Substituted benzamides as modulators of TREX1

Abstract

Provided are compounds of formula (I): (I) and pharmaceutically acceptable salts thereof and compositions thereof, wherein the compounds and pharmaceutically acceptable salts and compositions thereof can be used to treat various conditions associated with TREX1.

Description

[0001] Related Applications

[0002] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 030,590, filed on May 27, 2020, the entire contents of which are incorporated herein by reference. Background Art

[0003] Cancers related to non-self innate immune system recognition require potential immunotherapy, and potential immunotherapy is needed for detecting and preventing potential dangers. Cancer cells are different from their normal counterparts in antigens and send danger signals similar to viral infections to alert the immune system. These signals containing damage-associated molecular patterns (DAMPs) and pathogen-associated molecular patterns (PAMPs) further activate the innate immune system, thereby protecting the host from various threats (Front. Cell Infect. Microbiol. 2012, 2, 168).

[0004] Ectopically expressed single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) are known PAMPs and / or DAMPs that are recognized by cyclic GMP-AMP synthase (cGAS), a nucleic acid sensor (Nature 2011, 478, 515-518). Upon sensing cytosolic DNA, cGAS catalyzes the production of the cyclic dinucleotide 2',3'-cGAMP, a potent second messenger and activator of the ER transmembrane adaptor protein stimulator of interferon genes (STING) (Cell Rep. 2013, 3, 1355-1361). STING activation triggers phosphorylation of IRF3 via TBK1, which in turn leads to the production of type I interferons and activation of interferon-stimulated genes (ISGs); this is a prerequisite for activating innate immunity and initiating adaptive immunity. Therefore, the production of type I interferon constitutes a key bridge between innate immunity and adaptive immunity (Science 2013, 341, 903-906).

[0005] Excessive type I IFNs can be harmful to the host and induce autoimmunity, therefore, there is a negative feedback mechanism that inhibits type I IFN-mediated immune activation. 3' repair exonuclease I (TREX1) is a 3'-5' DNA exonuclease responsible for the removal of ectopically expressed ssDNA and dsDNA, and is therefore a key repressor of the cGAS / STING pathway (Proceedings of the National Academy of Sciences (PNAS) 2015, 112, 5117-5122).

[0006] Type I interferon and downstream proinflammatory cytokine responses are crucial for the development of immune response and its effectiveness. Type I interferon enhances dendritic cells and macrophages to absorb, process antigens, present and cross-present antigens to T cells by inducing the increase of costimulatory molecules such as CD40, CD80 and CD86, and the dendritic cells and macrophages stimulate the effectiveness of T cells (Journal of Experimental Medicine (J.Exp.Med.) 2011,208,2005-2016). Type I interferon also binds to its own receptor and activates interferon response genes, which help activate the cells involved in adaptive immunity (EMBO Rep. 2015,16,202-212).

[0007] From a therapeutic point of view, type I interferons and compounds that can induce the production of type I interferons have the potential to be used to treat human cancer (Nat. Rev Immunol. 2015, 15, 405-414). Interferons can directly inhibit the proliferation of human tumor cells. In addition, type I interferons can enhance anti-tumor immunity by triggering the activation of cells from both the innate and adaptive immune systems. Importantly, the anti-tumor activity of PD-1 blockade requires pre-existing intratumoral T cells. By converting cold tumors into hot tumors and thereby inducing spontaneous anti-tumor immunity, type I IFN induction therapy has the potential to expand the patient population that responds to anti-PD-1 therapy and enhance the effectiveness of anti-PD1 therapy.

[0008] Human and mouse genetic studies suggest that TREX1 inhibition may be amenable to systemic delivery approaches and thus TREX1 inhibitory compounds may play an important role in the field of anti-tumor therapy. TREX1 is a key determinant of the limited immunogenicity of cancer cells in response to radiation therapy [Trends in Cell Biol., 2017, 27(8), 543-4; Nature Commun., 2017, 8, 156-18]. TREX1 is induced by genotoxic stress and is involved in protecting glioma and melanoma cells from the effects of anti-cancer drugs [Biochim. Biophys. Acta, 2013, 1833, 1832-43]. STACT-TREX1 therapy has shown robust anti-tumor efficacy in a variety of murine cancer models [Glickman et al., Poster P235, 33rd Annual Meeting of the Society for Immunotherapy of Cancer (33 rdAnnual Meeting of Society for Immunotherapy of Cancer), Washington DC, November 7-11, 2018]. (TREX1) expression is associated with cervical cancer cell growth in vitro and disease progression in vivo [Scientific Reports 1019, 9, 351]. In addition to oncology, there is also support that IFN pathway agonists can be used for antiviral therapy, such as STING agonists that induce innate antiviral immune responses to hepatitis B virus by stimulating the IFN pathway and upregulating the expression of ISGs [Antimicrob. Agents Chemother. 2015, 59: 1273–1281], and TREX1 inhibits the innate immune response to HIV type 1 [Nature Immunology, 2010, 11(11), 1005]. Summary of the invention

[0009] Provided herein are compounds having Formula I:

[0010]

[0011] and pharmaceutically acceptable salts thereof and compositions thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , m, n and p are as described herein. The disclosed compounds of Formula I, pharmaceutically acceptable salts thereof and compositions thereof modulate TREX1 and can be used in a variety of therapeutic applications, for example, to treat cancer. Therefore, the use thereof for treating diseases that respond to inhibition of TREX1 is included.

[0012] In addition to the use of compounds of formula I, compounds and compositions having formula VI are provided, in particular for use in treating diseases responsive to inhibition of TREX1:

[0013]

[0014] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 2a , R 3a , R 4a , R 5a , R 6a , m1, n1 and p1 are as described herein. DETAILED DESCRIPTION

[0015] 1. Compound review

[0016] In a first embodiment, provided herein is a compound of formula I:

[0017]

[0018] or a pharmaceutically acceptable salt thereof, wherein

[0019] R 1 is halogen, hydroxy, (C1-C4)alkyl, halo-(C1-C4)alkyl, (C1-C4)alkoxy or halo-(C1-C4)alkoxy;

[0020] R 2 is hydrogen, (C1-C4)alkyl, halogenated (C1-C4)alkyl, -(C1-C4)alkyl, OR a 、-C(O)R b 、-C(O)OR b 、-C(O)NR b R c 、-C(O)NR b R c , 5 to 7 membered heteroaryl or 4 to 7 membered heterocyclyl, wherein the heteroaryl and heterocyclyl are each optionally selected from R 6 substituted with 1 to 3 groups;

[0021] R 3 is halogen, (C1-C4) alkyl, halogenated (C1-C4) alkyl, (C1-C4) alkoxy, halogenated (C1-C4) alkoxy, -NR b R c or CN;

[0022] R 4 is halogen, hydroxy, (C1-C4) alkyl, halogenated (C1-C4) alkyl, (C1-C4) alkoxy, halogenated (C1-C4) alkoxy, CN, -NR b R c 、-C(O)R b 、-C(O)OR b 、-C(O)NR b R c 、-SR b 、-C(O)NR b R c 、-S(O)2R b 、-S(O)R b 、-NR b C(O)R c 、-NR b C(O)OR c 、-NR b C(S)OR cand-NR b C(O)N c R d ;

[0023] Ring A is a nitrogen-containing 5- to 7-membered heteroaryl group;

[0024] R 5 is halogen, hydroxy, (C1-C4) alkyl, halo (C1-C4) alkyl, (C1-C4) alkoxy, halo (C1-C4) alkoxy, oxo, -(C1-C4) alkyl, OR d 、-(C1-C4)alkylC(O)R d 、-(C1-C4)alkylC(O)OR e 、-(C1-C4)alkylC(O)NR d R e 、-C(O)R d 、-C(O)OR d 、-C(O)NR d R e 、-C(O)NR d SO2(C1-C4)alkyl, CN, -NR d R e 、-C(O)NR d SO3H, -NR d C(O)R e 、-NR d C(O)OR e 、-NR d C(S)OR e 、-NR d C(O)N e R f 、-NR d C(S)NR e R c 、-NR d S(O)2NR e R f 、-C(S)R d 、-S(O)2R d 、-S(O)R d 、-C(S)OR d 、-C(S)NR d R e 、-NR d C(S)R e 、-SR d 、-(C1-C4)alkylC(O)NR d R e 、-(C1-C4)alkylC(O)NR dSO2(C1-C4)alkyl, -(C1-C4)alkylNR d R e 、-(C1-C4)alkylC(O)NR d SO3H, -(C1-C4)alkyl NR d C(O)R e 、-(C1-C4)alkylNR d C(O)OR e 、-(C1-C4)alkylNR d C(S)OR e 、-(C1-C4)alkylNR d C(O)N e R f 、-(C1-C4)alkylNR d C(S)NR e R f 、-(C1-C4)alkylNR d S(O)2NR e R f 、-(C1-C4)alkylC(S)R d 、-(C1-C4)alkylS(O)2R d 、-(C1-C4)alkylS(O)R d 、-(C1-C4)alkylC(S)OR d 、-(C1-C4)alkylC(S)NR d R e 、-(C1-C4)alkylNRd b C(S)R e 、-(C1-C4)alkyl SR d , 5 to 7 membered heteroaryl or 4 to 7 membered heterocyclyl, wherein the heteroaryl and heterocyclyl are each optionally selected from R 7 substituted with 1 to 3 groups;

[0025] R a , R b , R c , R d , R e and R f are each independently hydrogen, (C1-C4)alkyl or halogenated (C1-C4)alkyl;

[0026] R 6 and R 7 Each is independently halo, hydroxy, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, CN or oxo;

[0027] m and n are each independently 0, 1, 2 or 3; and

[0028] p is 0 or 1.

[0029] In a second embodiment, provided herein are compounds of formula VI, inter alia, for use in treating diseases responsive to inhibition of TREX1:

[0030]

[0031] or a pharmaceutically acceptable salt thereof, wherein

[0032] R 1a , R 2a , R 4a and R 5a Each is independently halogen, hydroxy, (C1-C4) alkyl, halo (C1-C4) alkyl, (C1-C4) alkoxy, halo (C1-C4) alkoxy, CN, -NR b1 R c1 、-C(O)R b1 、-C(O)OR b1 、-C(O)NR b1 R c1 、-SR b1 、-C(O)NR b1 R c1 、-S(O)2R b1 、-S(O)R b1 、-NR b1 C(O)R c1 、-NR b1 C(O)OR c1 、-NR b1 C(S)OR c1 and-NR b1 C(O)N c1 R d1 ;

[0033] R 3a is hydrogen, (C1-C4)alkyl, halogenated (C1-C4)alkyl, -(C1-C4)alkyl, OR a 、-C(O)R b 、-C(O)OR b 、-C(O)NR b R c 、-C(O)NR b R c , 5 to 7 membered heteroaryl or 5 to 7 membered heterocyclyl, wherein the heteroaryl and heterocyclyl are each optionally selected from R 7a substituted with 1 to 3 groups;

[0034] Ring A is a heteroaryl group;

[0035] R6a is halogen, hydroxy, (C1-C4) alkyl, halo (C1-C4) alkyl, (C1-C4) alkoxy, halo (C1-C4) alkoxy, oxo, -(C1-C4) alkyl, OR d1 、-(C1-C4)alkylC(O)R d1 、-(C1-C4)alkylC(O)OR e1 、-(C1-C4)alkylC(O)NR d1 R e1 、-C(O)R d1 、-C(O)OR d1 、-C(O)NR d1 R e1 、-C(O)NR d1 SO2(C1-C4)alkyl, CN, -NR d1 R e1 、-C(O)NR d1 SO3H, -NR d1 C(O)R e1 、-NR d1 C(O)OR e1 、-NR d1 C(S)OR e1 、-NR d1 C(O)N e1 R f1 、-NR d1 C(S)NR e1 R c1 、-NR d1 S(O)2NR e1 R f1 、-C(S)R d1 、-S(O)2R d1 、-S(O)R d1 、-C(S)OR d1 、-C(S)NR d1 R e1 、-NR d1 C(S)R e1 、-SR d1 、-(C1-C4)alkylC(O)NR d1 R e1 、-(C1-C4)alkylC(O)NR d1 SO2(C1-C4)alkyl, -(C1-C4)alkylNR d1 R e1 、-(C1-C4)alkylC(O)NR d1 SO3H, -(C1-C4)alkyl NR d1 C(O)R e1、-(C1-C4)alkylNR d1 C(O)OR e1 、-(C1-C4)alkylNR d1 C(S)OR e1 、-(C1-C4)alkylNR d1 C(O)N e1 R f1 、-(C1-C4)alkylNR d1 C(S)NR e1 R f1 、-(C1-C4)alkylNR d1 S(O)2NR e1 R f1 、-(C1-C4)alkylC(S)R d1 、-(C1-C4)alkylS(O)2R d1 、-(C1-C4)alkylS(O)R d1 、-(C1-C4)alkylC(S)OR d1 、-(C1-C4)alkylC(S)NR d1 R e1 、-(C1-C4)alkylNR b1 C(S)R e1 、-(C1-C4)alkyl SR d1 , 5 to 7 membered heteroaryl or 4 to 7 membered heterocyclyl, wherein the heteroaryl and heterocyclyl are each optionally selected from R 8a substituted with 1 to 3 groups;

[0036] R a1 , R b1 , R c1 , R d1 , R e1 and R f1 are each independently hydrogen, (C1-C4)alkyl or halogenated (C1-C4)alkyl;

[0037] R 7a and R 8a are each independently halo, hydroxy, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, CN or oxo; and

[0038] m1, n1 and p1 are each independently 0, 1, 2 or 3.

[0039] 2. definition

[0040] When used conjunctively to describe a chemical group that may have multiple points of attachment, the hyphen (-) indicates the point of attachment of the group to the variable defining it. For example, -NRb C(O)OR c and-NR b C(S)OR c This means that the point of attachment of this group occurs on the nitrogen atom.

[0041] The terms "halo" and "halogen" refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br) and iodine (iodo, -I).

[0042] The term "alkyl" when used alone or as part of a larger moiety such as "haloalkyl", etc., means a saturated straight or branched chain monovalent hydrocarbon radical.

[0043] "Alkoxy" means an alkyl group represented by -O-alkyl, which is linked through an oxygen linking atom. For example, "(C1-C4)alkoxy" includes methoxy, ethoxy, propoxy and butoxy.

[0044] The term "haloalkyl" embraces monohaloalkyl, polyhaloalkyl and perhaloalkyl wherein the halogen is independently selected from fluorine, chlorine, bromine and iodine.

[0045] "Haloalkoxy" is a haloalkyl group attached to another moiety through an oxygen atom, for example, -OCHF2 or -OCF3.

[0046] The term oxo refers to the group =0.

[0047] The term "heteroaryl", used alone or as part of a larger moiety, refers to a 5 to 12-membered (e.g., 4 to 6-membered) aromatic group containing 1-4 selected from N, O, and S. The heteroaryl group can be monocyclic or bicyclic, as size permits. Monocyclic heteroaryl includes, for example, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, triazinyl, tetrazinyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, etc. Bicyclic heteroaryl includes groups in which a monocyclic heteroaromatic ring is fused to one or more aryl or heteroaromatic rings. Non-limiting examples include indolyl, imidazopyridyl, benzoxazolyl, benzoxadiazole, indazolyl, benzimidazolyl, benzothiazolyl, quinolyl, quinazolinyl, quinoxalinyl, pyrrolopyridyl, pyrrolopyrimidinyl, pyrazolopyridyl, thienopyridyl, thienopyrimidinyl, indolizinyl, purinyl, naphthyridinyl and pteridinyl. It should be understood that when specified, the optional substituent on the heteroaryl can be present in any substitutable position and include, for example, the position where the heteroaryl is connected.

[0048] The term "heterocyclyl" means a 4- to 12-membered saturated or partially unsaturated heterocyclic ring containing 1 to 4 heteroatoms independently selected from N, O and S. The heterocyclic ring can be connected to its side group at any heteroatom or carbon atom forming a stable structure. The heterocyclic group can be monocyclic or bicyclic. Examples of monocyclic saturated heterocyclic groups or partially unsaturated heterocyclic groups include but are not limited to tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxacycloalkyl, dioxolanyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridyl, tetrahydropyridyl, dihydropyrimidinyl and tetrahydropyrimidinyl. Bicyclic heterocyclyls include, for example, unsaturated heterocyclyls fused to another unsaturated heterocyclyl, cycloalkyl, or aromatic or heteroaryl rings, for example, benzodioxolyl, dihydrobenzoxazinyl, dihydrobenzodioxinyl, 6,7-dihydro-5H-pyrrolo[2,1-c][1,2,4]triazolyl, 5,6,7,8-tetrahydroimidazo[1,2-a]pyridinyl, 1,2-dihydroquinolinyl, dihydrobenzofuranyl, tetrahydronaphthyridine, indolinone, dihydropyrrolotriazole, quinolinone, chromanyl, and dioxaspirodecane. It is also understood that, when specified, optional substituents on the heterocyclyl may be present at any substitutable position and include, for example, the position at which the heterocyclyl is attached.

[0049] The term "spirocycle" refers to two rings that share one ring atom (eg, carbon).

[0050] The term "fused" refers to two rings that share two adjacent ring atoms with each other.

[0051] The term "bridged" refers to two rings that share three ring atoms with each other.

[0052] The term "TREX1" refers to 3' repair exonuclease 1 or DNA repair exonuclease 1, which is an enzyme encoded by the TREX1 gene in humans. Mazur DJ, Perrino FW (August 1999). “Identification and expression of the TREX1 and TREX2 cDNA sequences encoding mammalian 3'-->5' exonucleases.” J Biol Chem. 274(28):19655–60. doi:10.1074 / jbc.274.28.19655. PMID 10391904; Hoss M, Robins P, Naven TJ, Pappin DJ, Sgouros J, Lindahl T (August 1999). “A human DNA editing enzyme homologous to the Escherichia coli DnaQ / MutD protein.” TREX1 protein". EMBO J. 18(13):3868-75. doi:10.1093 / emboj / 18.13.3868.PMC 1171463.PMID10393201. This gene encodes the major 3'->5' DNA exonuclease in human cells. The protein is a non-processive exonuclease that can provide proofreading functions to human DNA polymerase. The protein is also a component of the SET complex and is used to rapidly degrade the 3' ends of nicked DNA during granzyme A-mediated cell death. Cells lacking functional TREX1 exhibit chronic DNA damage checkpoint activation and extranuclear accumulation of endogenous single-stranded DNA substrates. It appears that the TREX1 protein normally acts on single-stranded DNA polynucleotide species generated by processing aberrant replication intermediates. This action of TREX1 attenuates DNA damage checkpoint signaling and prevents pathological immune activation. TREX1 metabolizes reverse transcribed single-stranded DNA from endogenous retroviral elements according to cell-intrinsic antiviral surveillance, resulting in a potent type I IFN response. TREX1 helps HIV-1 evade cytoplasmic sensing by degrading viral cDNA in the cytoplasm.

