278.1


 


D50








296.1


 


D51








312.1


 


D52








296.1













Step 1: Synthesis of phenyl N-(3-chloro-4-cyano-phenyl)carbamate




Dissolve 4-amino-2-chloro-benzonitrile (500 mg, 3.3 mmol) and pyridine (518 mg, 6.6 mmol) in THF (5 mL), cool to 0° C. and add slowly phenyl chloroformate (616 mg, 3.9 mmol). After addition, stir the reaction at room temperature for 2 hrs. TLC (EtOAc:PE=1:3) shows the reaction is complete. Add water, extract with DCM, separate the organic layer. Wash the organic layer with 1M HCl solution, brine sequentially, dry over anhydrous Na2SO4, and concentrate under reduced pressure to give the target compound (820 mg, 91.7%). MS: (M+1): 273.
Intermediate D54 can be synthesized with similar method (Table D11).




Step 1: Synthesis of 3-cyclopropyl-4-fluoro-aniline




Mix 3-bromo-4-fluoro-aniline (415 mg, 2.18 mmol), cyclopropylboronic acid (244 mg, 2.84 mmol), K3PO4 (1.62 g, 7.64 mmol), triphenylphosphine (61 mg, 0.22 mmol), Pd(OAc)2 (25 mg, 0.11 mmol), toluene (12 mL) and water (1 mL) under N2, stir at 100° C. for 16 hrs. Cool the reaction to 25° C., add water (10 mL), extract with EtOAc (10 mL×2). Combine the organic layers; wash with brine (15 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, PE:EtOAc=2:1) affords the target compound (200 mg, 61%).
Step 2: Synthesis of phenyl N-(3-cyclopropyl-4-fluoro-phenyl)carbamate




Add 3-cyclopropyl-4-fluoro-aniline (600 mg, 3.97 mmol), phenyl chloroformate (620 mg, 3.97 mmol) and pyridine (314 mg, 3.97 mmol) in DCM (30 mL), stir at room temperature (25° C.) for 4 hrs. After the reaction is complete, add water (15 mL), extract with EtOAc (15 mL×2), combine the organic layers, wash with 1M HCl solution (15 mL) and brine (15 mL) sequentially, dry over anhydrous Na2SO4. Concentrate under reduced pressure to give a crude product (350 mg) which is used directly without further purification.




Step 1: Synthesis of 3-(4-nitrophenoxy)propanoic acid




Dissolve p-nitrophenol (11 g, 100 mmol), 3-chloropropanoic acid (14 g, 100 mmol) in a mixture of ethanol (100 mL) and 20% aqueous KOH solution (100 mL). Heat to reflux for 2 hrs, then adjust pH=1-3, extract with EtOAc (300 mL×3), combine the organic layers and wash with brine (300 mL×2), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:4) affords the title compound (7.5 g, 14.9%). MS: (M+1): 212.1.
Step 2: Synthesis of 6-nitrochroman-4-one




Mix 3-(4-nitrophenoxy)propanoic acid (6.5 g, 30 mmol) and concentrated H2SO4 (30 mL). Once dissolved, add P2O5, stir at 65° C. for 3 hrs. Cool to room temperature, pour slowly the reaction mixture to ice water (50 mL), stir for 15 min, and extract with EtOAc (100 mL×3). Combine the organic layers, wash with brine (100 mL×2), dry over anhydrous Na2SO4, concentrate to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (4.6 g, 77.8%). MS: (M−1): 192.2.
Step 3: Synthesis of 4,4-difluoro-6-nitro-chromane




Add 6-nitrochroman-4-one (3 g, 15.7 mmol), bis-(2-methoxyethyl)aminosulfur trifluoride (Deoxo-Fluor®, BAST, 14 g, 62.8 mmol) and dry DCM (10 mL) in tube, seal the tube and heat at 50° C. for 12 hrs. Cool to room temperature, add dropwise methanol (5 mL), pour the mixture to ice water (50 mL), extract with DCM (100 mL×2), combine the organic layers, wash with brine (100 mL×2), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:9) affords the title compound (2.6 g, 77.2%).
Step 4: Synthesis of 4,4-difluorochroman-6-amine




Dissolve 4,4-difluoro-6-nitro-chromane (2.5 g, 11.6 mmol) in methanol (100 mL), add zinc powder (2.5 g, 34.8 mmol), then add dropwise a 15.5% aqueous solution of ammonium chloride (40 mL, 139 mmol), stir at room temperature for 5 hrs. Filter off the solid, add water (50 mL) to the filtrate, extract with EtOAc (50 mL×2). Combine the organic layers, wash with brine (50 mL×2), dry over anhydrous Na2SO4. Concentrate under reduced pressure to get a crude residue. Purification by chromatography (silica gel, EtOAc:PE=1:3) affords the title compound (1.8 g, 83.7%). MS: (M+1): 186.2.
Data on Intermediates D53-56 are summarized in Table D11:







TABLE D11







Intermediates D53-D56










Number
Starting material
Intermediate
MS [M + 1]+





D53








273.1


 


D54








266.1


 


D55








272.2


 


D56








306.2









Intermediates D57-62 can be synthesized with similar method (Table D12)







TABLE D12







Intermediates D57-D62










Number
Starting material
Intermediate
MS [M + 1]+













D57








316.1


 