[0053] The term "TREX2" refers to 3' repair exonuclease 2, which is an enzyme encoded by the TREX2 gene in humans. This gene encodes a nuclear protein with 3' to 5' exonuclease activity. The encoded protein is involved in double-strand DNA break repair and can interact with DNA polymerase δ. Enzymes with this activity are involved in DNA replication, repair and recombination. TREX2 is a 3'-exonuclease that is mainly expressed in keratinocytes and helps the epidermis respond to UVB-induced DNA damage. TREX2 biochemical and structural properties are similar to TREX1, but the two are not identical. The two proteins share a dimer structure and can be expressed in vitro with almost identical k cat TREX2 is a member of the cytoplasm of the mitochondria, and TREX1 is a member of the mitochondria. TREX2 is a member ...

[0054] The terms "subject" and "patient" are used interchangeably and refer to mammals in need of treatment, e.g., companion animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, pigs, horses, sheep, goats, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). Typically, the subject is a human in need of treatment.

[0055] The terms "inhibit", "inhibition" and "inhibiting" encompass a decrease in the baseline activity of a biological activity or process.

[0056] As used herein, the term "treatment (treatment, treat and treating)" refers to reversing, alleviating, delaying the onset of a disease or disorder as described herein or one or more of its symptoms or inhibiting its progress. In some aspects, treatment can be administered after one or more symptoms occur, i.e., therapeutic treatment. In other aspects, treatment can be administered in the absence of symptoms. For example, treatment can be administered to susceptible individuals before the onset of symptoms (e.g., according to symptom history and / or according to exposure to a specific organism or other susceptible factors), i.e., preventive treatment. Treatment can also be continued after the symptoms have subsided, for example, to delay its recurrence.

[0057] The term "pharmaceutically acceptable carrier" refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound formulated with it. Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances, such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulosic substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene glycol-polyoxypropylene block polymers, polyethylene glycol and lanolin.

[0058] For use in medicine, the salts of the compounds described herein refer to non-toxic "pharmaceutically acceptable salts". Pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic / anionic or basic / cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include, for example, salts of inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid and sulfuric acid) and salts of organic acids (such as acetic acid, benzenesulfonic acid, benzoic acid, methanesulfonic acid and p-toluenesulfonic acid). Compounds of the present teachings having acidic groups such as carboxylic acids can form pharmaceutically acceptable salts with pharmaceutically acceptable bases. Suitable pharmaceutically acceptable basic salts include, for example, ammonium salts, alkali metal salts (such as sodium and potassium salts) and alkaline earth metal salts (such as magnesium and calcium salts). Compounds having quaternary ammonium groups also contain counter anions such as chloride, bromide, iodide, acetate, perchlorate, etc. Other examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, benzoates, and salts with amino acids such as glutamic acid.

[0059] The term "effective amount" or "therapeutically effective amount" refers to the amount of a compound described herein that will elicit a desired or beneficial biological or medical response in a subject, for example, a dosage between 0.01-100 mg / kg body weight / day.

[0060] 3. Compound

[0061] In a second embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof wherein R 2 is hydrogen, (C1-C4)alkyl, halogenated (C1-C4)alkyl, -(C1-C4)alkyl, OR a , 5 to 7 membered heteroaryl or 4 to 7 membered heterocyclyl, wherein the heteroaryl or heterocyclyl is each independently selected from R 6 substituted with 1 to 3 groups; R 3is halo, (C1-C4)alkyl, halo(C1-C4)alkyl or CN; and m and n are each independently 0, 1 or 2, wherein the remaining variables are as described above for Formula I.

[0062] In a third embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof wherein R 3 wherein the remaining features are as described above for Formula I or the second embodiment. Alternatively, as part of the third embodiment, R in the compound of Formula I or a pharmaceutically acceptable salt thereof is 3 is halo or halo(C1-C4)alkyl, wherein the remaining features are as described above for Formula I or the second embodiment. In another alternative, as part of the third embodiment, R in the compound of Formula I or a pharmaceutically acceptable salt thereof is 3 is fluorine, chlorine, methyl, methoxy or CF3, wherein the remaining features are as described above for Formula I or the second embodiment. In another alternative, as part of the third embodiment, R in the compound of Formula I or a pharmaceutically acceptable salt thereof 3 is fluorine, chlorine or CF3, wherein the remaining features are as described above for Formula I or the second embodiment.

[0063] In a fourth embodiment, the compound of Formula I has Formula II:

[0064]

[0065] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I or the second embodiment or the third embodiment.

[0066] In a fifth embodiment, R in the compound of Formula I or II or a pharmaceutically acceptable salt thereof 2 is hydrogen or (C1-C4)alkyl, wherein the variables are as described above for Formula I or the second embodiment or the third embodiment.

[0067] In a sixth embodiment, the compound of Formula I has Formula III:

[0068]

[0069] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I or the second or third embodiment.

[0070] In a seventh embodiment, R in the compound of Formula I, II or III or a pharmaceutically acceptable salt thereof 4is halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy or halo(C1-C4)alkoxy or CN, wherein the remaining variables are as described above for Formula I or the second, third or fifth embodiment. Alternatively, as part of the seventh embodiment, R in the compound of Formula I, II or III or a pharmaceutically acceptable salt thereof is 4 is chloro, methyl or methoxy, wherein the remaining variables are as described above for Formula I or the second, third or fifth embodiment. In another alternative, as part of the seventh embodiment, R in the compound of Formula I, II or III or a pharmaceutically acceptable salt thereof is 4 is halo, wherein the remaining variables are as described above for Formula I or the second, third or fifth embodiment. In another alternative, as part of the seventh embodiment, R in the compound of Formula I, II or III or a pharmaceutically acceptable salt thereof is 4 is chloro, wherein the remainder of the variables are as described above for Formula I or the second, third or fifth embodiment.

[0071] In an eighth embodiment, m in a compound of Formula I, II or III, or a pharmaceutically acceptable salt thereof, is 0, 1 or 2, wherein the remaining variables are as described above for Formula I or the second, third, fifth or seventh embodiment. Alternatively, as part of the eighth embodiment, m in a compound of Formula I, II or III, or a pharmaceutically acceptable salt thereof, is 0 or 1, wherein the remaining variables are as described above for Formula I or the second, third, fifth or seventh embodiment.

[0072] In a ninth embodiment, the compound of Formula I has Formula IV:

[0073]

[0074] or a pharmaceutically acceptable salt thereof, wherein the remaining variables are as described above for Formula I or the second, third, fifth, seventh or eighth embodiment.

[0075] In a tenth embodiment, Ring A in a compound of Formula I, II, III or IV, or a pharmaceutically acceptable salt thereof, is pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl or pyrazolyl, wherein the remaining variables are as described above for Formula I or the second, third, fifth, seventh or eighth embodiment. Alternatively, as part of the tenth embodiment, Ring A in a compound of Formula I, II, III or IV, or a pharmaceutically acceptable salt thereof, is pyridinyl, wherein the remaining variables are as described above for Formula I or the second, third, fifth, seventh or eighth embodiment.

[0076] In an eleventh embodiment, the compound of Formula I has Formula V:

[0077]

[0078] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula I or the second or third embodiment.

[0079] In a twelfth embodiment, R in the compound of Formula I, II, III, IV or V or a pharmaceutically acceptable salt thereof is 5 is located in the para position relative to the point of attachment to Ring A, wherein the remainder of the variables are as described above for Formula I or the second, third, fifth, seventh, eighth or tenth embodiment.

[0080] In the thirteenth embodiment, R in the compound of Formula I, II, III, IV or V or a pharmaceutically acceptable salt thereof 5 is halogen, oxo, (C1-C4)alkyl, halogenated (C1-C4)alkyl, -C(O)OR b 、-NR b R c 、-C(O)NR b R c 、-(C1-C4)alkylOR b 、-(C1-C4)alkylC(O)R b 、-(C1-C4)alkylC(O)NR b R c 、-C(O)NR b SO2(C1-C4)alkyl, 5 to 6 membered heteroaryl, wherein the heteroaryl is optionally selected from R 7 wherein the remaining variables are as described above for Formula I or the second, third, fifth, seventh, eighth, tenth or twelfth embodiment. Alternatively, as part of the thirteenth embodiment, R in the compound of Formula I, II, III, IV or V or a pharmaceutically acceptable salt thereof is 5 is oxo, CF3, CH3, C(O)OCH3, C(O)OH, CH2COOH, NH2, CONH2, CH2CONH2, CONHCH3, CH2OH, CH2CH2OH, CONHSO2CH3, tetrazolyl, pyrazolyl, triazolyl or pyrazolyl, wherein the pyrazolyl is optionally substituted with hydroxy, and wherein the remaining variables are as described above for Formula I or the second, third, fifth, seventh, eighth, tenth or twelfth embodiment.

[0081] In a fourteenth embodiment, n in the compound of Formula I, II, III, IV or V, or a pharmaceutically acceptable salt thereof, is 0, 1 or 2, wherein the remaining variables are as described above for Formula I or the second, third, fifth, seventh, eighth, tenth, twelfth or thirteenth embodiment. Alternatively, as part of the fourteenth embodiment, n in the compound of Formula I, II, III, IV or V, or a pharmaceutically acceptable salt thereof, is 1 or 2, wherein the remaining variables are as described above for Formula I or the second, third, fifth, seventh, eighth, tenth, twelfth or thirteenth embodiment. In another alternative, as part of the fourteenth embodiment, n in the compound of Formula I, II, III, IV or V, or a pharmaceutically acceptable salt thereof, is 2, wherein the remaining variables are as described above for Formula I or the second, third, fifth, seventh, eighth, tenth, twelfth or thirteenth embodiment.

[0082] In the fifteenth embodiment, the compound of Formula VI or a pharmaceutically acceptable salt thereof wherein R 3a is hydrogen, (C1-C4)alkyl, halogenated (C1-C4)alkyl, -(C1-C4)alkyl, OR a , 5 to 7 membered heteroaryl or 4 to 7 membered heterocyclyl, wherein the heteroaryl and heterocyclyl are each optionally selected from R 7a and m1, n1 and p1 are each independently 0, 1 or 2, wherein the remaining variables are as described above for Formula VI.

[0083] In the seventeenth embodiment, the compound of Formula VI or a pharmaceutically acceptable salt thereof, wherein R 1a is halo, wherein the remaining variables are as described above for Formula VI or the fifteenth embodiment.

[0084] In a seventeenth embodiment, the compound of Formula VI has Formula VII:

[0085]

[0086] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula VI or the fifteenth or sixteenth embodiment.

[0087] In the eighteenth embodiment, R in the compound of Formula VI or VII or a pharmaceutically acceptable salt thereof 2a is halo, hydroxy, (C1-C4)alkyl, halo(C1-C4)alkyl, or (C1-C4)alkoxy, wherein the variables are as described above for Formula VI or the fifteenth or sixteenth embodiment. Alternatively, as part of the eighteenth embodiment, R in the compound of Formula VI or VII, or a pharmaceutically acceptable salt thereof, is 2a is halo, wherein the variables are as described above for Formula VI or the fifteenth or sixteenth embodiment.

[0088] In a nineteenth embodiment, the compound of formula VI has formula VIII:

[0089]

[0090] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth or eighteenth embodiment.

[0091] In a twentieth embodiment, the compound of formula VI has formula IX:

[0092]

[0093] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth or eighteenth embodiment.

[0094] In the twenty-first embodiment, R in the compound of Formula VI, VII, VIII or IX or a pharmaceutically acceptable salt thereof is 4a is halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, or halo(C1-C4)alkoxy, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, or eighteenth embodiments. Alternatively, as part of the twenty-first embodiment, R in the compound of Formula VI, VII, VIII, or IX, or a pharmaceutically acceptable salt thereof, is 4a is fluoro, chloro, methyl, methoxy or CF3, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth or eighteenth embodiment. In another alternative, R in the compound of Formula VI, VII, VIII or IX as part of the twenty-first embodiment or a pharmaceutically acceptable salt thereof is 4a is halo(C1-C4)alkyl or halo, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth or eighteenth embodiment. In another alternative, R in the compound of Formula VI, VII, VIII or IX as part of the twenty-first embodiment or a pharmaceutically acceptable salt thereof is 4a is fluoro, chloro or CF3, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth or eighteenth embodiment.

[0095] In the twenty-second embodiment, R in the compound of Formula VI, VII, VIII or IX or a pharmaceutically acceptable salt thereof is 3a is hydrogen or (C1-C4)alkyl, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth or twenty-first embodiment.

[0096] In a twenty-third embodiment, the compound of Formula VI has Formula X:

[0097]

[0098] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth or eighteenth embodiment.

[0099] In the twenty-fourth embodiment, R in the compound of Formula VI, VII, VIII, IX or X or a pharmaceutically acceptable salt thereof 5a is halo or (C1-C4)alkoxy, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth or twenty-first embodiments. Alternatively, as part of the twenty-fourth embodiment, R in the compound of Formula VI, VII, VIII, IX or X, or a pharmaceutically acceptable salt thereof, is 5a is chloro, methyl or methoxy, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth or twenty-first embodiment. In another alternative, as part of the twenty-fourth embodiment, R in the compound of Formula VI, VII, VIII, IX or X or a pharmaceutically acceptable salt thereof is 5a is halo, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth or twenty-first embodiments. In another alternative, as part of the twenty-fourth embodiment, R in the compound of Formula VI, VII, VIII, IX or X, or a pharmaceutically acceptable salt thereof, is 5a is chloro, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth or twenty-first embodiment.

[0100] In the twenty-fifth embodiment, m1 in the compound of Formula VI, VII, VIII, IX or X, or a pharmaceutically acceptable salt thereof, is 0, 1 or 2, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth, twenty-first or twenty-fourth embodiment.

[0101] In the twenty-sixth embodiment, ring A in the compound of formula VI, VII, VIII, IX or X, or a pharmaceutically acceptable salt thereof, is a 5- to 7-membered heteroaryl, wherein the variables are as described above for formula VI or the fifteenth, sixteenth, eighteenth, twenty-first, twenty-fourth or twenty-fifth embodiment. Alternatively, as part of the twenty-fourth embodiment, ring A in the compound of formula VI, VII, VIII, IX or X, or a pharmaceutically acceptable salt thereof, is a nitrogen-containing 5- to 7-membered heteroaryl, wherein the variables are as described above for formula VI or the fifteenth, sixteenth, eighteenth, twenty-first, twenty-fourth or twenty-fifth embodiment. In another alternative, as part of the twenty-fourth embodiment, ring A in the compound of formula VI, VII, VIII, IX or X, or a pharmaceutically acceptable salt thereof, is pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl or pyrazolyl, wherein the variables are as described above for formula VI or the fifteenth, sixteenth, eighteenth, twenty-first, twenty-fourth or twenty-fifth embodiment. In another alternative, as part of the twenty-fourth embodiment, Ring A in the compound of Formula VI, VII, VIII, IX or X, or a pharmaceutically acceptable salt thereof, is pyridinyl, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth, twenty-first, twenty-fourth or twenty-fifth embodiment.

[0102] In a twenty-seventh embodiment, the compound of Formula VI has Formula XI:

[0103]

[0104] or a pharmaceutically acceptable salt thereof, wherein the variables are as described above for Formula VI or the twenty-first, twenty-fourth, twenty-fifth or twenty-sixth embodiment.

[0105] In the twenty-eighth embodiment, R in the compound of Formula VI, VII, VIII, IX, X or XI or a pharmaceutically acceptable salt thereof 6a is halogen, oxo, (C1-C4)alkyl, halogenated (C1-C4)alkyl, -C(O)OR b1 、-NR b1 R c1 、-C(O)NR b1 R c1 、-(C1-C4)alkylOR b1 、-(C1-C4)alkylC(O)R b1 、-(C1-C4)alkylC(O)NR b1 R c1 、-C(O)NR b1 SO2(C1-C4)alkyl, 5 to 6 membered heteroaryl, wherein the heteroaryl is optionally selected from R 7awherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth, twenty-first, twenty-fourth, twenty-fifth or twenty-sixth embodiment. Alternatively, as part of the twenty-eighth embodiment, R in the compound of Formula VI, VII, VIII, IX, X or XI or a pharmaceutically acceptable salt thereof is 6a is oxo, CF3, CH3, C(O)OCH3, C(O)OH, CH2COOH, NH2, CONH2, CH2CONH2, CONHCH3, CH2OH, CH2CH2OH, CONHSO2CH3, tetrazolyl, pyrazolyl, triazolyl or pyrazolyl, wherein the pyrazolyl is optionally substituted with hydroxy, and wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth, twenty-first, twenty-fourth, twenty-fifth or twenty-sixth embodiment.

[0106] In the twenty-ninth embodiment, n1 in the compound of Formula VI, VII, VIII, IX, X or XI, or a pharmaceutically acceptable salt thereof, is 0, 1 or 2, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth, twenty-first, twenty-fourth, twenty-fifth, twenty-sixth or twenty-eighth embodiment. Alternatively, as part of the twenty-ninth embodiment, n1 in the compound of Formula VI, VII, VIII, IX, X or XI, or a pharmaceutically acceptable salt thereof, is 1 or 2, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth, twenty-first, twenty-fourth, twenty-fifth, twenty-sixth or twenty-eighth embodiment. In another alternative, as part of the twenty-ninth embodiment, n1 in the compound of Formula VI, VII, VIII, IX, X or XI, or a pharmaceutically acceptable salt thereof, is 0 or 1, wherein the variables are as described above for Formula VI or the fifteenth, sixteenth, eighteenth, twenty-first, twenty-fourth, twenty-fifth, twenty-sixth or twenty-eighth embodiment.

[0107] Compounds of Formula I and VI are further disclosed in the Examples and are included in the present disclosure, including pharmaceutically acceptable salts and neutral forms thereof.

[0108] 4. Use, preparation and application

[0109] The compounds and compositions described herein are generally useful for modulating the activity of TREX1. In some aspects, the compounds and pharmaceutical compositions described herein inhibit active TREX1.

[0110] In some aspects, the compounds and pharmaceutical compositions described herein can be used to treat disorders related to TREX1 function. Therefore, provided herein is a method for treating disorders related to TREX1 function, the method comprising administering a therapeutically effective amount of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a disclosed compound or a pharmaceutically acceptable salt thereof to a subject in need thereof. Also provided is a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a disclosed compound or a pharmaceutically acceptable salt thereof for the manufacture of a drug for treating disorders related to TREX1 function. Also provided is a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a disclosed compound or a pharmaceutically acceptable salt thereof for the treatment of disorders related to TREX1 function.

[0111] In some aspects, the compounds and pharmaceutical compositions described herein can be used to treat cancer.

[0112] In some aspects, the cancer treated by the compounds and pharmaceutical compositions described herein is selected from colon cancer, gastric cancer, thyroid cancer, lung cancer, leukemia, pancreatic cancer, melanoma, multiple melanomas, brain cancer, CNS cancer, kidney cancer, prostate cancer, ovarian cancer, leukemia and breast cancer.

[0113] In some aspects, the cancer treated by the compounds and pharmaceutical compositions described herein is selected from lung cancer, breast cancer, pancreatic cancer, colorectal cancer, and melanoma.

[0114] In some aspects, pharmaceutical compositions described herein are formulated to be used for patients who need such compositions. Pharmaceutical compositions described herein can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implantable reservoir. Term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. In certain embodiments, the compositions are administered orally, intraperitoneally or intravenously. The sterile injectable form of pharmaceutical compositions described herein can be aqueous or oily suspensions. These suspensions can be formulated using suitable dispersants or wetting agents and suspending agents according to technology known in the art.

[0115] In some aspects, the pharmaceutical composition is administered orally.

[0116] The specific dosage and treatment regimen for any particular patient will depend on various factors, including the activity of the specific compound employed, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination and the judgment of the treating physician and the severity of the specific disease being treated. The amount of the compound described herein in the composition will also depend on the specific compound in the pharmaceutical composition.

[0117] example

[0118] Chemical synthesis

[0119] The following representative examples are intended to help illustrate the present disclosure and are not intended to, and should not be construed to, limit the scope of the invention.

[0120] Unless otherwise noted, general starting materials used were obtained from commercial sources or prepared in other examples.

[0121] Abbreviations

[0122] ACN Acetonitrile

[0123] AcOH Acetic acid

[0124] AIBN α,α'-Azoisobutyronitrile

[0125] DCE 1,2-Dichloroethane

[0126] DCM Dichloromethane (Dichloromethane / methylene chloride)

[0127] DEA Diethylamine

[0128] DIEA N,N-Diisopropylethylamine

[0129] DMA N,N-Dimethylacetamide

[0130] DMAP N,N-Dimethylpyridin-4-amine

[0131] DMC 2-Chloro-1,3-dimethylimidazolinium chloride

[0132] DMF N,N-Dimethylformamide

[0133] DMSO Dimethyl sulfoxide

[0134] EtOAc Ethyl acetate

[0135] HATU 2-(3H-[1,2,3]triazolo[4,5-b]pyridin-3-yl)-1,1,3,3-tetramethyl-isourea hexafluorophosphate HCl Hydrogen chloride

[0136] HPLC High Performance Liquid Chromatography

[0137] IPA 2-Propanol

[0138] LAH Lithium Aluminum Hydride

[0139] LC / MS Liquid chromatography / mass spectrometry

[0140] K2CO3 Potassium carbonate

[0141] MeOH Methanol

[0142] MS mass spectrometry

[0143] NaBH(OAc)3 Sodium triacetoxyborohydride

[0144] NaCNBH4 Sodium cyanoborohydride

[0145] NaOH Sodium hydroxide

[0146] NaHCO3 Sodium bicarbonate

[0147] Na2SO4 Sodium sulfate

[0148] RT Room temperature

[0149] TEA Triethylamine

[0150] TFA Trifluoroacetic acid

[0151] THF Tetrahydrofuran

[0152] The progress of the reaction is usually monitored by TLC or LC-MS. LC-MS is recorded using one of the following methods.

[0153] LCMS Method-1:

[0154]

[0155]

[0156] LCMS Method-2:

[0157]

[0158] LCMS Method-3:

[0159]

[0160] LCMS Method-4:

[0161]

[0162] LCMS Method-5:

[0163]

[0164] LCMS Method-6:

[0165]

[0166] NMR was recorded at room temperature unless otherwise noted on a Varian Inova 400 or 500 MHz spectrometer (solvent peak used as reference) or on a Bruker 300 or 400 MHz spectrometer (TMS peak used as internal reference).

[0167] General key intermediates procedures

[0168] Synthesis of Intermediate 1[(3,5-dichlorophenyl)methyl][(pyridin-3-yl)methyl]amine hydrochloride

[0169]

[0170] Step 1. Under an inert atmosphere, 1-(pyridin-3-yl)methylamine (4.70 mL, 46.2 mmol) was combined with 3,5-dichlorobenzaldehyde (4.04 g, 23.1 mmol) in methanol (83 mL). Sodium cyanoborohydride (2.90 g, 46.2 mmol) and acetic acid (2.63 mL, 46.2 mmol) were added. The reaction mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated, taken up in ethyl acetate, and washed twice with saturated NaHCO3 solution. The organic phase was concentrated under vacuum. The residue was dissolved in THF, and HCl (4 M in 1,4-dioxane) was then added. The precipitated HCl salt was filtered, washed with THF, and dried under vacuum to give [(3,5-dichlorophenyl)methyl][(pyridin-3-yl)methyl]amine hydrochloride (5.859 g, 40.5%) as a white solid. MSES m / z: 267.1 [M+H] + .

[0171] Synthesis of Intermediate 2[(2,4-dichlorophenyl)methyl][(pyridin-3-yl)methyl]amine hydrochloride

[0172]

[0173] Step 1. Under an inert atmosphere of N2, 1-(pyridin-3-yl)methylamine (4.70 mL, 46.2 mmol) was dissolved in methanol (83 mL), followed by the addition of 2,4-dichlorobenzaldehyde (4.04 g, 23.1 mmol), sodium cyanoborohydride (2.90 g, 46.2 mmol) and acetic acid (2.63 mL, 46.2 mmol). The reaction was stirred at room temperature for 4 hours. The reaction mixture was concentrated, taken up in ethyl acetate, and washed twice with saturated NaHCO3 solution. The organic phase was concentrated. The residue was dissolved in THF, followed by the addition of HCl (4 M in 1,4-dioxane). The precipitated HCl salt was filtered, washed with THF, and dried under vacuum to give [(2,4-dichlorophenyl)methyl][(pyridin-3-yl)methyl]amine hydrochloride (5.692 g, 39.4%) as a white solid. MS ES m / z: 267.1 [M+H] + .

[0174] Synthesis of Intermediate 3 (1-(2,3-dichlorophenyl)propan-1-amine)

[0175]

[0176] Step 1. EtMgBr (1.55 g, 11.628 mmol) was added dropwise to a stirred solution of 2,3-dichlorobenzonitrile (1.00 g, 5.814 mmol) and THF (10.00 mL) at -50 ° C under an argon atmosphere. The resulting mixture was stirred at room temperature for 1 hour under an argon atmosphere. The reaction was quenched with saturated NH4Cl (aqueous solution) at 0 ° C. The aqueous layer was extracted with EtOAc (3x20 mL). The resulting mixture was concentrated under vacuum to obtain (1- (2,3-dichlorophenyl) propan-1-imine) (930 mg, 79%) as a yellow oil, and was used directly in the next step.

[0177]

[0178] Step 2. To a stirred solution of (1-(2,3-dichlorophenyl)propan-1-imine) (900 mg, 4.454 mmol) and EtOH (60.00 mL) was added NaBH4 (336.99 mg, 8.907 mmol) in portions at 0°C. The resulting mixture was stirred at 0°C for 2 hours. The reaction was quenched with saturated NH4Cl (aq) at 0°C. The aqueous layer was extracted with EtOAc (3 x 100 mL). The resulting mixture was concentrated under vacuum to give (1-(2,3-dichlorophenyl)propan-1-amine) (832 mg, 91%) as a yellow solid. ESMS m / z: 204.0 [M+H] + .

[0179] Synthesis of 1-(2,4-dichlorophenyl)-N-(pyridin-3-ylmethyl)-1-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methanamine (Intermediate 4)

[0180]

[0181] Step-1: 2,4-dichlorobenzoic acid methyl ester: Concentrated sulfuric acid (1ml) is added to a stirred solution of 2,4-dichlorobenzoic acid (1.5g, 7.85mmol) in MeOH (10ml), and the reaction mixture is heated at 80°C for 16 hours. After the reaction is completed (monitored by TLC), the reaction mixture is quenched with water (100ml) and extracted with ethyl acetate (3x100ml). The combined organic layer is washed with brine (100ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude compound is purified by using combi-flash chromatography to obtain the pure title compound (1.4g, 87%). 1 H NMR (400MHz, DMSO-d6): δ3.88 (s, 3H), 7.57 (dd, J = 8.4Hz, J = 2.0Hz, 1H), 7.79 (d, J = 1.6Hz, 1H), 7.86 (d, J = 8.4Hz, 1H).

[0182] Step-2: (2,4-Dichlorophenyl)(1H-pyrazol-4-yl)methanone: 2(g) Under atmosphere at -78 ° C, n-BuLi (6.3ml, 16.44mmol) is added to a solution of 4-bromo-1H-pyrazole (1.0g, 6.85mmol) in THF (10ml), and the reaction mixture is stirred for 5-10 minutes at -78 ° C. The reaction mixture is stirred at room temperature for 1 hour. After 1 hour, the reaction mixture is cooled to -78 ° C, and 2,4-dichlorobenzoic acid methyl esters (1.4g, 6.85mmol) are added at -78 ° C and allowed to reach room temperature, and stirred at room temperature for 1 hour. After the reaction is completed (monitored by TLC), the reaction mixture is quenched with saturated NH4Cl solution (100ml) and extracted with ethyl acetate (3x 100ml). The combined organic layer is washed with brine (100ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude compound is purified by using combi-flash chromatography to obtain pure title compound (0.6g, 51.3%). ES MS m / z:239.0[M+H] + . 1 H NMR (400MHz, DMSO-d6): δ7.56(bs,2H),7.91(s,1H),8.29(s,1H),13.65(s,1H).

[0183] Step-3: (2,4-dichlorophenyl)(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methanone: To a solution of (2,4-dichlorophenyl)(1H-pyrazol-4-yl)methanone (0.900 g, 3.73 mmol) in toluene (10 ml) was added 3,4-dihydropyran (0.470 g, 5.6 mmol) and then TEA (0.1 ml) was added at room temperature. The reaction mixture was heated at 100 °C for 48 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under vacuum to give a crude compound, which was used in the next step without further purification (0.780 g, crude).

[0184] Step-4: 1-(2,4-Dichlorophenyl)-N-(pyridin-3-ylmethyl)-1-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methanamine: To a solution of (2,4-dichlorophenyl)(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)methanone (0.780 g, 2.4 mmol) in toluene (10 ml) was added pyridin-3-ylmethanamine (0.389 g, 3.6 mmol) followed by titanium (IV) isopropoxide (1.4 g, 4.8 mmol) and the reaction mixture was heated at 90°C for 10 hours. NaCNBH3 (0.223 g, 3.6 mmol) and MeOH (20 ml) were added and the reaction mixture was heated at 80°C for additional 16 hours. After reaction is completed (monitored by TLC), water (50ml) is added to the reaction mixture reaction mixture and stirred for 10 minutes. After 10 minutes, the reaction mixture is filtered, and washed with DCM (2x 50ml). The filtrate is concentrated to obtain crude product. Crude compound is used for next step (0.650g, thick) without further purification.

[0185] Synthesis of methyl 2-(5-(((2,4-dichlorobenzyl)amino)methyl)pyridin-2-yl)acetate (Intermediate 5)

[0186]

[0187] Step-1: 2-(5-cyanopyridin-2-yl) dimethyl malonate: CsCO (53.40 g, 163.9 mmol) was added to a stirred solution of 6-bromonicotinoonitrile (10.0 g, 54.641 mmol) and dimethyl malonate (10.82 g, 81.9 mmol) in DMF (100 ml) at room temperature. The reaction mixture was stirred overnight at room temperature. After the reaction was completed (monitored by TLC), the reaction mixture was poured into ice-cold water (100 ml) and extracted with ethyl acetate (3 x 100 ml). The combined organic layer was washed with brine (100 ml) and dried over NaSO. It was evaporated under vacuum to obtain a crude compound. The crude compound was purified using column chromatography to obtain a pure compound (9.2 g, 71%). 1 H NMR (400MHz, DMSO-d6): δ3.81 (s, 6H), 5.04 (s, 1H), 7.68 (d, J = 8.0Hz, 1H), 8.01 (d, J = 6.4Hz, 1H), 8.85 (s, 1H).

[0188] Step-2: 2-(5-cyanopyridine-2-yl) methyl acetate: To a stirred solution of 2-(5-cyanopyridine-2-yl) dimethyl malonate (8.0 g, 34.2 mmol) in DMSO (80 ml) at room temperature, an aqueous solution of NaCl (2.19 g, 37.5 mmol) was added. The reaction mixture was stirred overnight at 130 ° C. After the reaction was completed (monitored by TLC), the reaction mixture was poured into ice-cold water (100 ml) and extracted with ethyl acetate (3 x 100 ml). The combined organic layer was washed with brine (100 ml), and dried over Na2SO4, and evaporated under vacuum to obtain a crude compound. The crude compound was purified using column chromatography to obtain a pure compound (2.7 g, 44%). 1 H NMR (400MHz, DMSO-d6): δ3.78 (s, 3H), 3.97 (s, 2H), 7.49 (d, J = 8.0 Hz, 1H), 7.97 (d, J = 8.0 Hz, 1H), 8.87 (s, 1H).

[0189] Step-3: Methyl 2-(5-(aminomethyl)pyridin-2-yl)acetate: To a stirred solution of methyl 2-(5-cyanopyridin-2-yl)acetate (0.50 g, 2.8 mmol) in MeOH (5 ml) was added NiCl2 6H2O (0.067 g, 0.28 mmol) in portions at 0°C followed by NaBH4 (0.750 g, 19.4 mmol). The reaction mixture was stirred at room temperature overnight. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through celite and the filtrate was evaporated under vacuum to obtain the crude compound. The crude compound was purified using RP-HPLC to obtain the pure compound (0.400 g, 78%). ES MS m / z: 181.1 [M+H] + .

[0190] Step-4: Methyl 2-(5-(((2,4-dichlorobenzyl)amino)methyl)pyridin-2-yl)acetate: To a stirred solution of methyl 2-(5-(aminomethyl)pyridin-2-yl)acetate (0.300 g, 1.66 mmol) and 2,4-dichlorobenzaldehyde (0.291 g, 1.66 mmol) in 1,2-dichloroethane (3 ml) was added AcOH (0.3 ml) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. Sodium cyanoborohydride (0.156 g, 2.49 mmol) and methanol (0.3 ml) were added and the reaction mixture was stirred at room temperature for another 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (20 ml) and extracted with ethyl acetate (2 x 20 ml). The combined organic layer was washed with brine (30 ml) and dried over Na2SO4. Evaporated under vacuum to obtain the crude compound. The crude compound was purified using column chromatography to obtain the pure compound (0.200 g, 35%). ES MS m / z: 339.4 [M+H] + .

[0191] Synthesis of ethyl 3-((tert-butoxycarbonyl)amino)-5-(((2-chlorobenzyl)amino)methyl)picolinate (Intermediate 6)

[0192]

[0193] Step-1: Ethyl 3-((tert-butoxycarbonyl)amino)-5-(((2-chlorobenzyl)amino)methyl)picolinate: To a stirred solution of ethyl 3-((tert-butoxycarbonyl)amino)-5-formylpicolinate (0.6 g, 2.04 mmol) and (2-chlorophenyl)methylamine (0.35 g, 2.45 mmol) in 1,2-dichloroethane (6.0 mL) was added AcOH (0.6 mL) and the reaction mixture was stirred at room temperature for 1 hour. NaCNBH (0.190 g, 3.06 mmol) and MeOH (0.6 mL) were added and stirred at room temperature for another 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with saturated NaHCO solution (50 ml) and extracted with dichloromethane (3 x 75 ml). The combined organic layers were washed with brine (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give a crude product. The crude compound was used in the next step without further purification. ES MS m / z: 420.35 [M+H] + .

[0194] Synthesis of methyl 5-(((2-chlorobenzyl)amino)methyl)pyrimidine-2-carboxylate (Intermediate 7)

[0195]

[0196] Step-1: 5-(bromomethyl)pyrimidine-2-carboxylic acid methyl ester: A stirred solution of 5-methylpyrimidine-2-carboxylic acid methyl ester (0.2 g, 1.31 mmol), N-bromosuccinimide (0.268 g, 1.51 mmol) and AIBN (0.086 g, 0.52 mmol) in CCl4 (2 ml) was refluxed for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated to give a crude compound. The crude compound was diluted with water (10 ml) and extracted with ethyl acetate (2x15 ml). The combined organic layers were washed with brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude compound was purified by using combi-flash chromatography to obtain the title compound (0.075 g, 24%). LCMS: m / z 231.1 [M+H] + .

[0197] Step-2: Methyl 5-(((2-chlorobenzyl)amino)methyl)pyrimidine-2-carboxylate: To a stirred solution of methyl 5-(bromomethyl)pyrimidine-2-carboxylate (0.2 g, 0.86 mmol) in DCM (2 ml) was added (2-chlorophenyl)methanamine (0.134 g, 0.95 mmol). The reaction mixture was stirred at room temperature for 72 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated. The crude compound was purified by using combi-flash chromatography to obtain the title compound (0.060 g, 23%). LCMS: m / z 292.3[M+H] + . 1 H NMR (400MHz, CDCl3): δ3.98(s,2H),4.00(s,2H),4.12(s,3H),7.28-7.32(m,2H),7.41-7.43(m,2H),9.00(s,2H).

[0198] Synthesis of methyl 5-(((1-(3-methoxy-2-(trifluoromethyl)phenyl)propyl)amino)methyl)picolinate (Intermediate 8)

[0199]

[0200] Step-1: (E)-N-(3-methoxy-2-(trifluoromethyl)benzylidene)-2-methylpropane-2-sulfinamide: To a stirred solution of 3-methoxy-2-(trifluoromethyl)benzaldehyde (0.3 g, 1.47 mmol) in dichloromethane (3 ml) was added 2-methylpropane-2-sulfinamide (0.267 g, 2.20 mmol) and copper sulfate (0.469 g, 2.93 mmol), and heated to reflux for 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with water (5 ml) and extracted with ethyl acetate (3 x 10 ml). The combined organic layers were washed with brine (10 ml), dried over Na2SO4, and concentrated under reduced pressure. The crude compound was purified by using combi-flash chromatography to obtain the title compound (0.3 g, 66%). LCMS: m / z 308.4 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ1.19 (s, 9H), 3.94 (s, 3H), 7.48-7.51 (m, 2H), 7.73 (t, J = 8.4Hz, 1H), 8.82 (dd, J = 7.2Hz, J = 3.6Hz, 1H).

[0201] Step-2: N-(1-(3-methoxy-2-(trifluoromethyl)phenyl)propyl)-2-methylpropane-2-sulfinamide: To a solution of (E)-N-(3-methoxy-2-(trifluoromethyl)benzylidene)-2-methylpropane-2-sulfinamide (0.3 g, 0.976 mmol) in tetrahydrofuran (3 ml) was added ethylmagnesium bromide (1.0 M in THF) (1.46 ml, 1.464 mmol) at 0°C and the reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with saturated ammonium chloride solution (5 ml) and extracted with ethyl acetate (3 x 20 ml). The combined organic layers were washed with brine (20 ml), dried over Na2SO4, and concentrated under reduced pressure. The crude compound was purified by using combi-flash chromatography to obtain the title compound (0.3 g, 91%). LCMS: m / z 338.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ0.88(t,3H),1.07(s,9H),1.60-1.67(m,1H),1.76-1.84(m,1H),3.85(s,3H),4.56-4.61(m,1H),5.57(d, J =6.8Hz,1H),7.09(d, J =8.4Hz,1H),7.28(d,J=8.4Hz,1H),7.56-7.60(t,J=8.0Hz,1H).