D58








359.1


 


D59








NA


 


D60








395.2


 


D61








NA


 


D62








NA









EXAMPLES
Example 1
Synthesis of N-[6-[4-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]-3-methyl-phenoxy]pyrimidin-4-yl]cyclopropanecarboxamide




Dissolve N-[6-(4-amino-3-methyl-phenoxy)pyrimidin-4-yl]cyclopropanecarboxamide (1.8 g, 6.3 mmol) in tetrahydrofuran (50 mL) and dichloromethane (50 mL), add slowly 4-chloro-3-(trifluoromethyl)phenyl isocynate (1.4 g, 6.3 mmol), stir the reaction at room temperature for 15 hrs. Collect the white solid formed by filtration (2.0 g), recover from the mother liquor to give another 1.0 g (total 3.0 g, 93.7% yield). MS: (M+1): 506.2.
When the crude product contains a small amount of impurities, the product may be purified by recrystallization in ethanol/tetrahydrofuran as the following: add the crude product (8.2 g) to ethanol (120 mL) in a 250 mL flask, heat to reflux, add slowly tetrahydrofuran (about 50 mL) to dissolve the solid. Then cool slowly, filter and collect the white solid to give the pure product (6.2 g, 75.6% recovery yield).
Example 2
Synthesis of N-[6-[3-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoyl-amino]-4-fluoro-phenoxy]pyrimidin-4-yl]cyclopropanecarboxamide




Dissolve N-[6-(3-amino-4-fluoro-phenoxy)pyrimidin-4-yl]cyclopropanecarboxamide (400 mg, 1.4 mmol) in THF (4 mL) and DCM (10 mL), add 4-chloro-3-(trifluoromethyl)phenyl isocynate (1.4 g, 6.3 mmol). Stir the reaction at room temperature overnight. Filter and collect the solid to get the title compound (545 mg, 77%). MS: (M+1): 510.2.
Examples 1-31 are prepared with similar method (Table 1).







TABLE 1







Examples 1-31









Example




Number
Structure
MS [M + 1]+












Example 1




506.2


 


Example 2




510.2


 


Example 3




524.2


 


Example 4




502.3


 


Example 5




510.2


 


Example 6




517.2


 


Example 7




492.2


 


Example 8




510.1


 


Example 9




510.2


 


Example 10




506.2


 


Example 11




507.2


 


Example 12




492.2


 


Example 13




510.2


 


Example 14




506.2


 


Example 15




493.2


 


Example 16




507.2


 


Example 17




506.2


 


Example 18




506.2


 


Example 19




526.1


 


Example 20




492.1


 


Example 21




492.1


 


Example 22




509.2


 


Example 23




505.2


 


Example 24




509.1


 


Example 25




510.1


 


Example 26




516.2


 


Example 27




508.2


 


Example 28




523.3


 


Example 29




509.2


 


Example 30




505.2


 


Example 31




505.3









Example 32
Synthesis of N-[6-[4-[[3-(isopropylamino)-5-(trifluoromethyl)phenyl]carbamoylamino]-3-methyl-phenoxy]pyrimidin-4-yl]cyclopropanecarboxamide




Dissolve phenyl N-[4-[6-(cyclopropanecarbonylamino)pyrimidin-4-yl]oxy-2-methyl-phenyl]carbamate (343 mg, 1.57 mmol), N1-isopropyl-5-(trifluoromethyl)benzene-1,3-diamine (700 mg, 1.73 mmol) and DIEA (608 mg, 4.72 mmol) in THF (4 mL), heat the reaction at 80° C. for 3 hrs. After the reaction is complete, remove the volatiles under reduced pressure. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (450 mg, 83%). MS: (M+1): 529.3.
Examples 33-66 are prepared with similar method (Table 2).







TABLE 2







Examples 32-66









Example




Number
Structure
MS [M + 1]+












Example 32




529.3


 


Example 33




472.3


 


Example 34




491.3


 


Example 35




489.3


 


Example 36




572.3


 


Example 37




553.2


 


Example 38




574.3


 


Example 39




561.2


 


Example 40




600.3


 


Example 41




574.2


 


Example 42




558.2


 


Example 43




562.2


 


Example 44




532.2


 


Example 45




601.3


 


Example 46




576.3


 


Example 47




588.3


 


Example 48




587.3


 


Example 49




530.3


 


Example 50




618.3


 


Example 51




574.3


 


Example 52




558.2


 


Example 53




605.3


 


Example 54




601.3


 


Example 55




587.3


 


Example 56




472.3


 


Example 57




447.2


 


Example 58




465.2


 


Example 59




464.3


 


Example 60




598.3


 


Example 61




475.2


 


Example 62




500.2


 


Example 63




525.1


 


Example 64




569.2


 


Example 65




454.3


 


Example 66




484.3









Example 67
Synthesis of N-[4-[2-fluoro-4-[[3-(1-piperazin-1-ylethyl)-5-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-2-pyridyl]cyclopropanecarboxamide




Step 1
Synthesis of tert-butyl 4-[1-[3-[[4-[[2-(cyclopropanecarbonylamino)-4-pyridyl]oxy]-3-fluoro-phenyl]carbamoylamino]-5-(trifluoromethyl)phenyl]ethyl]piperazine-1-carboxylate