[0202] Step-3: 1-(3-methoxy-2-(trifluoromethyl)phenyl)propan-1-amine HCl salt: To a solution of N-(1-(3-methoxy-2-(trifluoromethyl)phenyl)propyl)-2-methylpropane-2-sulfinamide (0.3 g, 0.89 mmol) in dichloromethane (3 ml) was added 1,4-dioxane (1.5 ml) containing HCl, and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The crude compound was triturated with n-pentane to obtain the title compound (0.230 g, 96%). LCMS: m / z 234.0 [M+H] +. 1H NMR (400MHz, DMSO-d6): δ0.78(t,J=7.6Hz,3H),1.78-1.85(m,1H),1.96-2.03(m,1H),3.89(s,3H),4 .50-4.54(m,1H),7.30(d,J=8.4Hz,1H),7.41(d,J=8.0Hz,1H),7.72(t,J=8.0Hz,1H),8.66(bs,2H).

[0203] Step-4: Methyl 5-(((1-(3-methoxy-2-(trifluoromethyl)phenyl)propyl)amino)methyl)picolinate: To a mixture of 1-(3-methoxy-2-(trifluoromethyl)phenyl)propan-1-amine HCl salt (0.20 g, 0.741 mmol) in 1,2-dichloroethane (2 ml) was added triethylamine (0.4 ml, 2.96 mmol) and stirred at room temperature for 30 minutes. Then molecular sieves, methyl-5-formyl pyridine carboxylate (0.122 g, 0.741 mmol) and acetic acid (0.5 ml, 8.89 mmol) were added and RM was stirred for 1 hour. Then NaCNBH3 (0.116 g, 1.85 mmol) was added and the reaction mixture was stirred at room temperature for 45 minutes. After the reaction was completed (monitored by TLC), the reaction mixture was filtered through a celite bed, and saturated sodium bicarbonate solution (5 ml) was added to the filtrate and extracted with ethyl acetate (2 x 20 ml). The combined organic layers were washed with brine (20 ml), dried over Na2SO4, and concentrated under reduced pressure. The crude product was purified by column chromatography to give the pure title compound (0.140 g, 49%). LCMS: m / z 383.6 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ0.86(t,J=7.2Hz,3H),1.50-1.66(m,2H),3.04(bs,1H),3.44(d,J=14.8Hz,1H),3.60(d,J=14.8Hz,1H),3.86(s,3H),3.88( s,3H),3.98(bs,1H),7.11(d,J=8.4Hz,1H),7.48(d,J=8.0Hz,1H),7.60(t ,J=8.0Hz,1H),7.87(d,J=7.6Hz,1H),7.99(d,J=8.0Hz,1H),8.57(s,1H).

[0204] Synthesis of 1-(6-bromopyridin-3-yl)-N-(2-chlorobenzyl)methylamine (Intermediate 64)

[0205]

[0206] Step-1: 1-(6-Bromopyridin-3-yl)-N-(2-chlorobenzyl)methylamine: A mixture of 6-bromonicotinaldehyde (0.75 g, 4.07 mmol), (2-chlorophenyl)methylamine (0.574 g, 4.07 mmol) and acetic acid (1.16 ml, 20.35 mmol) in 1-2 dichloroethane (10 ml) was stirred with argon at room temperature for 2 hours. NaCNBH3 (0.384 g, 6.105 mmol) was added and the reaction mixture was stirred for another 16 hours at room temperature. After completion of the reaction (monitored by TLC), water (20 ml) was added and the reaction mixture was extracted with dichloromethane (2 x 30 ml). The combined organic layers were washed with brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under vacuum. The crude product was purified by column chromatography to obtain the pure title compound (0.85 g, 67%) as a white solid. LCMS: m / z 311.2 [M+H] + .

[0207] Synthesis of methyl 5-(((1-(2-chloro-3,4-dimethoxyphenyl)propyl)amino)methyl)picolinate (Intermediate 9)

[0208]

[0209] Step-1: 2-Chloro-N,3,4-trimethoxy-N-methylbenzamide: To a stirred solution of 2-chloro-3,4-dimethoxybenzoic acid (3.0 g, 13.85 mmol) in dry DMF (30 ml) was added HATU (7.894 g, 20.77 mmol) at room temperature. Then DIPEA (4.903 ml, 27.70 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 1 hour. N,O-dimethylhydroxylamine hydrochloride (2.026 g, 20.77 mmol) was added at room temperature, and the reaction mixture was stirred at room temperature for 4 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with ice-cold water (40 ml), and the aqueous layer was extracted with ethyl acetate (2 x 80 ml). The combined organic layers were washed with brine (40 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude compound was purified by Combi flash column chromatography to obtain the pure title compound (2.5 g, 69.5%). LCMS: m / z 260.27 [M+H] + .

[0210] Step-2: 1-(2-Chloro-3,4-dimethoxyphenyl)propan-1-one:

[0211] To a stirred solution of 2-chloro-N,3,4-trimethoxy-N-methylbenzamide (1.0 g, 3.85 mmol) in dry THF (10 ml) was added ethylmagnesium bromide (6.40 ml, 3M solution in diethyl ether, 19.25 mmol) dropwise at 0 ° C under a nitrogen atmosphere, and the reaction mixture was stirred at room temperature for 1.5 hours. After the reaction was completed (monitored by TLC), the reaction mixture was slowly quenched with saturated aqueous ammonium chloride solution (10 ml). The water layer was extracted with ethyl acetate (3x20 ml). The combined organic layers were washed with brine (10 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude compound. The crude compound was purified by Combi flash column chromatography to obtain the pure title compound (0.600 g, 68%). LCMS: m / z 229 [M+H] + . 1 H NMR (400MHz, CDCl3): δ1.21(t,J=7.2Hz,3H),2.97(q,J=7.2Hz,2H),3.89(s,3H),3.98(s,3H),6.87(d,J=8.8Hz,1H),7.33(d,J=8.8Hz,1H).

[0212] Step-3: Methyl 5-(((1-(2-chloro-3,4-dimethoxyphenyl)propyl)amino)methyl)picolinate: To a stirred solution of methyl 5-(aminomethyl)picolinate dihydrochloride (0.250 g, 1.04 mmol) in DCE (2.5 ml) was added dropwise Et3N (0.436 ml, 3.13 mmol) under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 hours to release the free amine. 1-(2-chloro-3,4-dimethoxyphenyl)propan-1-one (0.191 g, 0.83 mmol), glacial acetic acid (0.1 ml) and 4A molecular sieves (0.500 g) were added and the reaction mixture was stirred at 60 ° C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature. Sodium cyanoborohydride (0.131 g, 2.09 mmol) was added and the reaction mixture was stirred at 60 ° C for 3 hours. After the reaction was completed (monitored by TLC), the reaction mixture was quenched by saturated aqueous sodium bicarbonate solution (5 ml). The aqueous layer was extracted with ethyl acetate (3 x 10 ml). The combined organic layers were washed with brine (5 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the crude compound. The crude compound was purified by Combi flash column chromatography to give the pure title compound (0.170 g, 53.8%). LCMS: m / z 379.3 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ0.80(t,J=7.4Hz,3H),1.53-1.61(m,2H),2.95(bs,1H),3.49(d,J=14.4Hz,1H),3.64(d,J=15.6Hz,1H),3.72(s,3H), 3.82(s,3H),3.86(s,3H),5.76(s,1H),7.08(d,J=8.4Hz,1H),7.32(d,J =8.8Hz,1H),7.87(d,J=7.6Hz,1H),7.99(d,J=7.6Hz,1H),8.56(s,1H).

[0213] Synthesis of methyl 5-(((1-(4-chlorobenzo[d][1,3]dioxol-5-yl)propyl)amino)methyl)picolinate (Intermediate 10)

[0214]

[0215] Step-1: 1-(2-Chloro-3,4-dihydroxyphenyl)propan-1-one: 1-(2-Chloro-3,4-dimethoxyphenyl)propan-1-one (1.00 g, 4.37 mmol) was dissolved in DCM (10 ml) and 1 M DCM containing BBr3 (8.74 ml, 1.0 M in DCM, 8.75 mmol) was added dropwise at -78°C. The reaction mixture was then stirred at -78°C for 1 hour and then at room temperature for 2.5 hours. The reaction was monitored by TLC (5% methanol in DCM). After completion of the reaction, the reaction mixture was quenched with 30% aqueous ammonia and concentrated. Methanol (1 ml) was added and the reaction mixture was concentrated to dryness. The reaction mixture was azeotroped twice with methanol (1 ml). The crude compound was loaded on celite and purified by RP Gold column chromatography using acetonitrile and water containing 0.1% formic acid to obtain the pure product (0.420 g, 47.9%). LCMS: m / z 199.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ1.04(t,J=7.0Hz,3H),2.86(q,J=7.2Hz,2H),6.79(d,J=8.4Hz,1H),7.07(d,J=8.4Hz,1H),9.49(s,1H),10.19(s,1H).

[0216] Step-2: 1-(4-Chlorobenzo[d][1,3]dioxol-5-yl)propan-1-one: To a stirred solution of 1-(2-chloro-3,4-dihydroxyphenyl)propan-1-one (0.40 g, 1.98 mmol) in dry DMF (6.0 ml) was added potassium fluoride (0.578 g, 9.96 mmol). The reaction mixture was stirred at room temperature for 30 minutes. Diiodomethane (0.20 ml, 2.38 mmol) was then added dropwise at room temperature and the reaction mixture was stirred at 100 °C for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water (10 ml) and the aqueous layer was extracted with ethyl acetate (2 x 20 ml). The combined organic layers were dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by Combi flash column chromatography to obtain the pure title compound (0.200 g, 47.2%). LCMS: m / z 213.12 [M + +1] and 215.1[M+2H] + . 1 H NMR (400MHz, DMSO-d6): δ1.04(t,J=7.2Hz,3H),2.93(q,J=6.8Hz,2H),6.21(s,2H),7.00(d,J=8.4Hz,1H),7.39(d,J=8.4Hz,1H).

[0217] Step-3: Methyl 5-(((1-(4-chlorobenzo[d][1,3]dioxol-5-yl)propyl)amino)methyl)picolinate: To a stirred solution of methyl 5-(aminomethyl)picolinate dihydrochloride (0.185 g, 0.78 mmol) in DCE (4 ml) was added Et3N (0.323 mL, 2.32 mmol) dropwise under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 hours to release the free amine. 1-(4-chlorobenzo[d][1,3]dioxol-5-yl)propan-1-one (0.132 g, 0.62 mmol), glacial acetic acid (0.1 mL) and 4A molecular sieves (0.300 g) were added and the reaction mixture was stirred at 60 °C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature. Sodium cyanoborohydride (0.097 g, 1.55 mmol) and methanol (5 drops) were added. The reaction mixture was stirred at 60 ° C for 3 hours. After the reaction was completed (monitored by TLC), the reaction mixture was quenched with saturated sodium bicarbonate aqueous solution (5 ml). The aqueous layer was extracted with ethyl acetate (3 x 10 ml). The combined organic layers were washed with brine (5 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude compound. The crude compound was purified by Combi flash column chromatography to obtain the pure title compound (0.088 g, 39.1%). LCMS: m / z 363.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ0.81(t,J=7.2Hz,3H),1.56-1.64(m,2H),3.51(d,J=14.8Hz,1H),3.67(d,J=13.6Hz,1H),3.84(s,1H),3 .88(s,3H),6.13(s,2H),6.96(d,J=8.0Hz,1H),7.11(d,J=8.4Hz,1H),7.89(d,J=8.0Hz,1H),8.01(d,J=7.6Hz,1H),8.58(s,1H).

[0218] Synthesis of methyl 5-(((1-(2-chloro-3-ethoxyphenyl)propyl)amino)methyl)picolinate (Intermediate 11)

[0219]

[0220] Step-1: 2-Chloro-3-ethoxybenzaldehyde: To a stirred solution of 2-chloro-3-hydroxybenzaldehyde (1.00 g, 6.38 mmol) in DMF (15 ml) was added potassium carbonate (1.76 g, 12.77 mmol) at room temperature. The reaction mixture was stirred at room temperature for 30 minutes. Then iodoethane (1.99 g, 12.77 mmol) was added dropwise, and the reaction mixture was stirred at room temperature for 1.5 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with ethyl acetate (50 ml), and washed with ice-cold water (50 ml), followed by washing with brine (30 ml). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by Combi flash column chromatography to obtain the pure title compound (1.18 g, 100%). 1 H NMR (400MHz, DMSO-d6): δ1.43 (t, J=7.0Hz, 3H), 4.19 (q, J=7.2Hz, 2H), 7.42-7.48 (m, 3H), 10.38 (s, 1H).

[0221] Step-2: 1-(2-Chloro-3-ethoxyphenyl)propan-1-ol: To a stirred solution of 2-chloro-3-ethoxybenzaldehyde (1.10 g, 5.95 mmol) in dry THF (25 ml) was added ethylmagnesium bromide (2.38 ml, 3M solution in diethyl ether, 7.14 mmol) dropwise at -78 °C under nitrogen atmosphere. The reaction mixture was stirred at -78 °C for 2 hours and continued to stir at room temperature for 13 hours. After completion of the reaction (monitored by TLC), the reaction mixture was slowly quenched by saturated aqueous ammonium chloride solution (20 ml). The aqueous layer was extracted with ethyl acetate (3 x 30 ml). The combined organic layers were washed with brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the crude compound. The crude compound was purified by Combi flash column chromatography to obtain the pure title compound (0.698 g, 54%). 1 H NMR (400MHz, DMSO-d6): δ0.88(t,J=7.4Hz,3H),1.35(t,J=7.0Hz,3H),1.43-1.52(m,1H),1.60-1.67(m,1H),4.09(q,J=6 .8Hz,2H),4.80-4.84(m,1H),5.27(d,J=4.4Hz,1H),6.98(d,J=7.2Hz,1H),7.14(d,J=7.2Hz,1H),7.27(d,J=7.8Hz,1H).

[0222] Step-3: 1-(2-Chloro-3-ethoxyphenyl)propan-1-one: To a stirred solution of 1-(2-chloro-3-ethoxyphenyl)propan-1-ol (0.690 g, 3.21 mmol) in dry DCM (15 ml) was added pyridinium chlorochromate (1.380 g, 6.43 mmol) at 0 °C under nitrogen atmosphere and the reaction mixture was stirred at room temperature for 13 hours. After completion of the reaction (monitored by TLC), the reaction mixture was filtered through a celite bed and the filtrate was concentrated under reduced pressure to get the crude compound. The crude compound was purified by Combi flash column chromatography to get the pure title compound (0.681 g, 99%). LCMS: m / z 213.2 [M + +1] and 215.3[M+2H] + . 1 H NMR (400MHz, DMSO-d6): δ1.06(t,J=7.2Hz,3H),1.36(t,J=7.0Hz,3H),2.87(q,J=7.2Hz,2H) ,4.14(q,J=6.8Hz,2H),7.08(d,J=6.8Hz,1H),7.23(d,J=7.6Hz,1H),7.37(d,J=7.8Hz,1H).

[0223] Step-4: Methyl 5-(((1-(2-chloro-3-ethoxyphenyl)propyl)amino)methyl)picolinate: To a stirred solution of methyl 5-(aminomethyl)picolinate dihydrochloride (0.365 g, 1.53 mmol) in DCE (4 ml) was added Et3N (0.48 ml, 3.53 mmol) dropwise under nitrogen atmosphere. The reaction mixture was stirred at room temperature for 3 hours to release the free amine. 1-(2-chloro-3-ethoxyphenyl)propan-1-one (0.250 g, 1.17 mmol), glacial acetic acid (0.28 ml) and 4A molecular sieves (0.50 g) were added and the reaction mixture was stirred at 70°C for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was cooled to room temperature. Sodium cyanoborohydride (0.147 g, 2.35 mmol) was added and the reaction mixture was stirred at 70°C for 16 hours. After the reaction was completed (monitored by TLC), the reaction mixture was quenched by saturated aqueous sodium bicarbonate solution (5 ml). The aqueous layer was extracted with ethyl acetate (3 x 10 ml). The combined organic layers were washed with brine (5 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the crude compound. The crude compound was purified by Combi flash column chromatography to give the pure title compound (0.240 g, 56%). LCMS: m / z 363.35 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ0.81(t,J=7.4Hz,3H),1.36(t,J=6.8Hz,3H),1.52-1 .67(m,2H),3.49(d,J=14.8Hz,1H),3.65(d,J=14.8Hz,1H),3.87(s,3H),4.00( bs,1H),4.07(q,J=6.8Hz,2H),6.99(d,J=8.0Hz,1H),7.21(d,J=7.6Hz,1H),7. 30(d,J=8.0Hz,1H),7.88(d,J=7.6Hz,1H),8.00(d,J=7.6Hz,1H),8.57(s,1H).

[0224] Synthesis of methyl 5-(((1-(2-chloro-3-(oxetan-3-yloxy)phenyl)propyl)amino)methyl)picolinate (Intermediate 12)

[0225]

[0226] Step-1: 2-chloro-3-(oxetane-3-yloxy)benzaldehyde: Cesium carbonate (4.16 g, 12.77 mmol) and NaI (0.478 g, 3.19 mmol) were added to a stirred solution of 2-chloro-3-hydroxybenzaldehyde (1.0 g, 6.38 mmol) in dry DMF (10 ml) at room temperature. After 15 minutes, DMF (5 ml) containing 3-iodooxetane (4.70 g, 25.54 mmol) was added, and the reaction mixture was heated at 60 ° C for 16 hours. After the reaction was completed (monitored by TLC), water (50 ml) was added, and the reaction mixture was extracted with ethyl acetate (2 x 100 ml). The combined organic layer was washed with brine (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude compound was purified by using combi-flash chromatography to obtain the pure title compound (0.4 g, 29%). 1 H NMR (400MHz, DMSO-d6): δ4.60-4.63(m,2H),4.97-5.00(m,2H),5.43-5.45(m,1H),7.11-7.14(m,1H),7.43-7.50(m,2H),10.39(s,1H).

[0227] Step-2: 1-(2-Chloro-3-(oxetane-3-yloxy)phenyl)propan-1-ol: To a stirred solution of 2-chloro-3-(oxetane-3-yloxy)benzaldehyde (1.0 g, 4.71 mmol) in dry THF (25 ml) was added ethylmagnesium bromide solution (2.04 ml, 3M in diethyl ether, 6.13 mmol) dropwise at 0°C and the reaction mixture was stirred at 0°C for 3 hours. After completion of the reaction (monitored by TLC), water (20 ml) was added and the reaction mixture was extracted with ethyl acetate (2 x 50 ml). The combined organic layers were washed with brine (50 ml), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude compound was purified by using combi-flash chromatography to obtain the pure title compound (0.6 g, 52%). 1 H NMR (400MHz, DMSO-d6): δ0.90(t,J=7.6Hz,3H),1.46-1.52(m,1H),1.62-1.69(m,1H),4.55-4.61(m, 2H),4.82-4.87(m,1H),4.94-4.97(m,2H),5.31-5.35(m,2H),6.63-6.65(m,1H),7.19-7.28(m,2H).

[0228] Step-3: 1-(2-Chloro-3-(oxetan-3-yloxy)phenyl)propan-1-one: To a stirred solution of 1-(2-chloro-3-(oxetan-3-yloxy)phenyl)propan-1-ol (0.606 g, 2.49 mmol) in dry DCM (9 ml) was added PCC (1.07 g, 4.99 mmol) at 0°C. The reaction mixture was stirred at room temperature for 6 hours. After completion of the reaction (monitored by TLC), water (20 ml) was added and the reaction mixture was extracted with ethyl acetate (2 x 50 ml). The combined organic layers were washed with brine (50 ml), dried over anhydrous sodium sulphate and concentrated under reduced pressure. The crude compound was purified by using combi-flash chromatography to obtain the pure title compound (0.5 g, 83%). LCMS: m / z 241.1[M+H] + . 1 H NMR (400MHz, DMSO-d6): δ1.07(t,J=7.2Hz,3H),2.89(t,J=7.2Hz,2H),4.56-4.62(m,2H),4.94- 4.99(m,2H),5.36-5.41(m,1H),6.88(d,J=8.8Hz,1H),7.11-7.16(m,1H),7.35(d,J=8.0Hz,1H).

[0229] Step-4: Methyl 5-(((1-(2-chloro-3-(oxetan-3-yloxy)phenyl)propyl)amino)methyl)picolinate: To a stirred solution of methyl 5-(aminomethyl)picolinate dihydrochloride (0.079 g, 0.33 mmol) in DCE (2 ml) was added TEA (0.139 ml, 0.99 mmol) and the reaction mixture was stirred for 2 hours. To this was added 1-(2-chloro-3-(oxetan-3-yloxy)phenyl)propan-1-one (0.080 g, 0.26 mmol), acetic acid (0.15 ml) and molecular sieves (200 mg). The reaction mixture was heated at 60°C for 16 hours. The reaction mixture was cooled to room temperature and sodium cyanoborohydride (0.041 g, 0.66 mmol) was added. The reaction mixture was heated at 60°C for 3 hours. After the reaction was completed (monitored by TLC), saturated NaHCO3 solution (5 ml) was added and the reaction mixture was extracted with ethyl acetate (2 x 10 ml). The combined organic layer was washed with brine (10 ml), dried over Na2SO4, and evaporated under vacuum to obtain the crude compound. The crude compound was purified using column chromatography to obtain the pure compound (60 g, 46%). LCMS: m / z 391.6 [M+H] + .

[0230] Synthesis of methyl 5-(((2,3-dichlorobenzyl)amino)methyl)-4-methoxypicolinate (Intermediate 13)

[0231]

[0232] Step-1: 5-cyano-4-methoxypicolinic acid methyl ester: To a stirred solution of 5-bromo-4-methoxypicolinic acid methyl ester (1.0 g, 4.06 mmol) in NMP (10 ml) was added Zn(CN)2 (1.19 g, 10.10 mmol) and the mixture was stirred with Ar (g) Degas for 10 min. To the above mixture was added Pd(dba)2 (0.233 g, 0.40 mmol) followed by Pd(dppf)Cl2 and the reaction mixture was heated with Ar (g) The mixture was stirred for 10 minutes. The reaction mixture was heated in an oil bath at 100 ° C for 4 hours. After the reaction was completed (monitored by TLC), the reaction mixture was diluted with ethyl acetate (30ml), and cold water was added. The reaction mixture was extracted with ethyl acetate (2x 30ml). The combined organic layer was washed with brine (50ml), dried over Na2SO4, and concentrated under reduced pressure to obtain the desired compound in a light brown oil. The crude compound was purified by combi flash chromatography to obtain a pure compound (0.24g, 30%). LCMS: m / z 193.1[M+H]+ . 1 H NMR (400MHz, CDCl3): δ8.80(s,1H),7.80(s,1H),4.12(s,3H),4.07(s,3H).

[0233] Step-2: Methyl 5-(aminomethyl)-4-methoxypicolinate: To a stirred solution of methyl 5-cyano-4-methoxypicolinate (0.65 g, 3.3 mmol) in MeOH (10 ml) was added Pd / C (0.065 g) in an autoclave and the reaction mixture was heated with H 2(g) The mixture was purged. The mixture was sealed and stirred at room temperature for 16 hours. After the reaction was completed (monitored by TLC), the reaction mixture was filtered through a celite bed, washed with MeOH, and concentrated under reduced pressure to give a crude compound which was used in the next step without further purification (0.40 g, 60%). LCMS: m / z 197.1 [M+H] + .

[0234] Step-3: 5-(((2,3-dichlorobenzyl)amino)methyl)-4-methoxypicolinic acid methyl ester: To a stirred solution of 5-(aminomethyl)-4-methoxypicolinic acid methyl ester (0.08g, 0.40mmol) and 2,3-dichlorobenzaldehyde (0.084g, 0.48mmol) in DCE (10ml) was added acetic acid (0.1ml) dropwise followed by powdered molecular sieves. The reaction mixture was stirred at room temperature for 1 hour. Sodium cyanoborohydride (0.05g, 0.80mmol) was added to the reaction mixture. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed (monitored by TLC), the reaction mixture was quenched by saturated sodium bicarbonate (5mL). The aqueous layer was extracted with ethyl acetate (2x 10mL). The combined organic layers were washed with brine (10ml), dried over Na2SO4, and concentrated under reduced pressure to obtain a crude compound. The crude compound was purified by combi flash chromatography to obtain pure ethyl 2-((4-cyano-2,6-difluorophenethyl)amino)-2-phenylacetate (0.082 g, 56%). 1 H NMR (400MHz, CDCl3): δ8.55 (s, 1H), 7.67 (s, 1H), 7.49-7.42 (m, 2H), 7.22 (t, J = 8Hz, 1H), 4.03-3.89 (m, 10H).

[0235] Synthesis of methyl 5-(((2,3-dichlorobenzyl)amino)methyl)-4-(dimethylamino)picolinate (Intermediate 14)

[0236]

[0237] Step-1: 4-(dimethylamino)picolinic acid methyl ester: To a stirred solution of 4-(dimethylamino)picolinic acid (3.0 g, 18.05 mmol) in methanol (60 ml) was added concentrated H2SO4 (1.5 ml) dropwise at room temperature. The reaction mixture was stirred at 60 ° C for 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure to obtain a residue, which was placed in dichloromethane (2X 50 ml), and the combined organic layers were washed with saturated sodium bicarbonate solution (50 ml) and subsequently washed with brine (50 ml). The organic layer was dried over anhydrous Na2SO4 and concentrated under reduced pressure to obtain a crude compound, which was triturated with n-pentane to obtain the title compound (2.2 g, 67%). LCMS: m / z181.0[M+H] + . 1H NMR (400MHz, DMSO-d6): δ3.02 (s, 6H), 3.84 (s, 3H), 6.78-6.80 (m, 1H), 7.24 (d, J = 2.4Hz, 1H), 8.20 (d, J = 5.6Hz, 1H).

[0238] Step-2: Methyl 5-bromo-4-(dimethylamino)picolinate: To a stirred solution of methyl 4-(dimethylamino)picolinate (2.2 g, 12.21 mmol) in DCE (22 ml) was added N-bromosuccinimide (2.17 g, 12.19 mmol) followed by AIBN (0.4 g, 2.44 mmol) and the reaction mixture was stirred at room temperature for 10 minutes and then heated at 90°C for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated to give the crude compound. The crude compound was purified by combi flash to obtain the title compound (2.2 g, 69%). LCMS: m / z 259.2 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ2.97(s,6H),3.86(s,3H),7.52(s,1H),8.54(s,1H).

[0239] Step-3: 4-(Dimethylamino)-5-vinylpicolinate: To a stirred solution of methyl 5-bromo-4-(dimethylamino)picolinate (2.2 g, 8.49 mmol) in DMSO (22 ml) was added trifluorovinylborane potassium salt (3.41 g, 25.45 mmol) portionwise at room temperature. The reaction mixture was stirred with Ar. (g)Degas for 10 minutes. K2CO3 (3.51 g, 25.39 mmol) was added to the above mixture and the mixture was again heated with Ar. (g) Degas for 10 minutes, then add Pd (dppf) Cl2 (1.24g, 1.69mmol), and the reaction mixture is stirred at 80 ° C for 1 hour. After the reaction is completed (monitored by TLC), the reaction mixture is cooled to room temperature, diluted with cold water, and extracted with ethyl acetate (2x50ml). The combined organic layer is washed with brine (50ml), dried over anhydrous Na2SO4 and concentrated under reduced pressure to obtain a crude compound, which is purified by column chromatography to obtain the title compound (1.4g, 79%). LCMS: m / z 206.5 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ2.88(s,6H),3.85(s,3H),5.41-5.44(d,J=11.6Hz,1H),5.80-5.85(m,1H),6.75-6.82(m,1H),7.45(s,1H),8.44(s,1H).

[0240] Step-4: Methyl 4-(dimethylamino)-5-formylpicolinate: To a stirred solution of methyl 4-(dimethylamino)-5-vinylpicolinate (1.4 g, 6.79 mmol) in 1,4-dioxane (42 ml) was added RuCl3.3H2O (0.017 g, 0.067 mmol) at 0°C followed by dropwise addition of water (14 mL) containing sodium periodate solution (5.78 g, 27.16 mmol) and the reaction was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with water and extracted with ethyl acetate (2 x 50 ml). The combined organic layers were washed with brine (50 ml), dried over anhydrous Na2SO4 and concentrated under reduced pressure to obtain the crude compound which was purified by column chromatography to obtain the title compound (0.6 g, 42%). LCMS: m / z 209.0 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ3.08(s,6H),3.88(s,3H),7.49(s,1H),8.69(s,1H),10.04(s,1H).

[0241] Step-5: Methyl 5-(((2,3-dichlorobenzyl)amino)methyl)-4-(dimethylamino)picolinate: To a solution of methyl 4-(dimethylamino)-5-formylpicolinate (0.600 g, 2.88 mmol) in 1,2-dichloroethane (9 ml) was added molecular sieves followed by (2,3-dichlorophenyl)methylamine (0.507 g, 2.88 mmol) and acetic acid (2.0 ml, 34.56 mmol) and stirred in RM for 2 hours. Sodium cyanoborohydride (0.271 g, 4.32 mmol) was then added followed by methanol (6 ml) and the reaction mixture was stirred at room temperature for 24 hours. After completion of the reaction (monitored by TLC), the reaction mixture was passed through a bed of celite. Saturated sodium bicarbonate solution (10 ml) was added to the filtrate and extracted with ethyl acetate (2 x 20 ml). The combined organic layers were washed with brine (20 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound (0.450 g, 42%). LCMS: m / z 368.3 [M+H] + The crude intermediate was used in the next step without further purification.

[0242] Synthesis of 3-(2,3-dichlorophenyl)-3-((pyridin-3-ylmethyl)amino)propionitrile (Intermediate 15)

[0243]

[0244] Step-1: (R,E)-N-(2,3-dichlorobenzylidene)-2-methylpropane-2-sulfenamide: To a stirred solution of 2,3-dichlorobenzaldehyde (3.0 g, 17.14 mmol) in THF (60 ml) was added 2-methylpropane-2-sulfenamide (2.49 g, 20.53 mmol) at room temperature and the reaction mixture was stirred for 15 minutes, then Ti(O-iPr)4 (9.15 g, 32.22 mmol) was slowly added at 0°C. The reaction mixture was stirred at room temperature under N 2(g) The mixture was stirred for 16 hours under an atmosphere of 4% paraformaldehyde (2% paraformaldehyde, 1 ... + .

[0245] Step-2: (R)-N-(2-Cyano-1-(2,3-dichlorophenyl)ethyl)-2-methylpropane-2-sulfenamide: To a stirred solution of n-BuLi (2.5 M in THF) (7.4 mL, 18.60 mmol) was added acetonitrile (0.682 g, 16.54 mmol) dropwise at -78 °C and the reaction mixture was stirred under N 2(g) Atmosphere stirred for 1 hour. (R, E) -N- (2,3- dichlorobenzylidene) -2- methylpropane -2- sulfenamide (2.3g, 8.27mmol) in tetrahydrofuran (60ml) solution was added at -78 ° C, and the reaction mixture was further stirred for 1.5 hours. After the reaction was completed (monitored by TLC), the reaction mixture was quenched with dilute hydrochloric acid solution (5ml) and extracted with ethyl acetate (3x 50ml). The combined organic layer was washed with brine (50ml), dried over Na2SO4, and concentrated under reduced pressure. The crude compound was purified by using reverse phase combi-flash chromatography using ACN and water containing 0.1% formic acid to obtain the title compound (1.8g, 68%). LCMS: m / z 319.2 [M + H] + . 1 H NMR (400MHz, DMSO-d6): δ1.26 (s, 9H), 2.96-3.08 (m, 2H), 4.97-5.03 (m, 1H), 6.24 (d, J = 9.2Hz, 1H), 7.44 (t, J = 8.0Hz, 1H), 7.63-7.67 (m, 2H).

[0246] Step-3: 3-Amino-3-(2,3-dichlorophenyl)propionitrile HCl salt: To a stirred solution of (R)-N-(2-cyano-1-(2,3-dichlorophenyl)ethyl)-2-methylpropane-2-sulfinamide (1.8 g, 5.64 mmol) in dichloromethane (20 ml) was added 4 M HCl in 1,4-dioxane (8.5 ml, 33.84 mmol), and the reaction mixture was stirred at room temperature for 2 hours. After completion of the reaction (monitored by TLC), the reaction mixture was concentrated under reduced pressure. The crude compound was triturated with n-pentane to obtain the title compound (1.4 g, 98%). LCMS: m / z 214.9 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ3.41-3.43(m,2H),5.08-5.12(m,1H),7.56(t,J=8.0Hz,1H),7.78-7.81(dd,J=1.2Hz,8.0Hz,1H),7.90-7.92(dd,J=1.2H Z,8.0Hz,1H),9.14(bs,3H).

[0247] Step-4: 3-(2,3-dichlorophenyl)-3-((pyridin-3-ylmethyl)amino)propionitrile: To a mixture of 3-amino-3-(2,3-dichlorophenyl)propionitrile HCl salt (1.6 g, 7.44 mmol) in 1,2-dichloroethane (20 ml) and methanol (1 mL) was added triethylamine (3.1 ml, 22.32 mmol) and stirred at room temperature for 30 minutes. Then molecular sieves (3.2 g), nicotinaldehyde (0.796 g, 7.44 mmol) and acetic acid (1.78 g, 29.76 mmol) were added and the reaction mixture was stirred for 2 hours. NaCNBH3 (0.701 g, 11.16 mmol) was added and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was passed through a celite bed. Saturated sodium bicarbonate solution (10 ml) was added to the filtrate and extracted with ethyl acetate (2 x 20 ml). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by RP-purification using ACN and water containing 0.1% formic acid to obtain the title compound (1.55 g, 68%). LCMS: m / z 306.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ2.87-2.99 (m, 2H), 3.49 (d, J = 14Hz, 1H), 3.64 (d, J = 13.6Hz, 1H), 4.38 (t, J = 6.0Hz, 1H), 7.34-7. 37(m,1H),7.47(t,J=8.0Hz,1H),7.62(dd,J=1.2Hz,8.0Hz,1H),7.72-7.75(m,2H),8.45(d,J=3.6Hz,1H),8.48(bs,1H).

[0248] Synthesis of 4-(2-chlorophenyl)-4-((pyridin-3-ylmethyl)amino)butyronitrile (Intermediate 16)

[0249]

[0250] Step-1: (E)-4-(2-chlorophenyl)-4-((pyridin-3-ylmethyl)amino)butyronitrile: To a solution of 4-(2-chlorophenyl)-4-oxobutyronitrile (0.3 g, 1.54 mmol) in THF (6 ml) was added pyridin-3-ylmethylamine (0.217 g, 2.01 mmol) at room temperature followed by dropwise addition of titanium (IV) isopropoxide (0.827 g, 2.91 mmol) at 0 °C. The reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was quenched with saturated NH4Cl (15 ml) and diluted with ethyl acetate (20 ml). The reaction mixture was filtered through a celite bed and washed with ethyl acetate (20 ml). The organic layer was separated and the aqueous layer was extracted with ethyl acetate (2 x 20 ml). The combined organic layers were washed with brine (50 ml), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude compound was used in the next step without further purification (0.5 g, crude). LCMS: m / z 284.3 [M+H] + .

[0251] Step-2: 4-(2-Chlorophenyl)-4-((pyridin-3-ylmethyl)amino)butyronitrile: To a stirred solution of (E)-4-(2-chlorophenyl)-4-((pyridin-3-ylmethyl)amino)butyronitrile (0.5 g, 1.76 mmol) in 1,2-dichloroethane (6 ml) and ethanol (2 ml) was added molecular sieves (1 g). The reaction mixture was stirred at room temperature for 10 minutes. Sodium cyanoborohydride (0.166 g, 2.64 mmol) was added at 0°C and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was diluted with 10% methanol in DCM (50 ml) and filtered through a celite bed. The filtrate was concentrated to get the crude compound, which was used in the next step without further purification. LCMS: m / z 286.3 [M+H] + .

[0252] Synthesis of ethyl 3-((tert-butoxycarbonyl)amino)-5-(((1-(2,3-dichlorophenyl)propyl)amino)methyl)picolinate (Intermediate 17)

[0253]

[0254] Step-1: Ethyl 3-amino-5-bromopicolinate: A solution of 3-amino-5-bromopicolinic acid (10 g, 46.08 mmol), potassium carbonate (6.36 g, 46.08 mmol) and iodoethane (3.70 ml, 46.08 mmol) in dimethylacetamide (100 ml) was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water, filtered and dried to obtain a pure solid compound (7 g, 62%). LCMS: m / z 244.9 [M+H] + .

[0255] Step-2: Ethyl 5-bromo-3-((tert-butoxycarbonyl)amino)picolinate: To a stirred solution of ethyl 3-amino-5-bromopicolinate (3 g, 12.24 mmol) in dichloromethane was added triethylamine (5.11 ml, 36.72 mmol) followed by di-tert-butyl dicarbonate (4.0 g, 18.36 mmol) and DMAP (0.299 g, 2.45 mmol) at 0°C. The reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (30 ml) and extracted with dichloromethane (2 x 30 ml). The combined organic layers were washed with brine (50 ml) and dried over Na2SO4. Evaporated under vacuum to obtain the crude compound. The crude compound was purified using column chromatography to obtain the pure compound (2.8 g, 66%). LCMS: m / z 345.3 [M+H] + .

[0256] Step-3: Ethyl 3-((tert-butoxycarbonyl)amino)-5-vinylpicolinate: To a stirred solution of ethyl 5-bromo-3-((tert-butoxycarbonyl)amino)picolinate (2.6 g, 7.53 mmol) and potassium trifluoro(vinyl)borane (3.02 g, 22.55 mmol) in dimethyl sulfoxide (26 ml) was added potassium carbonate (3.12 g, 22.6 mmol) followed by PdCl2(dppf) (1.10 g, 1.50 mmol) at room temperature. The reaction mixture was stirred for 2 h with Ar. (g) Purge and stir at 80°C for 1 hour. After the reaction is complete (monitored by TLC), the reaction mixture is poured into ice-cold water (30 ml) and extracted with ethyl acetate (2 x 30 ml). The combined organic layer is washed with brine (50 ml) and dried over Na2SO4. Evaporate under vacuum to obtain the crude compound. The crude compound is purified by column chromatography to obtain the pure compound (1.8 g, 81%). LCMS: m / z 293.0 [M+H] + . 1HNMR (400MHz, DMSO-d6): δ1.34(t,J=7.2Hz,3H),1.50(s,9H),4.34(q,J=6.8Hz,2H),5.57(d,J= 10.8Hz,1H),6.05(d,J=17.6Hz,1H),6.83-6.90(m,1H),8.49(s,1H),8.54(s,1H),10.02(s,1H).