Add phenyl N-[4-[[2-(cyclopropanecarbonylamino)-4-pyridyl]oxy]-3-fluoro-phenyl]carbamate (220 mg, 0.53 mmol), tert-butyl 4-[1-[3-amino-5-(trifluoromethyl)phenyl]-ethyl]piperazine-1carboxylate (165 mg, 0.442 mmol) and DIEA (120 mg, 0.88 mmol) in THF (10 mL), stir the reaction under N2 at 80° C. for 12 hrs. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the title compound (230 mg, 75.6%).
Step 2
Synthesis of N-[4-[2-fluoro-4-[[3-(1-piperazin-1-ylethyl)-5-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-2-pyridyl]cyclopropanecarboxamide




Dissolve the compound obtained in Step 1 (230 mg, 0.33 mmol) in DCM (15 mm), add trifluoroacetic acid (400 mg), stir at room temperature for 2 hrs. Remove the volatiles, partition the residue in EtOAc and saturated NaHCO3 solution, and separate the organic layer, dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=3:1) affords the title compound (173 mg, 88.3%). MS: (M+1): 588.
Examples 68-69 are prepared with similar method (Table 3).







TABLE 3







Examples 67-69









Example




number
Structure
MS [M + 1]+












Example 67




588


 


Example 68




518.2


 


Example 69




573.3









Example 70
Synthesis of N-[6-[4-[[4-fluoro-3-(trifluoromethyl)phenyl]carbamoylamino]-3-methyl-phenoxy]pyrimidin-4-yl]cyclopropanecarboxamide




Add N-[6-(4-amino-3-methyl-phenoxy)pyrimidin-4-yl]cyclopropanecarboxamide (99 mg, 0.35 mmol), phenyl N-[4-fluoro-3-(trifluoromethyl)phenyl]carbamate (100 mg, 0.35 mmol) and DIEA (135 mg, 1.05 mmol) in THF (4 mL), stir at 80° C. for 2 hrs, Remove the volatiles, suspend the residue in DCM, filter and collect the solid to get the target compound (132 mg, 77%). MS: (M+1): 490.3.
Examples 71-236 are prepared with similar method (Table 4)







TABLE 4







Examples 70-236









Example




number
Structure
MS [M + 1]+





Example 70 




490.3


 


Example 71 




486.3


 


Example 72 




486.3


 


Example 73 




569.4


 


Example 74 




472.3


 


Example 75 




506.2


 


Example 76 




584.4


 


Example 77 




541.3


 


Example 78 




584.4


 


Example 79 




488.3


 


Example 80 




506.2


 


Example 81 




506.3


 


Example 82 




522.2


 


Example 83 




486.3


 


Example 84 




468.3


 


Example 85 




502.3


 


Example 86 




463.2


 


Example 87 




557.3


 


Example 88 




510.2


 


Example 89 




510.2


 


Example 90 




510.2


 


Example 91 




520.2


 


Example 92 




490.3


 


Example 93 




502.2


 


Example 94 




472.3


 


Example 95 




488.3


 


Example 96 




502.3


 


Example 97 




494.2


 


Example 98 




494.2


 


Example 99 




506.2


 


Example 100




476.2


 


Example 101




476.2


 


Example 102




476.2


 


Example 103




472.3


 


Example 104




490.3


 


Example 105




506.2


 


Example 106




490.3


 


Example 107




494.2


 


Example 108




476.2


 


Example 109




490.2


 


Example 110




506.2


 


Example 111




490.3


 


Example 112




472.3


 


Example 113




488.2


 


Example 114




522.2


 


Example 115




486.2


 


Example 116




486.3


 


Example 117




502.2


 


Example 118




486.3


 


Example 119




476.2


 


Example 120




458.2


 


Example 121




474.2


 


Example 122




488.2


 


Example 123




489.3


 


Example 124




490.2


 


Example 125




502.2


 


Example 126




530.3


 


Example 127




490.2


 


Example 128




486.2


 


Example 129




559.3


 


Example 130




536.2


 


Example 131




570.3


 


Example 132




591.2


 


Example 133




483.2


 


Example 134




555.3


 


Example 135




458.2


 


Example 136




536.2


 


Example 137




491.2


 


Example 138




475.2


 


Example 139




482.2


 


Example 140




471.2


 


Example 141




475.3


 


Example 142




491.2


 


Example 143




554.3


 


Example 144




457.2


 


Example 145




535.2


 


Example 146




473.3


 


Example 147




454.3


 


Example 148




471.3


 


Example 149




487.2


 


Example 150




471.2


 


Example 151




481.3


 


Example 152




482.3


 


Example 153




493.2


 


Example 154




489.2


 


Example 155




493.1


 


Example 156




493.3


 


Example 157




509.2


 


Example 158




476.2


 


Example 159




552.2


 


Example 160




509.2


 


Example 161




587.3


 


Example 162




574.3


 


Example 163




544.2


 


Example 164




588.3


 


Example 165




568.2


 


Example 166




608.2


 


Example 167




587.3


 


Example 168




587.3


 


Example 169




476.2


 


Example 170




477.2


 


Example 171




574.2


 


Example 172




574.3


 


Example 173




588.3


 


Example 174




561.2


 


Example 175




491.2


 


Example 176




509.2


 


Example 177




509.2


 