[0257] Step-4: Ethyl 3-((tert-butoxycarbonyl)amino)-5-formylpicolinate: To a stirred solution of ethyl 3-((tert-butoxycarbonyl)amino)-5-vinylpicolinate (1.8 g, 6.16 mmol) in 1,4-dioxane (18 ml), ruthenium (III) chloride (0.012 g, 0.06 mmol) was added, followed by dropwise addition of a solution of sodium periodate (5.26 g, 24.56 mmol) in water (54 ml) at 0 °C. The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (20 ml) and extracted with ethyl acetate (2 x 20 ml). The combined organic layers were washed with brine (30 ml) and dried over Na2SO4. Evaporated under vacuum to obtain the crude compound. The crude compound was purified using column chromatography to obtain the pure compound (0.600 g, 33%). LCMS: m / z 295.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ1.34(t,J=6.8Hz,3H),1.50(s,9H),4.35(q,J=6.8Hz,2H),8.79(s,1H),8.80(s,1H),10.02(s,1H),10.16(s,1H).

[0258] Step-5: Ethyl 3-((tert-Butoxycarbonyl)amino)-5-(((1-(2,3-dichlorophenyl)propyl)amino)methyl)picolinate: To a stirred solution of 1-(2,3-dichlorophenyl)propan-1-amine hydrochloride (0.490 g, 2.04 mmol) in 1,2-dichloroethane (6 ml) was added triethylamine (0.71 ml, 5.093 mmol) and stirred at room temperature for 1 hour. A solution of ethyl 3-((tert-Butoxycarbonyl)amino)-5-formylpicolinate (0.500 g, 1.69 mmol) in 1,2-dichloroethane (6 ml) and acetic acid (0.1 ml) was added to the reaction mixture and stirred for another 1 hour at room temperature. Sodium cyanoborohydride (0.160 g, 2.55 mmol) and methanol (2 ml) were added and the reaction mixture was stirred for another 1 hour at room temperature. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (20 ml) and extracted with ethyl acetate (2 x 20 ml). The organic layer was washed with brine (30 ml), and the combined organic layers were evaporated under vacuum to obtain the crude compound. The crude compound was purified using column chromatography to obtain the pure compound (0.500 g, 61%). LCMS: m / z 484.0 [M+2H] + . 1 H NMR (400MHz, DMSO-d6): δ0.85(t,J=7.6Hz,3H),1.32(t,J=8Hz,3H),1.49(s,9H),3.49(d,J=14.8Hz,1H),3.66(d,J=14.4Hz,1H) ,4.32(q,J=6.8Hz,2H),7.39(t,J=3.2Hz,1H),7.52(d,J=6.4Hz,1H),7.63(d,J=6Hz,1H),8.21(s,1H),8.43(s,1H),9.99(s,1H).

[0259] Synthesis of ethyl 3-((tert-butoxycarbonyl)amino)-5-(((2,3-dichlorobenzyl)amino)methyl)picolinate (Intermediate 18)

[0260]

[0261] Step-1: Ethyl 3-amino-5-bromopicolinate: A solution of 3-amino-5-bromopicolinic acid (10 g, 46.08 mmol), potassium carbonate (6.36 g, 46.08 mmol) and iodoethane (3.70 ml, 46.08 mmol) in dimethylacetamide (100 ml) was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water, filtered and dried to obtain a pure solid compound (7 g, 62%). LCMS: m / z 244.9 [M+H] + .

[0262] Step-2: Ethyl 5-bromo-3-((tert-butoxycarbonyl)amino)picolinate: To a stirred solution of ethyl 3-amino-5-bromopicolinate (3 g, 12.24 mmol) in dichloromethane was added triethylamine (5.11 ml, 36.722 mmol) followed by di-tert-butyl dicarbonate (4.0 g, 18.36 mmol) and DMAP (0.299 g, 2.45 mmol) at 0°C. The reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (30 ml) and extracted with dichloromethane (2 x 30 ml). The combined organic layers were washed with brine (50 ml) and dried over Na2SO4. Evaporated under vacuum to obtain the crude compound. The crude compound was purified using column chromatography to obtain the pure compound (2.8 g, 66%). LCMS: m / z 345.3 [M+H] + .

[0263] Step-3: Ethyl 3-((tert-butoxycarbonyl)amino)-5-vinylpicolinate: To a stirred solution of ethyl 5-bromo-3-((tert-butoxycarbonyl)amino)picolinate (2.6 g, 7.53 mmol) and potassium trifluoro(vinyl)borane (3.02 g, 22.55 mmol) in dimethyl sulfoxide (26 ml) was added potassium carbonate (3.12 g, 22.59 mmol) followed by PdCl2(dppf) (1.10 g, 1.50 mmol) at room temperature. The reaction mixture was stirred for 2 h with Ar. (g) Purge and stir at 80°C for 1 hour. After the reaction is complete (monitored by TLC), the reaction mixture is poured into ice-cold water (30 ml) and extracted with ethyl acetate (2 x 30 ml). The combined organic layer is washed with brine (50 ml) and dried over Na2SO4. Evaporate under vacuum to obtain the crude compound. The crude compound is purified by column chromatography to obtain the pure compound (1.8 g, 81%). LCMS: m / z 293.0 [M+H] + . 1HNMR (400MHz, DMSO-d6): δ1.34(t,J=7.2Hz,3H),1.50(s,9H),4.34(q,J=6.8Hz,2H),5.57(d,J= 10.8Hz,1H),6.05(d,J=17.6Hz,1H),6.83-6.90(m,1H),8.49(s,1H),8.54(s,1H),10.02(s,1H).

[0264] Step-4: Ethyl 3-((tert-butoxycarbonyl)amino)-5-formylpicolinate: To a stirred solution of ethyl 3-((tert-butoxycarbonyl)amino)-5-vinylpicolinate (1.8 g, 6.16 mmol) in 1,4-dioxane (18 ml), ruthenium (III) chloride (0.012 g, 0.057 mmol) was added, followed by dropwise addition of a solution of sodium periodate (5.26 g, 24.56 mmol) in water (54 ml) at 0 ° C. The reaction mixture was stirred at room temperature for 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (20 ml) and extracted with ethyl acetate (2 x 20 ml). The combined organic layers were washed with brine (30 ml) and dried over Na2SO4. Evaporated under vacuum to obtain the crude compound. The crude compound was purified using column chromatography to obtain the pure compound (0.600 g, 33%). LCMS: m / z 295.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ1.34(t,J=6.8Hz,3H),1.50(s,9H),4.35(q,J=6.8Hz,2H),8.79(s,1H),8.80(s,1H),10.02(s,1H),10.16(s,1H).

[0265] Step-5: Ethyl 3-((tert-butoxycarbonyl)amino)-5-(((2,3-dichlorobenzyl)amino)methyl)picolinate: To a stirred solution of ethyl 3-((tert-butoxycarbonyl)amino)-5-formylpicolinate (0.200 g, 0.68 mmol) and (2,3-dichlorophenyl)methylamine (0.143 g, 0.81 mmol) in 1,2-dichloroethane (5 ml) was added acetic acid (0.05 ml). The reaction mixture was stirred at room temperature for 1 hour. Sodium cyanoborohydride (0.064 g, 1.02 mmol) and methanol (1 ml) were added and the reaction mixture was stirred at room temperature for another 1 hour. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ice-cold water (20 ml) and extracted with ethyl acetate (2 x 20 ml). The combined organic layer was washed with brine (30 ml) and dried over Na2SO4. Evaporated under vacuum to obtain the crude compound. The crude compound was purified using column chromatography to obtain the pure compound (0.190 g, 61%). LCMS: m / z 454.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ0.87(t,J=7.2Hz,3H),1.49(s,9H),3.83(s,4H),4.33(q,2H,J=7. 2Hz),7.37(t,J=7.6Hz,1H),7.55(t,J=6.4Hz,2H),8.32(s,1H),8.53(s,1H),10.00(s,1H).

[0266] The following compounds in Table 1 were prepared in a manner similar to the procedures for Intermediates I-III:

[0267] Table 1

[0268]

[0269]

[0270]

[0271]

[0272]

[0273]

[0274] General Library Program I

[0275] Synthesis of N-[(3,5-dichlorophenyl)methyl]-N-[(pyridin-3-yl)methyl]benzamide (Compound 1)

[0276]

[0277] Step 1. Benzoic acid (30 mg, 245 μmol) and HATU (111 mg, 294 μmol) were combined in DMF and stirred at room temperature for 10 minutes. This solution was added to a flask containing Intermediate I [(3,5-dichlorophenyl)methyl][(pyridin-3-yl)methyl]amine hydrochloride (89.2 mg, 294 μmol) and then N,N-diisopropylethylamine (211 μL, 1.22 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was directly subjected to mass-directed preparative HPLC to give N-[(3,5-dichlorophenyl)methyl]-N-[(pyridin-3-yl)methyl]benzamide (14.9 mg, 16.1%). MS ES m / z: 371.0 [M+H] + .

[0278] The following compounds in Table 2 were prepared in a manner similar to General Library Procedure I:

[0279] Table 2

[0280]

[0281]

[0282]

[0283]

[0284]

[0285]

[0286]

[0287]

[0288]

[0289]

[0290]

[0291]

[0292]

[0293]

[0294]

[0295]

[0296]

[0297] General Procedure I

[0298] Synthesis of N-(4-chlorobenzyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 138)

[0299]

[0300] Step 1 Synthesis of N-(4-chlorobenzyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide. 3-Fluorobenzoic acid (30 mg, 214 μmol) and HATU (97.3 mg, 256 μmol) were combined in DMF and stirred at room temperature for 10 minutes. This solution was then added to a flask containing [(4-chlorophenyl)methyl][(pyridin-3-yl)methyl]amine (59.5 mg, 256 μmol) and then N,N-diisopropylethylamine (185 μL, 1.07 mmol). The reaction was stirred at room temperature overnight. The reaction mixture was directly subjected to mass-directed preparative HPLC to obtain N-[(4-chlorophenyl)methyl]-3-fluoro-N-[(pyridin-3-yl)methyl]benzamide (46.9 mg, 60%). MS ES m / z: 355.1 [M+H] + .

[0301] The following compounds in Table 3 were prepared in a manner similar to the General Procedure I described above:

[0302] Table 3

[0303]

[0304]

[0305] General Procedure II

[0306] Synthesis of methyl 5-({[(2,4-dichlorophenyl)methyl]amino}methyl)pyridine-2-carboxylate (Compound 142).

[0307]

[0308] Step 1. 5-({N-[(2,4-dichlorophenyl)methyl]-1-(3-fluorophenyl)formamido}methyl)pyridine-2-carboxylic acid methyl ester (Compound 141, 64 mg, 143 umol) was dissolved in 5:1 THF:MeOH (500 uL). 1N NaOH (1.43 mL) aqueous solution was added and the reaction was stirred at room temperature for 4 hours. The reaction was acidified to pH~3 with 1N HCl and 4x5 mL CH2Cl2 was extracted. The organic layers were combined and concentrated to give 5-({N-[(2,4-dichlorophenyl)methyl]-1-(3-fluorophenyl)formamido}methyl)pyridine-2-carboxylic acid (56 mg, 82% yield) as a white solid. MS ES m / z: 433.0 [M+H] + .

[0309] The following compounds in Table 4 were prepared in a manner similar to the General Procedure II described above:

[0310] Table 4

[0311]

[0312]

[0313]

[0314]

[0315]

[0316]

[0317]

[0318]

[0319] General Procedure III

[0320] Synthesis of 5-({N-[(2,4-dichlorophenyl)methyl]-1-(3-fluorophenyl)formamido}methyl)pyridine-2-carboxamide (Compound 143).

[0321]

[0322] Step 1. 5-({N-[(2,4-dichlorophenyl)methyl]-1-(3-fluorophenyl)formamide}methyl)pyridine-2-carboxylic acid (Compound 142, 15 mg, 34 umol), NH4 (7M in MeOH; 10 uL, 69 umol) and Et3N (14 uL, 69 umol) were combined in anhydrous DCE (1.5 mL). DMC (6 mg, 35 umol) was added and the reaction was stirred at room temperature for 4 hours. The reaction was quenched with 1 mL of H2O, and the organic layer was separated and concentrated to dryness. Purification by reverse phase chromatography using Biotage (Sfar C18 12 g, 90% H2O / 10% CH3CN and 0.1% formic acid to 10% H2O / 90% CH3CN and 0.1% formic acid). The desired fractions were combined and concentrated to give 5-({N-[(2,4-dichlorophenyl)methyl]-1-(3-fluorophenyl)formamido}methyl)pyridine-2-carboxamide (8.9 mg, 60% yield). MS ES m / z: 432.1 [M+H] + .

[0323] The following compounds in Table 5 were prepared in a manner similar to the General Procedure III described above:

[0324] Table 5

[0325]

[0326] General Procedure IV

[0327] Synthesis of 5-({N-[(2,4-dichlorophenyl)methyl]-1-(3-fluorophenyl)formamido}methyl)-N-methylpyridine-2-carboxamide (Compound 144).

[0328]

[0329] Step 1. 5-({N-[(2,4-dichlorophenyl)methyl]-1-(3-fluorophenyl)formamide}methyl)pyridine-2-carboxylic acid (Compound 142, 15 mg, 35 umol), methylamine HCl (4.7 mg, 69 umol) and Et3N (14 uL, 103 umol) were combined in DCE (1.5 mL). DMC (6 mg, 35 umol) was added and the reaction was stirred at room temperature for 4 hours. The reaction was quenched with 1 mL of H2O, and the organic layer was separated and concentrated to dryness. Biotage was used to purify by reverse phase chromatography (Sfar C18 12 g, 90% H2O / 10% CH3CN and 0.1% formic acid to 10% H2O / 90% CH3CN and 0.1% formic acid). The desired fractions were combined and concentrated to give 5-({N-[(2,4-dichlorophenyl)methyl]-1-(3-fluorophenyl)formamido}methyl)-N-methylpyridine-2-carboxamide (12.5 mg, 81% yield). MS ES m / z: 446.1 [M+H] + .

[0330] General Procedure V

[0331] Synthesis of N-(2,4-dichlorobenzyl)-3-fluoro-N-((6-(hydroxymethyl)pyridin-3-yl)methyl)benzamide (Compound 145).

[0332]

[0333] Step 1. A solution of 5-({N-[(2,4-dichlorophenyl)methyl]-1-(3-fluorophenyl)formamido}methyl)pyridine-2-carboxylic acid methyl ester (compound 141, 48.7mg, 109umol) in anhydrous THF was added dropwise to a 2M solution of LAH (4.13uL, 218umol) in THF at 0°C under N2. The reaction was stirred at 0°C and slowly warmed to room temperature over 90 minutes. The reaction was quenched with 1mL of water, diluted with 5mL of Et2O and another 1mL of water. The reaction was stirred at room temperature for 10 minutes. The organic layer was separated and concentrated to dryness. Purification was performed by mass-directed preparative purification to obtain N-(2,4-dichlorobenzyl)-3-fluoro-N-((6-(hydroxymethyl)pyridin-3-yl)methyl)benzamide (2.12mg, 4.2% yield). 1H NMR (400MHz, DMSO-d6) δ=8.21(br s,1H),7.69(br s,1H),7.56(br s,2H),7.49-7.34(m,5H),7.29(brd,J=8.3Hz,3H),5.37(br s,1H),4.59(br s,3H),4.52(s,4H),3.29(br s,5H),0.94(t,J=7.1Hz,1H).MS: (ES,m / z):419.0[M+H] + .

[0334] General Program VI

[0335] Synthesis of N-[(6-aminopyridin-3-yl)methyl]-N-[(2,4-dichlorophenyl)methyl]-3-fluorobenzamide (Compound 146).

[0336]

[0337] Synthesis of tert-butyl N-[5-({[(2,4-dichlorophenyl)methyl]amino}methyl)pyridin-2-yl]carbamate

[0338]

[0339] Step 1. Combine 2,4-dichlorobenzaldehyde (194 mg, 1.11 mmol) with tert-butyl (5-(aminomethyl)pyridin-2-yl)carbamate (250 mg, 1.11 mmol) and 10 uL acetic acid in DCE (20 mL). Finally, add NaBH(OAc)3 (587 mg, 2.77 mmol). The reaction is stirred at room temperature overnight. The reaction is quenched with brine (15 mL). The organic layer is separated, dried over Na2SO4, filtered and concentrated. Purify by reverse phase chromatography using Biotage (Snap Ultra 30 g, 10% CH3CN / 90% H2O and 0.1% formic acid to 90% CH3CN / 10% H2O and 0.1% formic acid gradient). The desired fractions were combined and lyophilized to give tert-butyl N-[5-({[(2,4-dichlorophenyl)methyl]amino}methyl)pyridin-2-yl]carbamate (159.6 mg, 38% yield).

[0340]

[0341] Step 2. tert-Butyl N-[5-({[(2,4-dichlorophenyl)methyl]amino}methyl)pyridin-2-yl]carbamate (159.6mg, 415umol), 3-fluorobenzoic acid (64mg, 456umol), Et3N (115uL, 830umol) and HATU (146mg, 456umol) were combined in DMA (4mL). The reaction was stirred at room temperature overnight. The reaction was diluted with 5mL H2O and extracted with 3x8mL EtOAc. The organic layers were combined and concentrated. The residue was dissolved in EtOAc (2mL) and 5 equivalents of HCl (4M in 1,4-dioxane) and stirred at room temperature for 1 hour. The reaction was concentrated to dryness. Biotage was used to purify by normal phase column chromatography (ZIP Sphere 10g, 10% EtOAc / heptane to 100% EtOAc). The desired fractions were combined and concentrated to give N-[(6-aminopyridin-3-yl)methyl]-N-[(2,4-dichlorophenyl)methyl]-3-fluorobenzamide (32.1 mg, 17% yield) as a white solid. MS ES m / z: 404.0 [M+H] + .

[0342] The following compounds in Table 6 were prepared in a manner similar to General Procedure VI described above:

[0343] Table 6

[0344]

[0345] Preparation of N-((6-acetylaminopyridin-3-yl)methyl)-N-(2-chloro-3-methoxybenzyl)-3-fluorobenzamide (Compound 213)

[0346]

[0347] Step 1: N-((6-acetylaminopyridin-3-yl)methyl)-N-(2-chloro-3-methoxybenzyl)-3-fluorobenzamide. To a stirred solution of 1-(pyridin-3-yl)ethan-1-amine (0.1 g, 239 umol) in CH2Cl2 (2 ml), Et3N (20 uL, 263 umol) was added, followed by acetyl chloride (7 uL, 263 umol). The reaction mixture was stirred at room temperature for 8 hours. After completion of the reaction, the reaction mixture was washed with water (3 mL). The organic layer was removed, washed with brine (3 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by column chromatography to obtain the title compound (0.085 g, 77%). MS m / z 460.8 [M+1] + .

[0348] The following compounds in Table 7 were prepared in a manner similar to the procedures described above:

[0349] Table 7

[0350]

[0351] General ProcedureVII

[0352] Preparation of (R)-N-(2,4-dichlorobenzyl)-3-chloro-N-(1-(pyridin-3-yl)ethyl)benzamide (Compound 153) and (S)-N-(2,4-dichlorobenzyl)-3-chloro-N-(1-(pyridin-3-yl)ethyl)benzamide (Compound 154).

[0353]

[0354] Step 1: Synthesis of N-(2,4-dichlorobenzyl)-1-(pyridin-3-yl)ethyl-1-amine. To a stirred solution of 1-(pyridin-3-yl)ethyl-1-amine (1 g, 8.18 mmol) in DMF (5 ml) was added potassium carbonate (2.26 g, 16.4 mmol). DMF (5 ml) containing 1-(bromomethyl)-2,4-dichlorobenzene (2.55 g, 10.6 mmol) was added at room temperature. The reaction mixture was heated to 55 ° C for 2 hours. After the reaction was completed, the reaction mixture was diluted in ethyl acetate (50 ml) and washed with cold water (3 x 50 ml). The organic layer was washed with brine (50 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by column chromatography to obtain the title compound (0.6 g, 26%). MS m / z 281.46[M+1] + .

[0355] Step 2: Synthesis of N-(2,4-dichlorobenzyl)-3-fluoro-N-(1-(pyridin-3-yl)ethyl)benzamide.

[0356] To a stirred mixture of N-(2,4-dichlorobenzyl)-1-(pyridin-3-yl)ethyl-1-amine (0.6 g, 2.13 mmol) in DCM (6 ml) was added Et3N (0.54 g, 5.33 mmol) under a nitrogen atmosphere. 3-Fluorobenzoyl chloride (0.51 g, 3.20 mmol) was added dropwise at room temperature, and the reaction mixture was stirred at room temperature for 45 minutes. After the reaction was complete, the reaction mixture was quenched with cold water (12 ml) and extracted with DCM (2 x 20 ml). The combined organic layers were washed with brine (20 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by combi flash chromatography to obtain the title compound (0.2 g, 23%) as a racemic mixture. MS: racemic m / z 404.4 [M+H] + Chiral preparative HPLC: Shimadzu LC-20AP and UV detector; column: CHIRALPAK IB-N (250*21) mm, 5u; mobile phase A: 0.1% DEA / hexane, mobile phase B: 0.1% DEA / propan-2-ol; isocratic gradient 85:15 (A:B), column flow rate: 18 ml / min. FR-1 (isomer 1): R T =18.98; FR-2 (isomer 2): R T =25.87.

[0357] Isomer 1 was arbitrarily designated as (R)-N-(2,4-dichlorobenzyl)-3-chloro-N-(1-(pyridin-3-yl)ethyl)benzamide (Compound 153) LCMS: m / z 404.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ1.58(d,J=6.8Hz,3H),4.26(bs,1H),4.59(d,J=16.8Hz,1H),5.15(bs,1H),7.34-7.73(m,9H),8.45(d,J=4.0Hz,2H).

[0358] Isomer 2 was arbitrarily designated as (S)-N-(2,4-dichlorobenzyl)-3-chloro-N-(1-(pyridin-3-yl)ethyl)benzamide (Compound 154) LCMS: m / z 404.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ1.58(d,J=6.8Hz,3H),4.27(bs,1H),4.59(d,J=16.8Hz,1H),5.15(bs,1H),7.33-7.72(m,9H),8.46(d,J=4.0Hz,2H).

[0359] General Procedure VIII

[0360] Preparation of (R)-N-(1-(2,4-dichlorophenyl)propyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 157) and (S)-N-(1-(2,4-dichlorophenyl)propyl)-3-fluoro-N-(pyridin-3-ylphenyl)benzamide (Compound 158).

[0361]

[0362] Step 1. Synthesis of 1-(2,4-dichlorophenyl)-N-(pyridin-3-ylmethyl)propan-1-amine. A mixture of 1-(2,4-dichlorophenyl)propan-1-one (0.500 g, 2.46 mmol), pyridin-3-ylmethylamine (0.32 g, 2.95 mmol) and acetic acid (0.5 ml) in MeOH (5 ml) was stirred at room temperature for 2 hours. Sodium cyanoborohydride (0.23 g, 3.69 mmol) and DCE (0.5 ml) were added, and the reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth. The filtrate was concentrated under reduced pressure to give the title compound (0.730 g, crude) as an oil. LCMS: m / z 295.2 [M+H] + .

[0363] Step 2. Synthesis of N-(1-(2,4-dichlorophenyl)propyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide. To a solution of 1-(2,4-dichlorophenyl)-N-(pyridin-3-ylmethyl)propan-1-amine (0.73g, 2.47mmol) in anhydrous DCM (7.6ml) under a nitrogen atmosphere, 3-fluorobenzoyl chloride (0.47g, 2.96mmol) was added. Triethylamine (0.38g, 3.71mmol) was added dropwise at 0°C. The resulting reaction mixture was placed at room temperature and stirred for another 4 hours. After the reaction was complete, water (30ml) was slowly added, and the reaction mixture was extracted with DCM (2x 100ml). The combined organic layer was washed with brine (20ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by Combi-flash chromatography (RP Gold column) by using acetonitrile and water containing 0.1% formic acid to give the pure title compound (0.160 g, 15.53%) as a solid. Chiral preparative HPLC: Water's PSFC-200 and UV detector; column: CHIRALCEL OX-H (250*21) mm, 5u; mobile phase A: liquid CO2, mobile phase B: 0.1% DEA / methanol; isocratic gradient 82:18 (A:B), column flow: 80 ml / min (100 bar). FR-1 (isomer 1): R T =5.58; FR-2 (isomer 2): R T =7.97.

[0364] Isomer 1 was arbitrarily designated as (R)-N-(1-(2,4-dichlorophenyl)propyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 157) LCMS: m / z 417.27 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ0.80 (t, J=6.8Hz, 3H), 2.07 -2.15(m,1H),2.21-2.23(m,1H),4.44-4.55(m,2H),5.35(bs,1H),7.13-7.16(m,1H),7.20–7.27(m,3H ),7.35(d,J=7.2Hz,2H),7.43-7.50(m,2H),7.62(d,J=8.8Hz,1H),8.14(s,1H),8.32(d,J=4.0Hz,1H).

[0365] Isomer 2 was arbitrarily designated as (S)-N-(1-(2,4-dichlorophenyl)propyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 158) LCMS: m / z 417.30 [M+H]+ . 1 H NMR (400MHz, DMSO-d6): δ0.80 (t, J=6.4Hz, 3H), 2.07 -2.15(m,1H),2.21-2.23(m,1H),4.43-4.55(m,2H),5.35(bs,1H),7.13-7.16(m,1H),7.20–7.27(m,3H ),7.35(d,J=8.0Hz,2H),7.43-7.50(m,2H),7.62(d,J=8.8Hz,1H),8.14(s,1H),8.32(d,J=4.4Hz,1H).

[0366] General Procedure IX

[0367] Preparation of (R)-N-(1-(2,4-dichlorophenyl)-2-methylpropyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 159) and (S)-N-(1-(2,4-dichlorophenyl)-2-methylpropyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 160).

[0368]

[0369] Step 1.2,4-dichloro-N-methoxy-N-methylbenzamide synthesis. At 0 ° C, 2,4-dichlorobenzoic acid (10g, 52.35mmol) was added to a stirred solution in dry DMF (100ml) N, O-dimethylhydroxylamine hydrochloride (6.12g, 62.74mmol), followed by HATU (29.0g, 76.4mmol). The reaction mixture was stirred at 0 ° C for 1 hour. DIPEA (20.3g, 157mmol) was added dropwise at 0 ° C, and the resulting reaction mixture was stirred at room temperature for 4 hours. After the reaction was completed, water (250ml) was slowly added, and the reaction was extracted with ethyl acetate (2x 100ml). The combined organic layer was washed with brine (200ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by column chromatography to obtain the title compound (8.0g, 65%) in an oily state. LCMS: m / z 234.1 [M+H] + .

[0370] Step 2.1-(2,4-dichlorophenyl)-2-methylpropane-1-one synthesis. A solution of 2,4-dichloro-N-methoxy-N-methylbenzamide (0.47 g, 2.01 mmol) in dry THF (5 ml) was cooled to 0 ° C under a nitrogen atmosphere. THF containing isopropylmagnesium bromide (5 ml, 5.04 mmol) was added dropwise at 0 ° C. The resulting reaction mixture was placed at room temperature and heated at 60 ° C for 8 hours. After the reaction was completed, a saturated ammonium chloride solution (5 ml) was slowly added, and the reaction mixture was extracted with ethyl acetate (2 x 25 ml). The combined organic layer was washed with brine (10 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by column chromatography to obtain the title compound (0.04 g, 9%) as an oil. 1 H NMR (400MHz, DMSO-d6): δ1.065(d,J=6.8Hz,6H),3.25-3.35(m,1H),7.55(dd,J=8.4Hz,1.6Hz,1H),7.67(d,J=8.4Hz,1H),7.75(d,J=1.6Hz,1H).

[0371] Step 3.1-(2,4-dichlorophenyl)-2-methyl-N-(pyridin-3-ylmethyl)propan-1-amine synthesis. A mixture of 1-(2,4-dichlorophenyl)-2-methylpropan-1-one (1 g, 4.6 mmol), pyridin-3-ylmethylamine (2 g, 18.5 mmol) and acetic acid (1 ml) in DCE (10 ml) was stirred at room temperature for 2 hours. Sodium cyanoborohydride (0.43 g, 6.84 mmol) and methanol (1 ml) were added, and the reaction mixture was heated at 60 ° C for 24 hours. After completion of the reaction, the reaction mixture was filtered through diatomaceous earth and the filtrate was concentrated under reduced pressure. The crude compound was purified by Combi flash column chromatography (RP-Gold column) using acetonitrile and water containing 0.1% formic acid to obtain the pure title compound (0.260 g, 18%) as a solid. LCMS: m / z 309.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ0.75(d,J=6.8Hz,3H),0.95(d,J=6.8Hz 3H),1.79-1.84(m,1H),3.28-3.31(m,1H),3.57(d,J=14Hz,1H),3.73-3.74(m,1H),7 .30-7.33(m,1H),7.48(d,J=7.2Hz,1H),7.55(s,1H),7.65-7.69(m,2H),8.44(s,2H).

[0372] Step 4. Synthesis of N-(1-(2,4-dichlorophenyl)-2-methylpropyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide. A solution of 1-(2,4-dichlorophenyl)-2-methyl-N-(pyridin-3-ylmethyl)propan-1-amine (0.26g, 0.84mmol) in DCM (2.6ml) was cooled to 0°C. 3-Fluorobenzoyl chloride (0.198g, 1.25mmol) was added under a nitrogen atmosphere, followed by dropwise addition of triethylamine (0.35g, 3.43mmol) at 0°C. The resulting reaction mixture was placed at room temperature and stirred for another 4 hours. After the reaction was complete, water (5ml) was slowly added, and the reaction mixture was extracted with ethyl acetate (2x 15ml). The combined organic layer was washed with brine (15ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by Combi flash column chromatography (RP Gold column) using acetonitrile and water containing 0.1% formic acid to give the pure title compound as a solid (0.040 g, 11%). Chiral preparative HPLC: Shimadzu LC-20AP and UV detector; CHIRALPAK IH 250*21 mm, 5u; mobile phase A: 0.1% DEA / hexane, mobile phase B: 0.1% DEA / propan-2-ol:methanol (50:50); isocratic gradient 90:10 (A:B), column flow: 18 ml / min (100 bar). FR-1 (isomer 1): R T =10.71; FR-2 (isomer 2): R T =17.03.

[0373] Isomer 1 was arbitrarily designated as (R)-N-(1-(2,4-dichlorophenyl)-2-methylpropyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 159) LCMS: m / z 431.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ0.81(d,J=6.4Hz,3H),1.20(d,J=6.4Hz,3H),2.80-2.86(m,1H),4.55-4.65(m,2H),5.55(bs,1H),7.1 0-7.19(m,3H),7.23-7.29(m,3H),7.43-7.48(m,1H),7.53(s,1H),7.77(d,J=8.4Hz,1H),8.03(bs,1H),8.30(d,J=3.6Hz,1H).

[0374] Isomer 2 was arbitrarily designated as (S)-N-(1-(2,4-dichlorophenyl)-2-methylpropyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 160) LCMS: m / z 431.4 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ0.81(d,J=6.4Hz,3H),1.20(d,J=6.4Hz,3H),2.80-2.86(m,1H),4.55-4.65(m,2H),5.55(bs,1H),7.1 0-7.19(m,3H),7.23-7.29(m,3H),7.43-7.48(m,1H),7.53(s,1H),7.77(d,J=8.4Hz,1H),8.03(bs,1H),8.30(d,J=3.6Hz,1H).

[0375] General ProcedureX

[0376] Preparation of (R)-N-(1-(2,4-dichlorophenyl)-2-hydroxyethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 161) and (S)-N-(1-(2,4-dichlorophenyl)-2-hydroxyethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 162).

[0377]

[0378] Step 1. Synthesis of methyl 2-bromo-2-(2,4-dichlorophenyl)acetate. To a stirred solution of methyl 2-(2,4-dichlorophenyl)acetate (0.5 g, 2.28 mmol) in carbon tetrachloride (5 ml) was added N-bromosuccinimide (0.41 g, 2.28 mmol) and AIBN (0.074 g, 0.45 mmol). The reaction mixture was heated at 90 ° C for 4 hours. After the reaction was completed, the solvent was evaporated and the crude product was purified by column chromatography to obtain the title compound (0.6 g, 88%). 1 H NMR (400MHz, CDCl3): δ3.83 (s, 3H), 5.88 (s, 1H), 7.34 (d, J = 8.8Hz, 1H), 7.44 (s, 1H), 7.76 (d, J = 8.4Hz, 1H).

[0379] Step 2. Synthesis of methyl 2-(2,4-dichlorophenyl)-2-((pyridin-3-ylmethyl)amino)acetate. To a solution of methyl 2-bromo-2-(2,4-dichlorophenyl)acetate (0.4 g, 1.34 mmol) in anhydrous acetonitrile (4 ml) was added triethylamine (0.26 ml, 1.85 mmol) and 3-picolylamine (0.116 mg, 1.07 mmol) under a nitrogen atmosphere. The resulting reaction mixture was placed at room temperature and stirred for 5 hours. After the reaction was complete, water (15 ml) was slowly added, and the reaction mixture was extracted with ethyl acetate (3x10 ml). The combined organic layers were washed with brine (10 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by combi-flash chromatography to give the title compound (0.280 g, 64%) as a gum. LCMS: m / z 325.2 [M + +1]. 1 H NMR (400MHz, CDCl3): δ3.74(s,3H),3.82(s,2H),4.86(s,1H),7.28-7.30(bs ,2H),7.39(d,J=8.0Hz,1H),7.45(s,1H),7.71(d,J=6.8Hz,1H),8.56(s,2H).

[0380] Step 3. Synthesis of 2-(2,4-dichlorophenyl)-2-(3-fluoro-N-(pyridin-3-ylmethyl)benzamido)acetic acid methyl ester. 2-(2,4-dichlorophenyl)-2-((pyridin-3-ylmethyl)amino)acetic acid methyl ester (0.28g, 0.86mmol), triethylamine (0.66ml, 2.06mmol), DMAP (0.02g, 0.16mmol) and DCM (3ml) were stirred at room temperature under nitrogen atmosphere for 5 minutes. 3-Fluorobenzoyl chloride (0.204g, 1.28mmol) was added dropwise to the reaction mixture. The reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the reaction mixture was poured into a saturated sodium bicarbonate (10ml) solution and extracted with dichloromethane (3x 10ml). The combined organic layer was washed with brine (10ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by column chromatography to give the title compound as a gum (0.23 g, 59%). LCMS: m / z 447.3 [M+H] + .

[0381] Step 4. Synthesis of N-(1-(2,4-dichlorophenyl)-2-hydroxyethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide. To a solution of methyl 2-(2,4-dichlorophenyl)-2-(3-fluoro-N-(pyridin-3-ylmethyl)benzamide)acetate (0.17 g, 0.39 mmol) in methanol (6 ml) was added sodium borohydride (0.298 g, 7.9 mmol) in batches at 0 ° C. The resulting reaction mixture was placed at room temperature and stirred for another 16 hours. After the reaction was complete, saturated ammonium chloride solution (50 ml) was slowly added, and the reaction mixture was extracted with dichloromethane (2 x 30 ml). The combined organic layers were washed with brine (20 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was purified by column chromatography to obtain the title compound (0.12 g, 75%) as a jelly. Chiral preparative HPLC: Waters PSFC-200 and UV detector; column: CHIRALPAK IH (250*21) mm, 5u; mobile phase A: liquid CO2, mobile phase B: 0.1% DEA / IPA:ACN (50:50); isocratic gradient 80:20 (A:B), column flow: 80 ml / min (100 bar). FR-1 (isomer 1): R T =4.51 min; FR-2 (isomer 2): R T =9.80 minutes.

[0382] Isomer 1 was arbitrarily designated as (R)-N-(1-(2,4-dichlorophenyl)-2-hydroxyethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 161) LCMS: m / z 419.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ3.92-3.97(m,1H),4.07-4.11(m,1H),4.49(d,J=16.4Hz,1H),4.62-4.66(m,1H),5.10(br s,1H),5.40(br s,1H),7.15-7.18(m,1H),7.25-7.47(m,7H),7.62(d,J=8.4Hz,1H),8.20(s,1H),8.32(d,J=4.0Hz,1H).

[0383] Isomer 2 was arbitrarily designated as (S)-N-(1-(2,4-dichlorophenyl)-2-hydroxyethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 162) LCMS: m / z 419.1 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ3.92-3.98(m,1H),4.08-4.10(m,1H),4.49(d,J=16.0Hz,1H),4.62-4.66(m,1H),5.11(br s,1H),5.40(br s,1H),7.15-7.18(m,1H),7.22-7.47(m,7H),7.62(d,J=8.0Hz,1H),8.20(s,1H),8.32(d,J=4.4Hz,1H).

[0384] General Procedure XI

[0385] Preparation of (R)-N-(1-(4-carbamoyl-2-chlorophenyl)ethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 163) and (S)-N-(1-(4-carbamoyl-2-chlorophenyl)ethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 164).

[0386]

[0387] Step 1.1-(4-bromo-2-chlorophenyl)eth-1-amine synthesis. Molecular sieves (20 g) were added to a stirred solution of 1-(4-bromo-2-chlorophenyl)eth-1-one (10 g, 42.82 mmol) in methanol (200 ml), followed by hydroxylamine hydrochloride (8.93 g, 128.48 mmol). The reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was filtered through diatomaceous earth and the filtrate was evaporated to dryness. The crude solid was ground with hexane (20 ml) containing 10% ethyl acetate, then ground with n-pentane (20 mL) and decanted to obtain 1-(4-bromo-2-chlorophenyl)eth-1-one oxime (10 g). LCMS: m / z 249.7 [M+H] + .