Example 178




491.2


 


Example 179




489.3


 


Example 180




471.2


 


Example 181




489.3


 


Example 182




505.3


 


Example 183




489.3


 


Example 184




459.1


 


Example 185




509.2


 


Example 186




552.2


 


Example 187




532.2


 


Example 188




588.3


 


Example 189




560.3


 


Example 190




500.2


 


Example 191




489.3


 


Example 192




489.3


 


Example 193




471.3


 


Example 194




487.3


 


Example 195




505.3


 


Example 196




505.3


 


Example 197




485.2


 


Example 198




485.3


 


Example 199




501.3


 


Example 200




485.3


 


Example 201




476.1


 


Example 202




490.2


 


Example 203




488.3


 


Example 204




474.3


 


Example 205




493.2


 


Example 206




500.2


 


Example 207




489.2


 


Example 208




493.3


 


Example 209




493.2


 


Example 210




509.2


 


Example 211




475.2


 


Example 212




566.2


 


Example 213




476.2


 


Example 214




509.3


 


Example 215




509.2


 


Example 216




472.3


 


Example 217




471.2


 


Example 218




489.3


 


Example 219




505.2


 


Example 220




489.2


 


Example 221




466.2


 


Example 222




465.2


 


Example 223




489.3


 


Example 224




489.3


 


Example 225




489.3


 


Example 226




471.3


 


Example 227




485.3


 


Example 228




501.3


 


Example 229




485.3


 


Example 230




489.3


 


Example 231




471.3


 


Example 232




487.2


 


Example 233




485.2


 


Example 234




485.3


 


Example 235




501.3


 


Example 236




485.3









Example 237
Synthesis of N-[4-[4-[[3-(azetidin-3-ylmethoxy)-5-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-2-pyridyl]cyclopropanecarboxamide




Step 1
Synthesis of tert-butyl 3-[[3-[[4-[[2-(cyclopropanecarbonylamino)-4-pyridyl]oxy]phenyl]carbamoylamino]-5-(trifluoromethyl)phenoxy]methyl]azetidine-1-carboxylate




Dissolve N-[4-(4-aminophenoxy)-2-pyridyl]cyclopropanecarboxamide (200 mg, 0.74 mmol) and tert-butyl 3-[[3-(phenoxycarbonylamino)-5-(trifluoromethyl)phenoxy]methyl]-azetidine-1-carboxylate (415 mg, 0.89 mmol) in THF (10 mL), add DIEA (200 mg, 1.48 mmol), stir under N2 at 70° C. for 12 hrs. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=2:1) affords the title compound (250 mg, 52.4%). MS: (M+1): 642.3.
Step 2
Synthesis of N-[4-[4-[[3-(azetidin-3-ylmethoxy)-5-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-2-pyridyl]cyclopropanecarboxamide




Dissolve the compound obtained in last step (250 mg, 0.39 mmol) in DCM (15 mL), add trifluoroacetic acid (6 mL), stir at room temperature for 2 hrs. Remove the volatiles, partition between EtOAc and saturated NaHCO3 solution. Separate the organic layer, dry over anhydrous Na2SO4, concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=3:1) affords the title compound (150 mg, 71%). MS: (M+1): 542.1.
Examples 238-240 are prepared with the above method (Table 5).







TABLE 5







Examples 237-240









Example




number
Structure
MS [M + 1]+





Example 237




542.1


 


Example 238




541.2


 


Example 239




490.2


 


Example 240




587.3









Example 241
Synthesis of N-[4-[4-[[3-(1-methylazetidin-3-yl)oxy-5-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-2-pyridyl]cyclopropanecarboxamide




Step 1
Synthesis of tert-butyl 3-[3-[[4-[[2-(cyclopropanecarbonylamino)-4-pyridyl]oxy]phenyl]carbamoylamino]-5-(trifluoromethyl)phenoxy]azetidine-1-carboxylate




Dissolve N-[4-(4-aminophenoxy)-2-pyridyl]cyclopropanecarboxamide (186 mg, 0.69 mmol), tert-butyl 3-[3-(phenoxycarbonylamino)-5-(trifluoromethyl)phenoxy]azetidine-1-carboxylate (312 mg, 0.69 mmol) and DMAP (17 mg, 0.14 mmol) in DMF (10 mL), stir at 100° C. for 16 hrs. Cool the reaction, pour the mixture to water (50 mL), adjust pH=7 with saturated NaHCO3 solution, extract with EtOAc (15 mL×3). Combine the organic layers, wash with brine (100 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, EtOAc:PE=1:1) affords the target compound (300 mg, 69%). MS: (M+1): 628.3.
Step 2
Synthesis of N-[4-[4-[[3-(azetidin-3-yloxy)-5-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-2-pyridyl]cyclopropanecarboxamide




Dissolve tert-butyl 3-[3-[[4-[[2-(cyclopropanecarbonylamino)-4-pyridyl]oxy]phenyl]-carbamoylamino]-5-(trifluoromethyl)phenoxy]azetidine-1-carboxylate (300 mg, 0.48 mmol) in DCM (4 mL), add slowly trifluoroacetic acid (2 mL), stir the reaction at room temperature for 2 hrs. After reaction, pour the mixture to water (50 mL), adjust pH=7 with saturated NaHCO3 solution, extract with EtOAc (15 mL×3). Combine the organic layers, wash with brine (100 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product (240 mg) which is used without further purification. MS: (M+1): 528.2.
Step 3
Synthesis of N-[4-[4-[[3-(1-methylazetidin-3-yl)oxy-5-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-2-pyridyl]cyclopropanecarboxamide




Dissolve N-[4-[4-[[3-(azetidin-3-yloxy)-5-(trifluoromethyl)phenyl]carbamoylamino]-phenoxy]-2-pyridyl]cyclopropanecarboxamide (210 mg, 0.4 mmol) in methanol (10 mL), add 35% aqueous formaldehyde solution (205 mg, 2.39 mmol), stir for 1.5 hrs. Add Na(CN)BH3 (150 mg, 2.39 mmol) and stir at room temperature for 3 hrs. After reaction, pour the reaction mixture to water (50 mL), adjust pH=7 with saturated NaHCO3 solution, extract with EtOAc (15 mL×3), combine the organic layers, wash with brine (100 mL), dry over anhydrous Na2SO4. Concentrate under reduced pressure to give the crude product. Purification by chromatography (silica gel, DCM:MeOH=20:1) affords the target compound (16 mg, 7%). MS: (M+1): 542.3.
Example 242 is prepared with similar method (Table 7).







TABLE 6







Examples 241-242









Example




number
Structure
MS [M + 1]+





Example 241




542.3


 


Example 242




615.3









EXAMPLES ON BIOLOGICAL ACTIVITIES
Biological Activity Example 1
Inhibitory Activity of Compounds at KDR Kinase


    
    
        Experimental method: Use Lance@Ultra UlightTM-TK assay kit from PerKinElmer to evaluate the inhibitory activity of compounds at KDR kinase.
        Instrument: PerKinElmer's ENVISION plate reader
        Materials: Optiplate-384 well plate (PerKinElmer), kinase buffer (50 mM Hepes pH7.5, 0.25 mM EGTA, 2 mM DTT, 0.01% Tween 20, 10 mM Mg2+, 0.5 mM Mn2+), KDR kinase (790-1356AA, Crown Bioscience), KDR kinase substrate (PerKinElmer catalogue #TRF0127-M), Lance@Eu-W 1024-anti-phosphotyrosine (PT66) (PerKinElmer, catalogue #AD0068), ATP (Invitrogen), DMSO (Sigma, catalogue #34869), purified water (Millipore, type: Milli-Q).
        Study conditions: Mix KDR (final concentration: 20 nM) and compound (final DMSO: 0.5%), pre-incubate for 20 min at 30° C.; then add ATP (final concentration: 90 μM) and the substrate (final concentration 50 μM). React for 2 hrs at 30° C. After the reaction, add antibody and reaction for 60 min at 30° C. Read the plate (615 nm, 665 nm), calculate the ratio of the value at 665 nm vs 615 nm and analyze the data.
        Samples: Compounds in the examples and Sorafenib (positive control)
        Data Analysis: Use CBIS data analysis software to calculate IC50 values
    
    


Biological Activity Example 2
Inhibitory Activity of Compounds at B-Raf Kinase


    
    
        Experimental method: Use ADP-GLOTM assay kit from Promega to evaluate the inhibitory activity of compounds at B-Raf
        Instrument: PerKinElmer's ENVISION plate reader
        Material: Optiplate-384 well plate (PerKinElmer), kinase buffer (50 mM Hepes pH7.5, 1 mM EGTA, 2 mM DTT, 10 mM Mg2+, 0.05% BSA), B-Rafkinase (Millipore, catalogue #14-530-K), GST-MEK1 substrate (Carna, catalogue #07-141-10), Super pure ATP (Promega), ADP-GLOTM assay kit (Promega, catalogue #V9102), DMSO (Sigma, catalogue #34869), purified water (Millipore, type: Milli-Q).
        Study conditions: Mix B-Raf kinase (final concentration: 5 nM) and compound (DMSO final concentration: 0.25%) and preincubate at 30° C. for 20 min, then add ATP (final concentration: 6 μM) and reaction substrate (final concentration: 30 nM), React for 2 hrs at 30° C. After reaction, add ADP-GLOTM reagent, react for 40 min at room temperature, then add ADP-GLOTM detecting agent, react for 30 min at room temperature. After that, use ENVSION to measure the fluorescence.
        Samples: Compounds in the examples and Sorafenib (positive control)
        Data analysis: Use CBIS data analysis software to calculate IC50 values
    
    


Biological Activity Example 3
Inhibitory Activity of Compounds at pERK


    
    