[0388] 1-(4-bromo-2-chlorophenyl)ethan-1-one oxime (10g, 40.24mmol) was dissolved in EtOH:AcOH (1:1, 200ml), and zinc powder (21.05g, 322mmol) was added in batches at room temperature (note: exothermic reaction). The reaction was stirred at room temperature for 18 hours. After the reaction was completed, the reaction mixture was filtered through diatomaceous earth and the filtrate was evaporated to dryness. The crude residue was neutralized with sodium bicarbonate solution to adjust the pH to 8. The product was extracted with EtOAc (3x 100ml). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was ground with n-pentane (2x 50ml) and decanted to obtain the title compound (5.05g, 50%, 2 steps). LCMS: m / z 235.8[M+H] + .

[0389] Step 2. Synthesis of 1-(4-bromo-2-chlorophenyl)-N-(pyridin-3-ylmethyl)eth-1-amine. To a stirred mixture of 1-(4-bromo-2-chlorophenyl)eth-1-amine (5.0 g, 21.32 mmol) in DMF (50 ml) was added potassium carbonate (8.84 g, 64.0 mmol). 3-(chloromethyl)pyridine HCl (4.54 g, 27.7 mmol) was added at room temperature. The reaction mixture was heated to 60°C for 16 hours. After completion of the reaction, the reaction mixture was quenched with cold water 150 ml and extracted with ethyl acetate (3 x 50 ml). The combined organic layers were washed with cold water (2 x 50 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The crude compound was purified by RP Gold column chromatography using acetonitrile and water containing 0.1% formic acid to obtain the pure title compound (3.5 g, 50%). LCMS: m / z 326.0 [M+H] + .

[0390] Step 3. Synthesis of N-(1-(4-bromo-2-chlorophenyl)ethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide. To a stirred mixture of 1-(4-bromo-2-chlorophenyl)-N-(pyridin-3-ylmethyl)ethyl-1-amine (3.5g, 10.7mmol) in DCM (35ml) was added Et at room temperature under a nitrogen atmosphere N (2.72g, 26.9mmol). The reaction mixture was cooled to 0°C, and 3-fluorobenzoyl chloride (2.55g, 16.1mmol) was added dropwise. The reaction mixture was stirred at 15°C for 1 hour. After the reaction was completed, the reaction mixture was quenched with sodium bicarbonate solution (35ml) and extracted with DCM (2x 35ml). The combined organic layer was washed with water (2x 50ml), then washed with brine (50ml), dried over NaSO, filtered and concentrated under reduced pressure. The crude compound was purified by Combi flash column chromatography to give the title compound (3.0 g, 62%). LCMS: m / z 447.0 [M+H] + .

[0391] Step 4. Synthesis of methyl 3-chloro-4-(1-(3-fluoro-N-(pyridin-3-ylmethyl)benzamido)ethyl)benzoate. In a hydrogenator, sodium acetate (0.531 g, 6.48 mmol), palladium acetate (0.050 g, 0.23 mmol) and Pd(dppf)Cl2 (0.163 g, 0.23 mmol) were added to a stirred mixture of N-(1-(4-bromo-2-chlorophenyl)ethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (1 g, 2.23 mmol) in MeOH (20 ml) at room temperature. Carbon monoxide (g) pressure (125 psi) was applied and the hydrogenator was heated to 70 °C for 16 hours. After completion of the reaction, the reaction mixture was filtered through diatomaceous earth and the filtrate was concentrated. The crude compound was purified by Combi flash column chromatography to obtain the title compound (0.625 g, 65%). LCMS: m / z 427.0 [M+H] + .

[0392] Step 5. N-(1-(4-carbamoyl-2-chlorophenyl)ethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide. A stirred solution of methyl 3-chloro-4-(1-(3-fluoro-N-(pyridin-3-ylmethyl)benzamido)ethyl)benzoate (0.625 g, 1.46 mmol) in MeOH (12.5 ml) was purged with ammonia gas for 15-30 minutes at -78°C. The reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete, the reaction mixture was concentrated. The crude compound was purified by RP Gold column chromatography using acetonitrile and water containing 0.1% formic acid to give the pure title compound (0.160 g, 26%). Chiral preparative HPLC: Waters PSFC-200 and UV detector; column: CHIRALPAK IB-N (250*21) mm, 5u; mobile phase A: liquid CO2, mobile phase B: 0.1% DEA / IPA:ACN (50:50); isocratic gradient 78:22 (A:B), column flow: 80 ml / min (100 bar). FR-1 (isomer 1): R T =5.93 minutes; FR-2 (isomer 2): R T =6.28 minutes.

[0393] Isomer 1 was arbitrarily designated as (R)-N-(1-(4-carbamoyl-2-chlorophenyl)ethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 163) LCMS: m / z 412.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ1.64(d,J=7.2Hz,3H),4.49-4.59(m,2H),5.50(bs,1H),7.17-7.29(m,4H) ,7.44-7.50(m,2H),7.67(d,J=8.0Hz,1H),7.80-7.83(m,2H),8.23(bs,1H),8.33(d,J=4.0Hz,1H).

[0394] Isomer 2 was arbitrarily designated as (S)-N-(1-(4-carbamoyl-2-chlorophenyl)ethyl)-3-fluoro-N-(pyridin-3-ylmethyl)benzamide (Compound 164) LCMS: m / z 412.0 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ1.64 (d, J=7.2Hz, 3H), 4.49-4.58 (m, 2H), 5.50 (bs, 1H), 7.17-7.28 (m, 4H) ,7.45-7.50(m,2H),7.67(d,J=8.0Hz,1H),7.80-7.83(m,2H),8.23(bs,1H),8.33(d,J=4.0Hz,1H).

[0395] The following compounds in Table 8 were prepared in a manner similar to the General Procedures VII-XI described above:

[0396] Table 8

[0397]

[0398]

[0399] Synthesis of 2-(4-((N-(2,4-dichlorobenzyl)-3,5-difluorobenzamido)methyl)-1H-pyrazol-1-yl)acetic acid (Compound 165).

[0400]

[0401] Step 1. 1H-pyrazole-4-carboxaldehyde (1 g, 10.4 mmol) and 1-(2,4-dichlorophenyl)methanamine (2.00 g, 11.4 mmol) were combined in DCM (25 mL) and stirred at room temperature before the addition of sodium triacetoxyborohydride (3.30 g, 15.6 mmol). The mixture was stirred overnight under a nitrogen atmosphere. The reaction was quenched with saturated NaHCO3 solution and the product was extracted with DCM. The combined organic extracts were dried over Na2SO4, filtered and concentrated. The residue was purified by reverse phase chromatography (Biotage 60 g C18 column; 5-50% CH3CN / H2O and 0.1% formic acid gradient) to give N-((1H-pyrazol-4-yl)methyl)-1-(2,4-dichlorophenyl)methanamine. ES m / z 256.0 [M+H] + .

[0402]

[0403] Step 2. N-((1H-pyrazol-4-yl)methyl)-1-(2,4-dichlorophenyl)methanamine (925 mg, 3.6 mmol) and 3,5-difluorobenzoic acid (573 mg, 3.6 mmol), Et3N (1.00 mL, 7.2 mmol) and HATU (1.64 g, 4.32 mmol) were combined in DMF (25 mL) and stirred overnight at room temperature. The reaction was quenched with saturated NaHCO3 solution and the product was extracted with DCM. The combined organic extracts were dried over Na2SO4, filtered and concentrated. The residue was purified by reverse phase chromatography (Biotage 60 g C18 column; 5-50% CH3CN / H2O and 0.1% formic acid gradient) to give N-((1H-pyrazol-4-yl)methyl)-N-(2,4-dichlorobenzyl)-3,5-difluorobenzamide. ES m / z 396.0[M+H] + .

[0404]

[0405] Step 3. N-((1H-pyrazol-4-yl)methyl)-N-(2,4-dichlorobenzyl)-3,5-difluorobenzamide (461 mg, 1.1 mmol) and K2CO3 (322 mg, 2.2 mmol) were combined in DMF (15 mL). After 15 minutes, ethyl 2-bromoacetate (183 mg, 1.1 mmol) was added. The reaction was stirred at room temperature overnight. The reaction was quenched with saturated brine solution and the product was extracted with DCM. The combined organic extracts were dried over Na2SO4, filtered and concentrated. The residue was purified by reverse phase chromatography (Biotage 60g C18 column; 5-50% CH3CN / H2O and 0.1% formic acid gradient) to give ethyl 2-(4-((N-(2,4-dichlorobenzyl)-3,5-difluorobenzamide)methyl)-1H-pyrazol-1-yl)acetate. ES m / z482.1[M+H] + .

[0406]

[0407] Step 4. 40 mg of ester (1 eq), 2 eq of 1 M NaOH, ethyl 2-(4-((N-(2,4-dichlorobenzyl)-3,5-difluorobenzamido)methyl)-1H-pyrazol-1-yl)acetic acid (40 mg, 0.083 mmol) were dissolved in MeOH (1 mL) and 1 M aqueous NaOH (166 uL, 0.166 mmol) was added. The reaction was stirred at room temperature for 6 hours. The residue was purified by reverse phase chromatography (Biotage 60 g C18 column; 5-50% CH3CN / H2O and 0.1% formic acid gradient) to give 2-(4-((N-(2,4-dichlorobenzyl)-3,5-difluorobenzamido)methyl)-1H-pyrazol-1-yl)acetic acid. ES m / z 455.0 [M+H] + .

[0408] The following compounds in Table 9 were prepared in a manner similar to the methods and procedures described above:

[0409] Table 9

[0410]

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417]

[0418]

[0419]

[0420]

[0421]

[0422] Biochemical assays

[0423] 1. Silencing TREX1 in tumor cells

[0424] Activation of the cGAS / STING pathway can occur in both tumor cells and innate immune cells, particularly dendritic cells, following sensing of cytoplasmic DNA and subsequent type I IFN production. To evaluate whether TREX1 prevents type I IFN production in a well-described cold syngeneic tumor model that undergoes immune-mediated rejection following type I IFN activation by a STING agonist, TREX1 was knocked down in B16F10 tumor cells using CRISPR. Accumulation of cytoplasmic DNA by DNA transfection of tumor cells resulted in an approximately 5-fold increase in IFNβ production by TREX1 knockout B16F10 cells relative to parental tumor cells, indicating that TREX1 attenuates cGAS / STING pathway activation in B16F10 tumor cells.

[0425] 2. Growth of TREX1-competent and -deficient B16F10 tumor cells in vivo

[0426] Assessed the growth of TREX1 competent and defective B16F10 tumor cells in vivo. C57BL / 6J mice were inoculated with 300,000 B16F10 tumor cells of parent or TREX1 knockout subcutaneously in the right flank. Body weight was collected twice a week, and tumor measurements were performed two to three times a week, starting when tumor became measurable and continuing in the remaining duration of research. Compared with parental B16F10 tumor, TREX1 was presented significantly smaller volume by the silent tumor.

[0427] Tumors were collected on the 19th day after the study was terminated, and digested into single cell suspensions to enable the flow cytometry quantification of the immune population of tumor infiltration. It was found that TREX1 knocked out B16F10 tumors and showed a significant increase in overall immune cells, which reflected the increase in tumor infiltration CD4 and CD8 T cells and plasmacytoid dendritic cells (pDC) quantity. Known pDC plays a central role in inducing antigen-specific anti-tumor immune responses, and known T cells are the main effectors of anti-tumor efficacy in mice and humans. Therefore, the profound changes in the immune infiltration of the tumor lacking TREX1 show that the inhibition of the latter's tumor growth is at least partially immune-mediated.

[0428] TREX1 biochemical assay

[0429] Compound efficacy was assessed by fluorescence assay, which measures degradation of custom dsDNA substrates with fluorophore-quencher pairs on the opposite strands. Degradation of dsDNA releases free fluorophores to produce fluorescent signals. Specifically, 7.5 μL of reaction buffer (50 mM Tris (pH 7.4), 150 mM NaCl, 2 mM DTT, 0.1 mg / mL BSA, 0.01% (v / v) Tween-20 and 100 mM MgCl2) containing full-length human TREX1 (expressed in Escherichia coli (E. coli) and purified internally) with His-Tev tags at the N-terminus was added to 384-well Black ProxiPlate Plus (Perkin Elmer (Perkin Elmer)) containing different concentrations of compounds (150 nL) in DMSO as 10-point dose responses. To this was added 7.5 μL of reaction buffer containing dsDNA substrate (Chain A: 5'TEX615 / GCT AGG CAG 3'; Chain B: 5'CTG CCT AGC / IAbRQSp (Integrated DNA Technologies)). The final concentration was 150 pM TREX1, 60 nM dsDNA substrate in reaction buffer with 1.0% DMSO (v / v). After 25 minutes at room temperature, the reaction was quenched by adding 5 μL of stop buffer (same as reaction buffer plus 200 mM EDTA). The final concentration in the quenched reaction was 112.5 pM TREX1, 45 nM DNA and 50 mM EDTA in a volume of 20 μL. After incubation at room temperature for 5 minutes, the plate was read in a laser source Envision (PerkinElmer) and fluorescence at 615 nm was measured after excitation with 570 nm light. IC was calculated by comparing the ratio of fluorescence measured at 615 nm to control wells prequenched with stop buffer (100% inhibition) and controls without inhibitor (0% inhibition) using nonlinear least squares four-parameter fitting and Genedata or GraphPad Prism (GraphPad Software, Inc.). 50 value.

[0430] TREX2 biochemical assay

[0431] Compound efficacy was assessed by fluorescence assay, which measures the degradation of custom dsDNA substrates with fluorophore-quencher pairs on the opposite strands. The degradation of dsDNA releases free fluorophores to produce fluorescent signals. Specifically, 7.5 μL of reaction buffer (50 mM Tris (pH 7.4), 150 mM NaCl, 2 mM DTT, 0.1 mg / mL BSA, 0.01% (v / v) Tween-20 and 100 mM MgCl2) containing human TREX2 (residues M44-A279, expressed in Escherichia coli and purified internally) with His-Tev tags at the N-terminus was added to a 384-well Black ProxiPlate Plus (Perkin Elmer) containing compounds (150 nL) of varying concentrations in DMSO as a 10-point dose response. To this was added 7.5 μL of reaction buffer containing dsDNA substrate (Chain A: 5'TEX615 / GCT AGG CAG 3'; Chain B: 5'CTG CCT AGC / IAbRQSp(IDT). The final concentrations were 2.5 nM TREX2, 60 nM dsDNA substrate in reaction buffer with 1.0% DMSO (v / v). After 25 minutes at room temperature, the reaction was quenched by adding 5 μL of stop buffer (same as reaction buffer plus 200 mM EDTA). The final concentrations in the quenched reaction mixture were 1.875 pM TREX2, 45 nM DNA, and 50 mM EDTA in a volume of 20 μL. After incubation for 5 minutes at room temperature, the plate was read in a laser source Envision (PerkinElmer) and fluorescence at 615 nm was measured after excitation with 570 nm light. The fluorescence was determined by using a nonlinear least squares four-parameter fit and Genedata or GraphPad Prism (GraphPad Software, Inc. IC was calculated by comparing the ratio of fluorescence measured at 615 nm to control wells prequenched with stop buffer (100% inhibition) and controls without inhibitor (0% inhibition). 50 value.

[0432] The results are shown in Table 10. TREX1 EC 50 : A=<0.1 μM; B=0.1 to 1 μM; C=1 to 10 μM; D=>10 μM. TREX2 EC 50 : A=<1μM; B=1 to 10μM; C=10 to 100μM; D=>100μM.

[0433] Table 10

[0434]

[0435]

[0436]

[0437]

[0438]

[0439]

[0440]

[0441] Although multiple embodiments have been described, it is apparent that the basic examples can be modified to provide other embodiments utilizing the compounds and methods of the invention. It should therefore be understood that the scope of the invention is to be determined by the appended claims rather than by the specific embodiments that have been shown by way of example.

[0442] The contents of all references cited in this application (including literature references, granted patents, published patent applications and co-pending patent applications) are expressly incorporated herein by reference in their entirety. Unless otherwise defined, all technical terms and scientific terms used herein are consistent with the meanings commonly known to those of ordinary skill in the art.

Claims

1. A compound having formula II: or a pharmaceutically acceptable salt thereof, wherein R 2 is hydrogen or (C1-C4) alkyl; R 3 is halogen, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy or halo(C1-C4)alkoxy; R 4 is halogen, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy or CN; Ring A is a nitrogen-containing 5- to 7-membered heteroaryl group; R 5 is halogen, oxo, (C1-C4)alkyl, halogenated (C1-C4)alkyl, -C(O)OR b 、-NR b R c 、-C(O)NR b R c 、-(C1-C4)alkylOR b 、-(C1-C4)alkylC(O)R b 、-(C1-C4)alkylC(O)NR b R c 、-C(O)NR b SO2(C1-C4)alkyl, 5 to 6 membered heteroaryl, wherein the heteroaryl is optionally selected from R 7 substituted with 1 to 3 groups; R b and R c are each independently hydrogen, (C1-C4)alkyl or halogenated (C1-C4)alkyl; R 7 is halo, hydroxy, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, CN or oxo; m is 0, 1 or 2; and n is 0, 1, 2 or 3.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 3 It is a halogen group or a halogenated (C1-C4) alkyl group.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 3 is fluorine, chlorine, methyl, methoxy or CF3.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 3 It is fluorine, chlorine or CF3.

5. The compound according to claim 1, wherein the compound has formula III: or a pharmaceutically acceptable salt thereof.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 4 It is a halogen group.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein R 4 For chlorine. 8 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein m is 0 or 1.

9. The compound according to claim 1, wherein the compound has formula IV: or a pharmaceutically acceptable salt thereof. 10 . The compound according to claim 1 , or a pharmaceutically acceptable salt thereof, wherein Ring A is pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl or pyrazolyl. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ring A is pyridyl.

12. The compound of claim 1, wherein the compound has Formula V: or a pharmaceutically acceptable salt thereof.

13. The compound according to claim 12, wherein R 5 is located in the para position relative to the point of attachment of ring A, and wherein n is 1.

14. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 5 is oxo, CF3, CH3, C(O)OCH3, C(O)OH, NH2, CONH2, CH2CONH2, CONHCH3, CH2OH, CH2CH2OH, CONHSO2CH3, tetrazolyl, triazolyl or pyrazolyl, wherein the pyrazolyl is optionally substituted with hydroxyl.

15. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein n is 0, 1 or 2.

16. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein n is 2.

17. A compound selected from any one of the following: or a pharmaceutically acceptable salt thereof.

18. A pharmaceutical composition comprising the compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

19. Use of a compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof or a composition according to claim 18 in the preparation of a medicament for treating a disease in a subject that is responsive to inhibition of TREX1.

20. The use according to claim 19, wherein the disease is cancer.

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