        Experimental method: Use Acumen (TPP) to measure In Cell Western Blot results
        Instrument: Acumen (TPP), Centrifuge (Thermo Scientific, Sorvall ST16R)
        Material: MDA-MB-231 cells (ATCC), 96 well plate (BD, #356640), DMEM medium(Gibco, #11965-092), PBS (Invitrogen, #10010023), fetal bovine serum (Gibco, #16000044), BSA (Sigma, #A7030), DMSO (Sigma, #D2650); formaldehyde (Sinopharm), methanol (Sinopharm), 1st antibody (anti-pERK antibody, Cell Signaling, #CST 4307s), 2nd antibody (FITC Donkey anti-rabbit IgG, Biolegend, #406403), Propidium Iodide (Invitrogen, #P3566), purified water (Millipore, type: Milli-Q).
        Protocol: Grow MDA-MB-231 cells in DMEM medium (contains 10% FBS). When the cells reach 80% confluency, collect cells by trypsinization and centrifugation at 800 rpm for 3 minutes. Resuspend cells with appropriate volume of medium and count cell numbers. Adjust cell concentration to 70000 cells/mL, and split 100 μL cell suspension per well into 96-well plates. Transfer the plates into cell culture incubator (37° C., 5% CO2) and incubate cells overnight. Discard the medium, wash with PBS once, Prepare compound serial dilutions in culture medium with total 8 compound concentrations at 3-fold serial dilutions (the final top concentration starts from 30 μM). Dispense 75 μL DMEM medium (with 0.1% BSA) and 25 μL of each test compound dilution into corresponding wells with DMSO at 0.3% final concentration. Continue incubating cells in cell culture incubator for additional 2 hrs at 37° C., 5% CO2. Discard culture medium, and wash cells once with PBS. Add 100 μL of 4% formaldehyde solution to each well and fix for 20 min. Discard the formaldehyde, then add 100 μL ice-cold 100% methanol at 4° C. for 20 min. Discard methanol, wash with PBS three times. Add to each well 100 μL of 2% BSA solution, seal for 30 min at room temperature. Discard the BSA solution, add to each well 50 μL of the 1st antibody solution (prepared with 2% BSA, 1:250 dilution), incubate at 4° C. overnight. Discard the 1st antibody solution, wash with PBS four times. Add to each well 50 μL the 2nd antibody solution (prepared with 1% BSA, 1:1000 dilution), incubate at room temperature for 1 hr. Discard the 2nd antibody solution, wash with PBS four times, add to each well 100 μL PI solution (1.5 μM), incubate at room temperature for 30 min. Use Acumen (TPP) to measure the fluorescence.
        Samples: Compounds in the examples and sorafenib (positive control)
        Data Analysis: Use CBIS data analysis software to calculate IC50 values
    
    


Biological Activity Example 4
Inhibitory Activity of Compounds to the Proliferation of PLC-PRF-5 Cells


    
    
        experimental Method: Use PerKinElmer's ATPliteTM Luminescence ATP Detection Assay System to evaluate the inhibitory activity of compounds to the proliferation of PLC-PRF-5 cells.
        Instruments: ENVISION plate reader (PerKinElmer) and centrifuge (Thermo Scientific, Sorvall ST16R)
        material: PLC-PRF-5 cells (ATCC), 96-well plate (Nunc, #165305), DMEM medium (Gibco, #11965-092), PBS (Invitrogen, #10010023), fetal bovine serum (Gibco, #16000044), DMSO (Sigma, #D2650), ATPliteTM Luminescence ATP Detection Assay System (PerkinElmer, #6016949), purified water (Millipore, type: Milli-Q).
        Protocol: Grow PLC-PRF-5 cells in DMEM supplemented with 10% FBS. When the cells reach 80% confluency, collect cells by trypsinization and centrifugation at 800 rpm for 3 min. Resuspend cells with appropriate volume of medium and count cell numbers. Adjust cell concentration to 50000 cells/mL, and split 160 μL cell suspension per well into 96-well plates. Transfer the plates into cell culture incubator (37° C., 5% CO2) and incubate cells for 2 hrs. Prepare compound serial dilutions in culture medium with total 8 compound concentrations at 3-fold serial dilutions (the final top concentration starts from 30 μM). Dispense 40 μL of each test compound dilution into corresponding wells with DMSO at 0.3% final concentration. Continue incubating cells in cell culture incubator for additional 96 hrs at 37° C., 5% CO2. Discard culture medium, and wash cells once with PBS. Add 50 μL cell lysis buffer per well and shake plates for 5 min at room temperature to ensure complete lysis of cells. Then add 50 μL of substrate solution and shake plates for 1 min at room temperature. Allow the plates to incubate at room temperature for 5 min, followed by recording the luminescence signals using Envision, and subsequent data analysis.
        Samples: Compounds in the examples and Sorafenib (positive control)
        Data analysis: Use CBIS data analysis software to calculate IC50 values

In Vitro Activity Summary for Example 1-242 (IC50, nM)

    
    



















MDA-M
PLC-PRF-5


Example


B-231
cell


Number
KDR
B-Raf 
pERK
proliferation



















Exe. 001
10.9
39
31
6900


Exe. 002
5.4

15.5
3400


Exe. 003
16.7

87
11500


Exe. 004
292

74
21900


Exe. 005
27

40.6
6000


Exe. 006
25

281
5380


Exe. 007
5.7
24.8
46
7240


Exe. 008
5.7
24.9
124
4550


Exe. 009
8.9
26
20.5
3650


Exe. 010
13.7
97
43.1
4350


Exe. 011
13.4

261
19500


Exe. 012
9.9

71
10300


Exe. 013
14.2

50
16100


Exe. 014 
7

48.5
8700


Exe. 015
4.2

94
6500


Exe. 016
9.4

55
20100


Exe. 017
29.2
15.7
18.8
2820


Exe. 018
9.4

4.6
4040


Exe. 019
22.3
18.1
172
840


Exe. 020
5.8
19.7
290.6
1990


Exe. 021
7.5
24.7
10.2
600


Exe. 022
4.3
3.6
19.5
1710


Exe. 023
10.5
15
6.9
4170


Exe. 024
33.8
5.1
19.5
4240


Exe. 025
14.9
9.1
71.6
371


Exe. 026
18.2

39
900


Exe. 027
11.5

66.5
2390


Exe. 028
12.7
21
391
2490


Exe. 029
3.9
7.3
10.1
790


Exe. 030
12.4
18
23.2
2800


Exe. 031
14.5
25
33.8
1300


Exe. 032
26

115
20000


Exe. 033
5.6

5.0
35


Exe. 034
10.2
9.4
4.7
219


Exe. 035
7.8
8.9
9.3
259


Exe. 036
15.6

290
2400


Exe. 037
13.5
26
33.9
768


Exe. 038
8.2
5.3
23.5
130


Exe. 039
14.2

54.4
500


Exe. 040
15.2

134
640


Exe. 041
4.2
37.5
18.1
358


Exe. 042
11.5

94
3560


Exe. 043
18.1

30
604


Exe. 044
14.2
13
29.8
752


Exe. 045
15.4

40
108


Exe. 046
16.6

58.2
640


Exe. 047
6.2
73.6
53.3
682


Exe. 048
7.3
11.0
25
233


Exe. 049
9.9

51.3
642


Exe. 050
12.8

110
2230


Exe. 051
5.2
23.6
33.9
749


Exe. 052
15.5
13
22
360


Exe. 053
13

65.1
686


Exe. 054
15.3

97.6
340


Exe. 055
14.5

284
420


Exe. 056
17.5
65
39.7
110


Exe. 057
12.0

85
250


Exe. 058
13.7
116
30
43


Exe. 059
8.2
90
19.4
643


Exe. 060
34.3

324
1510


Exe. 061
3.73
6.3
26.3
211


Exe. 062
7.18
7.0
29.4
3050


Exe. 063
5.5
14.9
29.1
2330


Exe. 064
11.9
4.6

610


Exe. 065
6.5
25
46.5
162


Exe. 066
8.0

16
2500


Exe. 067
26.6

75.9
235


Exe. 068
15.7

8.4
325


Exe. 069
17.1

337
10500


Exe. 070
11.0
145
63.5
9300


Exe. 071
13.3

100
20500


Exe. 072
8.9

12
7000


Exe. 073
14

14
2970


Exe. 074
10.5
146
77.5
6070


Exe. 075
13.1

420
20000


Exe. 076
12.5

83
5600


Exe. 077
14.3

169
16500


Exe. 078
13.3

28
879


Exe. 079
14

80
6000


Exe. 080
8.1

29
3040


Exe. 081
13.3

104
16000


Exe. 082
6.8

36
22500


Exe. 083
14.3

609
20400


Exe. 084
12.2

122
5860


Exe. 085
10.7
45
19.3
11000


Exe. 086
8.3

226
19300


Exe. 087
12.9

36
5050


Exe. 088
11.4

106
2050


Exe. 089
15.2

19
6870


Exe. 090
12.2

42
2800


Exe. 091
12.8

53
7600


Exe. 092
192

57
17200


Exe. 093
200

244
17300


Exe. 094
200

163
19000


Exe. 095
434

82
19200


Exe. 096
624

28
21000


Exe. 097
62.8

137
9100


Exe. 098
105

752
8400


Exe. 099
41

37
15000


Exe. 100
9

300
2420


Exe. 101
12.5

682
2600


Exe. 102
15

294
936


Exe. 103
12.4

300
373


Exe. 104
10.2

252
100


Exe. 105
11.3
153
85.5
1900


Exe. 106
10.4

465
850


Exe. 107
13.4
31
20.2
324


Exe. 108
8.6
41
20
1610


Exe. 109
10.9
32
9.8
274


Exe. 110
7.3
24
13.4
1580


Exe. 111
7

41
2630


Exe. 112
14.5

85
2000


Exe. 113
8.1

54
781


Exe. 114
9.8

37
4600


Exe. 115
9.5

300
1310


Exe. 116
7.9
111
30.4
125


Exe. 117
12

27
2930


Exe. 118
7.4
293
55.3
706


Exe. 119
14.9

184
18800


Exe. 120
12

617
>30000


Exe. 121
11.5

252
>30000


Exe. 122
8.7

133
22000


Exe. 123
7.4

703
8380


Exe. 124
13.8
10.4
19.4
160


Exe. 125
8.7
8.6
4.7
1720


Exe. 126
30.2
61.4
24.3
2440


Exe. 127
11.8
7.2
11.6
642


Exe. 128
9.8
6.2
38.0
819


Exe. 129
14.2

105
2260


Exe. 130
4.6
29.7
725
4990


Exe. 131
6.1
12.5
>1000
10500


Exe. 132
10
39
341
5910


Exe. 133
9.1
25
23.6
1020


Exe. 134
7.8

33
199


Exe. 135
5.8

208
100


Exe. 136
6.6
29
46
470


Exe. 137
10.2
11.3
7.8
5960


Exe. 138
10.5
26
9.1
1510


Exe. 139
10
13
10.9
654


Exe. 140
10.0

10.6
230


Exe. 141
7.1
125
40.4
642


Exe. 142
10.4
39
13.9
642


Exe. 143
9.5
5.5
13.4
892


Exe. 144
11.4
30
6.0
403


Exe. 145
7.6
5.1
20.7
1670


Exe. 146
7.8
14
11.4
689


Exe. 147
11.1
15
23.8
1160


Exe. 148
6.2
16
7.7
240


Exe. 149
25.4
38
25.4
800


Exe. 150
11.5

20.4
737


Exe. 151
4.5
3.3
3.6
608


Exe. 152
7.1
72
33
390


Exe. 153
15.2
32
23
735


Exe. 154
11.3

14.9
130


Exe. 155
12.1

195
571


Exe. 156
8.3
37
37.7
52


Exe. 157
5.9
43
34.7
156


Exe. 158
8.3

81.9
1040


Exe. 159
8.0
35.4
44.3
642


Exe. 160
7.8

186
4100


Exe. 161
12.2
7.9
86.9
1340


Exe. 162
6.8
22.5
30.6
330


Exe. 163
11.6
24
32.9
998


Exe. 164
30.7
71.5
198
2950


Exe. 165
5.5
6.6
41.0
1070


Exe. 166
11.5
27
45.5
1640


Exe. 167
16.1
23.8
262
1230


Exe. 168
12.0
7.9
11.6
31


Exe. 169
14.6

43
340


Exe. 170
5.6
96
31.6
230


Exe. 171
12.4

30.2
1980


Exe. 172
14.8

74
197


Exe. 173
22.5

80
254


Exe. 174
5.4

60.6
240


Exe. 175
4.9
9.8
30.5
270


Exe. 176
5.7

40
60


Exe. 177
9.5

37
690


Exe. 178
5.3
34.7
159
4880


Exe. 179
10.5

51.5
206


Exe. 180
9.8

31.7
152


Exe. 181
7.1
4.8
25.7
23


Exe. 182
14.7
32
17.0
752


Exe. 183
13.6
56
15.7
230


Exe. 184
7.6
73.4
310
610


Exe. 185
12.3

46.4
1480


Exe. 186
5.0
27.5
53.3
640


Exe. 187
6.8
34.3
63.8
638


Exe. 188
17.2
19
23.5
34


Exe. 189
10.7
29.8
19.3
260


Exe. 190
11.9
87
74.3
875


Exe. 191
11.8
10
9.4
2530


Exe. 192
17.5

20
2910


Exe. 193
10.9
6.8
7.4
2300


Exe. 194
10.5
8.8
9.5
2320


Exe. 195
6.1
13
10.1
790


Exe. 196
11.8
13
14.6
2500


Exe. 197
5.7

14.2
3290


Exe. 198
8.3

5.7
740


Exe. 199 
14
11.7
11
3700


Exe. 200 
10

5.5
3180


Exe. 201
11.4

326
239


Exe. 202
12.7

246
66


Exe. 203
16.2

61
539


Exe. 204
9.3

72
1850


Exe. 205
25.1
13
9.2
1090


Exe. 206
14.6

8.2
1650


Exe. 207
10.2

8.3
71


Exe. 208
14.9
37
9.8
640


Exe. 209
8.4
9.0
5.9
51


Exe. 210
11.8
19
17
636


Exe. 211
7.8
6.4
2.3
190


Exe. 212
5.5
20.1
21.1
177


Exe. 213
10.7

7.2
79


Exe. 214
9.4

11.5
180


Exe. 215
13.2
19
14.5
618


Exe. 216
10.6
10
11.8
536


Exe. 217
9.9
8.9
2.3
178


Exe. 218
9.1
7.8
3.1
21


Exe. 219
11
8.3
8.8
632


Exe. 220
14.9
7.1
4.6
240


Exe. 221
6.7
11
7.8
1270


Exe. 222
8.7

17.3
490


Exe. 223
6.9

9.0
3430


Exe. 224
9.5
29
22.4
4700


Exe. 225
10.7
11
12
5720


Exe. 226
9.1

11
3530


Exe. 227
17.2
27
34.4
17800


Exe. 228
14.7

12
3330


Exe. 229
13.4

6.9
3170


Exe. 230
11.7
20.7
12
830


Exe. 231
11.6
12.5
7.1
310


Exe. 232
11
14
10.9
418


Exe. 233
8.1

18.5
559


Exe. 234
9.9
16
6.8
63


Exe. 235
13.4
22
12.2
1290


Exe. 236
9.5
21
7.1
404


Exe. 237
13.9
3.7
730
14900


Exe. 238
15.6

57.3
2220


Exe. 239
9.7
7.0
27.2
930


Exe. 240
19.1

27.2
1040


Exe. 241
17.9
9.1
61.5
1770


Exe. 242
16.7

372
4630

## Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof



wherein:
R1 is hydrogen;
M is N;
L is O;
A is CR5;
W is CR6;
R2, R5 and R6 are independently hydrogen, C1-6 alkyl, or halo;
X, Y, and Z are CH; and
R3 and R4 are independently hydrogen, halo, C1-6 haloalkyl or C1-6 haloalkoxy.

2. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.

3. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.

4. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.

5. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.

6. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.

7. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.

8. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.

9. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.

10. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.

11. The compound according to claim 1, having following structure:




or pharmaceutically acceptable salts thereof.